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1.
Article in English | MEDLINE | ID: mdl-38713196

ABSTRACT

The genus Exophiala is polymorphic, able to transition between yeast, hyphal and pseudohyphal forms. Species of the genus Exophiala are ubiquitous fungi that are distributed in various environments around the world. During a survey of fungal diversity in the gut of amphipods (Floresorchestia amphawaensis and undescribed Dogielinotid amphipods) from the Amphawa estuary, Samut Songkhram province, Thailand, five black yeast strains (DMKU-MG01, DMKU-MG07, DMKU-MG08, DMKU-HG10 and DMKU-FG04) were identified as representing a novel taxon on the basis of a combination of morphological and molecular phylogenetic features. The five strains did not produce filamentous hyphae or pseudohyphae. Only budding yeast cells were observed. On the basis of the phenotypic characteristics and the results of molecular analyses of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region, the five strains were identified as representing a novel species via applied nucleotide pairwise analysis. They differed from the most closely related species Exophiala alcalophiala by 3.54 % nucleotide substitutions (20 nucleotide substitutions in 572 bp) in the D1/D2 domains of the LSU rRNA gene. Moreover, the sequences of the ITS region of the five strains differed from those of the most closely related species E. alcalophiala, by 7.44-9.62 % nucleotide substitutions, and Exophiala halophiala, by 7.2-7.53 % nucleotide substitutions. The results of phylogenetic analyses based on the concatenated sequences of the ITS regions and the D1/D2 domains of the LSU rRNA gene confirmed that the five black yeast strains represented a single novel species of the genus Exophiala. In this study, Exophiala amphawaensis sp. nov. is proposed to accommodate these strains. The holotype is TBRC 15626T and the isotype is PYCC9020. The MycoBank accession number of the novel species is MB 851477.


Subject(s)
Amphipoda , DNA, Fungal , DNA, Ribosomal Spacer , Exophiala , Phylogeny , Sequence Analysis, DNA , Animals , Thailand , Amphipoda/microbiology , DNA, Fungal/genetics , DNA, Ribosomal Spacer/genetics , Exophiala/genetics , Exophiala/isolation & purification , Exophiala/classification , Mycological Typing Techniques , Gastrointestinal Tract/microbiology
2.
J Invertebr Pathol ; 200: 107970, 2023 09.
Article in English | MEDLINE | ID: mdl-37422088

ABSTRACT

Biological invasions may act as conduits for pathogen introduction. To determine which invasive non-native species pose the biggest threat, we must first determine the symbionts (pathogens, parasites, commensals, mutualists) they carry, via pathological surveys that can be conducted in multiple ways (i.e., molecular, pathological, and histological). Whole animal histopathology allows for the observation of pathogenic agents (virus to Metazoa), based on their pathological effect upon host tissue. Where the technique cannot accurately predict pathogen taxonomy, it does highlight pathogen groups of importance. This study provides a histopathological survey of Pontogammarus robustoides (invasive amphipod in Europe) as a baseline for symbiont groups that may translocate to other areas/hosts in future invasions. Pontogammarus robustoides (n = 1,141) collected throughout Poland (seven sites), were noted to include a total of 13 symbiotic groups: a putative gut epithelia virus (overall prevalence = 0.6%), a putative hepatopancreatic cytoplasmic virus (1.4%), a hepatopancreatic bacilliform virus (15.7%), systemic bacteria (0.7%), fouling ciliates (62.0%), gut gregarines (39.5%), hepatopancreatic gregarines (0.4%), haplosporidians (0.4%), muscle infecting microsporidians (6.4%), digeneans (3.5%), external rotifers (3.0%), an endoparasitic arthropod (putatively: Isopoda) (0.1%), and Gregarines with putative microsporidian infections (1.4%). Parasite assemblages partially differed across collection sites. Co-infection patterns revealed strong positive and negative associations between five parasites. Microsporidians were common across sites and could easily spread to other areas following the invasion of P. robustoides. By providing this initial histopathological survey, we hope to provide a concise list of symbiont groups for risk-assessment in the case of a novel invasion by this highly invasive amphipod.


