Towards European harmonisation of contagious equine metritis diagnosis through interlaboratory trials

2018 ◽  
Vol 183 (3) ◽  
pp. 96-96
Author(s):  
Sandrine Petry ◽  
Marie-France Breuil ◽  
Fabien Duquesne ◽  
Claire Laugier
Keyword(s):  
Culture Media ◽  
Culture Method ◽  
Contagious Equine Metritis ◽  
National Reference ◽  
Bacteriological Diagnosis ◽  
Specificity And Sensitivity ◽  
Interlaboratory Trial ◽  
Culture Performance ◽  
European Harmonisation ◽  
Better Than

The performance of culture and PCR methods routinely used to diagnose contagious equine metritis (CEM) was evaluated and compared by two interlaboratory trials involving a total of 24 European laboratories, including 22 National Reference Laboratories for CEM. Samples were swab specimens artificially contaminated with bacteria present in the genital tract of Equidae, some with and some without Taylorella equigenitalis, the causative agent of CEM, and T asinigenitalis, responsible for possible misidentification as T equigenitalis. Throughout both interlaboratory trials, PCR performed better in terms of specificity and sensitivity than the culture method, supporting the assertion that PCR should be accepted for CEM diagnosis. However, the culture performance during the second interlaboratory trial was better than during the first one, suggesting that the expertise of participants improved. This reveals the advantage of regular interlaboratory trials to constantly improve the expertise of laboratories. It also shows the need to develop new culture media that are more selective and/or better geared to the metabolism of T equigenitalis in order to improve the bacteriological diagnosis of CEM.

scholarly journals Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 20
Author(s):  
Luigi De Grossi ◽  
Davide Santori ◽  
Antonino Barone ◽  
Silvia Abbruzzese ◽  
Matteo Ricchi ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Liquid Culture ◽  
Culture Media ◽  
Small Ruminants ◽  
Culture Method ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Culture Methods ◽  
Specific Pcr ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is a chronic disease of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Since isolation of MAP type I (S) is rarely reported in Italy, our research was aimed at isolating, by an inexpensive liquid culture manual method, this type of MAP isolates. At first, we used an ELISA to point out to serologically positive samples from five flocks. Secondly, we used a fecal direct IS900-qPCR on the ELISA positive samples, in order to detect shedder animals. Feces from IS900-qPCR positive samples were inoculated in solid and liquid culture media. IS900-qPCR was further used to test the growth of MAP isolates in liquid medium, which were further confirmed by f57-qPCR and submitted to typing by specific PCR in order to identify the MAP type. Twenty-eight samples (24 fecal and four tissutal samples) were processed by culture methods, resulting in the isolation of six type I MAP field isolates. Notably, no isolates were recovered by solid media, underlining the utility of this liquid method. Few data about this type of MAP are currently available in Italy, and further analyses should be carried out in order to study the origin and epidemiology of type I strains circulating in Italy.

Croissance et fructification de Ceratocystis spp. avec attention particulière à C. piceae et C. fimbriata, en fonction du substrat

1982 ◽  
Vol 60 (4) ◽  
pp. 358-363
Author(s):  
A. Thuillier ◽  
P. Neumann
Keyword(s):  
Nutrient Solution ◽  
Culture Media ◽  
Basal Medium ◽  
Malt Agar ◽  
Fruiting Bodies ◽  
Ceratocystis Fimbriata ◽  
Good Production ◽  
Wood Extracts ◽  
Standard Culture ◽  
Better Than

Ceratocystis coerulescens, C. fimbriata, C. ips, and C. minor were tested for production of sexual fruiting bodies, and C. penicillata and C. piceae for asexual fruiting bodies. Ceratocystis fimbriata produced perithecia easily on standard culture media, but there were marked differences between the two strains tested (503, 560). Strain 503 had a good production of fruiting bodies on malt agar (M) and a basal nutrient solution (N). Strain 560 fared better than 503 on Leonian agar (L), but did not fructify on M and N. Supplementing media with various wood extracts produced better results. M + maple sapwood extracts and L + poplar sapwood extracts gave the best results with strain 503, and L + pine sapwood extracts was the best with strain 560.Production of coremia was also influenced by the basal medium and the kind of extracts added as supplements. Fir and maple extracts stimulated the production of fruiting bodies, whereas pine and poplar extracts had no or very little stimulating effects. In every other species tested, the production of fruiting bodies was none or very irregular. [Journal translation]

