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comparison of plastid 16s rrna (rrn16) genes from helicosporidium spp.: evidence supporting the reclassification of helicosporidia as green algae (chlorophyta).the helicosporidia are invertebrate pathogens that have recently been identified as non-photosynthetic green algae (chlorophyta). in order to confirm the algal nature of the genus helicosporidium, the presence of a retained chloroplast genome in helicosporidia cells was investigated. fragments homologous to plastid 16s rrna (rrn16) genes were amplified successfully from cellular dna extracted from two different helicosporidium isolates. the fragment sequences are 1269 and 1266 bp long, are very ...200314657099
diagnosis of clinical bovine mastitis by fine needle aspiration followed by staining and scanning electron microscopy in a prototheca zopfii outbreak.biopsy by fine needle aspiration together with microbiological examination and scanning electron microscopy were evaluated in diagnosis of clinical bovine mastitis in a prototheca zopfii outbreak. fine needle aspiration was performed in 21 mammary quarters from ten holstein cows presenting clinical mastitis caused by p. zopfii. the algae were previously identified in the microbiological examination of milk collected from these cows. material aspirated from these 21 mammary glands was submitted t ...200415487325
first large-scale isolation of prototheca zopfii from milk produced by dairy herds in italy.a total of 1045 milk samples collected from infected and non infected quarters of 269 cows were investigated. this study showed that 4.7% of samples possessed cells of prototheca spp. (10(6)cells/ml). the presence of other pathogenic microorganisms was also monitored. prototheca spp. isolates were classified on the basis of current taxonomic guidelines and identified as p. zopfii. susceptibility tests carried out in vitro by using 25 antibiotic compounds revealed that the strains of p. zopfii. w ...200415630551
urinary tract manifestations of protothecosis in dogs.records of 13 dogs with systemic infection with prototheca sp. from 3 veterinary teaching hospitals were reviewed. acute renal failure secondary to disseminated infection with prototheca zopfii was diagnosed in 2 dogs. in 1 dog, acute renal failure developed during administration of immunosuppressive drugs for treatment of anterior uveitis. during diagnostic evaluation of this dog, prototheca sp. organisms were noted in urine sediment and renal biopsy specimens. in the 2nd dog, acute renal failu ...200515715059
protothecosis in a dog.a case of a disseminated algal infection is reported in a young rough-coated collie dog with progressive neurologic deficits, blindness, and hemorrhagic diarrhea. prototheca zopfii organisms were cultured from feces, urine, and blood. at necropsy, granulomas containing typical organisms were identified within the proximal colon, heart, kidneys, and eyes.201425320382
optimization of heterotrophic culture conditions for n-alkane utilization and phylogenetic position based on the 18s rdna sequence of a thermotolerant prototheca zopfii strain.this study reports on the optimization of the culture conditions for a thermotolerant eukaryotic algal strain, prototheca zopfii rnd16, which can effectively degrade and assimilate n-alkanes at elevated temperatures. rnd16 was able to grow on 1% (v/v) n-alkanes (c14-c17) at temperatures up to 38 degrees c. this ability differs from a previous finding that p. zopfii did not grow on hydrocarbons under temperature conditions above 25 degrees c. increasing the temperature from 25 degrees c to 30-35 ...200216233286
immunohistopathology of prototheca wickerhamii in cutaneous lesions of protothecosis.protothecosis is a rare infection caused by pathogenic algae of the genus prototheca. prototheca wickerhamii causes cutaneous/subcutaneous opportunistic infections in humans and small animals. the diagnosis of protothecosis is based on histopathological examination of this organism, which can be confused with other fungi and inflammatory cells in infected tissues. in this study, immunohistopathological investigation was made of infected cutaneous human and animal tissues exhibiting protothecosis ...201424682095
microbicidal action of indole-3-acetic acid combined with horseradish peroxidase on prototheca zopfii from bovine mastitis.we investigated the toxic effect of indole-3-acetic acid (iaa) combined with horseradish peroxidase (hrp) on prototheca zopfii from bovine mastitis. p. zopfii isolates were identified and characterized by morpho-physiological parameters; presences of p. zopfii genotype 2 were also investigated. subsequently, p. zopfii was incubated in the absence (control) or presence of iaa/hrp and examined for: (i) cell viability; (ii) colonies number formation; (iii) antioxidant enzyme activity; and (iv) dna ...201019707886
nature of the growth factor for the colorless alga prototheca zopfii.barker's study on the nutritive requirements of prototheca zopfii indicated that this colorless alga fails to grow in the absence of small amounts of yeast extract. a study of the growth factor requirements of prototheca has shown that the active constituent of yeast extract necessary for the growth of this organism is thiamin (vitamin b(1)). thiamin can fully replace the complex yeast material and allows, in the basal medium used, a maximum cell yield in concentrations of 1-3 x 10(-7)m. thiamin ...194519873421
