Cankered, dying seedlings of Juglans regia were observed in Shaanxi province in the northwest region of China. Neofusicoccum parvum was isolated from these cankered tissues, with the identification based on mor- pholo...Cankered, dying seedlings of Juglans regia were observed in Shaanxi province in the northwest region of China. Neofusicoccum parvum was isolated from these cankered tissues, with the identification based on mor- phology and an ITS-nrDNA sequence. In order to demonstrate how cultures of N. parvum could cause the expected symptoms, artificial infection, using these isolates and re-isolation of the pathogen, was used. This is the first report on this taxon as a walnut canker pathogen in China.展开更多
Three new mycophenolic acid derivatives, penicacids E-G(1-3), together with three known analogues, mycophenolic acid(4), 4′-hydroxy-mycophenolic acid(5) and mycophenolic methyl ester(6), were isolated from a marine-d...Three new mycophenolic acid derivatives, penicacids E-G(1-3), together with three known analogues, mycophenolic acid(4), 4′-hydroxy-mycophenolic acid(5) and mycophenolic methyl ester(6), were isolated from a marine-derived fungus Penicillium parvum HDN17-478 from a South China Sea marine sediment sample. The structures of compounds 1-3 were elucidated by HRMS, NMR, and Mosher’s method. Among them, compounds 1 and 2 were the first examples of mycophenolic acid analogs with a double bond at C-3′/C-4′ position. The cytotoxicity of 1-6 was evaluated against the HCT-116, BEL-7402, MGC-803, SH-SY5 Y, HO-8910 and HL-60 cell lines, and compounds 4 and 6 showed potent cytotoxicity with IC50 values ranging from 1.69 to 12.98 μmol·L–1.展开更多
Neofusicoccum parvum,an endophytic fungus isolated from Elaeocarpus serratus(Ceylon Olive;family Elaeocarpaceae),was grown for 3 weeks in potato dextrose broth.Chromatographic separation of the ethyl acetate extracts ...Neofusicoccum parvum,an endophytic fungus isolated from Elaeocarpus serratus(Ceylon Olive;family Elaeocarpaceae),was grown for 3 weeks in potato dextrose broth.Chromatographic separation of the ethyl acetate extracts from the culture filtrate and mycelium over silica gel,Sephadex LH-20 and preparative thin layer chromatography furnished(R)-7-hydroxymellein(1),(3R,4R)-4-hydroxymellein(2),(3R,4S)-4-hydroxymellein(3),(R)-5-hydroxymellein(4),(R)-mellein(5),(3R,4R)-4,7-dihydroxymellein(6),(6R,7S)-dia-asperlin(7),CJ-14445(8)and 13,14,15,16-tetranorlabd-7-ene-19,6β:12,17-diolide(9).The structures of known compounds 1–9 were determined by spectroscopic methods and comparison with reported data.This is the first report of the isolation of an endophytic fungus from E.serratus,and the isolation of compounds 1,4,6,8 and 9 from N.parvum.It is important to note that compounds 1–7 are small molecules with an oxygen heterocyclic ring system.These compounds can be used as starting materials in the synthesis of pharmaceutically important large molecules with oxygen heterocyles.展开更多
The CP15/60 gene encoding the CP15/60 surface protein of sporozoites in Cryptosporidiumparvum was obtained by PCR so as to research the nucleic vaccine against C.parvum. Theeukaryotic expressing vector pcDNA3-15/60 wa...The CP15/60 gene encoding the CP15/60 surface protein of sporozoites in Cryptosporidiumparvum was obtained by PCR so as to research the nucleic vaccine against C.parvum. Theeukaryotic expressing vector pcDNA3-15/60 was constructed by inserting CP15/60 gene intopcDNA3 (+) in XhoⅠand EcoRⅠ. A vaccination protocol was the adult pregnant goatsinoculated intranasally with the pcDNA3-15/60 plasmid and their offspring were infectedwith C.parvum oocysts. The results showed that the pcDNA3-15/60 plasmid can induce theimmune response of goats and the vaccinated goats can transfer the immunity to offspringconferring protection against C.parvum infection. These suggested that the recombinantplasmid could be a DNA vaccine candidate.展开更多