Evaluation of Environmental Sampling Methods and Rapid Detection Assays for Recovery and Identification of Listeria spp. from Meat Processing Facilities

2009 ◽  
Vol 72 (4) ◽  
pp. 696-701 ◽  
Author(s):  
JOVANA KOVAČEVIĆ ◽  
VALERIE M. BOHAYCHUK ◽  
PABLO ROMERO BARRIOS ◽  
GARY E. GENSLER ◽  
DEANA L. ROLHEISER ◽  
...  
Keyword(s):  
Sampling Methods ◽  
Culture Method ◽  
Detection Methods ◽  
Lower Sensitivity ◽  
Meat Processing ◽  
Cotton Swab ◽  
Conventional Culture ◽  
Specificity And Sensitivity ◽  
Highly Sensitive ◽  
Labor Inputs

Studies that isolated Listeria spp. from the environment of two meat processing facilities were conducted. Samples were collected in the processing environment of the facilities with three different sampling methods (cotton swab, sterile sponge, and composite-ply tissues) to evaluate their ability to recover Listeria spp. A total of 240 samples for each sampling method were collected and tested. The cotton swab method of sampling was significantly (P < 0.01) less efficient in recovery of Listeria spp. than the sterile-sponge and composite-ply tissue methods, which were similar (P > 0.05) in their ability to recover Listeria spp. The specificity and sensitivity of four detection methods (conventional culture, Petrifilm Environmental Listeria Plates [ELP], lateral-flow immunoprecipitation [LFI], and automated PCR) were evaluated for identification of Listeria spp. Facilities were visited until a minimum of 100 positive and 100 negative samples per detection method were collected. The LFI and PCR methods were highly sensitive (95.5 and 99.1%, respectively) and specific (100%) relative to the culture method. The ELP method was significantly less efficient (P < 0.01) than LFI and PCR in detection of Listeria spp., with lower sensitivity (50.6%) and specificity (91.5%). Kappa values indicated excellent agreement of the LFI and PCR assays and moderate agreement of the ELP method to the culture method. Overall, ELP was easy to use but less efficient in detection of Listeria spp. from environmental samples, while the LFI and PCR methods were found to be excellent alternatives to culture, considering performance and time and labor inputs.

Effects of Herbicides on Diseases of Rice (Oryza sativa)

Weed Science ◽  
1977 ◽  
Vol 25 (5) ◽  
pp. 441-447 ◽  
Author(s):  
E. Inderawati ◽  
R. Heitefuss
Keyword(s):  
Oryza Sativa ◽  
Disease Severity ◽  
Culture Media ◽  
Direct Action ◽  
Culture Method ◽  
Xanthomonas Oryzae ◽  
Commercial Formulation ◽  
Rice Plants ◽  
Agar Media ◽  
Rice Leaves

Seven herbicides were tested for their effect on growth of Pyricularia oryzae Cavara, Hypochnus sasakii Shirai, and Xanthomonas oryzae Uyeda & Ishiyama Dowson on agar media and for subsequent influence on disease intensity on rice plants (Oryza sativa L.) in the greenhouse. The herbicides studied were: propanil 3′,4′-dichloro-propionanilide), NTN 5$006 [O-(2-nitro-4-methylphenyl)-O-ethyl-N-isopropyl-phosphor-amidothioate], simetryn [2,4-bis(ethylamino)-6-)methylthio)-s-triazine], terbutryn [2-(tert-butylamino)-4-(ethylamino)-6-(methylthio)-s-triazine], nitrofen (2,4-dichlorophenyl-p-nitrophenyl ether), molinate (S-ethyl hexahydro-1H-azepine-1-carbothioate), and aglypt (4-amino-3-methylthio-6-phenyl-1,2,4-triazine-5-on). The growth of P. oryzae, H. sasakii and X. oryzae on culture media containing 10 μg/ml commercial formulation of propanil was reduced to approximately 50% of the control. The other herbicides tested were less effective. Differences in disease severity produced on rice plants treated with the previously mentioned herbicides were in agreement with the results obtained by the culture method. The effect of simetryn and nitrofen on disease severity was stronger than expected from the small direct action on the pathogen in culture. It is suggested that the influence of these two compounds on the disease intensity is due to their effect on the host plant rather than the pathogen directly. Propanil was effective only if applied immediately or up to 1 day before inoculation, indicating that this herbicide is degraded on or within the rice leaves.

