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There are many resources available to mycobacterial researchers, including culture collections around the world that distribute biomaterials to the general scientific community, genomic and clinical databases, and powerful bioinformatics tools. However, many of these resources may be unknown to the research community. This review article aims to summarize and publicize many of these resources, thus strengthening the quality and reproducibility of mycobacterial research by providing the scientific community access to authenticated and quality-controlled biomaterials and a wealth of information, analytical tools and research opportunities. [ABSTRACT FROM AUTHOR]

Collection de cultures de microalgues d'eau douce de l'archipel des Açores : ressource pour la recherche taxonomique et la phycoprospection. Ces dernières années, un grand intérêt a été porté sur le potentiel biotechnologique des microalgues, notamment du fait de leur rapide croissance et de l'identification de certaines substances synthétisées par ces organismes. L'isolation puis la culture in vitro d'espèces de microalgues natives sont très importantes pour les études taxonomiques et les études de conservation. Elles représentent les premières étapes des études de faisabilité pour des productions commerciales locales. Dans cette étude, 114 microalgues ont été isolées à partir d'échantillons prélevés dans 23 stations de l'île de São Miguel, Açores. Parmi celles-ci, 60 espèces ont été identifiées dont 39 Chlorophytes, 10 Ochrophytes, 6 Cyanophytes, 3 Charophytes, 1 Euglénozoaire et 1 Cryptophyte. Dix-huit de ces identifications d'espèces constituent de nouveaux enregistrements pour l'île de São Miguel. In recent years, much interest has been focused on the biotechnological potential of microalgae, mainly due to their rapid growth and the identification of several substances synthesized by these organisms. The isolation and in vitro cultivation of native microalgae species is very important for taxonomic and conservation studies and is the first step towards feasibility studies of local commercial productions. In this study 114 isolates of microalgae were obtained from samples collected in 23 locations of the island of São Miguel, Azores. From them, 60 species were identified comprising 39 Chlorophyta, 10 Ochrophyta, 6 Cyanophyta, 3 Charophyta, 1 Euglenozoa and 1 Cryptophyta. Eighteen of the species identified constitute new records for the Island of São Miguel. [ABSTRACT FROM AUTHOR]

After the World Trade Organization’s agreement on Trade Related Aspect of Intellectual Property Right, patents have come into major play. Patenting of work related to live organisms that have medical, agricultural and other uses is always a tedious, complicated and controversial job. In view of this an agreement called as Budapest Treaty was passed in 1977 for deposition of microorganisms in culture collection centers for patent purpose. To make a culture collection center an IDA the culture center has to follow rules and regulations made in Budapest Treaty. Today several culture collection centers are working as International Depository Authority (IDA) in India and abroad that are storing microorganisms for patent purpose. India has two IDA units; one is Microbial Culture Collection in National Centre for Cell Science Pune, India, and second one is Microbial Type Culture Collection and Gene Bank at Institute of Microbial Technology Chandigarh, India. IDA units are important for valuable sample depositions, supply of cultures, characterization of samples and conservation of biological materials.

Fungal culture collections are important to biologists, microbiologists, epidemiologists and others involved in health and natural sciences. The improvement of techniques and methods for fungal isolation and preservation has contributed to maintain large microbial collections, which represent a rich source of biological sciences research, especially taxonomic, pathological and biodiversity studies as well as industrial applications. The collection centers are responsible for repository reference strains and for the maintenance of these microorganisms. The ITEM Microbial Culture Collection of ISPA (Institute of Sciences and of Food Production) includes more than 10,000 strains belonging to various agro-food microorganisms with phytopathological and toxicological significance. These microorganisms are mainly fungal pathogens belonging to toxigenic genera of Fusarium, Aspergillus, Alternaria, and Penicillium. This collection is a remarkable resource in the fight against mycotoxins: the increasing number of toxigenic fungi included in this collection ensures an original genetic source for biotechnological applications in several fields of research, contributing to knowledge improvement about fungal biology and strategies development for reducing mycotoxin contamination.

