scholarly journals Correlation of Lawsonia intracellularis positivity in quantitative PCR and herd factors in European pig herds

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Mirjam Arnold ◽  
Annelies Crienen ◽  
Hanny Swam ◽  
Stephan v. Berg ◽  
Rika Jolie ◽  
...  
Keyword(s):  
Direct Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Median Number ◽  
Growing Pigs ◽  
Economic Losses ◽  
Lawsonia Intracellularis ◽  
Poor Growth ◽  
Nursery Pigs ◽  
Potential Risk Factors ◽  
Polymerase Chain

Abstract Background Lawsonia intracellularis is causing diarrhea, poor growth and sudden death in pigs. It can be found in most pig populations leading to large economic losses worldwide. Many potential risk factors for the occurrence of disease or seropositivity have been described. The current study therefore focused on herd characteristics in European countries associated with direct detection of the pathogen determined by quantitative polymerase chain reaction. Results A median number of less than 30 nursery pigs per pen was correlated to less positive nursery pigs (p < 0.01) and generally less samples positive per herd (p < 0.05) as well as a lower median of genome equivalents determined per herd (p < 0.05). Routine use of zinc oxide at/ around weaning, which was mentioned by 41.0% of all farmers, was correlated to higher number of positive nursery pigs (p < 0.01) as well as higher median genome equivalents determined per herd (p < 0.05). Slatted flooring of more than 78.0% of the surface in nursery units was correlated to lower number of positive animals (p < 0.05) and a lower median of genome equivalents per herd (p < 0.05). A weight of more than 7.8 kg at weaning was correlated to a higher number of positive growing pigs (p < 0.05) as well as general higher number of positive samples/ herd (p < 0.01). Conclusions Weaning and subsequent accommodation of nursery pigs seem to be of particular importance in prevention of infection with Lawsonia intracellularis and the spread of the pathogen within the herd.

Detection ofLawsonia intracellularisin faeces of swine from the main producing regions in Brazil

1999 ◽  
Vol 45 (3) ◽  
pp. 230-234 ◽  
Author(s):  
Adriana E.C. Nascimento Chiriboga ◽  
Walter V Guimarães ◽  
Maria Cristina D. Vanetti ◽  
Elza F Araújo
Keyword(s):  
Polymerase Chain Reaction Method ◽  
Lawsonia Intracellularis ◽  
Specific Primers ◽  
Poor Growth ◽  
Mato Grosso ◽  
Proliferative Enteropathy ◽  
Total Incidence ◽  
Faecal Samples ◽  
Dna Fragment ◽  
Polymerase Chain

Swine proliferative enteropathy is an enteric disease caused by Lawsonia intracellularis which affects animals between 6 and 20 weeks of age, causing diarrhea, anorexia, and poor growth. The presence of L. intracellularis was evaluated in the faecal samples of 636 swine from 75 randomly chosen herds in the main swine-producing regions of Brazil. The pathogen was detected by the polymerase chain reaction method (PCR) using L. intracellularis specific primers. A 319-bp DNA fragment specific for L. intracellularis was produced on amplification of DNA from the faeces of pigs with proliferative enteropathy. Equal amounts of DNA extracted from the faeces of animals from the same herd were pooled together and, once L. intracellularis was detected, the faecal material of each animal was analyzed separately. The incidence of L. intracellularis was 33.4% in the state of Santa Catarina, 29.4% in Paraná, 26.3% in Minas Gerais, 16.7% in Mato Grosso, and 7.1% in São Paulo. The presence of the pathogenic agent was detected in samples from 15 farms, representing a total incidence of 20%. Although 46 animals (7.2%) were shown to be infected, 11% did not present any symptoms of swine proliferative enteropathy. The use of PCR allowed the detection of L. intracellularis in swine farms and the evaluation of the incidence of proliferative enteropathy in different regions of Brazil.Key words: proliferative enteropathy, diagnosis, Lawsonia intracellularis, PCR, incidence.

scholarly journals Torque teno sus virus 1 and 2 viral loads in faeces of porcine circovirus 2-positive pigs

2015 ◽  
Vol 84 (2) ◽  
pp. 91-95 ◽  
Author(s):  
Alessandra M. M. G. de Castro ◽  
Cintia M. Baldin ◽  
Cintia M. Favero ◽  
Priscilla F. Gerber ◽  
Rafael I. Cipullo ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Porcine Circovirus ◽  
Age Group ◽  
Nursery Pigs ◽  
Viral Loads ◽  
Faecal Samples ◽  
Disease Condition ◽  
Porcine Circovirus 2 ◽  
Polymerase Chain ◽  
Torque Teno Sus Virus

