scholarly journals Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

2013 ◽  
Vol 33 (2) ◽  
pp. 177-182 ◽  
Author(s):  
Thales Q. Furian ◽  
Karen A. Borges ◽  
Silvio L.S. Rocha ◽  
Everton E. Rodrigues ◽  
Vladimir P. do Nascimento ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
High Frequency ◽  
Respiratory Diseases ◽  
Pasteurella Multocida ◽  
Virulence Genes ◽  
Simultaneous Detection ◽  
Fowl Cholera ◽  
Avian Diseases ◽  
Genetic Profiles ◽  
Current Systems

The current systems of breeding poultry, based on high population density, increase the risk of spreading pathogens, especially those causing respiratory diseases and those that have more than one host. Fowl Cholera (FC) is one such pathogen, and even though it represents one of several avian diseases that should be considered in the differential diagnosis of notifiable diseases that present with sudden death, the pathogenesis and virulence factors involved in FC are still poorly understood. The objective of this study was to investigate twelve genes related to virulence in 25 samples of Pasteurella multocida isolated from FC cases in the southern region of Brazil through the development of multiplex PCR protocols. The protocols developed were capable of detecting all of the proposed genes. The ompH, oma87, sodC, hgbA, hgbB, exBD-tonB and nanB genes were present in 100% of the samples (25/25), the sodA and nanH genes were present in 96% (24/25), ptfA was present in 92% (23/25), and pfhA was present in 60% (15/25). Gene toxA was not identified in any of the samples studied (0/25). Five different genetic profiles were obtained, of which P1 (negative to toxA) was the most common. We concluded that the multiplex-PCR protocols could be useful tools for rapid and simultaneous detection of virulence genes. Despite the high frequency of the analyzed genes and the fact that all samples belonged to the same subspecies of P. multocida, five genetic profiles were observed, which should be confirmed in a study with a larger number of samples.

scholarly journals Development of a multiplex PCR for simultaneous detection of Pasteurella multocida, Mannheimia haemolytica and Trueperella pyogenes

2017 ◽  
Vol 65 (3) ◽  
pp. 327-339 ◽  
Author(s):  
Wenlong Zhang ◽  
Xiaodan Liu ◽  
Mengcheng Liu ◽  
Bo Ma ◽  
Li Xu ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Genomic Dna ◽  
Pasteurella Multocida ◽  
Bacterial Pathogens ◽  
Simultaneous Detection ◽  
Laboratory Diagnosis ◽  
Mannheimia Haemolytica ◽  
Bacterial Strains ◽  
Trueperella Pyogenes ◽  
Enrichment Technology

Pasteurella multocida, Mannheimia haemolytica and Trueperella pyogenes are three bacterial pathogens closely associated with the bovine respiratory disease complex (BRDC). In the current study, a multiplex PCR for the simultaneous detection of these three bacteria in cultures was established. After serial optimisation, the detection limit of the method for the genomic DNA of the three bacteria was 40 pg/μl. The method could detect the genomic DNA of these three bacteria but not the genomic DNA of seven other bacterial strains. Together with the bacterial enrichment technology, the multiplex PCR could be used for detecting the three bacteria in animal tissues. This method might be valuable for speeding up laboratory diagnosis and directing the treatment of BRDC to these three bacterial pathogens.

scholarly journals Assessment of FTA card employment for Pasteurella multocida DNA transport and detection of virulence-associated genes in strains isolated from fowl cholera in the United States

2018 ◽  
Vol 70 (6) ◽  
pp. 1855-1861 ◽  
Author(s):  
C.N. Almeida ◽  
T.Q. Furian ◽  
K.A. Borges ◽  
G. Perdoncini ◽  
M.J. Mauel ◽  
...  
Keyword(s):  
United States ◽  
Pasteurella Multocida ◽  
Virulence Genes ◽  
Genetic Material ◽  
Storage Period ◽  
The United States ◽  
Material Transport ◽  
Dna Transport ◽  
Fowl Cholera ◽  
Fta Cards

ABSTRACT Fowl Cholera (FC) is a disease caused by Pasteurella multocida. The severity of this disease is partly caused by virulence factors. Genes encoding fimbriae, capsule, sialidases and proteins for iron metabolism may be related to P. multocida’s ability to infect the host. Besides to examining DNA for the presence of virulence genes, DNA is essential for the diagnostic and FTA cards are an alternative for genetic material transport. The study aims to evaluate the viability of P. multocida DNA transport using the cards and to detect 14 virulence genes in 27 strains isolated from FC cases in the United States by multiplex-PCR. No growth was observed in any of the FTA cards, which was essential to assess the security. Furthermore, DNA detection was possible in 100% of the samples, independent of the storage period (7 to 35 days) and temperature (4°C and 37°C). ptfA, exbd-tonB, hgbA, nanB, oma87, hyaD-hyaC, sodC, hgbB, sodA, nanH and pfhA genes were detected in more than 80% of the samples. FTA cards have proven to be a viable and safe tool for DNA transport of P. multocida. A majority of genes showed a high frequency, which was similar to strains isolated from FC cases.

scholarly journals Virulence Genes and Antimicrobial Resistance Profiles ofPasteurella multocidaStrains Isolated from Rabbits in Brazil

