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Different types of yeasts and lactic acid bacteria dominate in spontaneously fermented products (food, beverages, and condiments) that are commonly consumed in Ethiopia. The aim of this study was to identify efficient fermentative yeasts from fermented foods, fermented beverages, honey and molasses using genotypic methods. Out of the 70 samples tested, 180 distinct wild yeast isolates were recovered. A total of 23 isolates were selected for genomic analysis based on their basis of biomass yield, fermentation capacity, and leavening performance. The nucleotide sequence analysis of the internal transcribed spacer ITS-5.8S rDNA region revealed that the indigenous yeast isolates had close relatedness to Saccharomycescerevisiae, Candida humilis, Kazachstania bulderi, Pichia fermentans and Pichia kudriavzevii with greater than 97% nucleotide similarity. The study shows a high diversity of indigenous wild yeasts in fermented products and that potent strains had higher biomass yield, good gas production and remarkable leavening capacity that indicates their inherent potential for use in the baking industry. [ABSTRACT FROM AUTHOR]

Two Pyrenopeziza species, P. protrusa and P. nervicola known to have similar morphology, were newly reported from Japan. This study clarified that P. protrusa morphologically differs from P. nervicola in (1) more protruded outermost cells in ectal excipulum, (2) smaller ascospores, and (3) longer and thinner asci. Pyrenopeziza protrusa produces chlamydospore-like brown cells under culture while P. nervicola forms more complex, brown bulbils. These species formed distinct clades based on ITS-5.8S rDNA in the phylogenetic analysis. The two species did not share the same host. • Two Pyrenopeziza species were newly reported from Japan. • The two species produce characteristic structures in culture previously unreported. • The two species were well-defined by barcoding sequences. [ABSTRACT FROM AUTHOR]

During a survey of trichodinids in Chongqing, China, two populations of Trichodina reticulata Hirschmann & Partsch, 1955 were isolated from gills of the freshwater fishes, Carassius auratus and Ctenopharyngodon idellus, and 11 molecular samples containing small subunit (SSU) rDNA and internal transcribed spacer (ITS)‐5.8S rDNA were newly sequenced. Both populations fell within the range of morphometry and corresponded well with the diagnostic morphological features of Trichodina reticulata Hirschmann & Partsch, 1955. It should be noted that one population possessed obvious central granules in the adhesive disc, while the other one did not. The detailed morphological redescription for these two populations is given in this work. Although some intraspecific differences were found, both populations were confirmed as Trichodina reticulata based on the morphological study and molecular data, including the sequence analysis of the primary and secondary structures of SSU rDNA, genetic distances and phylogenetic tree. Our study supports the assertion that central granules in the adhesive disc might be an auxiliary feature rather than a heritable character for determining taxonomic affiliations among trichodinids. [ABSTRACT FROM AUTHOR]

• Ostreopsis lenticularis has been reinvestigated from French Polyensia, its type locality. • 47 strains have been isolated and cultured from four archipelagos. • Morphology and LSU / ITS–5.8S rDNA sequences were identical for all strains and wild specimens. • From phylogenies, O. lenticularis clustered unambiguously within the clade Ostreopsis sp. 5. • No toxic effect was found on 19 strains tested using CBA-N2a and no PlTX-like molecules were detected in 4 analyzed strains. To date, the genus Ostreopsis comprises eleven described species, of which seven are toxigenic and produce various compounds presenting a major threat to human and environmental health. The taxonomy of several of these species however remains controversial, as it was based mostly on morphological descriptions leading, in some cases, to ambiguous interpretations and even possible misidentifications. The species Ostreopsis lenticularis was first described by Y. Fukuyo from French Polynesia using light microscopy observations, but without genetic information associated. The present study aims at revisiting the morphology, molecular phylogeny and toxicity of O. lenticularis based on the analysis of 47 strains isolated from 4 distinct locales of French Polynesia, namely the Society, Australes, Marquesas and Gambier archipelagos. Observations in light, epifluorescence and field emission scanning electron microscopy of several of these strains analyzed revealed morphological features in perfect agreement with the original description of O. lenticularis. Cells were oval, not undulated, 60.5–94.4 μm in dorso-ventral length, 56.1–78.2 μm in width, and possessed a typical plate pattern with thecal plates showing two sizes of pores. Phylogenetic analyses inferred from the LSU rDNA and ITS–5.8S sequences revealed that the 47 strains correspond to a single genotype, clustering with a strong support with sequences previously ascribed to Ostreopsis sp. 5. Clonal cultures of O. lenticularis were also established and further tested for their toxicity using the neuroblastoma cell-based assay and LC MS/MS analyses. None of the 19 strains tested showed toxic activity on neuroblastoma cells, while LC MS/MS analyses performed on the strains from Tahiti Island (i.e. type locality) confirmed that palytoxin and related structural analogs were below the detection limit. These findings allow to clarify unambiguously the genetic identity of O. lenticularis while confirming previous results from the Western Pacific which indicate that this species shows no toxicity, thus stressing the need to reconsider its current classification within the group of toxic species. [ABSTRACT FROM AUTHOR]