26s rdna-based phylogenetic investigation of japanese cattle-associated prototheca zopfii isolates.the study has been conducted to assess the applicability of partial 26s ribosomal dna sequence for the phylogenic analysis of p.zopfii and to arrive relationships between specific genotype of p.zopfii and clinical mastitis. the phylogenetic analysis indicated that all genotype 2 isolates were grouped into the cluster of type strain of p.zopfii. thus, all mastitis isolates in japan were identified as p.zopfii genotype 2 and were independent of the cluster of the genotype 1 isolates. in one clinic ...201019915324
phenotypic characterization of mastitic prototheca spp. isolates.bovine mastitis associated with prototheca is considered a rare pathology, but is increasing in prevalence all over the world and therefore becoming more relevant to the dairy industry. the biochemical characterization of 47 prototheca isolates retrieved from mastitic milk was performed in this study using api 20c aux and two bbl crystal kits, followed by an analysis with inforbio software. the usage of this methodology, allowed the identification of discriminative phenotypic characteristics for ...201020096906
an unusual case of granulomatous lymphadenitis due to prototheca zopfii var. portoricensis in an immunocompetent man in china.protothecosis is an uncommon human infection caused by prototheca. prototheca spp can be considered as saprophytes, and in spite of their frequency in the environment, they are of low virulence and may cause chronic infection with low-grade inflammation in humans. at present, only three species are recognized: prototheca wickerhamii, prototheca zopfii and prototheca stagnora. of these, the former two have been associated with human disease. this study was an investigation of the clinical and mic ...201020117953
experimental infection of bovine mammary gland with prototheca zopfii genotype 1.prototheca zopfii is divided into three genotypes, one of which, p. zopfii genotype 2, appears to be the main causative agent of bovine protothecal mastitis. however, the difference in pathogenicity between genotypes 1 and 2 has not been well investigated. in the present study, we experimentally infected normal bovine mammary gland with p. zopfii genotype 1 to investigate its pathogenicity. the mammary gland infected with p. zopfii genotype 1 showed no clinical signs. however, the histopathologi ...201120736514
effect of different heat treatments and disinfectants on the survival of prototheca zopfii.bovine mastitis caused by the yeast-like alga prototheca zopfii represents a serious veterinary problem and may result in heavy economic losses to particular dairy farms. the purpose of this study was to evaluate the survival of 50 isolates of p. zopfii in milk subjected to different heat treatments and the survival of further 106 p. zopfii isolates after exposure to three classes of teat disinfectants: iodine (dipal), quaternary ammonium compounds (teat), and dodecylbenzenesulphonic acid (blu-g ...201120853028
molecular characterization of prototheca strains isolated from italian dairy herds.one hundred sixty-one prototheca spp. strains isolated from composite milk and barn-surrounding environmental samples (bedding, feces, drinking, or washing water, surface swabs) of 24 italian dairy herds were characterized by genotype-specific pcr analysis. overall, 97.2% of strains isolated from composite milk samples were characterized as prototheca zopfii genotype 2, confirming its role as the main mastitis pathogen, whereas prototheca blaschkeae was only sporadically isolated (2.8%). regardi ...201020854996
a novel dna extraction and duplex polymerase chain reaction assay for the rapid detection of prototheca zopfii genotype 2 in milk.aims:ôçé to develop a pcr-based assay to detect prototheca zopfii (p.-ázopfii) and its mastitis-related subtype (genotype 2) directly from milk samples. methods and results:ôçé the dna extraction method herein is based on the lysing properties of chemical agents, mechanical grinding and dna-binding properties of silica particles; this method was developed to rapidly extract dna directly from p.-ázopfii in bovine milk. two pairs of primers specific for p.-ázopfii and genotype 2 were used in the d ...201121679201
in vitro algaecide effect of disinfectants on prototheca zopfii genotypes 1 and 2.bovine mastitis due to prototheca zopfii leads to reduced milk production and is difficult to cure. therefore, prevention is the best approach and this is best achieved through the use of effective disinfectants. the aim of this study was to evaluate the in vitro algaecide efficacy of conventional disinfectants against strains of p. zopfii genotype 1 and 2. the minimal algaecide concentration (mac) of alkyldiaminoethylglycine hydrochloride, chlorhexidine, dioxide chlorine, povidone iodine and so ...201121737962
cutaneous protothecosis in patient with diabetes mellitus and review of published case reports.protothecosis is an opportunistic infection caused by prototheca, usually called as saprophytes, and is frequently found in natural and living surroundings with low virulence, but may cause chronic infection in immunocompromised individuals. we report a case of cutaneous protothecosis with zopfii var. portoricensis infection in a 66-year-old diabetic woman following hand surgery on middle right finger. mycology study showed that smooth, creamy white, yeast-like colonies grown after necrotic tiss ...201121964624