Subject(s)
Amphipoda , Apicomplexa , Microsporidia , Parasites , Animals , Amphipoda/microbiology , Host-Parasite Interactions , United Kingdom , Introduced Species , Apicomplexa/physiology
3.
Mar Biotechnol (NY) ; 23(3): 463-471, 2021 Jun.
Article in English | MEDLINE | ID: mdl-34076776

ABSTRACT

Studies of invertebrates have shown that the internal environment of crustaceans is not always sterile in normal conditions, and in many species, it can be populated by microorganisms even in the absence of any visible pathological processes in the body. This observation raises the question of whether genetically modified indigenous hemolymph microorganisms can be used for biotechnological purposes inside the crustacean either as local producers of some compounds or as sensors to physiological parameters. In this study, we tested the ability of the bacteria isolated from the hemolymph of the amphipod Eulimnogammarus verrucosus to hide from the cellular immune response of the host as the most important feature for their potential long-term application in vivo. 16S rDNA amplicon sequencing revealed five common bacterial genera in all analyzed samples of the amphipod hemolymph, among which Pseudomonas is most easily subjected to genome modification and, thus, the most prospective for biotechnological application. Cultivation of Pseudomonas gave us a number of strains undoubtedly derived from the amphipod hemolymph, and one of them (belonging to the Pseudomonas fluorescens group) was chosen for further tests. The primary culture of amphipod hemocytes was used to analyze the immunogenicity of the strain and showed a pronounced reaction of the immune cells to a high amount of the bacteria within six hours. This result indicates that modulation of cellular immune response to metabolically active bacterial cells is not mandatory for the survival and wide distribution of these microorganisms in the hemolymph of numerous amphipod individuals.


Subject(s)
Amphipoda/immunology , Amphipoda/microbiology , Immunity, Cellular , Pseudomonas/physiology , Animals , Hemocytes , Hemolymph/cytology , Hemolymph/microbiology , Lakes , Siberia
4.
Sci Rep ; 10(1): 16842, 2020 10 08.
Article in English | MEDLINE | ID: mdl-33033309

ABSTRACT

Endosymbionts and intracellular parasites are common in arthropod hosts. As a consequence, (co)amplification of untargeted bacterial sequences has been occasionally reported as a common problem in DNA barcoding. While identifying amphipod species with universal COI primers, we unexpectedly detected rickettsial endosymbionts belonging to the Torix group. To map the distribution and diversity of Rickettsia species among amphipod hosts, we conducted a nationwide molecular screening of seven families of New Zealand freshwater amphipods. In addition to uncovering a diversity of Torix Rickettsia species across multiple amphipod populations from three different families, our research indicates that: (1) detecting Torix Rickettsia with universal primers is not uncommon, (2) obtaining 'Rickettsia COI sequences' from many host individuals is highly likely when a population is infected, and (3) obtaining 'host COI' may not be possible with a conventional PCR if an individual is infected. Because Rickettsia COI is highly conserved across diverse host taxa, we were able to design blocking primers that can be used in a wide range of host species infected with Torix Rickettsia. We propose the use of blocking primers to circumvent problems caused by unwanted amplification of Rickettsia and to obtain targeted host COI sequences for DNA barcoding, population genetics, and phylogeographic studies.


Subject(s)
Amphipoda/genetics , Amphipoda/microbiology , DNA Barcoding, Taxonomic , Electron Transport Complex IV/genetics , Host Microbial Interactions , Rickettsia/genetics , Animals , DNA, Bacterial/genetics , Genetic Variation , Genetics, Population , New Zealand , Phylogeny , Rickettsia/physiology , Symbiosis/genetics
5.
NPJ Biofilms Microbiomes ; 6(1): 23, 2020 06 05.
Article in English | MEDLINE | ID: mdl-32504001

ABSTRACT

Metaproteomics of gut microbiomes from animal hosts lacking a reference genome is challenging. Here we describe a strategy combining high-resolution metaproteomics and host RNA sequencing (RNA-seq) with generalist database searching to survey the digestive tract of Gammarus fossarum, a small crustacean used as a sentinel species in ecotoxicology. This approach provides a deep insight into the full range of biomasses and metabolic activities of the holobiont components, and differentiates between the intestine and hepatopancreatic caecum.


Subject(s)
Amphipoda/microbiology , Bacteria/classification , Proteogenomics/methods , Proteomics/methods , Animals , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/metabolism , DNA Barcoding, Taxonomic , Gastrointestinal Microbiome , Phylogeny , Sequence Analysis, RNA , Tandem Mass Spectrometry
6.
Parasitol Res ; 119(3): 915-923, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31970472

