2.5D transfer deep learning model for segmentation of contrast-enhancing lesions on brain magnetic resonance imaging of multiple sclerosis and neuromyelitis optica spectrum disorder.

Background: Multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) are the two mimic autoimmune diseases of the central nervous system, which are rare in East Asia. Quantitative detection of contrast-enhancing lesions (CELs) on contrast-enhancing T1-weighted magnetic resonance (MR) images is of great significance for assessing the disease activity of MS and NMOSD. However, it is challenging to develop automatic segmentation algorithms due to the lack of data. In this work, we present an automatic segmentation model of CELs based on Fully Convolutional with Attention DenseNet (FCA-DenseNet) and transfer learning strategy to address the challenge of CEL quantification in small-scale datasets. Methods: A transfer learning approach was employed in this study, whereby pretraining was conducted using 77 MS subjects from the open access datasets (MICCAI 2016, MICCAI 2017, ISBI 2015) for white matter hyperintensity segmentation, followed by fine-tuning using 24 MS and NMOSD subjects from the local dataset for CEL segmentation. The proposed FCA-DenseNet combined the Fully Convolutional DenseNet and Convolutional Block Attention Module in order to improve the learning capability. A 2.5D data slicing strategy was used to process complex 3D MR images. U-Net, ResUNet, TransUNet, and Attention-UNet are used as comparison models to FCA-DenseNet. Dice similarity coefficient (DSC), positive predictive value (PPV), true positive rate (TPR), and volume difference (VD) are used as evaluation metrics to evaluate the performances of different models. Results: FCA-DenseNet outperforms all other models in terms of all evaluation metrics, with a DSC of 0.661±0.187, PPV of 0.719±0.201, TPR of 0.680±0.254, and VD of 0.388±0.334. Transfer learning strategy has achieved success in building segmentation models on a small-scale local dataset where traditional deep learning approaches fail to train effectively. Conclusions: The improved FCA-DenseNet, combined with transfer learning strategy and 2.5D data slicing strategy, has successfully addressed the challenges in constructing deep learning models on small-scale datasets, making it conducive to clinical quantification of brain CELs and diagnosis of MS and NMOSD.

Transcriptional profile reveals the physiological responses to prey availability in the mixotrophic chrysophyte Poterioochromonas malhamensis.

Mixotrophic flagellates, which have diverse nutritional modes and play important roles in connecting the microbial loop with the classical food chain, are ideal models to study the mechanisms of adaptation between different nutritional modes in protists. In their natural ecosystems, mixotrophic flagellates may encounter microalgal prey of different digestibility, which may affect the carbon flow. To date, a molecular biological view of the metabolic processes in the mixotrophic flagellate Poterioochromonas malhamensis during nutritional adaptation and feeding on microalgal prey of different digestibility is still lacking. Accordingly, this study focused on the gene expression differences in P. malhamensis under autotrophy, being fed by the digestible microalga Chlorella sorokiniana GT-1, and being fed by the indigestible microalga C. sorokiniana CMBB-146. Results showed that the growth rate of P. malhamensis under autotrophy was much lower than that when fed by digestible microalgae. Addition of C. sorokiniana CMBB-146 could only increase the growth rate of P. malhamensis in the first 3 days, but the cell concentration of P. malhamensis started to decrease gradually after 4 days. Compared to autotrophic P. malhamensis, total 6,583 and 3,510 genes were significantly and differentially expressed in P. malhamensis fed by digestible microalgae and indigestible microalgae, respectively. Compared to autotrophic cells, genes related to the ribosome, lysosome, glycolysis, gluconeogenesis, TCA cycle, ß-oxidation, duplication, and ß-1,3-glucan in P. malhamensis grazing on digestible prey were up-regulated, while genes related to light harvesting and key enzymes referring to chlorophyll were down-regulated. Genes related to apoptosis and necrosis in P. malhamensis were up-regulated after grazing on indigestible microalgae compared to the autotrophic group, which we suggest is associated with the up-regulation of genes related to lysosome enzymes. This study provides abundant information on the potential intracellular physiological responses of P. malhamensis during the process of nutritional adaptation.

