Rapid Preparation of Filamentous Fungal DNA for PCR Analysis by Ultrasonic Treatment

A simple method to prepare of DNA template suitable for PCR amplification from filamentous fungi will be valuable for improving experimental efficiency.Here,a method was developed which just needed ultrasonic treatment of the mycelium at usual condition,and the produced solution could directly be used as DNA template for internal transcribed spacer(ITS)amplification successfully.The PCR could be improved by additional treatment of 60℃water baths,but was not centrifugation.When the template amount was 0.5-2μL and the ultrasonic time was 7-11 min,there was no distinctly influences on PCR.The method was commonly used for M.purpureus,I.cicadae,Lentinula sp.,Flammul sp.and Dictyophora sp.etc.to detect target sequences of ITS,hygromycin resistance gene(Hyg),CRISPR-associated protein 9(Cas9),Citrinin gene C(CtnC),Citrinin gene D(CtnD),large subunit rRNA gene(NL),and so on.The method could provide a simple,rapid,safe and economic approach to prepare the DNA template for large-scale PCR of the special filamentous fungi materials.

Diversity of filamentous fungi in organic layers of two forests in Zijin Mountain

A study was conducted to evaluate the cultivable filamentous fungal diversity in organic layers (L, F, and H layers) and A1 layer of two main forest types, Pinus massoniana and Liguidambar formasana mixed forest and Quercus variabilis forest, in Zijin Mountain(325?N, 11848?E), Nanjing, China. A total of 67 taxa comprising 56 Deuteromycetes, 3 Zygomycetes, 5 Asco-mycetes and 3 unidentified fungi were recognized from samples from the forest floor of the two forest types. The most abundant group was Deuteromycetes. The dominant genera in both forests were Alternaria sp., Aspergillus sp., Cladosporium sp., Mucor sp., Penicillium sp., Rhizopus sp., Gliocladium sp. and Trichoderma spp. The fungal diversity was higher in the mixed forest than that in Q. variabilis forest. For both forest types, the maximum fungal diversity was found in layer F and there existed significantly different in fungal diversity between layer F and layer L. In the mixed forest, richness of fungi isolated from needle litter (P. massoniana) was lower than that from leaf litter (L. formasana). The richness of fungi from needle litter increased with the in-crease of forest floor depth, but for leaf litter, the fungal diversity decreased with the depth of forest floor. The co-species of fungi from the two forest types, as well as from two kinds of litters in mixed forest, increased with the depth of the forest floor. The succession of fungi along with the process of decomposition was discussed here. The results also showed that litter quality was a critical factor affecting fungal diversity.

Study on Limited Filamentous Sludge Bulking Test in A/O Activated Sludge Process

[Objective] This study aimed to make use of the limited filamentous sludge bulking characteristics to purify water quality and save energy. [Method] The effects of dissolved oxygen （DO） on sludge bulking were explored by changing the DO con- centration in the aerobiotic pool of NO system from the low load （0.25 kg COD/（kg MLSS.d）） and the medium to high load （0.55 kgCOD/（kg MLSSod））. [Result] In the NO activated sludge system, when the sludge load was 0.25 kg COD/（kg MLSS.d）, DO=1.5 mg/L, the sludge index was at around 250, while when the sludge load- ing was 0.55 kg COD/（kg MLSS.d）, DO=1.5 mg/L, the sludge index was close to 300, occurring the limited filamentous bulking. The bulked sludge still showed high removal rates to COD, SS, nitrogen and phosphorus. [Conclusion] It could improve the oxygen transfer rate and reduce the aeration rate at low DO conditions to achieve energy-saving.

Approaches to functional genomics in filamentous fungi

The study of gene function in filamentous fungi is a field of research that has made great advances in very recent years. A number of transformation and gene manipulation strategies have been developed and applied to a diverse and rapidly expanding list of economically important filamentous fungi and oomycetes. With the significant number of fungal genomes now sequenced or being sequenced, functional genomics promises to uncover a great deal of new information in coming years. This review discusses recent advances that have been made in examining gene function in filamentous fungi and describes the advantages and limitations of the different approaches.