scholarly journals Tissue-Engineered Human Myobundle System as a Platform for Evaluation of Skeletal Muscle Injury Biomarkers

2020 ◽  
Vol 176 (1) ◽  
pp. 124-136
Author(s):  
Alastair Khodabukus ◽  
Amulya Kaza ◽  
Jason Wang ◽  
Neel Prabhu ◽  
Richard Goldstein ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Muscle Injury ◽  
Culture Media ◽  
Contractile Function ◽  
Significant Loss ◽  
Drug Induced ◽  
Skeletal Muscle Injury ◽  
Specificity And Sensitivity ◽  
Novel Biomarkers

Abstract Traditional serum biomarkers used to assess skeletal muscle damage, such as activity of creatine kinase (CK), lack tissue specificity and sensitivity, hindering early detection of drug-induced myopathies. Recently, a novel four-factor skeletal muscle injury panel (MIP) of biomarkers consisting of skeletal troponin I (sTnI), CK mass (CKm), fatty-acid-binding protein 3 (Fabp3), and myosin light chain 3, has been shown to have increased tissue specificity and sensitivity in rodent models of skeletal muscle injury. Here, we evaluated if a previously established model of tissue-engineered functional human skeletal muscle (myobundle) can allow detection of the MIP biomarkers after injury or drug-induced myotoxicity in vitro. We found that concentrations of three MIP biomarkers (sTnI, CKm, and Fabp3) in myobundle culture media significantly increased in response to injury by a known snake venom (notexin). Cerivastatin, a known myotoxic statin, but not pravastatin, induced significant loss of myobundle contractile function, myotube atrophy, and increased release of both traditional and novel biomarkers. In contrast, dexamethasone induced significant loss of myobundle contractile function and myotube atrophy, but decreased the release of both traditional and novel biomarkers. Dexamethasone also increased levels of matrix metalloproteinase-2 and -3 in the culture media which correlated with increased remodeling of myobundle extracellular matrix. In conclusion, this proof-of-concept study demonstrates that tissue-engineered human myobundles can provide an in vitro platform to probe patient-specific drug-induced myotoxicity and performance assessment of novel injury biomarkers to guide preclinical and clinical drug development studies.

Rapid Detection of Mycoplasma pneumoniae by Loop-Mediated Isothermal Amplification (LAMP) in Clinical Respiratory Specimens

2019 ◽  
Author(s):  
Maryam ARFAATABAR ◽  
Narjes NOORI GOODARZI ◽  
Davoud AFSHAR ◽  
Hamed MEMARIANI ◽  
Ghasem AZIMI ◽  
...  
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Rapid Detection ◽  
Lamp Assay ◽  
Culture Method ◽  
Isothermal Amplification ◽  
Lamp Method ◽  
Culture Methods ◽  
Loop Mediated Isothermal Amplification ◽  
Specificity And Sensitivity ◽  
Respiratory Specimens

  Background: Mycoplasma pneumoniae is a common cause of community-acquired pneumonia (CAP) worldwide, especially among children and debilitated populations. The present study aimed to investigate a loop-mediated isothermal amplification (LAMP) technique for rapid detection of M. pneumoniae in clini-cal specimens collected from patients with pneumonia. Methods: Throat swabs were collected from 110 outpatients who suffered from pneumonia. Throat swab samples were obtained from patients referred to the hospital outpatient clinics of Tehran University hospitals, Iran in 2017. The presence of M. pneumoniae in the clinical specimens was evaluated by LAMP, PCR and culture methods. Sensitivity and specificity of the LAMP and PCR assays were also determined. Results: Out of 110 specimens, LAMP assay detected M. pneumoniae in 35 specimens. Detection limit of the LAMP assay was determined to be 33fg /μL or ~ 40 genome copies/reaction. Moreover, no cross-reaction with genomic DNA from other bacteria was observed. Only 25 specimens were positive by the culture method. The congruence between LAMP assay and culture method was ‘substantial’ (κ=0.77). Specificity and sensitivity of LAMP assay were 88.2%, 100% in compare with culture. However, the con-gruence between LAMP assay and PCR assay was ‘almost perfect’ (κ=0.86). Specificity and sensitivity of LAMP assay were 92.5%, 100% in compare with PCR. Conclusion: Overall, the LAMP assay is a rapid and cost-efficient laboratory test in comparison to other methods including PCR and culture. Therefore, the LAMP method can be applied in identification of M. pneumoniae isolates in respiratory specimens.