A New Algivorous Heterolobosean Amoeba, Euplaesiobystra perlucida sp. nov. (Tetramitia, Discoba), Isolated from Pilot-Scale Cultures of Phaeodactylum tricornutum.

The diatom Phaeodactylum tricornutum is regarded as a prospective "cell factory" for the high-value products fucoxanthin and eicosapentaenoic acid (EPA). However, contamination with grazing protozoa is a significant barrier to its commercial cultivation. Here, we describe a new species of heterolobosean amoeba, Euplaesiobystra perlucida, which caused the loss of Phaeodactylum tricornutum in pilot-scale cultures. Morphological and molecular characteristics distinguish E. perlucida from the other species in the genus Euplaesiobystra. E. perlucida is 1.4 to 3.2 times larger than other Euplaesiobystra species in terms of average length/width and maximum length/width of the trophozoites. Unlike Euplaesiobystra salpumilio, E. perlucida has no cytostome; E. perlucida lacks a flagellate stage, whereas Euplaesiobystra hypersalinica and E. salpumilio both display a flagellate stage in their life cycle. The small-subunit rRNA gene sequence of E. perlucida shared only 88.02% homology with that of its closest relative, Euplaesiobystra dzianiensis, and had two distinctive regions. Its phylogenetic branch was clustered with one uncultured heterolobosean clone (bootstrap support/posterior probability = 100%/1.00). Results of feeding experiments demonstrated that E. perlucida could graze on various unicellular and filamentous eukaryotic microalgae (chlorophytes, chrysophytes, euglenids, and diatoms) and cyanobacteria. E. perlucida's ingestion rate declined exponentially with increasing size of unicellular prey, and E. perlucida attained the highest growth rates on P. tricornutum. On the basis of its strong ability to graze on microalgae, capacity to form large populations in a short period of time, and capacity to form resistant resting cysts, this contaminant has the potential to cause severe problems in large-scale microalgal culture and merits further attention. IMPORTANCE Heteroloboseans have garnered considerable interest because of their extraordinary ecological, morphological, and physiological diversity. Many heteroloboseans have adapted to various extensive habitats, including halophilic, acidophilic, thermophilic, psychrophilic, and anaerobic habitats. Most heteroloboseans are bacterivores, with a few algivorous species reported. In this study, a new species of algivorous heterolobosean amoeba, Euplaesiobystra perlucida, is described as a significant grazer that causes losses in outdoor industrial Phaeodactylum cultures. This study provides phenotypic, feeding, and genetic information on a previously unknown heterolobosean, emphasizes the impact of contaminating amoebae in commercial microalgal cultures, and will contribute to the management strategies for predicting this kind of contaminant in large-scale microalgal cultivation.

Application of microalgae Scenedesmus acuminatus enhances water quality in rice-crayfish culture.

Improper management of aquatic environments substantially restricts the development of the aquaculture industry. The industrialisation of the crayfish Procambarus clarkii, for example, is currently being limited by poor water quality. Research suggests that microalgal biotechnology has a great potential for water quality regulation. However, the ecological effects of microalgal applications on aquatic communities in aquaculture systems remain largely unknown. In the present study, 5 L Scenedesmus acuminatus GT-2 culture (biomass 120 g L-1) was added to an approximately 1,000 m2 rice-crayfish culture to examine the response of aquatic ecosystems to microalgal application. The total nitrogen content decreased significantly as a result of microalgal addition. Moreover, the microalgal addition changed the bacterial community structure directionally and produced more nitrate reducing and aerobic bacteria. The effect of microalgal addition on plankton community structure was not obvious, except for a significant difference in Spirogyra growth which was inhibited by 81.0% under microalgal addition. Furthermore, the network of microorganisms in culture systems with the added microalga had higher interconnectivity and was more complex, which indicating microalgal application enhance the stability of aquaculture systems. The application of microalgae was found to have the greatest effect on the 6th day of the experiment, as supported by both environmental and biological evidence. These findings can provide valuable guidance for the practical application of microalgae in aquaculture systems.