Granulation of filamentous microorganisms in a sequencing batch reactor with saline wastewater

Proliferation of filamentous microorganisms frequently leads to operational failure for activate sludge systems. In this study, it was found that filamentous microorganisms could grow in compact granular structure with 5% sodium chloride in the substrate. In the early period of experiment, coccoid and rode-like bacteria predominated in the yellowish-brown granules, and later the white and the black granules were developed by filamentous microorganisms. The filamentous granules exhibited low porosity and fast settling velocity, and were more compact even than bacteria granules. It was hypothesized that the elevated pH in the later period might be a possible reason for the compact growth of filamentous granules. However, the bacteria granules showed the high bioactivity in terms of specific oxygen utilizing rate, and comprised of a wider diversity of compounds based on the thermogravimetric evaluation. The findings in this study demonstrated that filamentous microbes could form compact granular structure, which may encourage the utilization of filamentous microorganisms rather than the inhibition of their growth, as the latter is frequently used for sludge bulking control.

A New Giant Compsognathid Dinosaur with Long Filamentous Integuments from Lower Cretaceous of Northeastern China

A new compsognathid dinosaur, Sinocalliopteryx gigas gen. et sp. nov., is erected based on a complete skeleton from the Early Cretaceous Yixian Formation of western Liaoning, northeastern China. It shares the features with Huaxiagnathus orientalis in having a manus as long as the humerus plus radius, very large and subequally long manual claws I and H, and reduced olecranon process on the ulna. But it differs from Huaxiagnathus orientalis in having the much large size, a very long maxillary process of premaxilla not extending the vertical level of the maxillary antorbital fossa, and the proportionally longer ulna and so on. Sinocalliopteryx gigas gen. et sp. nov. represents the largest species among the known compsognathid dinosaurs, suggesting the tendency of the body enlargement in compsognathids to some extent. The long filamentous integuments are attached to the whole body of this compsognathid, confirming that such integuments evolved firstly in the basal coelurosaurs. This new giant compsognathid was a fierce carnivorous theropod, as shown further by an incomplete dromaeosaurid leg inside its abdominal cavity.

Construction and Virulence of Filamentous Hemagglutinin Protein B1 Mutant of Pasteurella multocida in Chickens

Pasteurella multocida, a Gram-negative nonmotile coccobacillus, is the causative agent of fowl cholera, bovine hemorrhagic septicemia, enzoonotic pneumonia and swine atropic rhinitis. Two filamentous hemagglutinin genes, fhaB1 and JhaB2, are the potential virulence factors. In this study, an inactivationfhaB1 mutant ofP. multocida in avian strain C48-102 was constructed by a kanamycin-resistance cassette. The virulence of thefhaB1 mutant and the wild type strain was assessed in chickens by intranasal and intramuscular challenge. The inactivation offhaB1 resulted in a high degree of attenuation when the chickens were challenged intranasally and a lesser degree when challenged intramuscularly. ThefhaB1 mutant and the wild type strain were investigated their sensitivity to the antibody-dependent classical complement-mediated killing pathway in 90% convalescent chicken serum. ThefhaB1 mutant was serum sensitive as the viability has reduced between untreated serum and heat inactivated chicken serum （P〈0.007）. These results confirmed that FhaB1 played the critical roles in the bacterial pathogenesis and further studies were needed to investigate the mechanism which caused reduced virulence of the fhaB1 mutant.

Advanced Treatment of Wastewater Treatment Plant Effluent by Using Biofilms on Filamentous Bamboo

[ Objective ] The study aimed at treating wastewater treatment plant （WWTP） effluent by using bio-film reactor with filamentous bamboo as bio-carrier. [ Method] With the aid of a continuous flow reactor, a bio-film reactor using filamentous bamboo as bio-carrier was used to treat WWTP effluent with low C/N ratio, and the removal effects of CODc,, TN （total nitrogen）, and NO3--N in the wastewater were analyzed.[ Result ] The average removal rates of CODcr, TN, and NO3- -N reached 47.7%, 23.6% and 34.5% when the C/N ratio of influent was around 2. In addi- tion, a stable bio-film was formed very well in the secondary effluent with low C/N ratio and hardly degradable organic pollutants. The pollutants could be removed effectively because of the excellent surface characteristics and compositions of filamentous bamboo. [ Conclusion] The research provides a new method to treat WWTP effluent with low C/N ratio.