epidemiological analysis of worldwide bovine, canine and human clinical prototheca isolates by pcr genotyping and maldi-tof mass spectrometry proteomic phenotyping.this study presents information on the phenotypic and genotypic characterization of clinical prototheca spp. isolates obtained from different geographic regions. of 350 isolates studied, 342 came from cattle, six from canines and two from humans. phenotypic characterization was carried out by a matrix-assisted laser desorption/ionization-time of flight (maldi-tof) proteomic analysis. the peptide extraction that was used for this analysis included the additional steps of washing and sonication to ...201121905948
characterization of prototheca zopfii associated with outbreak of bovine clinical mastitis in herd of beijing, china.prototheca zopfii (p. zopfii) has become an important cause of bovine mastitis in many countries. in the present study, to better understand the occurrence of one clinical mastitis (cm) outbreak due to p. zopfii, the molecular characterization and resistance patterns of the microalga were described. p. zopfii strains were isolated from 17 of 23 quarters, which suffered cm in the outbreak, and 7 of 46 cm recovered quarters before the outbreak, as well as 2 of 75 environmental samples in the dairy ...201122160589
disseminated protothecosis diagnosed by evaluation of csf in a dog.a 5-year-old female spayed shetland sheepdog mix dog was evaluated for a history of recent seizure activity, progressive hind limb ataxia, polyuria, and polydipsia and no history of gastrointestinal signs. physical examination findings included conscious proprioceptive deficits, ataxia, and anterior uveitis along with a hypermature cataract in the right eye. results of a cbc, serum biochemical profile, urinalysis, and computed tomography scan of the brain were unremarkable. cerebrospinal fluid ( ...201222251039
well-known surface and extracellular antigens of pathogenic microorganisms among the immunodominant proteins of the infectious microalgae prototheca zopfii.microalgae of the genus prototheca (p.) are associated with rare but severe infections (protothecosis) and represent a potential zoonotic risk. genotype (gt) 2 of p. zopfii has been established as pathogenic agent for humans, dogs, and cattle, whereas gt1 is considered to be non-pathogenic. since pathogenesis is poorly understood, the aim of this study was to determine immunogenic proteins and potential virulence factors of p. zopfii gt2. therefore, 2d western blot analyses with sera and isolate ...201526484314
biosynthesis of dolichyl phosphate: characterization and site of synthesis in algae.this is the first report not only on the presence of polyprenyl phosphates and their site of synthesis in algae, but also on the formation of their sugar derivatives in this system.a glucose acceptor lipid was isolated from the nonphotosynthetic alga prototheca zopfii. the lipid was acidic and resistant to mild acid and alkaline treatments. the glucosylated lipid was labile to mild acid hydrolysis and resistant to phenol treatment and catalytic hydrogenation, as dolichyl phosphate glucose is. th ...197816660269
diversity within the current algal species prototheca zopfii: a proposal for two prototheca zopfii genotypes and description of a novel species, prototheca blaschkeae sp. nov.in order to clarify the intraspecies taxonomic position of the non-photosynthetic algal species prototheca zopfii, as well as the aetiology of bovine mammary protothecosis, a selection of p. zopfii strains isolated from clinical cases of protothecal mastitis and from various environmental habitats was characterized using a polyphasic molecular approach. based on sequence analysis of the 18s rrna gene, which showed distinct differences between the three currently known biotypes or 'variants', spe ...200616738123
the assimilation of acetate and propionate by prototheca zopfi.1. the tricarboxylic acid and glyoxylate cycles are of major importance in the assimilation of acetate and propionate by prototheca zopfii. the pattern of assimilation of [2-(14)c]acetate and [2-(14)c]propionate by whole cells growing with their respective substrates is similar except that, with propionate, beta-hydroxypropionate is the first labelled intermediate detected. 2. carbon dioxide fixation is of little quantitative importance for the growth of this organism with propionate. 3. the yie ...196516749100
effect of prototheca zopfii on neutrophil function from bovine milk.this study was carried to investigate neutrophil function in the presence of prototheca zopfii. for this purpose, bovine milk neutrophils were incubated in the absence (control) of and presence of p. zopfii, and then they were examined hydrogen peroxide (h(2)o(2)) production, antioxidant enzyme activities, and phagocytic capacity. milk was collected from negative "california mastitis test" (cmt) quarter from three lactating holstein cows after induction of leukocytosis with an intramammary infus ...200617146586
human protothecosis.human protothecosis is a rare infection caused by members of the genus prototheca. prototheca species are generally considered to be achlorophyllic algae and are ubiquitous in nature. the occurrence of protothecosis can be local or disseminated and acute or chronic, with the latter being more common. diseases have been classified as (i) cutaneous lesions, (ii) olecranon bursitis, or (iii) disseminated or systemic manifestations. infections can occur in both immunocompetent and immunosuppressed p ...200717428884