The supply of viable, authentic and secure microbial cultures is essential for obtaining agricultural products through biotechnology. The objective of this study was to establish, at the Institute of Animal Science, the methodology for organizing and managing the collection of microorganisms with potential as additives in animal nutrition. The establishment of the collection was developed in three stages: design of collection structure, definition of conservation methods for each type of microorganism and preparation of documents. Guidelines of the World Federation for Culture Collections, biological safety regulations, requirements of quality systems and requirements for lab competence were consulted. The establishment of a collection of microorganisms at the Institute of Animal Science, with appropriate conservation methods for each microbial group, guarantees the availability of strains for teaching, research and obtaining additives for animal feed. [ABSTRACT FROM AUTHOR]

After the World Trade Organization's agreement on Trade Related Aspect of Intellectual Property Right, patents have come into major play. Patenting of work related to live organisms that have medical, agricultural and other uses is always a tedious, complicated and controversial job. In view of this an agreement called as Budapest Treaty was passed in 1977 for deposition of microorganisms in culture collection centers for patent purpose. To make a culture collection center an IDA the culture center has to follow rules and regulations made in Budapest Treaty. Today several culture collection centers are working as International Depository Authority (IDA) in India and abroad that are storing microorganisms for patent purpose. India has two IDA units; one is Microbial Culture Collection in National Centre for Cell Science Pune, India, and second one is Microbial Type Culture Collection and Gene Bank at Institute of Microbial Technology Chandigarh, India. IDA unites are important for valuable sample depositions, supply of cultures, characterization of samples and conservation of biological materials. [ABSTRACT FROM AUTHOR]

Modern genomic and metabolomic tools have provided the possibility of generating and interrogating large datasets that can provide answers to previously imponderable taxonomic, evolutionary, ecological, and physiological questions. However, the curatorial tools needed to provide and maintain the relevant biological resources on which new knowledge can be built have not kept pace with this meteoric rise in scientific capacity, its associated activity, or the huge increase in published science. The availability of biological material of guaranteed identity and quality in Biological Resource Centers is fundamental for scientific research, but it crucially depends on there being adequate preservation/maintenance methods that are capable of ensuring phenotypic, genotypic, and functional security of the biological material(s). This article highlights the challenges to the long-term maintenance of genetic resources in general, focusing specifically on the issues associated with the maintenance of a large collection of strains of the ecologically significant diatom Skeletonema marinoi. This research collection, held at the Department of Marine Sciences, University of Gothenburg, has been systematically tested for its capacity to survive cryopreservation. A method, involving incubation in the dark for 20-24 hours before cryopreservation, followed by cryoprotection employing 10% dimethysulphoxide (DMSO) and conventional cooling in a passive cooler, before plunging into liquid nitrogen was successfully applied to ∼80% of the strains tested. In addition, the growth characteristics of exemplar strains were confirmed after storage. [ABSTRACT FROM AUTHOR]

Cyanobacterial blooms represent a health hazard worldwide due to their toxic potential. The CyanoSurvey project was set up to evaluate the risk associated with the presence of toxigenic cyanobacteria in Australian drinking water reservoirs. It mainly targeted members of the following genera: Dolichospermum, Sphaerospermopsis, Chrysosporum, Cylindrospermopsis and Microcystis. In 2010 and 2011, 64 samples were collected nationally and 196 isolates were purified using several culture media and a concentration technique. The strains obtained were then cryopreserved using three different cryoprotectants. The efficacy of each medium, the applicability of a pre-treatment to the sample and the various cryoprotective agents were compared for several isolates of cyanobacteria. This study highlighted that the ASM-1 medium, used routinely by many laboratories for the isolation and maintenance of cyanobacteria, had the lowest efficacy for the isolation of cyanobacteria from environmental samples. Moreover, the use of a range of media combined with a concentration technique can significantly improve the chances of isolating and purifying certain groups of cyanobacteria. The use of 5 % DMSO for cryopreservation was shown to be a preferable choice of cryopreservative for most strains tested. [ABSTRACT FROM AUTHOR]