Filamentous Green Algae Reduce Walleye Fingerling Production in Earthen-Substrate Ponds

Reduced walleye (Sander vitreus) fingerling production possibly related to nuisance filamentous green algae and overly stable water chemistry patterns is a concern in earthen-substrate ponds at Blue Dog State Fish Hatchery, South Dakota, USA. We describe the success of alfalfa (Medicago sativa) meal (AFM, n = 2), alfalfa meal plus soybean (Glycine max) meal (AFM + SBM, n = 1), and alfalfa meal plus liquid 28N:0P:0K (AFM + 28:0:0, n = 1) at achieving walleye production objectives (>104,000 walleye and 32 kilograms/hectare) as well as the occurrence of filamentous green algae and the associated patterns of ammonia-nitrogen, pH, and dissolved oxygen in earthen-substrate ponds. Walleye production objectives were only achieved when filamentous green algae were absent preceding harvest, which occurred in one pond that received AFM and in the pond that received AFM + 28:0:0. The presence of filamentous green algae preceding harvest was associated with higher dissolved oxygen and pH, whereas declines in these variables occurred when filamentous green algae were absent. Organic fertilizer alone exhibited low ammonia-nitrogen (<0.1 mg/L) despite the substitution of higher protein content soybean meal, but supplementation with 28:0:0 increased ammonia-nitrogen to 0.23 mg/L. These findings highlight the reduction in walleye fingerling production that occurred in the presence of filamentous green algae and the unpredictability of results when two earthen-substrate ponds are treated exactly the same with alfalfa meal. Achievement of walleye production objectives, lack of nuisance filamentous green algae, and promotion of favorable patterns in water chemistry warrant future experimentation with alfalfa meal supplemented with 28:0:0 in earthen-substrate ponds at this hatchery.

A Review of the Ubiquity of Ascomycetes Filamentous Fungi in Relation to Their Economic and Medical Importance

Filamentous fungi are found in different habitats in the environment including, air, water and soil. This group of fungi contains organisms from different classes under the sub-phylum Pezizomycotina. They occur in mixtures such that you find many genera of filamentous fungi dominating a particular habitat or substrate. The wide distribution of filamentous fungi has resulted in it being used for different purposes. This review aims to analyse the different genera of fungi species referred to as filamentous fungi and their relevance economically and medically.

Fractal characteristics of filamentous bacteria in activated sludge and its significance

On the basis of fractal theory, the fractal characteristics of filamentous bacteria in activated sludge are studied by image dissector system. The results show that the spatial distribution structure of filamentous bacteria in activated sludge has high self similarity in statistics and the filamentous bacteria have clear phenomenon of fractal growth. The critical fractal dimension values of filamentous bacteria bulking such as Eikelboom 021N, Eikelboom 1701, Sphaerotilus natans and Eikelboom 0041 are determined initially. The fractal dimensions can be taken as a parameter to describe filamentous bacteria of activated sludge quantitatively and bring to light the disorder in appearance and the rule in inherence of filamentous bacteria system.

A Population Morphologically Structured Model for Microscopic Growth of Filamentous Microorganism

The growth of filamentous microorganism is contributed by tip extension and branching. The microscopic growth of filamentous microorganism means the growth process from one or a few spores. In order to describe the microscopic process, a population morphologically structured model is proposed, in which three morphological compartment and their interactions were considered, and the heterogeneity of hyphal growth was included. The model was applied to describe the microscopic growth of Streptomyces tendae and Geotrichum candidum with good agreement. From model prediction, it is concluded that if the number of hyphae is large enough (macroscopic growth), the specific growth rate of filamentous microorganism and the ratio of morphological forms in hyphae will become constant.

pFTAA: a high affinity oligothiophene probe that detects filamentous tau in vivo and in cultured neurons

Tauopathies describe a group of neurodegenerative diseases in which the protein tau,encoded by the gene MAPT,is aberrantly misfolded,leading to tau aggregation,neural dysfunction,and cell death（Spillantini and Goedert,2013）.In Alzheimer＇s disease（AD）,tau forms the characteristic intracellular neurofibrillary tangles（NFTs）,which are thought to be the major cause of neurodegeneration（Bloom,2014）.In other tauopathies,including frontotemporal dementia with Parkinsonism linked to chromo- some 17 （FTDP-17T）, corticobasal degeneration and progressive supranuclear palsy, there are specific forms of tau aggregates and filaments without any amyloid pathology, demonstrating tau＇s po- tent disease-causing potential （Spillantini and Goedert, 2013）. Tau is a microtubule （MT） binding protein, which becomes abnormally hyperphosphorylated on several residues prior/during the process of aggregation, thereby causing loss of its MT binding activity （Mandelkow and Mandelkow, 2012）.