ABSTRACT

A new species and a new genus of a microsporidium Alternosema bostrichidis isolated from an adult Prostephanus truncatus in Mexico and from three species of the genus Dinoderus in Nigeria are described. The microsporidium is monomorphic, monoxenic, and develops in direct contact with host cell cytoplasm. The infection first appears with thoracic muscles, followed by a generalized invasion of the host. All developmental stages are diplokaryotic. Sporogony is disporoblastic. Mature spores are ovoid. Unfixed spores measure 3.7-4.2 × 2.0-2.6 µm, fixed and stained spores 3.5-5.0 × 2.4-2.8 µm. The polaroplast consists of dense lamellae and rare lamellae. The polar tube is slightly anisofilar, consisting of 11-17 coils, with 9-14 proximal (130 nm in diameter) and 2-3 distal coils (120 nm in diameter) arranged in one layer. Molecular phylogenetic analysis based upon a short portion of small-subunit ribosomal RNA gene (Genbank accession # KP455651) placed the new microsporidium within Liebermannia-Orthosomella lineage, which contains multiple undescribed parasites. In particular, A. bostrichidis showed maximal sequence similarity of 95% to Microsporidium sp. BBRE2 (# FJ755987) from Baikalian Diplacanthus brevispinus (Amphipoda: Acanthogammaridae) and Microsporidium sp. Comp CD Van 2 (# KC111784) from compost and soil in Canada. Frequent, devastating epizootics of laboratory cultures of A. bostrichidis support its potential as a biological control agent of grain borers.


Subject(s)
Amphipoda/microbiology , Biological Control Agents , Coleoptera/microbiology , Microsporidia, Unclassified/classification , Microsporidia, Unclassified/isolation & purification , Animals , Canada , Mexico , Microsporidia, Unclassified/genetics , Nigeria , Phylogeny , Prevalence , Spores, Fungal/classification
7.
Dis Aquat Organ ; 136(1): 3-36, 2019 Oct 02.
Article in English | MEDLINE | ID: mdl-31575832

ABSTRACT

With over 10000 species of Amphipoda currently described, this order is one of the most diverse groups of freshwater and marine Crustacea. Members of this group are globally distributed, and many are keystone species and ecosystem engineers within their respective ecologies. As with most organisms, disease is a key factor that can alter population size, behaviour, survival, invasion potential and physiology of amphipod hosts. This review explores symbiont diversity and pathology in amphipods by coalescing a range of current and historical literature to provide the first full review of our understanding of amphipod disease. The review is broken into 2 parts. The first half explores amphipod microparasites, which include data pertaining to viruses, bacteria, fungi, oomycetes, microsporidians, dinoflagellates, myxozoans, ascetosporeans, mesomycetozoeans, apicomplexans and ciliophorans. The second half reports the metazoan macroparasites of Amphipoda, including rotifers, trematodes, acanthocephalans, nematodes, cestodes and parasitic Crustacea. In all cases we have endeavoured to provide a complete list of known species that cause disease in amphipods, while also exploring the effects of parasitism. Although our understanding of disease in amphipods requires greater research efforts to better define taxonomic diversity and host effects of amphipod symbionts, research to date has made huge progress in cataloguing and experimentally determining the effects of disease upon amphipods. For the future, we suggest a greater focus on developing model systems that use readily available amphipods and diseases, which can be comparable to the diseases in other Crustacea that are endangered, economically important or difficult to house.


Subject(s)
Amphipoda/microbiology , Amphipoda/parasitology , Amphipoda/virology , Parasites , Animals , Disease Models, Animal , Ecosystem
8.
PLoS One ; 14(10): e0222527, 2019.
Article in English | MEDLINE | ID: mdl-31577799

ABSTRACT

Crangonyx islandicus is a groundwater amphipod endemic to Iceland, considered to have survived the Ice Ages in subglacial refugia. Currently the species is found in spring sources in lava fields along the tectonic plate boundary of the country. The discovery of a groundwater species in this inaccessible habitat indicates a hidden ecosystem possibly based on chemoautotrophic microorganisms as primary producers. To explore this spring ecosystem, we assessed its microbial diversity and analysed whether and how the diversity varied between the amphipods and the spring water, and if was dependent on environmental factors and geological settings. Isolated DNA from spring water and from amphipods was analysed using metabarcoding methods, targeting the 16S rRNA gene. Two genera of bacteria, Halomonas and Shewanella were dominating in the amphipod samples in terms of relative abundance, but not in the groundwater samples where Flavobacterium, Pseudomonas and Alkanindiges among others were dominating. The richness of the bacteria taxa in the microbial community of the groundwater spring sources was shaped by pH level and the beta diversity was shaped by geographic locations.