Chattonella species in a Mediterranean lagoon (Santa Giusta Lagoon, Sardinia, Italy) were identified by applying a molecular approach to fixed natural phytoplankton samples collected over the last two decades. Like the other raphidophytes, Chattonella cells are naked and lose their shape when fixed, making species identification difficult on the basis of their morphological characteristics. Employing species-specific primers (oBTG-005-F, oBTG-027-R, oBTG-028-R) for the amplification of the ITS-5.8S rDNA region, we established the occurrence of C. subsalsa in fixed natural phytoplankton samples collected in coincidence with fish death events. Additionally, we established the presence of the recently discovered C. cf. subsalsa Adriatic genotype by analysing cellular cultures obtained from the same lagoon in 2013. This is the second worldwide record of C. cf. subsalsa Adriatic genotype. Our results revealed that the species-specific primers oBTG-005-F and oBTG-028-R distinguished this new genotype only when present singularly. This study provides valuable data that increase knowledge of C. subsalsa genotypes and of the long-term occurrence of Chattonella blooms in a transitional ecosystem through the use of samples up to 20 years old.

A crown canker disease causing yellowing and wilting of the leaves was observed on muskmelon (Cucumis melo) plants in a number of greenhouses in Shouguang county, Shandong province, China, during April 2014 to July 2015, resulting in serious economic losses. Black sporodochia with white mycelial tufts emerged on surfaces of lesions at the base of the stem. Eleven fungal strains were isolated from infected tissues. On the basis of morphological features, ITS-5.8S rDNA and TEF1 sequence analysis, the isolates were identified asParamyrothecium roridum(Tode) L. Lombard & Crous (syn.Myrothecium roridumTode). The pathogenicity of the isolates was confirmed. The morphological characteristics of reisolated strains from inoculated muskmelon plants showing symptoms of crown canker were consistent with the original strains. A host range test indicated that strains had a wide range of hosts. To the best of our knowledge, this is the first report of muskmelon crown canker caused byP. roridumin China. This disease could have a significant economic impact on muskmelon production. D’avril 2014 à juillet 2015, un chancre du collet causant le jaunissement et le flétrissement des feuilles a été observé chez des plants de melon brodé (Cucumis melo) dans un certain nombre de serres dans le comté de Shouguang, dans la province du Shandong, en Chine, ce qui a occasionné d’importantes pertes financières. Des conidies noires accompagnées de touffes mycéliennes blanches émergeaient à la surface des lésions formées à la base de la tige. Onze souches fongiques ont été isolées à partir de tissus infectés. En se basant sur les traits morphologiques, l’ITS-5.8S de l’ADNr et l’analyse des séquences du TEF1, les isolats ont été identifiés en tant queParamyrothecium roridum(Tode) L. Lombard & Crous (syn.Myrothecium roridumTode). La pathogénicité des isolats a été confirmée. Les caractéristiques morphologiques des souches isolées de nouveau, issues de plants de melon brodé inoculés affichant des symptômes du chancre du collet, correspondaient à celles des souches originales. Un test de gamme d’hôtes a indiqué que les souches possédaient une vaste gamme d’hôtes. À notre connaissance, il s’agit de la première mention de chancre du collet chez le melon brodé causé parP. roridumen Chine. Cette maladie pourrait entraîner de fâcheuses conséquences financières pour la production de melon brodé. [ABSTRACT FROM AUTHOR]