The Occurrence and Prevention of Filamentous Algae in Ecological Purification System of Micro-polluted Water

[ Objective ] The aim was to study the influence of filamentous algae on the process of water ecological purification. [ Method ] The occurrence mechanism of filamentous algae and its ecological system were summarized and analyzed. Considering the ecological purification in north- ern Jiangsu, the occurrence and prevention of filamentous algae in water, the method to prevent filamentous algae in polluted water was discussed. [ Result] The results showed that by measures of improving planting density, regular harvesting, and water flow state control before the filamentous algae blooming period, together with improving local pH value, light interference and ecological control during the blooming period, can effectively control the filamentous algae blooming. [ Condusion] The study of the happening mechanism of filamentous algae provided theoretical references and the technical basis in the work of filamentous algae prevention and control.

Biocidal Effect of Gamma Radiation on the Ecology of Filamentous Fungal Populations Associated with Stone Deterioration

This study focused on the identification of the filamentous fungi associated with soapstone samples exposed to outdoor conditions and the biocidal effect of gamma radiations on isolated fungal populations in Minas Gerais State in Brazil. Two soapstone blocks were placed outdoors under tropical environmental conditions for 12 months. A total of 9 filamentous fungal populations were identified on their surfaces, namely Acremomium （cf.） alternatum, Alternaria alternata, Aspergillusfumigatus, Calcarisporium （cf.） arbuscula, Cladosporium cladosporioides, Curvularia lunata, Epicoccum nigrum, Fusarium equiseti and Penicillium citrinum. The gamma radiation assay was then carried out as a test of biocidal action by exposing all fungal populations to the ionizing radiation. The results showed that only the C. cladosporioides species was resistant to this biocidal agent, since it was able to increase its population post exposure. Scanning electron microscopy images identified the microbial colonization on the soapstone blocks and the stone elementar composition was analyzed by energy dispersive X-ray spectrometry. After treatment, there was no structural and aesthetic alteration in the soapstone samples, and evidencing that gamma radiation can be used as a biocidal agent. However, the resistance of the black fungal population indicates caution in the choice of gamma irradiation as biocidal treatment.

Isolation of Filamentous Fungi from the Caatinga Region and Production of Amylolytic Enzymes of Great Industrial Interest

The industrial importance and the high cost of the commercial amylase require the study of microorganisms that produce these enzymes. For this reason, the objective of this work was to isolate filamentous fungi from a region of the caatinga and evaluate their potential for the production of amylase. Four soil samples were collected from a deactivated dump located in the city of Diamantina, MG, in a region of the caatinga. The analysis of amylolytic production in a submerged medium at the ideal temperature of each microorganism was performed using the saccharification method, and the reducing sugars formed were quantified by DNS. Fourteen filamentous fungi were isolated, which had different morphological aspects. Regarding amylase production, a mean activity of 0.477 U·mL-1 was obtained with the isolates I 1.2.1 and I 4.4.1. These results bring important information regarding the biodiversity of the caatinga, in addition to the isolation of microorganisms that can be used as biological machinery to obtain metabolites with high biotechnological and industrial potential.

Diversity of Filamentous Fungi of Area from Brazilian Caatinga and High-Level Tannase Production Using Mango (<i>Mangifera indica</i>L.) and Surinam Cherry (<i>Eugenia uniflora</i>L.) Leaves under SSF

Tannase is a biotechnologically important enzyme that can be produced during fungal fermentation of organic matter. The Caatinga is an exclusive Brazilian ecosystem that has been largely unexplored by science, particularly its filamentous fungal diversity. This study evaluated the diversity of filamentous fungi in the Caatinga soils of Pernambuco, Brazil, and their potential for tannase production by solid-state fermentation (SSF) of mango (Mangifera indica L.) and Surinam cherry (Eugenia uniflora L.) leaves. A total of 4711 isolates were obtained, 2090 during the rainy seasonand 2621 during the dry season. The isolates belonged to 18 genera and 66 species, with Aspergillus and Penicillium having the highest species richness. The dry season had a higher diversity index. Aspergillus was the dominant genus, and A. flavus, A. sclerotiorum, and A. ochraceus the most abundant species. A representative of each species was tested for tannase production using dried mango and Surinam cherry leaves as substrates;the leaves contained 14.28 and 7.0 g/L tannin, respectively. Most fungal species produced tannase, but the highest yields were obtained when mango leaves were used as substrate for Penicillium restrictum (accession URM 6044), Aspergillus flavofurcatus (URM 6142), and A. stromatoides (URM 6609), which produced 104.16, 87.51, and 81.83 U/mL tannase, respectively. These yields exceeded previously published reports. Filamentous fungi from Caatinga soils have great potential for producing tannase by SSF, and low-cost mango leaves make excellent substrate.