repeated batch cultivation of the hydrocarbon-degrading, micro-algal strain prototheca zopfii rnd16 immobilized in polyurethane foam.this study reports on the stability of the cells of a heterotrophic green micro-algal strain prototheca zopfii rnd16 immobilized in polyurethane foam (puf) cubes during degradation of mixed hydrocarbon substrate, which was composed of n-alkanes and polycyclic aromatic hydrocarbons (pahs), in 5 successive cycles of repeated batch cultivation at 30 degrees c. both rnd16 cells and mixed hydrocarbon substrate components had been entrapped in puf cubes through cultivation. puf-immobilized rnd16 degra ...200818388973
large-scale screening of the in vitro susceptibility of prototheca zopfii towards polyene antibiotics.a large scale screening of the in vitro susceptibility of 105 strains of prototheca zopfii to a panel of polyene antibiotics (amphotericin b, nystatin, primaricin and filipin) was conducted. strains studied were isolated from dairy-associated environments in five different localities. groups 1-4 included strains recovered from four separate regions of italy, while group 5 included isolates from belgium. amphotericin b and primaricin exhibited the highest activity, with th mic90 ranging from 4 an ...200818608920
short communication: isolation of prototheca species strains from environmental sources in dairy herds.composite milk samples from 548 cows, and samples from feces, feed, bedding, water, liners (before and after milking), and the postdipping product were aseptically collected from 2 italian dairy herds from february to november of 2006. prototheca zopfii was isolated from 11.9% of milk samples, 15% of feces, and 33.3% of bedding samples. no viable cells of p. zopfii were observed in water before washing procedures, whereas 25 to 28.6% of samples from water used for washing both refrigeration tank ...200818765606
rapid identification of prototheca zopfii by nested polymerase chain reaction based on the nuclear small subunit ribosomal dna. 200919203861
the in vitro efficacy of essential oils and antifungal drugs against prototheca zopfii.the algae of the genus prototheca are environmental pathogens whose main reservoir is the habitat of cows. they can cause protothecosis in domestic and wild animals, as well as human beings, with the main etiological agents being prototheca zopfii in animals and prototheca wickerhamii in humans.201626940809
prototheca miyajii sp. nov., isolated from a patient with systemic protothecosis.species of the genus prototheca are achlorophyllous algae and ubiquitous in nature, and so far, six species have been listed in this genus: prototheca wickerhamii, prototheca zopfii, prototheca blaschkeae, prototheca cutis, prototheca stagnora and prototheca ulmea. a strain of the genus prototheca, ifm 53848t, was isolated in japan from a patient with systemic protothecosis and had been designated p. wickerhamii. our previous study, by using pcr analysis, revealed that its ssu rrna gene (rdna) w ...201626791403
achlorophyllous alga prototheca zopfii oxidizes n-alkanes with different carbon-chain lengths through a unique subterminal oxidation pathway.some prototheca spp. were previously reported to convert n-hexadecane to 5-hexadecanol and then to 5-hexadecanone through a unique subterminal oxidation pathway. further analysis of derivatives derived from n-hexadecane indicated that prototheca zopfii oxidized n-alkanes with c11 to c17 chain lengths at not only the 5th but also the 4th, 3rd and 2nd positions.201424099955
onychoprotothecosis: an uncommon presentation of protothecosis.onychomycosis is a fairly common condition seen in a dermatology clinic. dermatophytes trichophyton and epidermophyton are the known filamentous fungi implicated. the yeast-like fungi such as candida less commonly cause onychomycosis. the genus prototheca may on preliminary observation resemble yeast-like fungi but a detailed microscopy will reveal the absence of budding and presence of endospores. onychoprotothecosis is an uncommon presentation of human protothecosis. of the two prototheca spec ...201526068353
prototheca zopfii genotype 2-induced nasal dermatitis in a cat.few published cases of feline protothecosis exist; all of these were restricted to the skin and speciation of the causative organism revealed an infection with prototheca wickerhamii in each case. this report describes prototheca zopfii genotype 2-induced inflammation of the nasal skin and cutaneous mucosa of the right nostril in a 14-year-old neutered female domestic shorthair cat. microscopical examination revealed marked pyogranulomatous inflammation with numerous intralesional algae. these h ...201525814431
protothecosis in hematopoietic stem cell transplantation: case report and review of previous cases.prototheca species are achlorophyllus algae. prototheca wickerhamii and prototheca zopfii cause human disease. in immunocompetent individuals, they cause soft tissue infections and olecranon bursitis, but in transplant recipients, these organisms can cause disseminated disease. we report a fatal case of disseminated p. zopfii infection in an hematopoietic stem cell transplant (hsct) recipient with bloodstream infection and involvement of multiple soft tissue sites. we review all previous cases o ...201424797402
protothecosis.protothecosis is a rare infection caused by achlorophyllic algae that are members of the genus prototheca. they are ubiquitous in nature in organic material. the clinical manifestations can be acute or chronic and local or disseminated. the disease is classified as cutaneous, causing bursitis or disseminated/systemic, affecting both immunocompetent and immunosuppressed patients, with more severe and disseminated infections occurring in immunocompromised individuals. prototheca wickerhamii and pr ...201322682193