Overexpression of a novel gene (Pt2015) endows the commercial diatom Phaeodactylum tricornutum high lipid content and grazing resistance.

BACKGROUND: The marine diatom Phaeodactylum tricornutum is a commercially viable species due to its bioactive substances and lipid productivity. Increasing attention has been paid to the isolation or genetic modification of species or strains with a rapid growth rate and large quantities of lipids. Furthermore, contamination of microzooplankton has been one of the major constraints in P. tricornutum large-scale cultivation, which adversely affects growth and greatly impedes the course of biomass production industrialization. RESULTS: Here, based on our previous transcriptomics of P. tricornutum, we found a novel gene (ID: 7202015, hereafter called Pt2015) which affects morphotype of P. tricornutum. Pt2015 protein is located in the plastid, which is highly homologous to part of the sequences of exosome component. The morphotype of the Pt2015 knockout strain (termed 2015KO) using CRISPR/Cas9 method is fusiform, but the Pt2015 overexpression strain (termed oeT) demonstrates a majority triradiate morphotype (approximately 95%) which is stable and has been cultured for more than 200 generations. In addition, the oeT strain demonstrated a similar growth rate to the WT and simultaneously accumulated larger lipids droplets that increased by approximately 30% compared to that of the WT. More importantly, the grazing rate of the amoebae cultured in the oeT strain significantly decreased in comparison with that cultured in WT, suggesting that the oeT can effectively avoid being eaten by microzooplankton. CONCLUSIONS: Therefore, the oeT strain not only improves our understanding of morphotype conversion in diatoms but also demonstrates potential applications for large-scale cultivation of P. tricornutum.

Kinopus chlorellivorus gen. nov., sp. nov. (Vampyrellida, Rhizaria), a New Algivorous Protist Predator Isolated from Large-Scale Outdoor Cultures of Chlorella sorokiniana.

The large-scale culture of low-cost algal biomass can be significantly affected by microbial grazing on the algae. To minimize the impact, it is necessary to manage the predators. In this study, we describe a new genus and species of vampyrellid amoeba, Kinopus chlorellivorus, which caused the loss of Chlorella sorokiniana in large-scale cultures. We assigned it to the family Leptophryidae (Vampyrellida) based on morphology and small-subunit (SSU) rRNA gene sequence comparisons. Using transmission electron microscopy, we found spherical lucent inclusions, which have not been reported for any leptophryids or other vampyrellids. The gene sequence of SSU rRNA did not match any recognized genera or species and contained four characteristic regions. K. chlorellivorus preys on algae by engulfment. Laboratory feeding experiments confirmed that its grazing rate was as high as 131 Chlorella cells day-1 individual-1. Results of prey-range experiments demonstrated that it could consume other chlorophyte microalgae (e.g., Scenedesmus, Coelastrella, and Haematococcus) but with a strong feeding ability on Chlorella spp., with ingestion rates ranging from 2.67 to 3.15 prey predator-1 h-1 and growth rates of the amoeba ranging from 0.039 to 0.045 h-1. On the basis of its high grazing ability on Chlorella, capacity to form large populations in a short period of time, and capacity to form resistant resting stages, this contaminant has the potential to cause serious problems in large-scale Chlorella culture and should be of concern to operators of algal production facilities. IMPORTANCE The vampyrellids (Vampyrellida, Rhizaria) are a major group of predatory amoebae that have attracted significant attention because of their diversity of feeding strategies. The crucial roles they play in important processes such as suppressing soil disease and controlling aquatic algae, and as microbial contaminants in outdoor large-scale algal cultures, have also received increasing attention. In this study, a new genus and species of algivorous vampyrellid amoeba, Kinopus chlorellivorus, is described as a significant grazer responsible for losses in outdoor industrial Chlorella cultures. We found that the amoeba's detrimental effects on Chlorella cultures may be related to its specific feeding characteristics. This study provides phenotypic and genetic information on a previously unknown vampyrellid, emphasizes the impact of contaminating vampyrellids in commercial microalgal cultures, and will contribute to the development of management strategies for predicting this kind of contaminant in large-scale microalgal cultivation.