Recently, studies have suggested an association between the Torque teno sus virus (TTSuV) and the Porcine circovirus-2 (PCV2) in PCV2-associated disease cases. The aim of this study was to verify TTSuVs loads in pig faeces from PCV2-positive animals with and without diarrhea from PCVAD-affected and PCV2-unvaccinated herds. A total of 80 faecal samples were collected individually from nursery and grow-finish pigs with (n = 40) or without (n = 40) diarrhea. The samples were tested for PCV2 and TTSuVs by using DNA binding dye SYBR Green quantitative polymerase chain reaction (qPCR). Torque teno sus virus k2 (TTSuVk2) load in the faeces was significantly higher in the nursery pigs with diarrhea, and these pigs also exhibited significantly higher PCV2 (P < 0.01) faecal matter loads compared to the non-diarrheic animals from the same age group. Torque teno sus virus 1 (TTSuV1) viral loads were the same regardless of age group and disease condition. There were no correlations between PCV2 and TTSuV1 or TTSuVk2 and TTSuV1 viral loads; however, a weak correlation (r = 0.23, P = 0.03) was found between TTSuVk2 and PCV2 viral loads. In conclusion, TTSuVk2 viral loads were significantly higher in the diarrheic faeces from the nursery pigs. Additionally, the higher loads of PCV2 and TTSuVk2 in the nursery-diarrheic animals revealed that diarrhea might have an important role in the spread of both viruses in herds.

scholarly journals A Rapid and Reliable Liquid Chromatography/Mass Spectrometry Method for SARS-CoV-2 Diagnostics from Gargle Solutions and Saliva

2021 ◽  
Author(s):  
Marc Kipping ◽  
Dirk Taenzler ◽  
Andrea Sinz
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Direct Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Multiple Reaction Monitoring ◽  
Diagnostic Tools ◽  
Liquid Chromatography Mass Spectrometry ◽  
Clinical Environment ◽  
Chromatography Mass Spectrometry ◽  
Polymerase Chain

We describe a rapid liquid chromatography/mass spectrometry (LC/MS) method for the direct detection and quantitation of SARS-CoV-2 nucleoprotein in gargle solutions and saliva. The method is based on a multiple-reaction monitoring (MRM) mass spectrometry approach with a total cycle time of 5 minutes per analysis and allows the detection and accurate quantitation of SARS-CoV-2 nucleoprotein as low as 500 amol/ul. We improved the sample preparation protocol of our recent piloting SARS-CoV-2 LC/MS study regarding sensitivity, reproducibility, and compatibility with a complementary reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) analysis of the same sample. The aim of this work is to promote diagnostic tools that allow identifying and monitoring SARS-CoV-2 infections by LC/MS methods in a routine clinical environment.

scholarly journals A rapid and reliable liquid chromatography/mass spectrometry method for SARS-CoV-2 analysis from gargle solutions and saliva

2021 ◽  
Author(s):  
Marc Kipping ◽  
Dirk Tänzler ◽  
Andrea Sinz
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Direct Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Multiple Reaction Monitoring ◽  
Diagnostic Tools ◽  
Liquid Chromatography Mass Spectrometry ◽  
Clinical Environment ◽  
Liquid Chromatography Tandem Mass ◽  
Polymerase Chain

AbstractWe describe a rapid liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the direct detection and quantitation of SARS-CoV-2 nucleoprotein in gargle solutions and saliva. The method is based on a multiple-reaction monitoring (MRM) mass spectrometry approach with a total cycle time of 5 min per analysis and allows the detection and accurate quantitation of SARS-CoV-2 nucleoprotein as low as 500 amol/μL. We improved the sample preparation protocol of our recent piloting SARS-CoV-2 LC-MS study regarding sensitivity, reproducibility, and compatibility with a complementary reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) analysis of the same sample. The aim of this work is to promote diagnostic tools that allow identifying and monitoring SARS-CoV-2 infections by LC-MS/MS methods in a routine clinical environment. Graphical abstract

scholarly journals Exploring the Cause of Diarrhoea and Poor Growth in 8–11-Week-Old Pigs from an Australian Pig Herd Using Metagenomic Sequencing

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1608
Author(s):  
Tarka Raj Bhatta ◽  
Anthony Chamings ◽  
Soren Alexandersen
Keyword(s):  
Growing Pigs ◽  
Clinical Disease ◽  
Porcine Circovirus ◽  
Economic Losses ◽  
Porcine Parvovirus ◽  
Metagenomic Sequencing ◽  
Poor Growth ◽  
Dna Viruses ◽  
Porcine Astrovirus ◽  
Wide Range