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Thais Sebastiana Porfida Ferreira ◽  
Maria Roberta Felizardo ◽  
Débora Dirani Sena de Gobbi ◽  
Cleise Ribeiro Gomes ◽  
Pedro Henrique de Lima Nogueira Filsner ◽  
...  
Keyword(s):  
Respiratory Diseases ◽  
Pasteurella Multocida ◽  
Virulence Genes ◽  
Domestic Animals ◽  
The Other ◽  
Nasal Discharge ◽  
Capsular Type ◽  
Biochemical Tests ◽  
Paulo State ◽  
Wide Range

Pasteurella multocidais responsible for a wide range of diseases in domestic animals. In rabbits, the agent is related to nasal discharge, pneumonia, otitis media, pyometra, orchitis, abscess, and septicemia. One hundred and forty rabbits with respiratory diseases from four rabbitries in São Paulo State, Brazil were evaluated for the detection ofP. multocidain their nasal cavities. A total of twenty-nine animals were positive toP. multocidaisolation, and 46 strains were selected and characterized by means of biochemical tests and PCR.P. multocidastrains were tested for capsular type, virulence genes, and resistance profile. A total of 45.6% (21/46) of isolates belonged to capsular type A, and 54.34% (25/46) of the isolates were untypeable. None of the strains harbouredtoxAorpfhAgenes. The frequency of the other twenty genes tested was variable, and the data generated was used to build a dendrogram, showing the relatedness of strains, which were clustered according to origin. Resistance revealed to be more common against sulfonamides and cotrimoxazole, followed by erythromycin, penicillin, and amoxicillin.

scholarly journals Novel multi-strain probiotics reduces Pasteurella multocida induced fowl cholera mortality in broilers

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Rine Christopher Reuben ◽  
Shovon Lal Sarkar ◽  
Habiba Ibnat ◽  
Md. Ali Ahasan Setu ◽  
Pravas Chandra Roy ◽  
...  
Keyword(s):  
Growth Performance ◽  
Feed Efficiency ◽  
Biochemical Parameters ◽  
Pasteurella Multocida ◽  
White Blood Cells ◽  
Clinical Signs ◽  
Basal Diet ◽  
Prostaglandin E ◽  
Fowl Cholera ◽  
Anti Inflammatory

AbstractPasteurella multocida causes fowl cholera, a highly contagious poultry disease of global concern, causing significant ecological and economic challenges to the poultry industry each year. This study evaluated the effects of novel multi-strain probiotics consisting of Lactobacillus plantarum, L. fermentum, Pediococcus acidilactici, Enterococcus faecium and Saccharomyces cerevisiae on growth performance, intestinal microbiota, haemato-biochemical parameters and anti-inflammatory properties on broilers experimentally challenged with P. multocida. A total of 120 birds were fed with a basal diet supplemented with probiotics (108 CFU/kg) and then orally challenged with 108 CFU/mL of P. multocida. Probiotics supplementation significantly (P < 0.05) improved growth performance and feed efficiency as well as reducing (P < 0.05) the population of intestinal P. multocida, enterobacteria, and mortality. Haemato-biochemical parameters including total cholesterol, white blood cells (WBC), proteins, glucose, packed cell volume (PCV) and lymphocytes improved (P < 0.05) among probiotic fed birds when compared with the controls. Transcriptional profiles of anti-inflammatory genes including hypoxia inducible factor 1 alpha (HIF1A), tumor necrosis factor- (TNF) stimulated gene-6 (TSG-6) and prostaglandin E receptor 2 (PTGER2) in the intestinal mucosa were upregulated (P < 0.05) in probiotics fed birds. The dietary inclusion of the novel multi-strain probiotics improves growth performance, feed efficiency and intestinal health while attenuating inflammatory reaction, clinical signs and mortality associated with P. multocida infection in broilers.

A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species

Food Control ◽  
2009 ◽  
Vol 20 (4) ◽  
pp. 366-370 ◽  
Author(s):  
Weibin Bai ◽  
Wentao Xu ◽  
Kunlun Huang ◽  
Yanfang Yuan ◽  
Sishuo Cao ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Meat Species ◽  
Pcr Method

Use of a Multiplex PCR System for the Simultaneous Detection of Heat Labile Toxin I and Heat Stable Toxin II Genes of Enterotoxigenic Escherichia coli in Skim Milk and Porcine Stool

1998 ◽  
Vol 61 (2) ◽  
pp. 141-145 ◽  
Author(s):  
HAU-YANG TSEN ◽  
LIANG-ZHAO JIAN ◽  
WAN-RONG CHI
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Skim Milk ◽  
Pcr Primers ◽  
Enterotoxigenic Escherichia Coli ◽  
Farm Animals ◽  
Target Cells ◽  
Heat Stable ◽  
Heat Labile

Enterotoxigenic Escherichia coli (ETEC) strains which produce heat labile and/or heat stable toxins (LT and ST) may cause diarrhea in humans and farm animals. Using PCR primers specific for the LT I and ST II genes, a multiplex PCR system which allows detection of LT I- and ST II-producing ETEC strains was developed. When skim milk was used for a PCR assay, it was found that if target cells in the sample were precultured in MacConkey broth for 8 h prior to PCR as few as 100 cells per ml of the sample could be detected. Without the preculture step, 104 CFU of target cells per 0.2 g of porcine stool specimen were required to generate visible PCR products. The multiplex PCR System can be used for rapid testing of fecal specimens, food and possibly environmental samples for the presence of ETEC strains.

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