Assessment of Eicosapentaenoic Acid (EPA) Production from Filamentous Microalga Tribonema aequale: From Laboratory to Pilot-Scale Study.

It has long been explored to use EPA-rich unicellular microalgae as a fish oil alternative for production of the high-value omega-3 fatty acid eicosapentaenoic acid (EPA, 20:5, n-3). However, none of the efforts have ever reached commercial success. This study reported a filamentous yellow-green microalga Tribonema aequale that possesses the ability to grow rapidly and synthesize significant amounts of EPA. A series of studies were conducted in a glass column photobioreactor under laboratory culture conditions and in pilot-scale open raceway ponds outdoors. The emphasis was placed on the specific nutrient requirements and the key operational parameters in raceway ponds such as culture depth and mixing regimes. When optimized, T. aequale cells contained 2.9% of EPA (w/w) and reached a very high biomass concentration of 9.8 g L-1 in the glass column photobioreactor. The cellular EPA content was increased further to 3.5% and the areal biomass and EPA productivities of 16.2 g m-2 d-1 and 542.5 mg m-2 d-1, respectively, were obtained from the outdoor pilot-scale open raceway ponds, which were the record high figures reported thus far from microalgae-based EPA production. It was also observed that T. aequale was highly resistant to microbial contamination and easy for harvesting and dewatering, which provide two additional competitive advantages of this filamentous microalga over the unicellular counterparts for potential commercial production of EPA and other derived co-products.

Critical assessment of protozoa contamination and control measures in mass culture of the diatom Phaeodactylum tricornutum.

The diatom Phaeodactylum tricornutum is considered a promising "cell factory" for high-value products fucoxanthin and EPA. But its potential has not been realized due largely to microbial contamination. In this study, seven protozoan strains were identified, of which a heterolobosean amoeba was identified as the most frequently occurring and destructive predator in P. tricornutum culture. The addition of 400 mg L-1 NH4HCO3 inhibited amoeba proliferation with little impact on algal growth. Halting culture mixing at night induced a hypoxia environment that further inhibited amoeba growth. Regardless of culture systems employed, a periodical supply of proper amounts of NH4HCO3 alone or in combination with halting culture mixing at night may prevent or treat protozoa contamination in mass culture of P. tricornutum.

Effects of Nutritional Mode on the Physiological and Biochemical Characteristics of the Mixotrophic Flagellate Poterioochromonas malhamensis and the Potential Ecological Implications.

Mixotrophic flagellates play an important role in connecting the classical food chain and microbial food loop. The feeding characteristics of the mixotrophic flagellate Poterioochromonasmalhamensis have been well studied, but its role as a food source for other large zooplankton is less studied. This study focuses on the physiological and biochemical changes in P. malhamensis when using autotrophy, chemoheterotrophy, and phagotrophy, and the effect of these changes on the feeding ability of one of its predators, the ciliate Paramecium caudatum. The results showed that chemoheterotrophic P. malhamensis had a higher growth rate and larger cell size than autotrophic and phagotrophic P. malhamensis. The biochemical composition of P. malhamensis also varied greatly between the three nutritional modes. The protein, total absolute amino acid, and fucoxanthin contents were highest for autotrophic P. malhamensis, while chemoheterotrophic P. malhamensis had the highest contents of total sugar and total absolute fatty acid. The contents of most biochemical components in phagotrophic P. malhamensis fell between those in autotrophic and chemoheterotrophic P. malhamensis. A feeding experiment showed that the grazing ability of P. caudatum on chemoheterotrophic P. malhamensis was significantly higher than that on phagotrophic P. malhamensis and autotrophic P. malhamensis. This study showed that the transformation of nutritional modes can alter the biochemical composition of the mixotrophic flagellate P. malhamensis and, as a result, affect the grazing ability of its predator P. caudatum.