Formal living collections have unique characteristics that distinguish them from other types of biorepositories. Comprising diverse resources, microbe culture collections, crop and biodiversity plant germplasm collections, and animal germplasm repositories are commonly allied with specific research communities or stakeholder groups. Among living collections, microbial culture collections have very long and unique life histories, with some being older than 100 years. Regulatory, financial, and technical developments have impacted living collections in many ways. International treaty obligations and restrictions on release of genetically modified organisms complicate the activities of living collections. Funding for living collections is a continuing challenge and threatens to create a two-tier system where medically relevant collections are well funded and all other collections are underfunded and hence understaffed. Molecular, genetic, and whole genome sequence analysis of contents of microbes and other living resource collections bring additional value to living collections. [ABSTRACT FROM AUTHOR]

The unique metagenomic, metaviromic libraries and indigenous micro diversity within Southern Africa have the potential for global beneficiation in academia and industry. Microorganisms that flourish at high temperatures, adverse pH conditions, and high salinity are likely to have enzyme systems that function efficiently under those conditions. These attributes afford researchers and industries alternative approaches that could replace existing chemical processes. Thus, a better understanding of African microbial/genetic diversity is crucial for the development of "greener" industries. A concerted drive to exploit the potential locked in biological resources has been previously seen with companies such as Diversa Incorporated and Verenium (Badische Anilin-und SodaFabrik-BASF) both building business models that pioneered the production of high-performance specialty enzymes for a variety of different industrial applications. The market potential and accompanying industry offerings have not been fully exploited in South Africa, nor in Africa at large. Utilization of the continent's indigenous microbial repositories could create long-lasting, sustainable growth in various production sectors, providing economic growth in resource-poor regions. By bolstering local manufacture of high-value bio-based products, scientific and engineering discoveries have the potential to generate new industries which in turn would provide employment avenues for many skilled and unskilled laborers. The positive implications of this could play a role in altering the face of business markets on the continent from costly import-driven markets to income-generating export markets. This review focuses on identifying microbially diverse areas located in South Africa while providing a profile for all associated microbial/genetically derived libraries in this country. A comprehensive list of all the relevant researchers and potential key players is presented, mapping out existing research networks for the facilitation of collaboration. The overall aim of this review is to facilitate a coordinated journey of exploration, one which will hopefully realize the value that South Africa's microbial diversity has to offer., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Published by Elsevier Ltd.)

The microbiome in the rhizosphere-the region surrounding plant roots-plays a key role in plant growth and health, enhancing nutrient availability and protecting plants from biotic and abiotic stresses. To assess bacterial diversity in the tomato rhizosphere, we performed two contrasting approaches: culture-dependent and -independent. In the culture-dependent approach, two culture media (Reasoner's 2A agar and soil extract agar) were supplemented with 12 antibiotics for isolating diverse bacteria from the tomato rhizosphere by inhibiting predominant bacteria. A total of 689 bacterial isolates were clustered into 164 operational taxonomic units (OTUs) at 97% sequence similarity, and these were found to belong to five bacterial phyla ( Proteobacteria, Actinobacteria, Bacteroidetes, Acidobacteria, and Firmicutes). Of these, 122 OTUs were retrieved from the antibiotic-containing media, and 80 OTUs were recovered by one specific antibiotic-containing medium. In the culture-independent approach, we conducted Illumina MiSeq amplicon sequencing of the 16S rRNA gene and obtained 19,215 high-quality sequences, which clustered into 478 OTUs belonging to 16 phyla. Among the total OTUs from the MiSeq dataset, 22% were recovered in the culture collection, whereas 41% of OTUs in the culture collection were not captured by MiSeq sequencing. These results showed that antibiotics were effective in isolating various taxa that were not readily isolated on antibiotic-free media, and that both contrasting approaches provided complementary information to characterize bacterial diversity in the tomato rhizosphere. [ABSTRACT FROM AUTHOR]