Subject(s)
Amphipoda/microbiology , Bacteria/growth & development , Biodiversity , Groundwater/microbiology , Animals , Bacteria/genetics , Genetic Variation , Geography , Iceland , Linear Models , Microbiota , RNA, Ribosomal, 16S/genetics
9.
Appl Environ Microbiol ; 85(1)2019 01 01.
Article in English | MEDLINE | ID: mdl-30366990

ABSTRACT

Hadal environments sustain diverse microorganisms. A few studies have investigated hadal microbial communities consisting of free-living or particle-associated bacteria and archaea. However, animal-associated microbial communities in hadal environments remain largely unexplored, and comparative analyses of animal gut microbiota between two isolated hadal environments have never been done so far. In the present study, 228 Gb of gut metagenomes of the giant amphipod Hirondellea gigas from two hadal trenches, the Mariana Trench and Japan Trench, were sequenced and analyzed. Taxonomic analysis identified 49 microbial genera commonly shared by the gut microbiota of the two H. gigas populations. However, the results of statistical analysis, in congruency with the alpha and beta diversity analyses, revealed significant differences in gut microbial composition across the two trenches. Abundance variation of Psychromonas, Propionibacterium, and Pseudoalteromonas species was observed. Microbial cooccurrence was demonstrated for microbes that were overrepresented in the Mariana trench. Comparison of functional potential showed that the percentage of carbohydrate metabolic genes among the total microbial genes was significantly higher in the guts of H. gigas specimens from the Mariana Trench. Integrating carbon input information and geological characters of the two hadal trenches, we propose that the differences in the community structure might be due to several selective factors, such as environmental variations and microbial interactions.IMPORTANCE The taxonomic composition and functional potential of animal gut microbiota in deep-sea environments remain largely unknown. Here, by performing comparative metagenomics, we suggest that the gut microbial compositions of two Hirondellea gigas populations from the Mariana Trench and the Japan Trench have undergone significant divergence. Through analyses of functional potentials and microbe-microbe correlations, our findings shed light on the contributions of animal gut microbiota to host adaptation to hadal environments.


Subject(s)
Amphipoda/microbiology , Archaea/physiology , Bacterial Physiological Phenomena , Gastrointestinal Microbiome/physiology , Animals , Archaea/classification , Bacteria/classification , Hydrothermal Vents
10.
J Invertebr Pathol ; 156: 41-53, 2018 07.
Article in English | MEDLINE | ID: mdl-30017949

ABSTRACT

Invasive and non-native species can pose risks to vulnerable ecosystems by co-introducing bacterial pathogens. Alternatively, co-introduced bacterial pathogens may regulate invasive population size and invasive traits. We describe a novel candidate genus and species of bacteria ('Candidatus Aquirickettsiella gammari') found to infect Gammarus fossarum, from its native range in Poland. The bacterium develops intracellularly within the haemocytes and cells of the musculature, hepatopancreas, connective tissues, nervous system and gonad of the host. The developmental cycle of 'Candidatus Aquirickettsiella gammari' includes an elementary body (496.73 nm ±â€¯37.56 nm in length, and 176.89 nm ±â€¯36.29 nm in width), an elliptical, condensed spherical stage (737.61 nm ±â€¯44.51 nm in length and 300.07 nm ±â€¯44.02 nm in width), a divisional stage, and a spherical initial body (1397.59 nm ±â€¯21.26 nm in diameter). We provide a partial genome for 'Candidatus Aquirickettsiella gammari', which clades phylogenetically alongside environmental 16S rRNA sequences from aquatic habitats, and bacterial symbionts from aquatic isopods (Asellus aquaticus), grouping separately from the Rickettsiella, a genus that includes bacterial pathogens of terrestrial insects and isopods. Increased understanding of the diversity of symbionts carried by G. fossarum identifies those that might regulate host population size, or those that could pose a risk to native species in the invasive range. Identification of 'Candidatus Aquirickettsiella gammari' and its potential for adaptation as a biological control agent is explored.