detection of helicosporidium spp. in metagenomic dna.distinct isolates of the invertebrate pathogenic alga helicosporidium sp., collected from different insect hosts and different geographic locations, were processed to sequence the 18s rdna and β-tubulin genes. the sequences were analyzed to assess genetic variation within the genus helicosporidium and to design helicosporidium-specific 18s rdna primers. the specificity of these primers was demonstrated by testing not only on the helicosporidium sp. isolates, but also on two trebouxiophyte algae ...201222609409
in vitro algicidal effect of guanidine on prototheca zopfii genotype 2 strains isolated from clinical and subclinical bovine mastitis.prototheca species have increasingly been reported to be opportunistic pathogens that cause mastitis in dairy herds, and it poses an emergent problem because at present, there are no effective therapies for the treatment of protothecal mastitis. the present study investigated the in vitro algicidal effect of guanidine on 75 prototheca zopfii genotype 2 strains isolated from 75 cases of clinical and subclinical bovine mastitis. all strains were susceptible to guanidine in vitro with minimal algae ...201728349671
evidence of a prototheca zopfii genotype 2 disseminated infection in a dog with cutaneous lesions.protothecosis is a disease caused by saprophyte aerobic unicellular algae belonging to the genus prototheca. in dogs, it mainly occurs as a disseminated form, with initial clinical manifestations often referable to the gastrointestinal tract, followed by typical ocular and neurological signs. so far, prototheca zopfii genotype 2 infection has been reported in severe forms of disseminated protothecosis, while in dogs has never been associated with cutaneous forms. in this study, we describe a cas ...201628025757
isobaric tag for relative and absolute quantitation-based comparative proteomic analysis of human pathogenic prototheca zopfii genotype 2 and environmental genotype 1 strains.prototheca species are ubiquitous achlorophyllic microalgae belonging to the family chlorellaceae, which can cause a wide range of infections in humans and animals. mainly in individuals with immunologic defects or trauma, prototheca spp. can cause even lethal diseases. however, the exact pathogenic mechanism of prototheca in causing disease remains largely unknown. to investigate the differences between pathogenic and nonpathogenic prototheca spp. genotypes on proteome level, a nonpathogenic pr ...201627590985
short communication: molecular typing of prototheca zopfii from bovine mastitis in japan.prototheca zopfii causes bovine mastitis, resulting in reduced milk production and the secretion of thin watery milk with white flakes. prototheca zopfii has been biochemically and serologically divided into at least 2 genotypes, p. zopfii genotype 1 and p. zopfii genotype 2. the latter is known to be the main causative agent of bovine protothecal mastitis. prototheca zopfii was later reclassified into 5 varieties: var. zopfii (genotypes 1 and 2), var. 1 (formerly prototheca blaschkeae), var. 3 ...201222818457
an investigation of the innate immune response in bovine mammary epithelial cells challenged by prototheca zopfii.prototheca zopfii is an important bovine mastitis pathogen, which could result in severe mammary infection. however, the innate immune response in bovine mastitis associated with p. zopfii was not clear. therefore, the aim of this study is to investigate in vitro innate immune responses implicated by p. zopfii. bovine mammary epithelial cells (bmecs) were infected with 5.0 × 10(4) cells/ml p. zopfii genotypes i and ii independently, and the mrna expression of tlr-2, tlr-4, tnf-α, il-1β, il-8, no ...201627558223
prototheca species and pithomyces chartarum as causative agents of rhinitis and/or sinusitis in horses.pyogranulomatous rhinitis associated with an algal infection was diagnosed in a 25-year-old gelding and a 23-year-old mare had necrotizing sinusitis with intralesional algae and pigmented fungi. algae were identified immunohistochemically in both cases as prototheca spp. in the gelding, further characterization by polymerase chain reaction and sequencing revealed that the organism was prototheca zopfii genotype 2. fungi from the mare were identified as pithomyces chartarum by molecular analysis. ...201627394651
short communication: elisa system for screening of bovine mastitis caused by prototheca zopfii.prototheca zopfii is an achlorophyllic alga that causes bovine mastitis, resulting in a reduction in milk production and the secretion of thin, watery milk with white flakes. this study evaluated the use of an elisa system for distinguishing cows with mastitis due to p. zopfii genotype 2 from healthy cows and cows with chronic candidal mastitis. we also investigated the transitional changes of specific antibody titers in healthy cows injected with inactivated p. zopfii genotype 2 cells. the elis ...201627265167
molecular typing and differences in biofilm formation and antibiotic susceptibilities among prototheca strains isolated in italy and brazil.bovine mastitis caused by prototheca is a serious and complex problem that accounts for high economic losses in the dairy industry. the main objective of this study was to identify and characterize at genetic level different prototheca strains and provide the most complete data about protothecal antibiotic resistance. the study involves 46 isolates from italian (13 strains) and brazilian (33 strains) mastitic milk. these strains were identified by multiplex pcr and single strand conformation pol ...201627236754