With the discovery that the coccidian parasite Cryptosporidium sp. can cause severe symptoms in humans, many diagnostic techniques were quickly implemented such as microscopic visualization, immunofluorescence and PCR...With the discovery that the coccidian parasite Cryptosporidium sp. can cause severe symptoms in humans, many diagnostic techniques were quickly implemented such as microscopic visualization, immunofluorescence and PCR. Currently, there is no effective drug treatment and none of the current diagnostic methods is 100% accurate. In this study, a BALB/C mouse was subcutaneously immunized with Cryptosporidium parvum oocysts extract. The spleen was removed and the splenocytes were fused with SP2/0 myeloma cells in order to obtain hybridoma cells secreting antibodies specific to C. parvum antigens. Human and cattle fecal samples previously characterized by microscopy [Ziehl-Neelsen staining (ZN) and Lugol] and PCR for the presence of C. parvum and Giardia duodenalis, were analyzed by indirect immunofluorescence, using the developed hybridomas supernatants. The study shows that the selected hybridomas supernatants identify C. parvum oocysts in fecal samples in correlation with C. parvum oocysts identified using ZN/PCR. No false positive results were obtained and the two best supernatants gave 20% -30% of false negative results. No cross reaction with G. duodenalis was observed. By comparing our results with those obtained with an immunofluorescence commercial kit, it suggests the potential use of the monoclonal antibodies present in two of the hybridomas supernatants as a detection tool of C. parvum. With a reliability of 80.8% and 73.1% versus ZN and PCR methods for IFI, compared with a reliability of 76.9% and 92.3% versus ZN and PCR for commercial DIF kit, the supernatant 4.1D5 seems to be the most promising subject to further study its usefulness for C. parvum detection.展开更多
[Objectives]The study was to identify the causal agent of leaf spots of Camellia drupifera in Hainan Province,China.[Methods]Phylogenetic analyses,spore morphology and pathogenicity tests were adopted to identify the ...[Objectives]The study was to identify the causal agent of leaf spots of Camellia drupifera in Hainan Province,China.[Methods]Phylogenetic analyses,spore morphology and pathogenicity tests were adopted to identify the pathogen.[Results]The fungus was identified as Neofusicoccum parvum.The colonies were initially pale to white on PDA,with diffuse yellow pigment around the colonies in the agar after 5 d.After 10 d,the aerial mycelium became gray,and the substrate mycelium became black.Thirty days later,a large number of conidia were generated on OMA.Conidia were hyaline,nonseptate,and fusiform.The mean size of 100 conidia was(13.6-25.4)×(6.2-10.3)μm;the mean length/width ratio was(2.5±0.3)μm.[Conclusions]N.parvum was the causal agent of leaf spots of C.drupifera observed in Hainan Province,China.展开更多
Background:Access to clean and safe drinking water that is free from pathogenic protozoan parasites,especially Cryptosporidium parvum and Giardia lamblia that cause gastrointestinal illness in humans,is still an issue...Background:Access to clean and safe drinking water that is free from pathogenic protozoan parasites,especially Cryptosporidium parvum and Giardia lamblia that cause gastrointestinal illness in humans,is still an issue in Southeast Asia(SEA).This study is the first attempt to detect the aforementioned protozoan parasites in water samples from countries in SEA,using real-time polymerase chain reaction(qPCR)assays.Methods:A total of 221 water samples of 10 l each were collected between April and October 2013 from Malaysia(53),Thailand(120),the Philippines(33),and Vietnam(15).A physicochemical analysis was conducted.The water samples were processed in accordance with the US Environmental Protection Agency’s methods 1622/1623.1,microscopically observed and subsequently screened using qPCR assays.Results:Cryptosporidium oocysts were detected in treated water samples from the Philippines(1/10),with a concentration of 0.06±0.19 oocyst/L,and untreated water samples from Thailand(25/93),Malaysia(17/44),and the Philippines(11/23),with concentrations ranging from 0.13±0.18 to 0.57±1.41 oocyst/L.Giardia cysts were found in treated water samples from the Philippines(1/10),with a concentration of 0.02±0.06 cyst/L,and in untreated water samples from Thailand(20/93),Vietnam(5/10),Malaysia(22/44),and the Philippines(16/23),with concentrations ranging from 0.12±0.3 to 8.90±19.65 cyst/L.The pathogens C.parvum and G.lamblia were detected using using qPCR assays by targeting the 138-bp fragment