Subject(s)
Amphipoda/microbiology , Coxiellaceae/physiology , Animals , Coxiellaceae/classification , Gammaproteobacteria/classification , Gammaproteobacteria/physiology , Phylogeny , RNA, Ribosomal, 16S/genetics
11.
Sci Rep ; 8(1): 8945, 2018 06 12.
Article in English | MEDLINE | ID: mdl-29895884

ABSTRACT

Microsporidia are common parasites infecting animals and protists. They are specifically common pathogens of amphipods (Crustacea, Malacostraca), with Dictyocoela spp. being particularly frequent and highly prevalent, exhibiting a range of phenotypic and ecological effects. Until now, seven species of Dictyocoela were defined, predominantly based on the genetic distance. However, neither the taxonomic status of this provisionally erected genus (based on eight novel sequences and one micrograph of the spore), nor its internal phylogenetic relationships have been clearly revealed. The formal description of the genus and of most of the putative species are still lacking. Here we aimed to fill this gap and performed both ultrastructural and molecular studies (based on SSU, ITS and partial LSU) using different species delimitation methods. As a consensus of these results and following conservative data interpretation, we propose to distinguish five species infecting gammarid hosts, and to keep the names introduced by the authors of the type sequences: Dictyocoela duebenum, D. muelleri, D. berillonum and D. roeselum. We provide full descriptions of these species. Moreover, thanks to our extensive sampling, we extend the known host and geographic range of these Microsporidia.


Subject(s)
Amphipoda/microbiology , Microsporidia/physiology , Spores, Fungal/physiology , Animals , DNA, Fungal/chemistry , DNA, Fungal/genetics , Europe , Geography , Host-Parasite Interactions , Microscopy, Electron, Transmission , Microsporidia/classification , Microsporidia/genetics , Phylogeny , Sequence Analysis, DNA , Species Specificity , Spores, Fungal/genetics , Spores, Fungal/ultrastructure
12.
Sci Rep ; 8(1): 4691, 2018 03 16.
Article in English | MEDLINE | ID: mdl-29549322

ABSTRACT

Phylogenetically distant parasites often infect the same host. Indeed, co-infections can occur at levels greater than expected by chance and are sometimes hyperparasitic. The amphipod Echinogammarus marinus presents high levels of co-infection by two intracellular and vertically transmitted parasites, a paramyxid (Paramarteilia sp. Em) and a microsporidian strain (Dictyocoela duebenum Em). This co-infection may be hyperparasitic and result from an exploitative 'hitchhiking' or a symbiotic relationship between the parasites. However, the best-studied amphipod species are often collected from contaminated environments and may be immune-compromised. Immune-challenged animals frequently present co-infections and contaminant-exposed amphipods present significantly higher levels of microsporidian infection. This suggests the co-infections in E. marinus may result from contaminant-associated compromised immunity. Inconsistent with hyperparasitism, we find that artificial infections transmit Paramarteilia without microsporidian. Our population surveys reveal the co-infection relationship is geographically widespread but find only chance co-infection between the Paramarteilia and another species of microsporidian, Dictyocoela berillonum. Furthermore, we identify a haplotype of the Paramarteilia that presents no co-infection, even in populations with otherwise high co-infection levels. Overall, our results do not support the compromised-immunity hypothesis but rather that the co-infection of E. marinus, although non-hyperparasitic, results from a relationship between specific Paramarteilia and Dictyocoela duebenum strains.


Subject(s)
Amphipoda/microbiology , Amphipoda/parasitology , Microsporidiosis/diagnosis , Protozoan Infections, Animal/diagnosis , Animals , Coinfection , DNA, Fungal/genetics , DNA, Protozoan , Female , Host-Parasite Interactions , Male , Mass Screening/veterinary , Microsporidia/genetics , Microsporidia/isolation & purification , Microsporidiosis/transmission , Microsporidiosis/veterinary , Phylogeny , Protozoan Infections, Animal/transmission
13.
J Eukaryot Microbiol ; 65(3): 296-307, 2018 05.
Article in English | MEDLINE | ID: mdl-28922521

ABSTRACT

Five out of one hundred adults of Niphargogammarus intermedius caught at the Azov sea shore were found to be infected with microsporidia. The infection was found in the subcuticular fat body and myocytes. Parasites developed in direct contact with host cells, displayed a disporoblastic sporogony and diplokaryotic arrangement of nuclei at all stages. Spores were oval, 4.6-5.8 × 2.6-3.0 µm. Exospore appendages, vesicular-tubular secretions, and the anisofilar polar filament indicated a similarity to Anncaliia species. Sporont surfaces displayed ridges of amorphous material. Meronts and sporonts formed protoplastic extensions, similar to A. vesicularum and A. meligheti. Mature spores possessed a bipartite polaroplast. The polar tube was arranged in one row of 13-18 coils including 0-3 distal coils of lesser diameter. Partial sequencing of SSU, ITS, and LSU regions of rRNA gene (GenBank accessions: KY288064-KY288065) confirmed this new species to be congeneric with A. algerae (#AF069063) and A. meligheti (#AY894423). The SSU gene of this novel microsporidium shared 99.4% sequence similarity to A. algerae and 98.9% to A. meligheti. Genes for HSP70 and RPB1 amplified with primers designed for A. algerae orthologs displayed 99.7% and 97.4% similarity, respectively, between A. algerae and the novel microsporidium. A new species, Anncaliia azovica, is described based on morphological and molecular characterization.