characterization of prototheca zopfii genotypes isolated from cases of bovine mastitis and cow barns in china.protothecal mastitis, caused mostly by prototheca zopfii (p. zopfii), is increasing in dairy herds and is being reported globally. the present study was aimed at studying the epidemiology of mastitis and at molecular characterization of p. zopfii isolates from dairy herds and their surroundings in three provinces of china using microbiological, biochemical and molecular methods, and antibiotic susceptibility tests. samples from milk (n = 620) of mastitic cows and their barns sources (n = 410) in ...201626450620
in vitro photoinactivation of bovine mastitis related pathogens.bovine mastitis is considered the most important disease of worldwide dairy industry. treatment of this disease is based on the application intramammary antibiotic, which favors an increase in the number of resistant bacteria in the last decade. photodynamic inactivation (pdi) has been investigated in different areas of health sciences, and has shown great potential for inactivating different pathogens, without any selection of resistant microorganisms. the objective of this study was to investi ...201626315923
biofilm-producing ability and efficiency of sanitizing agents against prototheca zopfii isolates from bovine subclinical mastitis.the objectives of the present study were to evaluate (1) the capacity of the microalga prototheca zopfii isolated from subclinical bovine mastitis cases to form biofilms; and (2) the resistance of these isolates to sanitizing agents. ten isolates of p. zopfii from cows with subclinical mastitis (somatic cell count>200×10(3) cells/ml), distributed in 5 dairy farms, were evaluated for their capacity to form biofilms in polystyrene microplate assays and stainless steel coupons, at 25°c and 37°c±1°c ...201525841971
identification of immunodominant proteins of the microalgae prototheca by proteomic analysis.prototheca zopfii associated with bovine mastitis and human protothecosis exists as two genotypes, of which genotype 1 is considered as non-infectious and genotype 2 as infectious. the mechanism of infection has not yet been described. the present study was aimed to identify genotype 2-specific immunodominant proteins. prototheca proteins were separated using two-dimensional gel electrophoresis. subsequent western blotting with rabbit hyperimmune serum revealed 28 protein spots. matrix-assisted ...201525755891
internal transcribed spacer sequence-based rapid molecular identification of prototheca zopfii and prototheca blaschkeae directly from milk of infected cows.the increasing incidence of rare mastitis-causing pathogens has urged the implementation of fast and efficient diagnostic and control measures. prototheca algae are known to be associated with diseases in humans and animals. in the latter, the most prevalent form of protothecosis is bovine mastitis with prototheca zopfii and prototheca blaschkeae representing the most common pathogenic species. these nonphotosynthetic and colorless green algae are ubiquitous in different environments and are wid ...201525726118
induction of oxidative stress in prototheca zopfii by indole-3-acetic acid/hrp or 2,4-pentanedione/hrp systems and their oxidation products.we investigated the toxic effects on prototheca zopfii of indole-3-acetic acid (iaa) and 2,4-pentanedione (pd) combined with horseradish peroxidase (hrp) alongside the oxidation products of 3-methyl-2-oxindole (moi) and indole-3-carbinol (i3c) from the iaa/hrp system and methylglyoxal (mgo) from the pd/hrp system. the microorganism was incubated in the absence (control) or presence of iaa, pd, iaa/hrp, pd/hrp, moi, i3c and mgo and determined: (1) cytotoxicity by mtt (3-(4,5-dimethylthiazol-2-yl) ...201525173924
short communication: dairy bedding type affects survival of prototheca in vitro.protothecae are algal pathogens, capable of causing bovine mastitis, that are unresponsive to treatment; they are believed to have an environmental reservoir. the role of bedding management in control of protothecal mastitis has not been studied. the purpose of this study was to evaluate the growth of either environmental or mastitis-associated prototheca genotypes in dairy bedding materials that are commonly used in maine. prototheca zopfii genotypes 1 and 2 (gt1 and gt2) were inoculated into s ...201324119794
detection and identification of genotypes of prototheca zopfii in clinical samples by quantitative pcr analysis.in this study, a specific quantitative pcr system for the detection and identification of prototheca zopfii genotypes was developed using a taqman(®) mgb probe and resolight dye. the p. zopfii-specific primers 18pzf1 and 18pzr1 were generated on the basis of the alignment of the small subunit ribosomal dna domain base sequences of the genera chlorella and prototheca obtained from ddbj/embl/genbank, and the taqman(®) mgb probe pzp1 was designed corresponding to this amplification region. analysis ...201324047735
comparative proteomics lends insight into genotype-specific pathogenicity.comparative proteomic analyses have emerged as a powerful tool for the identification of unique biomarkers and mechanisms of pathogenesis. in this issue of proteomics, murugaiyan et al. utilize difference gel electrophoresis (dige) to examine differential protein expression between nonpathogenic and pathogenic genotypes of prototheca zopfii, a causative agent in bovine enteritis and mastitis. their findings provide insights into molecular mechanisms of infection and evolutionary adaptation of pa ...201323925996