In the last few decades, the frequency of the toxic benthic dinoflagellate Ostreopsis cf. ovata proliferation has increased in the Mediterranean Sea. These blooms are associated with harmful effects on human health and the environment. The present work provides the first long term study on the spatio-temporal distribution of O . cf. ovata in relation to physical parameters in the Gulf of Gabès coastal waters (south-eastern Mediterranean Sea), as well as its morphological, molecular and physiological features. The strains of O. cf. ovata were identified morphologically by light and epifluorescence microscopy. The morphology and the size range of cultured strains were similar to those described regarding O . cf. ovata isolated from the Mediterranean Sea. The ultrastructural analysis of O . cf. ovata cells using the transmission electron microscopy showed the presence of numerous vesicles (VE) containing spirally coiled fibers (SCFs) connected to the mucus canal (CH). The phylogenetic tree based on the internal transcribed spacer region containing the 5.8S rDNA (ITS-5.8S rDNA) revealed that O . cf. ovata strains were placed into the Mediterranean/Atlantic clade. In addition, O . cf. ovata toxicity was evaluated by the mouse bioassay and a dose level ≥ 4 × 10 4 cells was found to be lethal to mice. The examination of the O. cf. ovata occurrence in the Gulf of Gabès at a large temporal scale (1997–2012) revealed a clear seasonal pattern with dominance from midsummer (July) to late autumn (November). Furthermore, a positive correlation was found between the abundance of O. cf. ovata and salinity, whereas no correlation was found as regards temperature. The occurrence of O. cf. ovata was only detected at salinity above 35 and the highest concentrations were observed at 45. Laboratory experiments confirmed such a result and showed that isolated O . cf. ovata strains had optimal growth at salinity ranging between 35 and 45, with its peak at 40. [ABSTRACT FROM AUTHOR]

Stemphylium lycopersici (Enjoji) W. Yamam was initially described from tomato and has been reported to infect different hosts worldwide. Sequence analyses of the internal transcribed spacer (ITS) regions 1 and 2, including 5.8S rDNA (ITS-5.8S rDNA) and glyceraldehyde-3-phosphate dehydrogenase ( gpd) gene, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), as well as virulence studies were conducted to analyze 46 S. lycopersici isolates. Stemphylium lycopersici isolates used in this study were obtained from diseased tomato ( Solanum lycopersicum L.), eggplant ( Solanum melongena L.), pepper ( Capsicum annuum L.) and lettuce ( Lactuca sativa L.) from major vegetable growing regions of Malaysia, including the three states of Pahang, Johor and Selangor between 2011 and 2012. Phylogenetic analysis of a combined dataset of the ITS-5.8S rDNA and gpd regions indicated that all isolates were clustered in the sub-cluster that comprised S. lycopersici, and were distinguished from other Stemphylium species. Cluster analyses using the UPGMA method for both RAPD and ISSR markers grouped S. lycopersici isolates into three main clusters with similarity index values of 67 and 68 %. The genetic diversity data confirmed that isolates of S. lycopersici are in concordance to host plants, and not geographical origin of the isolates. All S. lycopersici isolates were pathogenic on their original host plants and showed leaf spot symptoms; however, virulence variability was observed among the isolates. In cross-inoculation assays, the representative isolates were able to cause leaf spot symptoms on eggplant, pepper, lettuce and tomato, but not on cabbage. [ABSTRACT FROM AUTHOR]

A leaf spot disease was observed on cabbage (Brassica oleraceaL. var.capitata) affecting 80% of plants growing in greenhouses and fields in Serdang, Selangor, Malaysia. Symptomatic leaf samples were collected from infected plants and isolations made on agar medium. Single-spore isolates from resulting colonies were identified based on cultural and morphological characteristics asPithomyces chartarum. Morphological identification was confirmed by sequence analysis of the internal transcribed spacer (ITS) regions 1 and 2, including 5.8S rDNA (ITS1-5.8S-ITS2). Pathogenicity tests indicated thatP. chartarumcauses leaf spot on cabbage. This is the first report of leaf spot caused byP. chartarumon cabbage in Malaysia. [ABSTRACT FROM PUBLISHER]

Seventy-eight marine fungal strains were isolated from sediment samples collected off the coast of Nanji Island, Wenzhou, Zhejiang Province, China. Antibacterial screening using the agar disc method showed that 19 of the isolated strains could inhibit at least one pathogenic V ibrio from P seudosciaena crocea. Subsequent screening confirmed that nine strains produced antibacterial metabolites that had activity against one or several types of pathogenic V ibrio. Strain NJ0104 had the widest antimicrobial spectrum and strong activity, particularly against Vibrio parahaemolyticus-MM0810072. A preliminary study of NJ0104 antibacterial metabolites demonstrated that they had thermal stability up to 80°C, ultraviolet stability up to 40 min and pH stability between 4.0-7.0. In addition, the antibacterial metabolites were readily soluble in butanol. To identify the specific strain, the ITS-5.8S rDNA regions of NJ0104 were PCR amplified and sequenced. Based on the combination of phenotypic and genotypic data, the strain was identified as Arthrinium sp. [ABSTRACT FROM AUTHOR]