Analysis and Calculation of Stability Coefficients of Cross-Laminated Timber Axial Compression Member.

Cross-laminated timber (CLT) elements are becoming increasingly popular in multi-storey timber-based structures, which have long been built in many different countries. Various challenges are connected with constructions of this type. One such challenge is that of stabilizing the structure against vertical loads. However, the calculations of the stability bearing capacity of the CLT members in axial compression in the structural design remains unsolved in China. This study aims to determine the stability bearing capacity of the CLT members in axial compression and to propose the calculation method of the stability coefficient. First, the stability coefficient calculation theories in different national standards were analyzed, and then the stability bearing capacity of CLT elements with four slenderness ratios was investigated. Finally, based on the stability coefficient calculation formulae in the GB 50005-2017 standard and the regression method, the calculation method of the stability coefficient for CLT elements was proposed, and the values of the material parameters were determined. The result shows that the average deviation between fitting curve and calculated results of European and American standard is 5.43% and 3.73%, respectively, and the average deviation between the fitting curve and the actual test results was 8.15%. The stability coefficients calculation formulae could be used to predict the stability coefficients of CLT specimens with different slenderness ratios well.

Gluing Techniques on Bond Performance and Mechanical Properties of Cross-Laminated Timber (CLT) Made from Larix kaempferi.

Previous studies have proved that Larix kaempferi is a good material for preparing cross-laminated timber (CLT), but under bending shear stress, CLT made by Larix kaempferi is prone to the phenomenon of bonding face cracking, which seriously affects the shear performance of CLT. To solve this problem, this paper took Larix kaempferi as raw material, conducted experiments on the surface sanding conditions, gluing pressure and adhesive types of sawing timber, and explored the influence of these three factors on the bonding quality of CLT. The microscopic characteristics of the bonding layer were further studied. The results showed that for Larix kaempferi with a density of 0.68 g/cm3 used in this experiment, a high bonding pressure is required. Among the three cold curing adhesives selected in the experiment, emulsion polymer isocyanate (EPI) adhesive needs 1.5 MPa bonding pressure to ensure the bonding quality, while for polyurethane (PUR) and phenol resorcinol formaldehyde (PRF), 1.2 MPa can meet the need of adhesive pressure. This is concerned with the permeability of different adhesives under different pressures. The microscopic results of the bonding layer show that EPI adhesives have poor permeability, so it requires high bonding pressure. The influence of sanding surface of different sand-belt on block shear strength (BSS) and wood failure percentage (WFP) is not obvious, while the durability of bonding layer is better when sanding mesh number is 100. Hence, a high pressure should be used for CLT industrial production when the laminate density is higher, especially when the adhesive has poor permeability. Reasonable sanding surface treatment can be used in laminate surface treatment to improve the durability of CLT.

Assessment of a Novel Oleaginous Filamentous Microalga Klebsormidium sp. Lgx80 (Streptophyta, Klebsormidiales) for Biomass and Lipid Production1.