Equivalency testing of TTC Tergitol 7 agar (ISO 9308-1:2000) with five culture media for the detection of E. coli in water samples in Greece

2010 ◽  
Vol 61 (1) ◽  
pp. 67-76 ◽  
Author(s):  
A. Mavridou ◽  
E. Smeti ◽  
G. Mandilara ◽  
P. Boufa ◽  
M. Vagiona-Arvanitidou ◽  
...  
Keyword(s):  
Water Samples ◽  
Culture Media ◽  
Reference Method ◽  
Relative Difference ◽  
Alternative Methods ◽  
Lauryl Sulfate ◽  
Counting Problems ◽  
E Coli ◽  
Better Than ◽  
The Eu

In this study ten laboratories in Greece compared the performance of reference method TTC Tergitol 7 Agar (with the additional test of β-glucuronidase production) with five alternative methods, to detect E. coli in water, in line with European Water Directive recommendations. The samples were prepared by spiking drinking water with sewage effluent following a standard protocol. Chlorinated and non-chlorinated samples were used. The statistical analysis was based on the mean relative difference of confirmed counts and was performed in line with ISO 17994. The results showed that in total, three of the alternative methods (Chromocult Coliform agar, Membrane Lauryl Sulfate agar and Trypton Bilex-glucuronidase medium) were not different from TTC Tergitol 7 agar (TTC Tergitol 7 agar vs Chromocult Coliform agar, 294 samples, mean RD% 5.55; vs MLSA, 302 samples, mean RD% 1; vs TBX, 297 samples, mean RD% −2.78). The other two alternative methods (Membrane Faecal coliform medium and Colilert 18/ Quantitray) gave significantly higher counts than TTC Tergitol 7 agar (TTC Tergitol 7 agar vs MFc, 303 samples, mean RD% 8.81; vs Colilert-18/Quantitray, 76 samples, mean RD% 18.91). In other words, the alternative methods generated performance that was as reliable as, or even better than, the reference method. This study will help laboratories in Greece overcome culture and counting problems deriving from the EU reference method for E. coli counts in water samples.

scholarly journals ACEI of Trichoderma reesei Is a Repressor of Cellulase and Xylanase Expression

2003 ◽  
Vol 69 (1) ◽  
pp. 56-65 ◽  
Author(s):  
Nina Aro ◽  
Marja Ilmén ◽  
Anu Saloheimo ◽  
Merja Penttilä
Keyword(s):  
Trichoderma Reesei ◽  
Culture Media ◽  
Carbon Sources ◽  
Cellulase Production ◽  
The Novel ◽  
Hypocrea Jecorina ◽  
Gene Encoding ◽  
Gene Coding ◽  
Better Than

ABSTRACT We characterized the effect of deletion of the Trichoderma reesei (Hypocrea jecorina) ace1 gene encoding the novel cellulase regulator ACEI that was isolated based on its ability to bind to and activate in vivo in Saccharomyces cerevisiae the promoter of the main cellulase gene, cbh1. Deletion of ace1 resulted in an increase in the expression of all the main cellulase genes and two xylanase genes in sophorose- and cellulose-induced cultures, indicating that ACEI acts as a repressor of cellulase and xylanase expression. Growth of the strain with a deletion of the ace1 gene on different carbon sources was analyzed. On cellulose-based medium, on which cellulases are needed for growth, the Δace1 strain grew better than the host strain due to the increased cellulase production. On culture media containing sorbitol as the sole carbon source, the growth of the strain with a deletion of the ace1 gene was severely impaired, suggesting that ACEI regulates expression of other genes in addition to cellulase and xylanase genes. A strain with a deletion of the ace1 gene and with a deletion of the ace2 gene coding for the cellulase and xylanase activator ACEII expressed cellulases and xylanases similar to the Δace1 strain, indicating that yet another activator regulating cellulase and xylanase promoters was present.

scholarly journals In VitroPollen Viability and Pollen Germination in Cherry Laurel (Prunus laurocerasusL.)