Abstract: Fibrocapsa japonica resting cysts were detected in coastal sediments of the Marche region (northern Adriatic Sea, Italy), where summer blooms regularly occur. Sampling was carried out along 18 transects (2 stations each) during May–June 2007. The identification and counting of raphidophyte cysts were carried out under the inverted microscope and molecular analyses were applied to sediment samples to confirm species-specific identification. Raphidophyte cysts were recorded in almost all stations with maximum abundance of 9±5 and 12±3cystsg−1 dw for cysts of F. japonica and undetermined Raphidophyceae, respectively. Cyst formation was induced in F. japonica monoclonal cultures under different conditions: shadow-optimal temperature and dark-low temperature treatment; both experiments were performed at two salinity values (30 and 35). Cyst formation (preliminary cysts) was observed in both experiments, although with different encystment rates. However, only a further permanence in the dark at 15°C in microplates led to the formation of mature cysts, probably supported by the possibility to adhere to solid surfaces. Before pre-cyst formation, vegetative cells showed a cytoplasmatic “brown body”. [ABSTRACT FROM AUTHOR]

Aspergillus genus is an ubiquitous fungal group that colonizes a wide range of substrates. A total of 100 Tunisian fungal strains isolated at harvest time from fresh and dried grapes were identified within the section Nigri and tested for their ochratoxin A (OTA) producing abilities. Of the isolates, 45% were identified as Aspergillus tubingensis, 34% as Aspergillus niger, 12% as Aspergillus japonicus and 9% as Aspergillus carbonarius. The OTA production was assessed using Czapeck yeast extract agar which revealed that 25% of the isolates belonging to the Aspergillus niger aggregate were OTA producers, while 97% were identified as belonging to the A. Carbonarius species. To distinguish between A. Niger and A. Tubingensis, PCR was used to amplify the ITS-5.8S rDNA for these two strains. The sequence analysis of the PCR products revealed a full similarity between the two species. Thus, a new molecular strategy has been developed based on the targeting of the gene from nitrate reductase ( niiA) and nitrite reductase gene ( niaD), as well as the intergenic region of the gene ( niiA-niaD). The subsequent screening revealed that two new specific primer pairs can be used to identify specifically A. Niger and A. Tubingensis. [ABSTRACT FROM AUTHOR]

Pythium insidiosum causes pythiosis, a life-threatening disease that occurs in tropical regions and affects man and animals. Although pythiosis in Brazil had been described in various animal species, the first human case was only recently reported. The present study aimed to characterize the morphologic and molecular characteristics of a new equine isolate of P. insidiosum and compare them with those of the first Brazilian human isolate. Both isolates were recovered from the same region of the country. Macroscopic and microscopic features were evaluated in two culture media. Sporangia formation and zoospore release were obtained after culturing the isolates with fragments of grasses and crops in an appropriate liquid induction medium. The molecular analysis of the isolates consisted of the complete sequencing of the ITS-5.8S rDNA region and sequences of both showed identical composition of 836 bp and 99% similarity with the isolates M16, 65, M12, 339 and 394 deposited at GenBank. Simple mycological procedures such as the production of sporangia and zoospores may distinguish P. insidiosum from zygomycetes. The rDNA sequencing indicates that, in Brazil, both humans and animals might be infected by a common genotype of the pathogen. [ABSTRACT FROM AUTHOR]