Commercial cultivation of eukaryotic microalgae has so far employed a unicellular form of species only (e.g., Chlorella pyrenoidosa, Dunaliella salina, and Haematococcus pluvialis). In this study, we assessed the feasibility of using the filamentous eukaryotic microalga Klebsormidium sp. LGX80 as a new cultivar for biomass and lipid production. The effects of different forms and concentrations of nitrogen on growth and lipid production of Klebsormidium sp. LGX80 were studied by using a glass column (ø4.5 × 60 cm) photobioreactor under laboratory conditions. Growth and lipid production of the new strain were further evaluated in an outdoor pilot-scale tubular photobioreactor. The results showed that when supplied with urea as a source of nitrogen Klebsormidium sp. LGX80 yielded a final biomass concentration of 8.49 ± 0.10 g · L-1 in which a cellular lipid content was 59.2 ± 0.4% DW. Under such conditions, the biomass and lipid productivities were 471.7 ± 5.9 and 248.1 ± 0.0 mg · L-1  · d-1 , respectively. Fatty acid analysis revealed that the main fatty acids of Klebsormidium sp. LGX80 were palmitic acid (C16:0), linoleic acid (C18:2ω6), and linolenic acid (C18:3ω3), of which linoleic acid (C18:2ω6) accounted for up to 67.5 ± 0.1% of total fatty acids. When grown outdoors in a 13,000-L tubular photobioreactor with an initial nitrogen concentration of 3 mM urea, Klebsormidium sp. LGX80 reached the highest biomass concentration of 2.63 ± 0.09 g · L-1 with the cells containing 38.0 ± 0.5% lipids (% DW), resulting in the volumetric biomass and lipid productivities of 147.2 ± 3.6 and 37.9 ± 0.9 mg · L-1  d-1 , respectively. The results of light:dark cycle experiment showed that a durative and prolonged light irradiation hindered the biosynthesis of chlorophyll a and chlorophyll b in the cells, but promoted the carotenoid accumulation. These results suggested that Klebsormidium sp. LGX80 can be a potential oleaginous filamentous microalga for commercial production of microalgal oils.

Factors Affecting the Mixotrophic Flagellate Poterioochromonas malhamensis Grazing on Chlorella Cells.

Grazing behaviour between protozoa and phytoplankton exists widely in planktonic ecosystems. Poterioochromonas malhamensis is a well-known and widespread mixotrophic flagellate, which is recognized to play an important role within marine and freshwater planktonic ecosystems and regarded as the greatest contamination threat for mass algal cultures of Chlorella. In this study, a comprehensive range of factors, including morphological characters, biochemical compositions, and specific growth rate of ten species or strains of Chlorella, were evaluated for their effect on the feeding ability of P. malhamensis, which was assessed by two parameters: the clearance rate of P. malhamensis on Chlorella spp. and the specific growth rate of P. malhamensis. The results showed that the clearance rate of P. malhamensis was negatively correlated with cell wall thickness and specific growth rate of Chlorella spp., while the specific growth rate of P. malhamensis was positively correlated with carbohydrate percentage and C/N ratio and negatively correlated with protein, lipid percentage, and nitrogen mass. In conclusion, the factors influencing feeding selectivity include not only the morphological character and chemical composition of Chlorella, but also its population dynamics. Our study provides useful insights into the key factors that affect the feeding selectivity of P. malhamensis and provides basic and constructive data to help in screening for grazing-resistant microalgae.

Taming chlorophylls by early eukaryotes underpinned algal interactions and the diversification of the eukaryotes on the oxygenated Earth.

Extant eukaryote ecology is primarily sustained by oxygenic photosynthesis, in which chlorophylls play essential roles. The exceptional photosensitivity of chlorophylls allows them to harvest solar energy for photosynthesis, but on the other hand, they also generate cytotoxic reactive oxygen species. A risk of such phototoxicity of the chlorophyll must become particularly prominent upon dynamic cellular interactions that potentially disrupt the mechanisms that are designed to quench photoexcited chlorophylls in the phototrophic cells. Extensive examination of a wide variety of phagotrophic, parasitic, and phototrophic microeukaryotes demonstrates that a catabolic process that converts chlorophylls into nonphotosensitive 132,173-cyclopheophorbide enols (CPEs) is phylogenetically ubiquitous among extant eukaryotes. The accumulation of CPEs is identified in phagotrophic algivores belonging to virtually all major eukaryotic assemblages with the exception of Archaeplastida, in which no algivorous species have been reported. In addition, accumulation of CPEs is revealed to be common among phototrophic microeukaryotes (i.e., microalgae) along with dismantling of their secondary chloroplasts. Thus, we infer that CPE-accumulating chlorophyll catabolism (CACC) primarily evolved among algivorous microeukaryotes to detoxify chlorophylls in an early stage of their evolution. Subsequently, it also underpinned photosynthetic endosymbiosis by securing close interactions with photosynthetic machinery containing abundant chlorophylls, which led to the acquisition of secondary chloroplasts. Our results strongly suggest that CACC, which allowed the consumption of oxygenic primary producers, ultimately permitted the successful radiation of the eukaryotes throughout and after the late Proterozoic global oxygenation.