Coccidioidomycosis is an emerging fungal disease in Brazil; adequate maintenance and authentication of Coccidioides isolates are essential for research into genetic diversity of the environmental organisms, as well as for understanding the human disease. Seventeen Coccidioides isolates maintained under mineral oil since 1975 in the Instituto de Medicina Tropical de São Paulo (IMTSP) culture collection, Brazil, were evaluated with respect to their viability, morphological characteristics and genetic features in order to authenticate these fungal cultures. Only five isolates were viable after almost 30 years, showing typical morphological characteristics, and sequencing analysis using Coi-F and Coi-R primers revealed 99% identity with Coccidioides genera. These five isolates were then preserved in liquid nitrogen and sterile water, and remained viable after two years of storage under these conditions, maintaining the same features.Coccidioidomicose é uma doença emergente no Brasil; a manutenção adequada e autenticação de isolados de Coccidioides spp são essenciais para a pesquisa em diversidade genética de micro-organismos, bem como para a compreensão da doença em humanos. Dezessete isolados de Coccidioides preservados em óleo mineral desde 1975 na coleção de culturas do Instituto de Medicina Tropical de São Paulo (IMTSP) foram avaliados com relação à viabilidade, características morfológicas e genéticas, com o objetivo de autenticação das culturas fúngicas. Dos 17 isolados, apenas cinco foram viáveis após quase 30 anos mantidos em óleo mineral, apresentando características morfológicas e moleculares típicas do gênero, o sequenciamento utilizando os oligonucleotídeos Coi-F e Coi-R revelou identidade de 99% com isolados de Coccidioides. Estes cinco isolados foram preservados em nitrogênio líquido e água destilada esterilizada, e permaneceram viáveis após dois anos de armazenamento sob estas condições, mantendo as mesmas características.

Cryopreservation, the use of very low temperatures to preserve structurally intact living cells and tissues, is a key underpinning technology for life science research and medicine. It is employed to ensure the stability of critical biological resources including viruses, bacteria, protists, animal cell cultures, plants, reproductive materials and embryos. Fundamental to ensuring this stability is assuring stability of cryogenic storage temperatures. Here we report the occurrence of a failure in refrigeration in a cryostat holding > 600 strains of cyanobacteria and eukaryotic microalgae. A strategic approach was adopted to assess viability across a cross-section of the biodiversity held, both immediately after the potentially damaging temperature shift and 10 years later, on subsequent cryostorage in liquid-phase nitrogen (∼−196 °C). Furthermore, the event was replicated experimentally and the effects on the viability of cryo-tolerant and cryo-sensitive strains monitored. Our results have significant implications to all users of this storage method and parallels have been drawn with the ongoing development in other fields and in particular, human cell therapy. Based on our practical experience we have made a series of generic recommendations for emergency, remedial and ongoing preventative actions. [ABSTRACT FROM AUTHOR]