Subject(s)
Amphipoda/microbiology , Microsporidia/classification , Microsporidiosis/microbiology , Animals , DNA, Fungal/genetics , DNA, Ribosomal/genetics , Microsporidia/genetics , Microsporidia/isolation & purification , Phylogeny , RNA, Fungal/genetics , RNA, Ribosomal/genetics , Spores, Fungal/physiology
14.
Environ Sci Pollut Res Int ; 24(29): 23417-23421, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28905182

ABSTRACT

Endocrine disruption compounds (EDCs) and parasitism can both interfere with the reproduction process of organisms. The amphipod Gammarus pulex is the host of the vertically transmitted microsporidia Dictyocoela duebenum, and this work was devoted to the investigation of the effect of an exposure to the anti-androgen compound, cyproterone acetate (CPA), and/or of the presence of D. duebenum on the spermatozoa production and length. Significant reduction of the spermatozoa production was observed when G. pulex males were uninfected and exposed to CPA. There also appeared a lower number of spermatozoa when D. duebenum infects G. pulex, whatever the exposure condition. Moreover, we highlighted that CPA has no effect on spermatozoa production when males are infected by D. duebenum, and no treatment has impacted the spermatozoa length. Our results suggest CPA and D. duebenum could impact the endocrine system of G. pulex and especially processes close to the spermatozoa production (e.g., androgenic gland, androgen gland hormone released, gonad-inhibiting hormone synthesized by X-organ). However, as no mechanism of action was highlighted, further testing need to be performed to improve the understanding of their impacts. Finally, results confirm that vertically transmitted microsporidia could be a confounding factor in the endocrine disruption assessments in Gammaridae.


Subject(s)
Amphipoda/microbiology , Cyproterone Acetate/toxicity , Endocrine Disruptors/toxicity , Infectious Disease Transmission, Vertical , Microsporidia/isolation & purification , Microsporidiosis/transmission , Spermatozoa/microbiology , Amphipoda/drug effects , Animals , Male , Microsporidia/physiology , Microsporidiosis/microbiology , Spermatozoa/drug effects
15.
Antonie Van Leeuwenhoek ; 110(12): 1593-1611, 2017 Dec.
Article in English | MEDLINE | ID: mdl-28721507

ABSTRACT

The emergence of pathogenic bacteria resistant to antibiotics increases the need for discovery of new effective antimicrobials. Unique habitats such as marine deposits, wetlands and caves or unexplored biological communities are promising sources for the isolation of actinobacteria, which are among the major antibiotic producers. The present study aimed at examining cultivated actinobacteria strains associated with endemic Lake Baikal deepwater amphipods and estimating their antibiotic activity. We isolated 42 actinobacterial strains from crustaceans belonging to Ommatogammarus albinus and Ommatogammarus flavus. To our knowledge, this is the first report describing the isolation and initial characterization of representatives of Micromonospora and Pseudonocardia genera from Baikal deepwater invertebrates. Also, as expected, representatives of the genus Streptomyces were the dominant group among the isolated species. Some correlations could be observed between the number of actinobacterial isolates, the depth of sampling and the source of the strains. Nevertheless, >70% of isolated strains demonstrated antifungal activity. The dereplication analysis of extract of one of the isolated strains resulted in annotation of several known compounds that can help to explain the observed biological activities. The characteristics of ecological niche and lifestyle of deepwater amphipods suggests that the observed associations between crustaceans and isolated actinobacteria are not random and might represent long-term symbiotic interactions.


Subject(s)
Actinobacteria/classification , Actinobacteria/physiology , Amphipoda/microbiology , Anti-Bacterial Agents/biosynthesis , Lakes/microbiology , Water Microbiology , Actinobacteria/genetics , Actinobacteria/isolation & purification , Animals , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
16.
Parasit Vectors ; 10(1): 193, 2017 Apr 20.
Article in English | MEDLINE | ID: mdl-28427445