Diarrhoea and poor growth among growing pigs is responsible for significant economic losses in pig herds globally and can have a wide range of possible aetiologies. Next generation sequencing (NGS) technologies are useful for the detection and characterisation of diverse groups of viruses and bacteria and can thereby provide a better understanding of complex interactions among microorganisms potentially causing clinical disease. Here, we used a metagenomics approach to identify and characterise the possible pathogens in colon and lung samples from pigs with diarrhoea and poor growth in an Australian pig herd. We identified and characterized a wide diversity of porcine viruses including RNA viruses, in particular several picornaviruses—porcine sapelovirus (PSV), enterovirus G (EV-G), and porcine teschovirus (PTV), and a porcine astrovirus (PAstV). Single stranded DNA viruses were also detected and included parvoviruses like porcine bocavirus (PBoV) and porcine parvovirus 2 (PPV2), porcine parvovirus 7 (PPV7), porcine bufa virus (PBuV), and porcine adeno-associated virus (AAV). We also detected single stranded circular DNA viruses such as porcine circovirus type 2 (PCV2) at very low abundance and torque teno sus viruses (TTSuVk2a and TTSuVk2b). Some of the viruses detected here may have had an evolutionary past including recombination events, which may be of importance and potential involvement in clinical disease in the pigs. In addition, our metagenomics data found evidence of the presence of the bacteria Lawsonia intracellularis, Brachyspira spp., and Campylobacter spp. that may, together with these viruses, have contributed to the development of clinical disease and poor growth.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

Molecular Detection of Bacterial Pathogens Directly from the Nasal Swabs and Lung Tissue of Sheep and Goats

2019 ◽  
Vol 15 (02) ◽  
pp. 22-25
Author(s):  
Sunaina Thakur ◽  
Subhash Verma ◽  
Prasenjit Dhar ◽  
Mandeep Sharma
Keyword(s):  
Pasteurella Multocida ◽  
Direct Detection ◽  
Bacterial Species ◽  
Economic Losses ◽  
Isolation And Identification ◽  
Sheep And Goats ◽  
Histophilus Somni ◽  
Nasal Swabs ◽  
Mycoplasma Spp

Respiratory infections of sheep and goats cause heavy morbidity and mortality, leading to huge economic losses. Conventional methods of diagnosis that include isolation and identification of incriminating microbes are time-consuming and fraught with logistic challenges. Direct detection of incriminating microbes using molecular tools is gaining popularity in clinical, microbiological settings. In this study, a total of 50 samples (44 nasal swabs and 6 lung tissues) from sheep and goats were screened for the detection of different bacterial species by in vitro amplification of genus or species-specific genes. Histophilus somni was detected in 2% goat samples, Trueperella pyogenes in 20% goat nasal swabs, whereas 22% goat nasal swab samples were found positive for Mycoplasma spp. None of the samples from sheep was detected positive for H. somni, T. pyogenes, Mycoplasma spp. Similarly, all samples, irrespective, whether from sheep or goats, showed negative results for Pasteurella multocida, Mannheimia haemolytica, and Corynebacterium pseudotuberculosis.

Pim-1 is up-regulated by constitutively activated FLT3 and plays a role in FLT3-mediated cell survival

Blood ◽  
2005 ◽  
Vol 105 (4) ◽  
pp. 1759-1767 ◽  
Author(s):  
Kyu-Tae Kim ◽  
Kristin Baird ◽  
Joon-Young Ahn ◽  
Paul Meltzer ◽  
Michael Lilly ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Quantitative Polymerase Chain Reaction ◽  
Colony Growth ◽  
Dominant Negative ◽  
Internal Tandem Duplication ◽  
Downstream Target ◽  
Before And After ◽  
Flt3 Expression ◽  
Polymerase Chain ◽  
Expression Microarrays

AbstractConstitutively activating internal tandem duplication (ITD) mutations of the receptor tyrosine kinase FLT3 (Fms-like tyrosine kinase 3) play an important role in leukemogenesis, and their presence is associated with poor prognosis in acute myeloid leukemia (AML). To better understand FLT3 signaling in leukemogenesis, we have examined the changes in gene expression induced by FLT3/ITD or constitutively activated wild-type FLT3 expression. Microarrays were used with RNA harvested before and after inhibition of FLT3 signaling. Pim-1 was found to be one of the most significantly down-regulated genes upon FLT3 inhibition. Pim-1 is a proto-oncogene and is known to be up-regulated by signal transducer and activator of transcription 5 (STAT5), which itself is a downstream target of FLT3 signaling. Quantitative polymerase chain reaction (QPCR) confirmed the microarray results and demonstrated approximately 10-fold decreases in Pim-1 expression in response to FLT3 inhibition. Pim-1 protein also decreased rapidly in parallel with decreasing autophosphorylation activity of FLT3. Enforced expression of either the 44-kDa or 33-kDa Pim-1 isotypes resulted in increased resistance to FLT3 inhibition-mediated cytotoxicity and apoptosis. In contrast, expression of a dominant-negative Pim-1 construct accelerated cytotoxicity in response to FLT3 inhibition and inhibited colony growth of FLT3/ITD-transformed BaF3 cells. These findings demonstrate that constitutively activated FLT3 signaling up-regulates Pim-1 expression in leukemia cells. This up-regulation contributes to the proliferative and antiapoptotic pathways induced by FLT3 signaling. (Blood. 2005;105: 1759-1767)

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