The twenty-first century has brought new opportunities and challenges to yeast culture collections, whether they are long-standing or recently established. Basic functions such as archiving, characterizing and distributing yeasts continue, but with expanded responsibilities and emerging opportunities. In addition to a number of well-known, large public repositories, there are dozens of smaller public collections that differ in the range of species and strains preserved, field of emphasis and services offered. Several collections have converted their catalogues to comprehensive databases and synchronize them continuously through public services, making it easier for users worldwide to locate a suitable source for specific yeast strains and the data associated with these yeasts. In-house research such as yeast taxonomy continues to be important at culture collections. Because yeast culture collections preserve a broad diversity of species and strains within a species, they are able to make discoveries in many other areas as well, such as biotechnology, functional, comparative and evolution genomics, bioprocesses and novel products. Due to the implementation of the Convention of Biological Diversity (CBD) and the Nagoya Protocol (NP), there are new requirements for both depositors and users to ensure that yeasts were collected following proper procedures and to guarantee that the country of origin will be considered if benefits arise from a yeast's utilization. Intellectual property rights (IPRs) are extremely relevant to the current access and benefit-sharing (ABS) mechanisms; most research and development involving genetic resources and associated traditional knowledge will be subject to this topic. Copyright © 2016 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Culture collections contain indispensable information about the microorganisms preserved in their repositories, such as taxonomical descriptions, origins, physiological and biochemical characteristics, bibliographic references, etc. However, information currently accessible in databases rarely adheres to common standard protocols. The resultant heterogeneity between culture collections, in terms of both content and format, notably hampers microorganism-based research and development (R&D). The optimized exploitation of these resources thus requires standardized, and simplified, access to the associated information. To this end, and in the interest of supporting R&D in the fields of agriculture, health and biotechnology, a pan-European distributed research infrastructure, MIRRI, including over 40 public culture collections and research institutes from 19 European countries, was established. A prime objective of MIRRI is to unite and provide universal access to the fragmented, and untapped, resources, information and expertise available in European public collections of microorganisms; a key component of which is to develop a dynamic Information System. For the first time, both culture collection curators as well as their users have been consulted and their feedback, concerning the needs and requirements for collection databases and data accessibility, utilised. Users primarily noted that databases were not interoperable, thus rendering a global search of multiple databases impossible. Unreliable or out-of-date and, in particular, non-homogenous, taxonomic information was also considered to be a major obstacle to searching microbial data efficiently. Moreover, complex searches are rarely possible in online databases thus limiting the extent of search queries. Curators also consider that overall harmonization-including Standard Operating Procedures, data structure, and software tools-is necessary to facilitate their work and to make high-quality data easily accessible to their users. Clearly, the needs of culture collection curators coincide with those of users on the crucial point of database interoperability. In this regard, and in order to design an appropriate Information System, important aspects on which the culture collection community should focus include: the interoperability of data sets with the ontologies to be used; setting best practice in data management, and the definition of an appropriate data standard. [ABSTRACT FROM AUTHOR]

The Cawthron Institute Culture Collection of Micro-algae (CICCM) is a unique, nationally and internationally significant collection of 450 strains of approximately 100 micro-algae and 50 cyanobacteria species, both living and cryopreserved. The collection comprises 13 taxonomic classes and underpins research into the ecology and taxonomy of the isolates and the biotoxins they produce and their toxicity. The CICCM expanded significantly in the 1990s after a major harmful algae bloom event in the Hauraki Gulf, New Zealand. Since 2000, it has underpinned development of molecular detection tools and enabled development of new chemical testing methods for biotoxins in seafood. The tropical collection allows research into potential issues for New Zealand as the oceans warm, for example, the risks of ciguatera fish poisoning and palytoxins in seafood. Research results generated from isolates in the cyanobacterial collection have assisted national risk management regarding drinking water and informed the development of the New Zealand guidelines for managing cyanobacteria in recreational fresh waters. The actual living and cryopreserved micro-algae and cyanobacteria collection (without infrastructure) has a current estimated value of NZ$1.6 million. [ABSTRACT FROM AUTHOR]

Millions of microbial taxa inhabit the rhizosphere and could be used as biofertilizers, biopesticides, and/or for bioremediation. Only a fraction of these microbes have been described and/or are being utilized. Most are dispersed in collections, but coordination of their accessibility and availability is challenging. Here, we present the Pan-European Rhizosphere Resource Network (PERN), which is a transnational repository of microorganisms whose objectives are to facilitate access to rhizosphere resources and information and help users with technical and legal issues. [ABSTRACT FROM AUTHOR]

All laboratories that do microbiological researches need culture collections because laboratories have toform their own collections and preserve on suitable conditions them for control of testing. Basic principle of culture preservation is to keep cells alive and in pure form for a long time without causing mutations and variations in microorganisms. Drying is the oldest method of culture preservation. Because it is an inexpensive and easy method, it is frequently used for the preservation of microbial cultures. Stages of all drying methods are generally similar to each other. Initially, culture is mixed with protectants and dried with a desired process. Then, dry culture is stored. When needed, cells are activated by rehydrating culture. This review explains simple drying methods like sand drying, drying on paper disc, drying with gelatin and silica gel, self-drying and alternative drying methods (except lyophilization) like spray drying, fluidised bed drying, foam drying and drying under vacuum. [ABSTRACT FROM AUTHOR]

