Poly-β-hydroxybutyrate accumulation in bacterial consortia from different environments

2012 ◽  
Vol 58 (5) ◽  
pp. 660-667 ◽  
Author(s):  
Rahela Carpa ◽  
Anca Butiuc-Keul ◽  
Iulia Lupan ◽  
Lucian Barbu-Tudoran ◽  
Vasile Muntean ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Flood Plain ◽  
Soil Samples ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Stressful Environment ◽  
Transmission Electron ◽  
Polymerase Chain

The aim of the present study was to examine soil samples from various vegetation zones in terms of physicochemical properties, microbial communities, and isolation and identification (by polymerase chain reaction and transmission electron microscopy) of bacteria producing poly-β-hydroxybutyrates (PHBs). Soil samples were analysed originating from zones with heterogeneous environmental conditions from the Romanian Carpathian Mountains (mountain zone with alpine meadow, karstic zone with limestone meadow, hill zone with xerophilous meadow, and flood plain zone with hygrophilic meadow). Different bacterial groups involved in the nitrogen cycle (aerobic mesophilic heterotrophs, ammonifiers, denitrifiers, nitrifiers, and free nitrogen-fixing bacteria from Azotobacter genus) were analysed. Soil biological quality was assessed by the bacterial indicator of soil quality, which varied between 4.3 and 4.7. A colony polymerase chain reaction technique was used for screening PHB producers. With different primers, specific bands were obtained in all the soil samples. Some wild types of Azotobacter species were isolated from the 4 studied sites. Biodegradable polymers of PHB were assessed by negative staining in transmission electron microscopy. The maximum PHB granules density was obtained in the strains isolated from the xerophilous meadow (10–18 granules/cell), which was the most stressful environment from all the studied sites, as the physicochemical and microbiological tests proved.

scholarly journals Comparison of Three Rapid Commercial Canine Parvovirus Antigen Detection Tests with Electron Microscopy and Polymerase Chain Reaction

2009 ◽  
Vol 21 (3) ◽  
pp. 344-345 ◽  
Author(s):  
Silke Schmitz ◽  
Christina Coenen ◽  
König Matthias ◽  
Thiel Heinz-Jürgen ◽  
Reto Neiger
Keyword(s):  
Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Chronic Diarrhea ◽  
Gastrointestinal Disease ◽  
Clinical Signs ◽  
Canine Parvovirus ◽  
Chain Reaction ◽  
Group A ◽  
Polymerase Chain ◽  
Group B

Different antibody-based tests for rapid detection of Canine parvovirus antigens in feces are commercially available, allowing quick diagnosis in a clinical setting. However, the diagnostic accuracy of these tests compared with standard methods has not been evaluated so far. In the current study, 3 commercial tests were compared with immune-electron microscopy (IEM) and polymerase chain reaction (PCR). Dogs were divided into 3 groups: group A, samples from dogs with acute hemorrhagic diarrhea ( n = 50); group B, dogs with chronic diarrhea ( n = 10); and group C, dogs with no evidence of gastrointestinal disease ( n = 40). Specificity of all 3 commercial tests versus PCR and IEM was good to excellent (92.2–100%). Sensitivity, in contrast, was poor: 15.8–26.3% versus PCR and 50–60% versus IEM. In group A, 10 dogs were positive by IEM and 24 dogs were positive by PCR. Positive PCR results were also obtained from animals in control groups (group B, 1 dog; group C, 5 dogs). No dog in group B or C was positive by IEM. In conclusion, the rapid tests are useful to diagnose canine parvoviral enteritis, but they do not rule out parvovirus infection in an animal with typical clinical signs. In addition, a small percentage of healthy dogs and dogs with chronic diarrhea showed positive PCR results; this may be due to asymptomatic/persistent infection or intestinal passage of virus. The significance of this finding remains unclear.

scholarly journals Identification of aflatoxigenic fungi using polymerase chain reaction-based assay

2014 ◽  
pp. 259-269 ◽  
Author(s):  
Vladislava Soso ◽  
Marija Skrinjar ◽  
Nevena Blagojev ◽  
Slavica Veskovic-Moracanin
Keyword(s):  
Polymerase Chain Reaction ◽  
Regulatory Gene ◽  
Control Point ◽  
Complex Mixture ◽  
Target Sequence ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Close Link ◽  
Critical Control Point ◽  
Polymerase Chain