Phylogenomics solves a long-standing evolutionary puzzle in the ciliate world: The subclass Peritrichia is monophyletic.

The phylum Ciliophora is one of the most broadly studied protozoan lineages. The era of molecular investigation has brought forth a major ongoing debate: is the subclass Peritrichia Stein, 1859 monophyletic? Numerous analyses mostly using the small subunit (SSU) rRNA gene have failed to recover the Mobilida and Sessilida, the two peritrich orders, as sister clades. Here we have sequenced five peritrich species - three sessilids and two mobilids. We constructed a supermatrix of 158 genes and 44,696 characters for 24 ciliate species, and as outgroup taxa, nine species from the Apicomplexa and four from the Dinophyceae. Our analyses using both maximum likelihood and Bayesian methods recover a monophyletic class Oligohymenophorea and two robust clades within it. The first clade is a monophyletic Peritrichia with the orders Sessilida and Mobilida maximally supported as sister clades. The second oligohymenophorean clade includes species of the subclasses Scuticociliatia and Hymenostomatia, which are sister clades. Our analyses resolve a long-standing debate in ciliate molecular phylogenetics and provide support for the classical view that the morphological features of the two peritrich orders Mobilida and Sessilida arose by descent from the same common ancestor and are not the result of convergence.

Vernalophrys algivore gen. nov., sp. nov. (Rhizaria: Cercozoa: Vampyrellida), a New Algal Predator Isolated from Outdoor Mass Culture of Scenedesmus dimorphus.

Microbial contamination is the main cause of loss of biomass yield in microalgal cultures, especially under outdoor environmental conditions. Little is known about the identities of microbial contaminants in outdoor mass algal cultures. In this study, a new genus and species of vampyrellid amoeba, Vernalophrys algivore, is described from cultures of Scenedesmus dimorphus in open raceway ponds and outdoor flat-panel photobioreactors. This vampyrellid amoeba was a significant grazer of Scenedesmus and was frequently associated with a very rapid decline in algal numbers. We report on the morphology, subcellular structure, feeding behavior, molecular phylogeny, and life cycle. The new amoeba resembles Leptophrys in the shape of trophozoites and pseudopodia and in the mechanism of feeding (mainly by engulfment). It possesses two distinctive regions in helix E10_1 (nucleotides 117 to 119, CAA) and E23_1 (nucleotides 522 and 523, AG) of the 18S rRNA gene. It did not form a monophyletic group with Leptophrys in molecular phylogenetic trees. We establish a new genus, Vernalophrys, with the type species Vernalophrys algivore. The occurrence, impact of the amoeba on mass culture of S. dimorphus, and means to reduce vampyrellid amoeba contamination in Scenedesmus cultures are addressed. The information obtained from this study will be useful for developing an early warning system and control measures for preventing or treating this contaminant in microalgal mass cultures.

Description of two new species of ectoparasitic Trichodina Ehrenberg, 1830 (Ciliophora: Trichodinidae) from freshwater fishes in the river Ganges, India.

Two new species of the genus Trichodina Ehrenberg, 1838, T. silondiata sp. nov. and T. pangasi sp. nov. from the gills of freshwater fish Silonia silondia (Hamilton 1822) and Pangasius pangasius (Hamilton-Buchanan) respectively from the river Ganges of West Bengal are described here. Wet smears of gills and skins were prepared in the field, air dried and impregnated with Klein's dry silver method. In case of S. silondia (Hamilton 1822) 24 out of 146 host fishes were parasitized on the gills. Infestation rate in case of P. pangasius (Hamilton-Buchanan) was not significant. From a total of 86 examined host fish, only seven were parasitized on the gills. The mean diameters of the body of the specimens of T. silondiata sp. nov. and T. pangasi sp. nov. were 32.7-60.6 (46.4 ± 6.3) µm and 38.9-54.1 (44.9 ± 3.0) µm respectively. Taxonomic and morphometric data for these ectoparasitic trichodinids based on wet silver nitrate impregnated specimens are presented.

