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.

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.

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.

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.

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.

Effect of nitrate concentration on filamentous bulking under low level of dissolved oxygen in an airlift inner circular anoxic-aerobic incorporate reactor

This laboratory research investigated a possible cause of filamentous bulking under low level of dissolved oxygen conditions （dissolved oxygen value in aerobic zone maintained between 0.6-0.8 mgO2 /L） in an airlift inner-circular anoxic-aerobic reactor. During the operating period, it was observed that low nitrate concentrations affected sludge volume index significantly. Unlike the existing hypothesis, the batch tests indicated that filamentous bacteria （mainly Thiothrix sp.） could store nitrate temporarily under carbon restricted conditions. When nitrate concentration was below 4 mg/L, low levels of carbon substrates and dissolved oxygen in the aerobic zone stimulated the nitrate-storing capacity of filaments. When filamentous bacteria riched in nitrate reached the anoxic zone, where they were exposed to high levels of carbon but limited nitrate, they underwent denitrification. However, when nonfilamentous bacteria were exposed to similar conditions, denitrification was restrained due to their intrinsic nitrate limitation. Hence, in order to avoid filamentous bulking, the nitrate concentration in the return sludge （from aerobic zone to the anoxic zone） should be above 4 mg/L, or alternatively, the nitrate load in the anoxic zone should be kept at levels above 2.7 mg NO-3N/g SS.

Numerical simulation on mixing dynamics of flexible filamentous particles in the transverse section of a rotary drum

Flexible filamentous particles are a special kind of particles and play a significant role in many industrial processes. The mixing dynamics of flexible filamentous particles in the transverse section of a rotary drum were analyzed numerically in two dimensions. First, a chain model of slender bodies was introduced for particle dynamic studies, and each individual particle as well as each segment of the particle was tracked during the process. Then, the bulk movement of particles in the transverse section of a rotary drum was explored numerically and mixing dynamics of the particles were further investigated with visual representation. To quantify the quality of mixing, the mixing rates were investigated to determine the mixing extent of particles in the rotary drum. Furthermore, the effects of rotational velocity, flight height and filling degree on mixing dynamics were examined in detail. Moreover, the numerical results were compared with experimental data, and reasonable agreements were obtained, The numerical analyses provide valuable insights into the mixing dynamics of flexible filamentous particles.

Genomics and proteomics of Apis mellifera filamentous virus isolated from honeybees in China

Apis mellifera filamentous virus(Am FV)is a large DNA virus that is endemic in honeybee colonies.The genome sequence of the Am FV Swiss isolate(Am FV CH–C05)has been reported,but so far very few molecular studies have been conducted on this virus.In this study,we isolated and purified Am FV(Am FV CN)from Chinese honeybee(Apis mellifera)colonies and elucidated its genomics and proteomics.Electron microscopy showed ovoid purified virions with dimensions of 300–500×210–285 nm,wrapping a 3165×40 nm filamentous nucleocapsid in three figure-eight loops.Unlike Am FV CH–C05,which was reported to have a circular genome,our data suggest that Am FV CN has a linear genome of approximately 493 kb.A total of 197 ORFs were identified,among which36 putative genes including 18 baculoviral homologs were annotated.The overall nucleotide similarity between the CN and CH–C05 isolates was 96.9%.Several ORFs were newly annotated in Am FV CN,including homologs of per os infectivity factor 4(PIF4)and a putative integrase.Phylogenomic analysis placed Am FVs on a separate branch within the newly proposed virus class Naldaviricetes.Proteomic analysis revealed 47 Am FV virionassociated proteins,of which 14 had over 50%sequence coverage,suggesting that they are likely to be main structural proteins.In addition,all six of the annotated PIFs(PIF-0–5)were identified by proteomics,suggesting that they may function as entry factors in Am FV infection.This study provides fundamental information regarding the molecular biology of Am FV.

Frontogenesis and Frontolysis of a Cold Filament Driven by the Cross-Filament Wind and Wave Fields Simulated by a Large Eddy Simulation

The variations of the frontogenetic trend of a cold filament induced by the cross-filament wind and wave fields are studied by a non-hydrostatic large eddy simulation. Five cases with different strengths of wind and wave fields are studied.The results show that the intense wind and wave fields further break the symmetries of submesoscale flow fields and suppress the levels of filament frontogenesis. The changes of secondary circulation directions—that is, the conversion between the convergence and divergence of the surface cross-filament currents with the downwelling and upwelling jets in the filament center—are associated with the inertial oscillation. The filament frontogenesis and frontolysis caused by the changes of secondary circulation directions may periodically sharpen and smooth the gradient of submesoscale flow fields.The lifecycle of the cold filament may include multiple stages of filament frontogenesis and frontolysis.

Plasma potential measurements using an emissive probe made of oxide cathode

A novel emissive probe consisting of an oxide cathode coating is developed to achieve a low operating temperature and long service life.The properties of the novel emissive probe are investigated in detail,in comparison with a traditional tungsten emissive probe,including the operating temperature,the electron emission capability and the plasma potential measurement.Studies of the operating temperature and electron emission capability show that the tungsten emissive probe usually works at a temperature of 1800 K-2200 K while the oxide cathode emissive probe can function at about 1200 K-1400 K.In addition,plasma potential measurements using the oxide cathode emissive probe with different techniques have been accomplished in microwave electron cyclotron resonance plasmas with different discharge powers.It is found that a reliable plasma potential can be obtained using the improved inflection point method and the hot probe with zero emission limit method,while the floating point method is invalid for the oxide cathode emissive probe.

Simple procedure for assessing diffuse subarachnoid hemorrhage successfully created using filament perforation method in mice

The murine model of subarachnoid hemorrhage(SAH)is a valuable experimental tool for investigating molecular and cellular mechanisms,and the endovascular filament perforation technique can be used to simulate prominent pathophysiological features observed after human SAH;however,current validation methods for assessing an appropriate SAH model are limited.Here,we introduce a simple procedure for se-lecting a mouse model of diffuse SAH.SAH was induced in 24 mice using a standard filament perforation method.After confirming survival at 24 h,SAH was scored 0-1 based on T2*-weighted images on whole-brain magnetic resonance imaging(MRI)and visual surveillance of the cisterna magna(CM)through the dura mater.The CM-based SAH grading correlated well with a reference parameter defined by extracted brain(r^(2)=0.53,p<0.0001).The receiver operating characteristic curve revealed a sensi-tivity of 85%and a specificity of 91%for detecting diffuse SAH,with a similar area under the curve(0.89±0.06[standard error of the mean])as the MRI-based grading(0.72±0.10,p=0.12).Our data suggest that confirming an SAH clot in the CM is a valuable way to select a clinically relevant diffuse SAH model that can be used in future experimental studies.