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Melekber Sulusoglu ◽  
Aysun Cavusoglu
Keyword(s):  
Pollen Germination ◽  
Culture Media ◽  
Pollen Viability ◽  
Second Step ◽  
Triphenyl Tetrazolium Chloride ◽  
Tetrazolium Chloride ◽  
Germinated Pollen ◽  
Cherry Laurel ◽  
Better Than

Pollen quality is important for growers and breeders. This study was carried out to determinein vitropollen viability and pollen germination in seven genotypes of cherry laurel (Prunus laurocerasusL.). Two pollen viability tests, TTC (2,3,5-triphenyl tetrazolium chloride) and IKI (iodine potassium iodide), were used. Pollen traits of genotypes were studied using anin vitromedium containing 0%, 5%, 10%, 15%, and 20% sucrose to determine the best sucrose concentrations for germination. In the second step, the germinated pollen was counted 1, 4, 6, 10, 12, 24, and 48 hours later until there was no further germination. The viability rates were different according to genotypes and tests used. The IKI and TTC staining tests and pollen germination had low correlation (r2= 0.0614 andr2= 0.0015, resp.). Painted pollen rate was higher and pollen was well-stained with IKI test and pollen viability estimated with TTC staining test was better than that estimated with the IKI staining test. 15% sucrose gave the best germination rates in most of the genotypes. Pollen germination rates were recorded periodically from one hour to 48 hours in 15% sucrose and the results showed that pollen germination rates increased after 6 hours of being placed in culture media.

Generation of parthenogenetic goat blastocysts: effects of different activation methods and culture media

Zygote ◽  
2014 ◽  
Vol 23 (3) ◽  
pp. 327-335 ◽  
Author(s):  
Hruda Nanda Malik ◽  
Dinesh Kumar Singhal ◽  
Shrabani Saugandhika ◽  
Amit Dubey ◽  
Ayan Mukherjee ◽  
...  
Keyword(s):  
Culture Media ◽  
Combined Treatment ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Cleavage Rate ◽  
Cell Stage ◽  
Quantitative Expression ◽  
Pattern Of Variation ◽  
Better Than

SummaryThe present study was carried out to investigate the effects of different activation methods and culture media on the in vitro development of parthenogenetic goat blastocysts. Calcium (Ca2+) ionophore, ethanol or a combination of the two, used as activating reagents, and embryo development medium (EDM), modified Charles Rosenkrans (mCR2a) medium and research vitro cleave (RVCL) medium were used to evaluate the developmental competence of goat blastocysts. Quantitative expression of apoptosis, stress and developmental competence-related genes were analysed in different stages of embryos. In RVCL medium, the cleavage rate of Ca2+ ionophore-treated oocytes (79.61 ± 0.86) was significantly (P < 0.05) higher than in ethanol (74.90 ± 1.51) or in the combination of both Ca2+ ionophore and ethanol. In mCR2a or EDM, hatched blastocyst production rate of Ca2+ ionophore-treated oocytes (8.33 ± 1.44) was significantly higher than in ethanol (6.46 ± 0.11) or in the combined treatment (6.70 ± 0.24). In ethanol, the cleavage, blastocyst and hatched blastocyst production rates in RVCL medium (74.90 ± 1.51, 18.30 ± 1.52 and 8.24 ± 0.15, respectively) were significantly higher than in EDM (67.81 ± 3.21, 14.59 ± 0.27 and 5.59 ± 0.42) or mCR2a medium (65.09 ± 1.57, 15.36 ± 0.52 and 6.46 ± 0.11). The expression of BAX, Oct-4 and GlUT1 transcripts increased gradually from 2-cell stage to blastocyst-stage embryos, whereas the transcript levels of Bcl-2 and MnSOD were significantly lower in blastocysts. In addition, different activation methods and culture media had little effect on the pattern of variation and relative abundance of the above genes in different stages of parthenogenetic activated goat embryos. In conclusion, Ca2+ ionophore as the activating agent, and RVCL as the culture medium are better than other tested options for development of parthenogenetic activated goat blastocysts.

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