two-dimensional proteome reference map of prototheca zopfii revealed reduced metabolism and enhanced signal transduction as adaptation to an infectious life style.biochemical, serological, and genetic analyses have identified two genotypes of prototheca zopfii, a unicellular microalga belonging to the family chlorellaceae. the p. zopfii genotype 1, abundantly present in cow barns and environment, remains nonpathogenic, while p. zopfii genotype 2 has been isolated from cows with bovine mastitis. the present study was carried out to identify the protein expression level difference between the pathogenic and nonpathogenic strains of p. zopfii. a total of 782 ...201323852777
treatment with gentamicin on a murine model of protothecal mastitis.the aim of this study was to establish a murine protothecal mastitis model and to evaluate the treatment efficiency of gentamicin. challenge routes were determined with a pathogenic prototheca zopfii genotype 2 (p. zopfii) strain. 25 balb/c mice were inoculated in mammary glands with graded dosages (10(3), 10(4), 10(5), 10(6), 10(7) cfu of p. zopfii) and killed on the 7th day. another 25 animals were also killed at 1, 3, 5, 7, and 9 days after inoculation of 1 × 10(6) cfu of p. zopfii, the milk ...201323463523
first outbreak of bovine mastitis caused by prototheca blaschkeae.the most important animal disease caused by yeast-like algae belonging to the genus prototheca is bovine mastitis. although the infection can be caused by both prototheca zopfii genotype 2 and prototheca blaschkeae, the bulk of prevalence of bovine protothecal mastitis has been so far attributed to the former, being p. blaschkeae only sporadically isolated. however, we report here the first outbreak of bovine mastitis caused by p. blaschkeae in an italian dairy herd. one hundred and four individ ...201323201242
identification of prototheca zopfii from bovine mastitis.the aim of this study was identification of the epidemiology of prototheca zopfii species from the milk samples of dairy cattle in isfahan, central iran.201223113230
herd characteristics and cow-level factors associated with prototheca mastitis on dairy farms in ontario, canada.prototheca spp. are algae that cause incurable acute or chronic mastitis in dairy cows. the aim of this case-control study was the identification of cow- and herd-level risk factors for this unusual mastitis pathogen. aseptically collected composite milk samples from 2,428 milking cows in 23 case and 23 control herds were collected between january and may 2011. a questionnaire was administered to the producers, and cow-level production and demographic data were gathered. in 58 of 64 isolates, pr ...201222884347
galactomannan enzymatic immunoassay cross-reactivity caused by prototheca species.we report a reactive aspergillus galactomannan enzymatic immunoassay against the serum of a patient with invasive prototheca zopfii infection. analysis of the supernatants of suspensions of p. zopfii and other prototheca isolates revealed positive results as well. these data suggest cross-reactivity with the serum aspergillus galactomannan assay in invasive protothecosis.201222837317
multicentre etest evaluation of in vitro activity of conventional antifungal drugs against european bovine mastitis prototheca spp. isolates.bovine mammary protothecosis is a serious pathology that entails high economic losses in the dairy industry. the disease, the frequency of which has recently been increasing worldwide, is caused by unicellular, achlorophyllous, yeast-like algae of two species: prototheca zopfii and prototheca blaschkeae. the objective of this study was to investigate the in vitro activity of a panel of conventional antifungal drugs against prototheca spp. isolates.201222523316
serial mri and csf analysis in a dog treated with intrathecal amphotericin b for protothecosis.a 3 yr old female spayed english setter mixed-breed dog presented with diarrhea, weight loss, side stepping to the right, and a right head tilt. rectal and cerebrospinal fluid cytology and culture confirmed a diagnosis of prototheca zopfii. mri of the brain showed inflammation of the brain and meninges, ventriculomegaly, and syringomyelia. treatment with prednisone, itraconazole, and amphotericin b lipid complex administered intrathecally yielded transient improvement. progressive brainstem sign ...201322267176
β-glucan biosynthesis in synchronous cells of prototheca zopfii.synchronous cultures of the colorless chlorophyte prototheca zopfii kruger were obtained by a shift-down of the basal medium plus addition of 1.5 mm ethylenediaminetetracetic acid for a 10-14-h period, followed by periodic dilutions with the basal medium. the cell cycle has a duration of 10-12 h at 25°c and an average of eight autospores are produced at the end of each cycle. the incorporation of [(14)c]glucose into β-glucans was determined in vivo. very high incorporation occurs between hours 3 ...198524241139
prototheca zopfii induced ultrastructural features associated with apoptosis in bovine mammary epithelial cells.prototheca zopfii infections are becoming global concerns in humans and animals. bovine protothecal mastitis is characterized by deteriorating milk quality and quantity, thus imparting huge economic losses to dairy industry. previous published studies mostly focused on the prevalence and characterization of p. zopfii from mastitis. however, the ultrastructural pathomorphological changes associated with apoptosis in bovine mammary epithelial cells (bmecs) are not studied yet. therefore, in this s ...201728752077