Abstract: The effects of temperature, salinity, inorganic and organic nutrients on the growth and physiological performance of Fibrocapsa japonica have been investigated in strains isolated from the northern Adriatic Sea, where intense and regular blooms have occurred since 1997 in coastal waters during summer. Strains isolated at different times and from different locations appeared homogeneous in terms of both physiological responses and molecular (ITS-5.8S rDNA) characteristics. Growth rates were higher at temperatures between 20 and 26°C and in a salinity range of 30–35 (0.7divday−1). The temperature of 16°C inhibited growth, more markedly at the lowest and highest salinity values, a result also confirmed by a lower photosynthetic efficiency and by an increase in cell volume due to impaired division. Higher cell concentrations were obtained with macronutrients at f/2 levels than in a fivefold diluted medium. Comparing the utilization of 200μM nitrate to that of different N sources (inorganic and organic), F. japonica showed an efficient growth with equivalent amounts of ammonia, urea and amino acids, such as glycine and tryptophan; glutamate was less effective, while methionine had toxic effects. Organic phosphate, administered as glycerophosphate, could also sustain F. japonica growth, probably on account of an alkaline phosphatase whose activity was enhanced in the presence of the organic form. Vitamins were necessary for growth, though no further stimulation was observed when a surplus of vitamin B12 (3nM) was added. The addition of 11.7μM iron instead of 2.3μM, as well as that of humic acid, with or without macronutrients, did not enhance algal growth either. These results led us to hypothesise that F. japonica blooms became more frequent due to a general seawater temperature increase and to the availability of organic forms which, in coastal anthropized areas, are especially abundant in summer periods. [Copyright &y& Elsevier]

Recurrent fish kills in the Spanish Alfacs Bay (NW Mediterranean) have been detected during winter seasons since 1994, and were attributed to an unarmored, ichthyotoxic, dinoflagellate, initially identified as Gyrodinium corsicum Paulmier, Berland, Billard, & Nezan. Several strains were isolated from the bay and their clonal cultures were compared by combined techniques, including light and electron microscopy, internal transcribed spacer and 5.8S rDNA nucleotide sequencing, and HPLC pigment analyses, together with studies of their photochemical performance, growth rates, and toxicity. Using phylogenetic analyses, all strains were identified as members of the genus Karlodinium, but they were separated into two genetically distinct groups. These groups, identified as Karlodinium veneficum (Ballantine) J. Larsen and K. armiger Bergholtz, Daugbjerg et. Moestrup, were also supported by the other techniques used. Detailed analyses of fine structural characteristics (including plug-like structures in amphiesma and a possible layer of semi-opaque material beneath the outer membrane) allowed discrimination of the mentioned two species. Specific differences in pigment patterns coincided with that expected for low- ( K. veneficum) and high-light ( K. armiger) adapted relatives. The higher photosynthetic efficiency of K. veneficum and the longer reactivation times of the PSII reaction centers observed for K. armiger were in agreement with this hypothesis. The two species differed in toxicity, but the strains used always induced mortality when incubated with bivalves, rotifers, and finfish. Compared with K. armiger, strains of K. veneficum yielded higher cell densities, but had lower growth rates. [ABSTRACT FROM AUTHOR]

Among dinoflagellates responsible for benthic harmful algal blooms, the genus Ostreopsis primarily described from tropical areas has been increasingly reported from subtropical and temperate areas worldwide. Several species of this toxigenic genus produce analogs of palytoxin, thus representing a major threat to human and environmental health. The taxonomy of several species needs to be clarified as it was based mostly on morphological descriptions leading in some cases to ambiguous interpretations and misidentifications. The present study aims at reporting a benthic bloom that occurred in April 2019 in Tahiti island, French Polynesia. A complete taxonomic investigation of the blooming Ostreopsis species was realized using light, epifluorescence and field emission electron microscopy and phylogenetic analyses inferred from LSU rDNA and ITS-5.8S rDNA regions. Toxicity of a natural sample and strains isolated from the bloom was assessed using both neuroblastoma cell-based assay and LC-MS/MS analyses. Morphological observations showed that cells were round to oval, large, 58.0-82.5 µm deep (dorso-ventral length) and 45.7-61.2 µm wide. The cingulum was conspicuously undulated, forming a 'V' in ventral view. Thecal plates possessed large pores in depressions, with a collar rim. Detailed observation also revealed the presence of small thecal pores invisible in LM. Phylogenetic analyses were congruent and all sequences clustered within the genotype Ostreopsis sp. 6, in a subclade closely related to sequences from the Gulf of Thailand and Malaysia. No toxicity was found on the field sample but all the strains isolated from the bloom were found to be cytotoxic and produced ostreocin D, a lower amount of ostreocins A and B and putatively other compounds. Phylogenetic data demonstrate the presence of this species in the Gulf of Thailand, at the type locality of O. siamensis, and morphological data are congruent with the original description and support this identification., (Copyright © 2020 Elsevier B.V. All rights reserved.)