Inorganic and Organic Compounds of Freshwater Filamentous Cyanobacteria under Normal and Salt Stress Conditions

The main aim of this work was to investigate the effects of salt stress conditions on the inorganic and organic compounds of extracts from six filamentous cyanobacteria： Arthospira platensis, Oscillatoria sp., Oscillatoria salina, Tolypothrix sp., Oscillatoria sp. SWU （Srinakharinwirot University）121 and Tolypothrix sp. SWU213. All cyanobacteria were cultures in BG （blue-green algae）11 medium： pH 7.5 at 35 ℃ for 30 days of sodium chloride in the culture medium presented 0-1.0 M. The cyanobacteria isolates grew well in BGH medium, nevertheless, growth of the majority of isolates was reduced by about 50% in the same medium containing 0.5 M NaCl. The inorganic compounds such as, Na^＋, K^＋, Ca^2＋, Mg^2＋, NH4^＋ and NO3^- were determined. All ions were reduced when NaCI was increased and Na^＋ showed highest amount in the medium followed with Mg^2＋, NH4^＋, NO3^- and Ca^2＋ in all cyanobaeteria. The organic compounds such as, betaine, proline and total lipid were determined under normal and salt stress conditions. We found that all cyanobacteria increased interesting organic compound under salt stress condition at least two folds to compare with normal condition. This is the first finding indicated that freshwater filamentous cyanobacteria could grow under salt stresses by accumulation of some organic compounds as osmoprotectants such as betaine and amino acids, being the reduction related to the amount of inorganies compounds present in cultures.

Spectrum of Filamentous Fungi from Sputum of Tuberculosis Relapse and Retreatment Patients in Tuberculosis Reference Facilities

Background: Fungal infections represent a significant cause of morbidity and mortality among immunocompromised individuals. Pulmonary fungal infection may be missed or misdiagnosed as tuberculosis (TB) hence complicating management of these patients. The current study reports the spectrum of filamentous fungi isolated from sputum of TB relapse and retreatment cases at selected reference facilities in Kenya. Methods: A total of 340 sputum samples collected during the period of June 2018 to June 2019 were subjected to mycological investigations. The samples were mucolysed and inoculated on sabourauds dextrose agar (SDA) and incubated at 30°C for 7 days and checked daily for fungal growth. Moulds were identified by macroscopic and microscopic morphological features and the species were confirmed by sequencing. Results: The diversity of fungi out of the 340 sputum samples analyzed was as follows;16% (n = 53) were positive for moulds with Aspergillus species being the predominant constituting 68 % (n = 36). Among the Aspergilli, A. flavus and A. niger were the most frequently isolated adding up to 23%, (n = 12) and 15% (n = 8) respectively. Additionally, Paecillomyces variotii (9%, n = 5), Scedosporium aspiospermum (6%, n = 3), Mucor racemosus (8%, n = 4) and Penicillium spp. (9%, n = 5) were also recovered. Conclusion: The isolated fungi represented potential respiratory pathogens that could be responsible for persistent TB like symptoms despite treatment that could be misdiagnosed as relapse requiring treatment. Fungal investigation of all presumptive TB relapse cases should be advocated before treatment. This will reduce unnecessary retreatment, delayed antifungal intervention and unwarranted morbidity and mortality associated with misdiagnosis.

Cladophora as ecological engineer:A new test from the largest lake of Qinghai-Tibet plateau with filamentous algal blooms

Filamentous algae blooms(FABs)have been increasing globally in recent years,and their presence can have both harmful and beneficial effects on aquatic ecosystems.As one of the most common FABs,Cladophora blooms have been reported in the lakes of the Qinghai-Tibet Plateau during the past few years.However,there have been few studies focused on how FABs impact other aquatic organisms,especially in alpine lakes since these are at the forefront of responding to global climate change.In this study,the phytoplankton communities in different regions of Qinghai Lake were profiled in different seasons using meta-barcode sequencing.The phytoplankton assemblages in areas with Cladophora blooms were compared to those without Cladophora.The phytoplankton community structure correlated with physicochemical properties including water temperature,electrical conductivity,nitrate,and the presence or absence of Cladophora blooms.The relative abundance of Bacillariophytes was found to be higher in zones with Cladophora blooms than in other regions.Significant seasonal changes in phytoplankton biomass andβdiversity were observed in zones with Cladophora blooms.Growth and microbial degradation of Cladophora can change the pH,dissolved oxygen,secchi depth,and nitrate.Together with seasonal temperature and electrical conductivity changes,Cladophora growth can significantly impact the phytoplankton biomass,community dissimilarity and assembly process.These results showed that Cladophora plays a key role in littoral aquatic ecosystem ecology.