From March 1990 to December 1992, the National Institute for Quality Control of Health-INCQS Research Collection received 1476 bacterial samples isolated from human cerebrospinal fluid of patients suspect of meningitis in Rio de Janeiro, from the São Sebastião State Institute of Infectious Diseases (IEISS). Neisseria meningitidis was found in most of these materials, followed in smaller number by Haemophilus sp. and Streptococcus pneumoniae. The great majority of N. meningitidis strains was serogroup B, followed by serogroup C and a few strains of serogroup W135. More than 50 of the isolated bacterial agents came from the predominant 0-4 years age group. The majority of the strains were from patients in the region known as "Baixada Fluminense" (Low Lands). The aim of the work presented here is to obtain samples of meningitis cases in at least 70 of the State of Rio de Janeiro and develop a collaborative research between INCQS-FIOCRUZ and the IEISS, in order to set up a collection of strains for future studies. However, despite work being carried out in a rather satisfactory way, difficulties still arise and have to be overcome, to survey data.

Culture collections are valuable resources for the sustainable use of microbial diversity and its conservation. Advances in biotechnology have further increased their importance and some of these have been recognized as International Depositary Authority (IDA) for the deposition of patent cultures. Microbial Culture Collection at National Centre for Cell Science was established by the Department of Biotechnology, Government of India is country's newest culture collection with largest holdings. It is recognized as an IDA under the Budapest Treaty and Designated National Repository under the Biodiversity Act 2002. This article describes its various service related activities. [ABSTRACT FROM AUTHOR]

Culture collections of microorganisms, including fungi, are strain deposits recognised as Biological Resource Centers (BRCs) with a great importance in science, industry and education. Their objective is to preserve the purity, viability and genomic integrity of every single strain as a member of such collection. Since improvement of molecular methods nowadays brought many novel approaches in manipulation with strains of microorganisms, they can also be useful for characterization of existing stored strains. ITS1 region in nuclear DNA is preferred barcoding marker for taxon identification, which can be explained by its great inter-species variability. This paper presents results from analysing ITS1 region sequences (17) obtained from fungal DNA of culture collection of autochthonous, lignicolous genera Piptoporus, Pleurotus, Ganoderma and Schizophyllum cultured on malt agar plates for 14 days at 25°C. BLAST (Basic Local Alignment Search Tool) was used for comparison with online databases, while alignment of sequences was made with MEGA 5.10 software. Morphological determination of species or genus was confirmed for 13 cultures, while the others were disproved. The resulting alignment indicated small intra-species variability of ITS1 region and pointed to it as an ideal marker for verification of fungal culture collections' authenticity. [ABSTRACT FROM AUTHOR]

The online community resource Phytophthora database (PD) was developed to support accurate and rapid identification of Phytophthora and to help characterize and catalog the diversity and evolutionary relationships within the genus. Since its release in 2008. the sequence database has grown to cover 1 to 12 loci for ≈ 2,600 isolates (representing 138 described and provisional species). Sequences of multiple mitochondrial loci were added to complement nuclear loci-based phylogenetic analyses and diagnostic tool development. Key characteristics of most newly described and provisional species have been summarized. Other additions to improve the PD functionality include: (i) geographic information system tools that enable users to visualize the geographic origins of chosen isolates on a global-scale map, (ii) a tool for comparing genetic similarity between isolates via microsatellite markers to support population genetic studies, (iii) a comprehensive review of molecular diagnostics tools and relevant references, (iv) sequence alignments used to develop polymerase chain reaction-based diagnostics tools to support their utilization and new diagnostic tool development, and (v) an online community forum for sharing and preserving experience and knowledge accumulated in the global Phytophthora community. Here we present how these improvements can support users and discuss the PD's future direction. [ABSTRACT FROM AUTHOR]