As the aflatoxins represent a health-risk for humans because of their proven carcinogenicity, food-borne fungi that produce them as secondary metabolites, mainly Aspergillus flavus and Aspergillus parasiticus, have to be isolated and identified. The best argument for identifying problem fungi is that it indicates control points within the food system as part of a hazard analysis critical control point (HACCP) approach. This assumes there is a close link between fungus and toxin. Conventional methods for isolation and identification of fungi are time consuming and require admirably dedicated taxonomists. Hence, it is imperative to develop methodologies that are relatively rapid, highly specific and as an alternative to the existing methods. The polymerase chain reaction (PCR) facilitates the in vitro amplification of the target sequence. The main advantages of PCR is that organisms need not be cultured, at least not for a long time, prior to their detection, target DNA can be detected even in a complex mixture, no radioactive probes are required, it is rapid, sensitive and highly versatile. The gene afl-2 has been isolated and shown to regulate aflatoxin biosynthesis in A. flavus. Also, the PCR reaction was targeted against aflatoxin synthesis regulatory gene (aflR1) since these genes are nearly identical in A. flavus and A. parasiticus in order to indicate the possibility of detection of both the species with the same PCR system (primers/reaction). [Projekat Ministarstva nauke Republike Srbije, br. III46009] <br><br><font color="red"><b> This article has been retracted. Link to the retraction <u><a href="http://dx.doi.org/10.2298/APT1647265E">10.2298/APT1647265E</a><u></b></font>

scholarly journals Laboratory capability in Europe for foodborne viruses

2002 ◽  
Vol 7 (4) ◽  
pp. 61-65 ◽  
Author(s):  
B A Lopman ◽  
Y van Duynhoven ◽  
F X Hanon ◽  
M Reacher ◽  
M Koopmans ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Human Serum ◽  
National Laboratory ◽  
National Database ◽  
Chain Reaction ◽  
Foodborne Viruses ◽  
Food Items ◽  
Polymerase Chain ◽  
Almost All

This report describes a survey of national laboratory capabilities of diagnostics and surveillance databases for foodborne viruses among the &quot;Foodborne Viruses in Europe&quot; consortium. All the countries have laboratories that can test for HAV antibody in human serum. Eight of the ten surveyed European countries maintain a national database of HAV cases. Food can be tested for the presence of HAV in Finland, Italy, Spain, France and Denmark. All surveyed countries have at least one laboratory that tests for Norwalk-like virus (NLV) by reverse transcriptase-polymerase chain reaction and all also have the capability to use electron microscopy. Five countries maintain a national database of NLV cases and nine maintain a national database of NLV outbreaks. Almost all participant countries have laboratories that can test for NLV in food items including shellfish.

scholarly journals Diversity ofLeptospiraspp. in Rats and Environment from Urban Areas of Sarawak, Malaysia

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Chai Fung Pui ◽  
Lesley Maurice Bilung ◽  
Kasing Apun ◽  
Lela Su’ut
Keyword(s):  
Polymerase Chain Reaction ◽  
Water Samples ◽  
Urban Areas ◽  
Environmental Samples ◽  
Soil Samples ◽  
Chain Reaction ◽  
Prevalence Studies ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Soil And Water

Various prevalence studies onLeptospirain animals and humans, as well as environmental samples, had been conducted worldwide, including Malaysia. However, limited studies have been documented on the presence of pathogenic, intermediate, and saprophyticLeptospirain selected animals and environments. This study was therefore conducted to detectLeptospiraspp. in rats, soil, and water from urban areas of Sarawak using the polymerase chain reaction (PCR) method. A total of 107 rats, 292 soil samples, and 324 water samples were collected from April 2014 to February 2015. PathogenicLeptospirawas present in 5.6% (6/107) of rats, 11.6% (34/292) of soil samples, and 1.9% (6/324) of water samples. IntermediateLeptospirawas present in 2.7% (8/292) of soil samples and 1.9% (6/324) of water samples. SaprophyticLeptospirawas present in 10.3% (11/107) of rats, 1.4% (4/292) of soil samples, and 0.3% (1/324) of water samples. From this study, 76Leptospiraspp. were isolated. Based on DNA sequencing, the dominantLeptospiraspp. circulating in urban areas of Sarawak are pathogenicLeptospira noguchii, intermediateLeptospira wolffiiserovar Khorat, and saprophyticLeptospira meyeri, respectively. Overall, this study provided important surveillance data on the prevalence ofLeptospiraspp. from rats and the environment, with dominant local serovars in urban areas of Sarawak.

Detection of Aspergillus fumigatus by Polymerase Chain Reaction (PCR)

2019 ◽  
Vol 10 (04) ◽  
pp. 655-661
Author(s):  
Zainab H Abood AL-Asadi
Keyword(s):  
Polymerase Chain Reaction ◽  
Aspergillus Fumigatus ◽  
Oligonucleotide Primer ◽  
Rrna Gene ◽  
Fungal Culture ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Internal Transcribed Spacer Region ◽  
Polymerase Chain ◽  
The Impact