a comparative study of the in vitro activity of iodopropynyl butylcarbamate and amphotericin b against prototheca spp. isolates from european dairy herds.the objective of this study was to assess the in vitro effect of iodopropynyl butylcarbamate (ipbc) and amphotericin b (amb) on prototheca zopfii genotype 2 and prototheca blaschkeae isolates recovered from dairy herds of belgium, france, italy, germany, and poland. the combination of ipbc with amb on prototheca isolates and toxicity of ipbc to the bovine mammary epithelial cells were also evaluated. the in vitro activity of ipbc and amb against 96 isolates of p. zopfii genotype 2 and 42 isolate ...201728711267
molecular characterization of prototheca strains isolated in china revealed the first cases of protothecosis associated with prototheca zopfii genotype 1.in this study, six strains of prototheca isolated in china from human patients diagnosed as protothecosis and cows with mastitis were characterized by polymerase chain reaction (pcr) and nucleotide sequencing of the ribosomal rna gene (rdna) and by matrix-assisted laser desorption/ionization time of flight mass spectrometry (maldi-tof ms). the results indicated that three strains isolated from the human patients were p. zopfii genotype 1, revealing the first cases of human protothecosis associat ...201728525645
mass spectrometry data from label-free quantitative proteomic analysis of harmless and pathogenic strains of infectious microalgae, prototheca spp.here, we provide the dataset associated with our research article 'label-free quantitative proteomic analysis of harmless and pathogenic strains of infectious microalgae, prototheca spp.' (murugaiyan et al., 2017) [1]. this dataset describes liquid chromatography-mass spectrometry (lc-ms)-based protein identification and quantification of a non-infectious strain, prototheca zopfii genotype 1 and two strains associated with severe and mild infections, respectively, p. zopfii genotype 2 and protot ...201728480323
prototheca zopfii isolated from bovine mastitis induced oxidative stress and apoptosis in bovine mammary epithelial cells.bovine protothecal mastitis results in considerable economic losses worldwide. however, prototheca zopfii induced morphological alterations and oxidative stress in bovine mammary epithelial cells (bmecs) is not comprehensively studied yet. therefore, the aim of this current study was to investigate the p. zopfii induced pathomorphological changes, oxidative stress and apoptosis in bmecs. oxidative stress was assessed by evaluating catalase (cat), superoxide dismutase (sod), glutathione peroxidas ...201728404882
the first case of human protothecosis caused by prototheca zopfii in japan.this report describes a fatal case prototheca zopfii genotype 2 infection in an immunosuppressed patient. the patient was a 62-year-old housewife who presented general malaise in april 2011. hairy cell leukemia was highly suspected. chemotherapy was started because the patient developed severe pancytopenia in october 2011. itraconazole capsules (100 mg/day) and trimethoprim (320 mg/day) plus sulfamethoxazole (1600 mg/day) combinations were orally administered for prophylaxis of fungal infections ...201425048395
paraparesis as initial manifestation of a prototheca zopfii infection in a dog.a case of protothecosis causing non-ambulatory paraparesis in a dog without clinical evidence of disseminated infection is described. a five-year-old female labrador retriever was referred with a 10-day history of progressive non-ambulatory paraparesis and lumbar pain as the only physical and neurological abnormalities. lumbar myelography revealed severe extradural spinal cord compression extending from l4 to l7 vertebrae, and a right hemilaminectomy was performed. surgical findings included an ...201424502403
human cutaneous protothecosis: report of a case and literature review.the prototheca species is achlorophyllic algae and rarely causes human infection. human protothecosis presents clinically as a cutaneous infection, olecranon bursitis, and disseminated systemic disease. we report a case of human cutaneous protothecosis involving the left wrist. a 68-year-old man presented with an ill-defined erythematous lesion with crust at the dorsal aspect of his left wrist. a punch biopsy was performed to reveal the histologic features of granulomatous inflammation with necr ...201324421852
antimicrobial activity of mentha piperita and saturenja hortensis in a murine model of cutaneous protothecosis.to date there is no defined pharmacologic treatment protocol available against cutaneous protothecosis, which is difficult to combat using conventional drugs.201424387811
rapid molecular typing of prototheca zopfii by high resolution melting real-time pcr (pcr-hrm).prototheca zopfii is an achlorophyllic alga that is ubiquitous around cow sheds. the alga is associated with bovine mastitis, which causes a reduction in milk production and secretion of thin watery milk containing white flakes. isolates of p. zopfii from bovine mastitis were almost all identified as p. zopfii genotype 2, suggesting that it is the main causative agent of bovine protothecal mastitis. the ability to differentiate between genotype 1 and genotype 2 is therefore very important for pr ...201324292136
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