Studies on Trichodinid and Chilodonellid Ciliophorans (Protozoa: Ciliophora) in the Indian freshwater and estuarine fishes with description of Trichodinella sunderbanensis sp. nov. and Trichodina nandusi sp. nov.

A comprehensive icthyoparasotological survey among estuarine and freshwater fishes of West Bengal, India revealed Trichodinella sunderbanensis sp. nov. and Trichodina acuta Lom, 1961 from an estuarine fish Mystus gulio (Hamilton, 1822) from the Vidyadhari river; Trichodina nandusi sp. nov. from a freshwater fish, Nandus nandus (Hamilton-Buchanan); Chilodonella hexasticha (Kiernik, 1909) Kahl, 1931 from freshwater fishes Labeo rohita (Hamilton, 1822); and Cyprinus carpio Linnaeus, 1758 from the Rupnarayan river. Taxonomic description of all the species based on wet silver nitrate impregnation method along with additional comments based on scanning electron microscopic descriptions of T. nandusi sp. nov. are also provided in this paper. Prevalence rate, morphometric parameters and comparisons with closely related species are also discussed.

Host species as a strong determinant of the intestinal microbiota of fish larvae.

We investigated the influence of host species on intestinal microbiota by comparing the gut bacterial community structure of four cohabitating freshwater fish larvae, silver carp, grass carp, bighead carp, and blunt snout bream, using denaturing gradient gel electrophoresis (DGGE) of the amplified 16S and 18S rRNA genes. Similarity clustering indicated that the intestinal microbiota derived from these four fish species could be divided into four groups based on 16S rRNA gene similarity, whereas the eukaryotic 18S rRNA genes showed no distinct groups. The water sample from the shared environment contained microbiota of an independent group as indicated by both 16S and 18S rRNA genes segments. The bacterial community structures were visualized using rank-abundance plots fitted with linear regression models. Results showed that the intestinal bacterial evenness was significantly different between species (P<0.05) and between species and the water sample (P<0.01). Thirty-five relatively dominant bands in DGGE patterns were sequenced and grouped into five major taxa: Proteobacteria (26), Actinobacteria (5), Bacteroidetes (1), Firmicutes (2), and Cyanobacterial (1). Six eukaryotes were detected by sequencing 18S rRNA genes segments. The present study suggests that the intestines of the four fish larvae, although reared in the same environment, contained distinct bacterial populations, while intestinal eukaryotic microorganisms were almost identical.

Occurrence of Trichodinid Ciliophorans (Ciliophora: Peritrichida) in the freshwater fishes of the river Churni with description of Trichodina glossogobae sp. nov. in West Bengal, India.

Between January 2011 and March 2011 occurrence of trichodinid ciliophorans were studied in the freshwater fishes of the river Churni. The biodiversity survey revealed presence of a new species of the genus Trichodina (Ehrenberg 1830) infesting Glossogobius giuris (Hamilton-Buchanan). Another freshwater fish Labeo bata (Hamilton 1822) were found to be infested with two species of the genus Tripartiella Lom 1959: Tripartiella bulbosa (Davis 1947) Lom 1959 and T. copiosa Lom 1959. Adhesive disc size of the new Trichodina species is small measuring 30.6-46.9 (36.2 ± 3.9) µm in diameter. The centre of the disc is finely granular and transforms into dark when impregnated with silver. The overall prevalence of this species is 15.6% (12/77). Tripartiella bulbosa and T. copiosa are very small sized ciliophorans, measuring 16.7-23.8 (20.6 ± 2.2) µm and 15.2-22.4 (19.7 ± 1.9) µm in diameter, respectively.