and the small subunit gene,respectively.C.parvum was detected in untreated water samples from the Philippines(1/23)and Malaysia(2/44),whilst,G.lamblia detected was detected in treated water samples from the Philippines(1/10)and in untreated water samples from Thailand(21/93),Malaysia(12/44),and the Philippines(17/23).Nitrate concentration was found to have a high positive correlation with(oo)cyst(0.993).Conclusion:The presence of(oo)cysts in the water samples means that there is potential risk for zoonotic disease transmission in the studied countries.Detection using qPCR is feasible for quantifying both pathogenic C.parvum and G.lamblia in large water samples.展开更多
The object of this study concentrated on investigating the antimicrobial susceptibilities of Ureaplasma parvum isolates to determine the most suitable antibiotic for treating the infection.In total,35 samples of Ureap...The object of this study concentrated on investigating the antimicrobial susceptibilities of Ureaplasma parvum isolates to determine the most suitable antibiotic for treating the infection.In total,35 samples of Ureaplasma parvum isolates were included in this study.Antibiotic susceptibility was studied by broth dilution method which was for the purpose of susceptibility testing of serovar isolates of Ureaplasma parvum against eight antibiotics.The results revealed the serovar 3 isolates were fully resistant(100%)to gentamicin,azithromycin and erythromycin while susceptible at the rates of 80%to doxycycline,60%to levofloxacin and 60%to clarithromycin.Serovar 14 isolate was revealed fully susceptible(100%)to clarithromycin,ciprofloxacin and doxycycline,while fully resistant(100%)to gentamicin and azithromycin.Serovar 1 and serovar 6 were showed to be fully resistant(100%)to azithromycin and gentamicin.Sevorar 1 was susceptible to at the rates of 70%to doxycycline,60%to tetracycline,90%to ciprofloxacin,70%to levofloxacin,70%to erythromycin and 70%to clarithromycin.Serovar 6 was susceptible at the rates of 80%to doxycycline,100%to tetracycline,100%to ciprofloxacin,80%to levofloxacin,80%to erythromycin and 80%to clarithromycin.These results evidently demonstrated that doxycycline,clarithromycin and levofloxacin should be the preferred drug when empirical treatment was required.展开更多
基金supported by the program for tacking key problem of Shaanxi agricultural scientific and technological extent(2015NY124)Project NSFC(31270690)+1 种基金Project PCSIRT(NO.IRT1035)special funding for basic S&T work of Ministry of Science and Technology(2009FY210100)of China
文摘Cankered, dying seedlings of Juglans regia were observed in Shaanxi province in the northwest region of China. Neofusicoccum parvum was isolated from these cankered tissues, with the identification based on mor- phology and an ITS-nrDNA sequence. In order to demonstrate how cultures of N. parvum could cause the expected symptoms, artificial infection, using these isolates and re-isolation of the pathogen, was used. This is the first report on this taxon as a walnut canker pathogen in China.
基金supported by the National Key R&D Program of China (No. 2018YFC0310800)the National Science and Technology Major Project for Significant New Drugs Development (No. 2018ZX09735004)+3 种基金the Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology (Qingdao)(Nos. 2018SDKJ0401-2 and 2016ASKJ05-04)the NSFC-Shandong Joint Fund (No. U1906212)the Fundamental Research Funds for the Central Universities (No. 201941001)the Taishan Scholar Youth Expert Program in Shandong Province (No. tsqn201812021)。
文摘Three new mycophenolic acid derivatives, penicacids E-G(1-3), together with three known analogues, mycophenolic acid(4), 4′-hydroxy-mycophenolic acid(5) and mycophenolic methyl ester(6), were isolated from a marine-derived fungus Penicillium parvum HDN17-478 from a South China Sea marine sediment sample. The structures of compounds 1-3 were elucidated by HRMS, NMR, and Mosher’s method. Among them, compounds 1 and 2 were the first examples of mycophenolic acid analogs with a double bond at C-3′/C-4′ position. The cytotoxicity of 1-6 was evaluated against the HCT-116, BEL-7402, MGC-803, SH-SY5 Y, HO-8910 and HL-60 cell lines, and compounds 4 and 6 showed potent cytotoxicity with IC50 values ranging from 1.69 to 12.98 μmol·L–1.