Improving the availability of representative isolates from the coral microbiome is essential for investigating symbiotic mechanisms and applying beneficial microorganisms to improve coral health. However, few studies have explored the diversity of bacteria which can be isolated from a single species. Here, we isolated a total of 395 bacterial strains affiliated with 49 families across nine classes from the coral Pocillopora damicornis. Identification results showed that most of the strains represent potential novel bacterial species or genera. We also sequenced and assembled the genomes of 118 of these isolates, and then the putative functions of these isolates were identified based on genetic signatures derived from the genomes and this information was combined with isolate-specific phenotypic data. Genomic information derived from the isolates identified putative functions including nitrification and denitrification, dimethylsulfoniopropionate transformation, and supply of fixed carbon, amino acids, and B vitamins which may support their eukaryotic partners. Furthermore, the isolates contained genes associated with chemotaxis, biofilm formation, quorum sensing, membrane transport, signal transduction, and eukaryote-like repeat-containing and cell-cell attachment proteins, all of which potentially help the bacterium establish association with the coral host. Our work expands on the existing culture collection of coral-associated bacteria and provides important information on the metabolic potential of these isolates which can be used to refine understanding of the role of bacteria in coral health and are now available to be applied to novel strategies aimed at improving coral resilience through microbiome manipulation. IMPORTANCE Microbes underpin the health of corals which are the building blocks of diverse and productive reef ecosystems. Studying the culturable fraction of coral-associated bacteria has received less attention in recent times than using culture-independent molecular methods. However, the genomic and phenotypic characterization of isolated strains allows assessment of their functional role in underpinning coral health and identification of beneficial microbes for microbiome manipulation. Here, we isolated 395 bacterial strains from tissues of Pocillopora damicornis with many representing potentially novel taxa and therefore providing a significant contribution to coral microbiology through greatly enlarging the existing cultured coral-associated bacterial bank. Through analysis of the genomes obtained in this study for the coral-associated bacteria and coral host, we elucidate putative metabolic linkages and symbiotic establishment. The results of this study will help to elucidate the role of specific isolates in coral health and provide beneficial microbes for efforts aimed at improving coral health.

Abstract: High quality 16S ribosomal RNA (rRNA) gene sequences from the type strains of all species with validly published names, as defined by the International Code of Nomenclature of Bacteria, are a prerequisite for their accurate affiliations within the global genealogical classification and for the recognition of potential new taxa. During the last few years, the Living Tree Project (LTP) has taken care to create a high quality, aligned 16S and 23S rRNA gene sequence database of all type strains. However, the manual curation of the sequence dataset and type strain information revealed that a total of 552 “orphan” species (about 5.7% of the currently classified species) had to be excluded from the reference trees. Among them, 322 type strains were not represented by an SSU entry in the public sequence repositories. The remaining 230 type strains had to be discarded due to bad sequence quality. Since 2010, the LTP team has coordinated a network of researchers and culture collections in order to improve the situation by (re)-sequencing the type strains of these “orphan” species. As a result, we can now report 351 16S rRNA gene sequences of type strains. Nevertheless, 201 species could not be sequenced because cultivable type strains were not available (121), the cultures had either been lost or were never deposited in the first place (66), or it was not possible due to other constraints (14). The International Code of Nomenclature of Bacteria provides a number of mechanisms to deal with the problem of missing type strains and we recommend that due consideration be given to the appropriate mechanisms in order to help solve some of these issues. [Copyright &y& Elsevier]

Coccidioidomycosis is an emerging fungal disease in Brazil; adequate maintenance and authentication of Coccidioides isolates are essential for research into genetic diversity of the environmental organisms, as well as for understanding the human disease. Seventeen Coccidioides isolates maintained under mineral oil since 1975 in the Instituto de Medicina Tropical de Sao Paulo (IMTSP) culture collection, Brazil, were evaluated with respect to their viability, morphological characteristics and genetic features in order to authenticate these fungal cultures. Only five isolates were viable after almost 30 years, showing typical morphological characteristics, and sequencing analysis using Coi-F and Coi-R primers revealed 99% identity with Coccidioides genera. These five isolates were then preserved in liquid nitrogen and sterile water, and remained viable after two years of storage under these conditions, maintaining the same features. [ABSTRACT FROM AUTHOR]