ABSTRACT

BACKGROUND: Whilst vastly understudied, pathogens of non-native species (NNS) are increasingly recognised as important threats to native wildlife. This study builds upon recent recommendations for improved screening for pathogens in NNS by focusing on populations of Gammarus roeselii in Chojna, north-western Poland. At this location, and in other parts of continental Europe, G. roeselii is considered a well-established and relatively 'low-impact' invader, with little understanding about its underlying pathogen profile and even less on potential spill-over of these pathogens to native species. RESULTS: Using a combination of histological, ultrastructural and phylogenetic approaches, we define a pathogen profile for non-native populations of G. roeselii in Poland. This profile comprised acanthocephalans (Polymorphus minutus Goese, 1782 and Pomphorhynchus sp.), digenean trematodes, commensal rotifers, commensal and parasitic ciliated protists, gregarines, microsporidia, a putative rickettsia-like organism, filamentous bacteria and two viral pathogens, the majority of which are previously unknown to science. To demonstrate potential for such pathogenic risks to be characterised from a taxonomic perspective, one of the pathogens, a novel microsporidian, is described based upon its pathology, developmental cycle and SSU rRNA gene phylogeny. The novel microsporidian Cucumispora roeselii n. sp. displayed closest morphological and phylogenetic similarity to two previously described taxa, Cucumispora dikerogammari (Ovcharenko & Kurandina, 1987), and Cucumispora ornata Bojko, Dunn, Stebbing, Ross, Kerr & Stentiford, 2015. CONCLUSIONS: In addition to our discovery extending the host range for the genus Cucumispora Ovcharenko, Bacela, Wilkinson, Ironside, Rigaud & Wattier, 2010 outside of the amphipod host genus Dikerogammarus Stebbing, we reveal significant potential for the co-transfer of (previously unknown) pathogens alongside this host when invading novel locations. This study highlights the importance of pre-invasion screening of low-impact NNS and, provides a means to document and potentially mitigate the additional risks posed by previously unknown pathogens.


Subject(s)
Amphipoda/parasitology , Host-Parasite Interactions , Amphipoda/microbiology , Amphipoda/pathogenicity , Animals , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/pathogenicity , Microsporidia/classification , Microsporidia/genetics , Microsporidia/isolation & purification , Microsporidia/pathogenicity , Parasites/classification , Parasites/genetics , Parasites/isolation & purification , Parasites/pathogenicity , Phylogeny , Poland , Species Specificity , Symbiosis , Trematoda/classification , Trematoda/genetics , Trematoda/isolation & purification , Trematoda/pathogenicity
17.
Int J Syst Evol Microbiol ; 67(4): 824-831, 2017 Apr.
Article in English | MEDLINE | ID: mdl-27902293

ABSTRACT

An obligately piezophilic strain was isolated from an amphipod crustacean obtained in the Challenger Deep region of the Mariana Trench during the DEEPSEA CHALLENGE expedition. The strain, MTCD1T, grew at extremely high hydrostatic pressures, with a growth range of 80-140 MPa (optimum, 120 MPa) at 6 °C. Phylogenetic analyses based on the 16S rRNA gene sequence indicate that it is closely affiliated with the genus Colwellia. Comparative 16S rRNA gene sequence analyses revealed 95.7, 95.5 and 95.2 % similarity to Colwellia maris ABE-1T, Colwellia piezophila Y233GT and Colwellia psychrerythraea ATCC 27364T, respectively. The major cellular fatty acids were C16 : 1, C16 : 0 and C22 : 6 (docosahexaenoic acid), and the sole isoprenoid quinone produced was ubiqinone-8. DNA G+C content was 48.6 mol%. The strain was positive for oxidase and catalase activities. Based on the results from this study, strain MTCD1T is a novel Gram-negative species of the genus Colwellia, and the name Colwellia marinimaniae sp. nov. (type strain MTCD1T=ATCC TSD-5T=JCM 30270T) is proposed. It is the most piezophilic organism yet described.


Subject(s)
Alteromonadaceae/classification , Amphipoda/microbiology , Phylogeny , Alteromonadaceae/genetics , Alteromonadaceae/isolation & purification , Animals , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , DNA, Ribosomal/genetics , Fatty Acids/chemistry , Hydrostatic Pressure , Nucleic Acid Hybridization , Pacific Ocean , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/chemistry
18.
Res Microbiol ; 168(1): 74-84, 2017 Jan.
Article in English | MEDLINE | ID: mdl-27531409