文摘Neofusicoccum parvum,an endophytic fungus isolated from Elaeocarpus serratus(Ceylon Olive;family Elaeocarpaceae),was grown for 3 weeks in potato dextrose broth.Chromatographic separation of the ethyl acetate extracts from the culture filtrate and mycelium over silica gel,Sephadex LH-20 and preparative thin layer chromatography furnished(R)-7-hydroxymellein(1),(3R,4R)-4-hydroxymellein(2),(3R,4S)-4-hydroxymellein(3),(R)-5-hydroxymellein(4),(R)-mellein(5),(3R,4R)-4,7-dihydroxymellein(6),(6R,7S)-dia-asperlin(7),CJ-14445(8)and 13,14,15,16-tetranorlabd-7-ene-19,6β:12,17-diolide(9).The structures of known compounds 1–9 were determined by spectroscopic methods and comparison with reported data.This is the first report of the isolation of an endophytic fungus from E.serratus,and the isolation of compounds 1,4,6,8 and 9 from N.parvum.It is important to note that compounds 1–7 are small molecules with an oxygen heterocyclic ring system.These compounds can be used as starting materials in the synthesis of pharmaceutically important large molecules with oxygen heterocyles.
基金supported by Natural Science Foundation of Jilin Province for Outstanding Young scientists Award,China(2002).
文摘The CP15/60 gene encoding the CP15/60 surface protein of sporozoites in Cryptosporidiumparvum was obtained by PCR so as to research the nucleic vaccine against C.parvum. Theeukaryotic expressing vector pcDNA3-15/60 was constructed by inserting CP15/60 gene intopcDNA3 (+) in XhoⅠand EcoRⅠ. A vaccination protocol was the adult pregnant goatsinoculated intranasally with the pcDNA3-15/60 plasmid and their offspring were infectedwith C.parvum oocysts. The results showed that the pcDNA3-15/60 plasmid can induce theimmune response of goats and the vaccinated goats can transfer the immunity to offspringconferring protection against C.parvum infection. These suggested that the recombinantplasmid could be a DNA vaccine candidate.
文摘With the discovery that the coccidian parasite Cryptosporidium sp. can cause severe symptoms in humans, many diagnostic techniques were quickly implemented such as microscopic visualization, immunofluorescence and PCR. Currently, there is no effective drug treatment and none of the current diagnostic methods is 100% accurate. In this study, a BALB/C mouse was subcutaneously immunized with Cryptosporidium parvum oocysts extract. The spleen was removed and the splenocytes were fused with SP2/0 myeloma cells in order to obtain hybridoma cells secreting antibodies specific to C. parvum antigens. Human and cattle fecal samples previously characterized by microscopy [Ziehl-Neelsen staining (ZN) and Lugol] and PCR for the presence of C. parvum and Giardia duodenalis, were analyzed by indirect immunofluorescence, using the developed hybridomas supernatants. The study shows that the selected hybridomas supernatants identify C. parvum oocysts in fecal samples in correlation with C. parvum oocysts identified using ZN/PCR. No false positive results were obtained and the two best supernatants gave 20% -30% of false negative results. No cross reaction with G. duodenalis was observed. By comparing our results with those obtained with an immunofluorescence commercial kit, it suggests the potential use of the monoclonal antibodies present in two of the hybridomas supernatants as a detection tool of C. parvum. With a reliability of 80.8% and 73.1% versus ZN and PCR methods for IFI, compared with a reliability of 76.9% and 92.3% versus ZN and PCR for commercial DIF kit, the supernatant 4.1D5 seems to be the most promising subject to further study its usefulness for C. parvum detection.
基金Hainan Provincial Natural Science Foundation of China(319MS081)Central Finance Forestry Science and Technology Extension Project(Qiong[2020]TG06).
文摘[Objectives]The study was to identify the causal agent of leaf spots of Camellia drupifera in Hainan Province,China.[Methods]Phylogenetic analyses,spore morphology and pathogenicity tests were adopted to identify the pathogen.[Results]The fungus was identified as Neofusicoccum parvum.The colonies were initially pale to white on PDA,with diffuse yellow pigment around the colonies in the agar after 5 d.After 10 d,the aerial mycelium became gray,and the substrate mycelium became black.Thirty days later,a large number of conidia were generated on OMA.Conidia were hyaline,nonseptate,and fusiform.The mean size of 100 conidia was(13.6-25.4)×(6.2-10.3)μm;the mean length/width ratio was(2.5±0.3)μm.[Conclusions]N.parvum was the causal agent of leaf spots of C.drupifera observed in Hainan Province,China.