Aspergillosis refers to fungi infections of the respiratory tract caused by Aspergillus species, especially Aspergillus fumigatus. Infection of A. fumigatus was increased in the last few years due to either resistances to antibiotics or the influence of other factors such as other fungal infections. The present study aimed to review the impact of Aspergillus fumigatus in Aspergillosis cases, and study the role of Singleplex PCR for amplification of ITS1, ITS4 of rRNA gene in the detection of fungal isolate. In this study, One hundred sputum samples were collected from patients admitted to the specialize chest and respiratory diseases center / Baghdad who were suffering from respiratory problems. During these studied, molds were isolation and identification based on Conventional method (Direct microscopy by using 10% KOH, and fungal culture was done on Sabouraud Dextrose agar supplemented with chloramphenicol and on Czapek-Dox agar incubated at 37°C and examined for 3-7 days then macroscopic, microscopic examination of the colony by(lactophenol cotton blue stain )and molecular methods by using Polymerase chain reaction (PCR)technique for identification. The 10% KOH examination was positive for 35 cases, while laboratory culturing was positive for 53 cases. Aspergillus sp were isolated from 44(83%) patients; A. fumigatus was isolated in 23 (42. 4%) patients while A. flavus, A. niger, and A. terreus were isolated from 11 (20. 08%), (13. 2%) and 3 (5. 7%) patients respectively, also isolated Penicillium spp. at percentage 1(1. 9%). In this study. The ages of participants ranged from 10-70years with a mean age of 34years, the males were more susceptible to fungal infection, were recorded 35/53 (66. 3), compared to females were 18/53 (33. 96). The infection of fungi was more prevalent in ages 30-40recorded 26(53. 06%) followed by ages 40-50, 13(26. 5), while the lowest infection recorded in the age group 10- 20 years was 2(2. 04%). DNA isolated from twenty-three A. fumigatus isolates was used as a template, and the specific of oligonucleotide primer sequences were used in conventional PCR to detect the presence of internal transcribed spacer region ( ITS) region of the rRNA gene for Aspergillus fumigates. The results of the PCR amplification of the rRNA gene showed that this gene was present in 19 samples out 23 positive samples which isolation with a PCR product size of approximated 385 bp, while 4 samples out 23 positive samples showed negative results for the presence of this gene as indicated by the absence of the PCR products in their relevant lanes. Statistical analysis revealed that the PCR to have a sensitivity of 95. 1% in the detection of Aspergillus fumigatus in Aspergillosis cases. Polymerase chain reaction (PCR) is a rapid, specific, and sensitive method to detect Aspergillus fumigatus in aspergillosis cases of humans.

Validation of droplet digital Polymerase Chain Reaction for the detection and absolute quantification of Taenia solium eggs in spiked soil samples

Acta Tropica ◽  
2019 ◽  
Vol 200 ◽  
pp. 105175
Author(s):  
Justine Daudi Maganira ◽  
Beda John Mwang'onde ◽  
Winifrida Kidima ◽  
Chacha John Mwita ◽  
Gamba Nkwengulila ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Taenia Solium ◽  
Absolute Quantification ◽  
Soil Samples ◽  
Chain Reaction ◽  
Spiked Soil ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

scholarly journals Encephalitozoon Cuniculi Placentitis and Abortion in a Quarterhorse Mare

2003 ◽  
Vol 15 (1) ◽  
pp. 57-59 ◽  
Author(s):  
J. C. Patterson-Kane ◽  
P. Caplazi ◽  
F. Rurangirwa ◽  
R. R. Tramontin ◽  
K. Wolfsdorf
Keyword(s):  
Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Epithelial Cells ◽  
Ribosomal Dna ◽  
Encephalitozoon Cuniculi ◽  
Chain Reaction ◽  
University Of Kentucky ◽  
Livestock Disease ◽  
Diagnostic Center ◽  
Polymerase Chain

Encephalitozoon cuniculi is a microsporidial parasite, which has rarely been reported to cause placentitis in animals. A late-term aborted fetus and placenta from a Quarterhorse were presented to the Livestock Disease Diagnostic Center, University of Kentucky, for diagnostic examination. There was a necrotizing placentitis, with distension of many chorionic epithelial cells by intracytoplasmic vacuoles containing 1–2-μm-diameter, elongated, gram-positive organisms. The organisms were identified as E. cuniculi by electron microscopy and by polymerase chain reaction using primers to microsporidial ribosomal DNA. Joints of the fetus were swollen, with gross and microscopic lesions of synovitis; however, E. cuniculi DNA was not detected.

Detection of BK polyomavirus after kidney transplantation: a comparison of urine electron microscopy with plasma polymerase chain reaction

2012 ◽  
Vol 27 (1) ◽  
pp. E42-E48 ◽  
Author(s):  
Justin D. Westervelt ◽  
Barbara D. Alexander ◽  
Sylvia F. Costa ◽  
Sara E. Miller ◽  
David N. Howell ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Kidney Transplantation ◽  
Chain Reaction ◽  
Bk Polyomavirus ◽  
Polymerase Chain
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