ABSTRACT

Talitrid amphipods (sandhoppers and beach fleas) are typical of the supralittoral zone. They are known to thrive on stranded materials, including detrital marine angiosperms and macroalgae, as well as occasional dead animals. In this work, the gut microbiota of five species of talitrid amphipods (Talitrus saltator, Talorchestia ugolinii, Sardorchestia pelecaniformis, Orchestia montagui and Orchestia stephenseni) collected in Sardinia (Italy) has been investigated through: i) metabarcoding analysis of the amplified 16S rRNA V4 region; and ii) quantification of family 48 glycosyl hydrolase genes (GHF48), involved in cellulose degradation. Results indicate that, though talitrid gut biodiversity is not directly related to taxon or sampling locality, the animals' digestive tracts may host species-specific bacterial communities. In particular, gut microbiota of O. montagui, an inhabitant of Posidonia banquettes and macro-algae mat, showed the greatest differences in taxonomic composition and the highest proportion of GHF48 genes with respect to 16S rRNA genes. These results suggest that the different talitrid species may host species-specific bacterial communities whose function could partially reflect the different microhabitats and food preferences of their host.


Subject(s)
Amphipoda/classification , Amphipoda/microbiology , Stramenopiles/isolation & purification , Animals , DNA Barcoding, Taxonomic , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Gastrointestinal Microbiome , Gastrointestinal Tract/microbiology , Glycoside Hydrolases/genetics , Italy , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Stramenopiles/metabolism
19.
Ecotoxicol Environ Saf ; 133: 188-94, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27454203

ABSTRACT

The protozoa Toxoplasma gondii and Cryptosporidium parvum are public health priorities because their oocysts can persist in recreational, surface, drinking, river, and sea water sources for a long time. To evaluate the capacity of the freshwater crustacean Gammarus fossarum to accumulate T. gondii and C. parvum oocysts, gammarids were exposed to 200, 2000 or 20,000 oocysts per gammarid and per day for 21 days followed by 5 days of depuration. C. parvum DNA was detected by qPCR in G. fossarum in only one out of four pools for the highest concentration and after 14 days of exposure, and T. gondii DNA was detected after 7 days of exposure to the two highest concentrations. Our results document the capacity of G. fossarum to accumulate T. gondii in its tissues proportionally to the ambient concentration; the maximum number of oocysts was detected in gammarid tissues after exposure to 20,000 oocysts per day. Mean values of 3.26 (±3), 21.71 (±15.18), and 17.41 (±10.89) oocysts were detected in gammarids after 7, 14, and 21 days, respectively, and after 5 days of depuration, T. gondii oocysts were still present in gammarid tissues. These results show for the first time that a freshwater crustacean can bioaccumulate T. gondii oocysts, suggesting that G. fossarum is a potential effective bioindicator of protozoan contamination in biomonitoring studies. Moreover, due to its key position in freshwater food webs, G. fossarum could also play a role in the trophic transfer of protozoa.


Subject(s)
Amphipoda/microbiology , Amphipoda/parasitology , Cryptosporidium , Environmental Monitoring/methods , Toxoplasma , Animals , Fresh Water , Oocysts , Real-Time Polymerase Chain Reaction , Rivers , Seawater , Shellfish , Surveys and Questionnaires
20.
Folia Microbiol (Praha) ; 61(2): 159-68, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26347255

ABSTRACT

The high demand for new antibacterials fosters the isolation of new biologically active compounds producing actinobacteria. Here, we report the isolation and initial characterization of cultured actinobacteria from dominant benthic organisms' communities of Lake Baikal. Twenty-five distinct strains were obtained from 5 species of Baikal endemic macroinvertebrates of amphipods, freshwater sponges, turbellaria worms, and insects (caddisfly larvae). The 16S ribosomal RNA (rRNA)-based phylogenic analysis of obtained strains showed their affiliation to Streptomyces, Nocardia, Pseudonocardia, Micromonospora, Aeromicrobium, and Agromyces genera, revealing the diversity of actinobacteria associated with the benthic organisms of Lake Baikal. The biological activity assays showed that 24 out of 25 strains are producing compounds active against at least one of the test cultures used, including Gram-negative bacteria and Candida albicans. Complete dereplication of secondary metabolite profiles of two isolated strains led to identification of only few known compounds, while the majority of detected metabolites are not listed in existing antibiotic databases.


Subject(s)
Actinobacteria/isolation & purification , Invertebrates/microbiology , Actinobacteria/chemistry , Actinobacteria/classification , Actinobacteria/metabolism , Amphipoda/microbiology , Animals , Anti-Infective Agents/chemistry , Anti-Infective Agents/metabolism , Bacteria/drug effects , Fungi/drug effects , Insecta/microbiology , Lakes/microbiology , Phylogeny , Porifera/microbiology
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