The availability of public genomics data has become essential for modern life sciences research, yet the quality, traceability, and curation of these data have significant impacts on a broad range of microbial genomics research. While microbial genome databases such as NCBI's RefSeq database leverage the scalability of crowd sourcing for growth, genomics data provenance and authenticity of the source materials used to produce data are not strict requirements. Here, we describe the de novo assembly of 1,113 bacterial genome references produced from authenticated materials sourced from the American Type Culture Collection (ATCC), each with full genomics data provenance relating to bioinformatics methods, quality control, and passage history. Comparative genomics analysis of ATCC standard reference genomes (ASRGs) revealed significant issues with regard to NCBI's RefSeq bacterial genome assemblies related to completeness, mutations, structure, strain metadata, and gaps in traceability to the original biological source materials. Nearly half of RefSeq assemblies lack details on sample source information, sequencing technology, or bioinformatics methods. Deep curation of these records is not within the scope of NCBI's core mission in supporting open science, which aims to collect sequence records that are submitted by the public. Nonetheless, we propose that gaps in metadata accuracy and data provenance represent an "elephant in the room" for microbial genomics research. Effectively addressing these issues will require raising the level of accountability for data depositors and acknowledging the need for higher expectations of quality among the researchers whose research depends on accurate and attributable reference genome data. IMPORTANCE The traceability of microbial genomics data to authenticated physical biological materials is not a requirement for depositing these data into public genome databases. This creates significant risks for the reliability and data provenance of these important genomics research resources, the impact of which is not well understood. We sought to investigate this by carrying out a comparative genomics study of 1,113 ATCC standard reference genomes (ASRGs) produced by ATCC from authenticated and traceable materials using the latest sequencing technologies. We found widespread discrepancies in genome assembly quality, genetic variability, and the quality and completeness of the associated metadata among hundreds of reference genomes for ATCC strains found in NCBI's RefSeq database. We present a comparative analysis of de novo -assembled ASRGs, their respective metadata, and variant analysis using RefSeq genomes as a reference. Although assembly quality in RefSeq has generally improved over time, we found that significant quality issues remain, especially as related to genomic data and metadata provenance. Our work highlights the importance of data authentication and provenance for the microbial genomics community, and underscores the risks of ignoring this issue in the future.

Nunney, L., Elfekih, S., and Stouthamer, R. 2012. The importance of multilocus sequence typing: Cautionary tales from the bacterium Xylella fastidiosa. Phytopathology 102:456-460. Microbial identification methods have evolved rapidly over the last few decades. One such method is multilocus sequence typing (MLST). MLST is a powerful tool for understanding the evolutionary dynamics of pathogens and to gain insight into their genetic diversity. We illustrate the importance of accurate typing by reporting on three problems that have arisen in the study of a single bacterial species, the plant pathogen Xylella fastidiosa. Two of these were particularly serious since they concerned contamination of important research material that has had detrimental consequences for Xylella research: the contamination of DNA used in the sequencing of an X. fastidiosa genome (Ann-l) with DNA from another X. fastidiosa strain, and the unrecognized mislabeling of a strain (Temeculal) distributed from a culture collection (ATCC). We advocate the routine use of MLST to define strains maintained in culture collections and emphasize the importance of confirming the purity of DNA submitted for sequencing. We also present a third example that illustrates the value of MLST in guiding the choice of taxonomic types. Beyond these situations, there is a strong case for MLST whenever an isolate is used experimentally, especially where genotypic differences are suspected to influence the outcome. [ABSTRACT FROM AUTHOR]

Abstract: Most fungal biology researchers depend on culture collections, or more aptly, ex situ fungal germplasm repositories, either for the materials upon which they work, or as a long-term home for their materials after their projects are finished. These collections are broadly distributed and typically supported by the local government. The large number of collections, notwithstanding, some collections have greater impact than others. This review will discuss the fungal germplasm repositories around the world with special attention paid to the Fungal Genetics Stock Center. To facilitate their activities collections have joined together in networks, both locally and internationally. Additional information on public policy and how it impacts collections will be presented and the impact of collections will be highlighted. [Copyright &y& Elsevier]