基金study was supported by the University of Malaya High Impact Research Grant(UM.C/625/1/HIR/MoHE/MED/23 and UM.0000041/HIR.C3)from the Ministry of Higher Education,Malaysia+1 种基金postgraduate research grants(PV 049/2011B and PV 014/2012A)University Malaya Research Grants(UMRG 544/14HTM and UMRG 362/15AFR).
文摘Background:Access to clean and safe drinking water that is free from pathogenic protozoan parasites,especially Cryptosporidium parvum and Giardia lamblia that cause gastrointestinal illness in humans,is still an issue in Southeast Asia(SEA).This study is the first attempt to detect the aforementioned protozoan parasites in water samples from countries in SEA,using real-time polymerase chain reaction(qPCR)assays.Methods:A total of 221 water samples of 10 l each were collected between April and October 2013 from Malaysia(53),Thailand(120),the Philippines(33),and Vietnam(15).A physicochemical analysis was conducted.The water samples were processed in accordance with the US Environmental Protection Agency’s methods 1622/1623.1,microscopically observed and subsequently screened using qPCR assays.Results:Cryptosporidium oocysts were detected in treated water samples from the Philippines(1/10),with a concentration of 0.06±0.19 oocyst/L,and untreated water samples from Thailand(25/93),Malaysia(17/44),and the Philippines(11/23),with concentrations ranging from 0.13±0.18 to 0.57±1.41 oocyst/L.Giardia cysts were found in treated water samples from the Philippines(1/10),with a concentration of 0.02±0.06 cyst/L,and in untreated water samples from Thailand(20/93),Vietnam(5/10),Malaysia(22/44),and the Philippines(16/23),with concentrations ranging from 0.12±0.3 to 8.90±19.65 cyst/L.The pathogens C.parvum and G.lamblia were detected using using qPCR assays by targeting the 138-bp fragment and the small subunit gene,respectively.C.parvum was detected in untreated water samples from the Philippines(1/23)and Malaysia(2/44),whilst,G.lamblia detected was detected in treated water samples from the Philippines(1/10)and in untreated water samples from Thailand(21/93),Malaysia(12/44),and the Philippines(17/23).Nitrate concentration was found to have a high positive correlation with(oo)cyst(0.993).Conclusion:The presence of(oo)cysts in the water samples means that there is potential risk for zoonotic disease transmission in the studied countries.Detection using qPCR is feasible for quantifying both pathogenic C.parvum and G.lamblia in large water samples.
文摘The object of this study concentrated on investigating the antimicrobial susceptibilities of Ureaplasma parvum isolates to determine the most suitable antibiotic for treating the infection.In total,35 samples of Ureaplasma parvum isolates were included in this study.Antibiotic susceptibility was studied by broth dilution method which was for the purpose of susceptibility testing of serovar isolates of Ureaplasma parvum against eight antibiotics.The results revealed the serovar 3 isolates were fully resistant(100%)to gentamicin,azithromycin and erythromycin while susceptible at the rates of 80%to doxycycline,60%to levofloxacin and 60%to clarithromycin.Serovar 14 isolate was revealed fully susceptible(100%)to clarithromycin,ciprofloxacin and doxycycline,while fully resistant(100%)to gentamicin and azithromycin.Serovar 1 and serovar 6 were showed to be fully resistant(100%)to azithromycin and gentamicin.Sevorar 1 was susceptible to at the rates of 70%to doxycycline,60%to tetracycline,90%to ciprofloxacin,70%to levofloxacin,70%to erythromycin and 70%to clarithromycin.Serovar 6 was susceptible at the rates of 80%to doxycycline,100%to tetracycline,100%to ciprofloxacin,80%to levofloxacin,80%to erythromycin and 80%to clarithromycin.These results evidently demonstrated that doxycycline,clarithromycin and levofloxacin should be the preferred drug when empirical treatment was required.
