Identification of the capsule type of Pasteurella multocida isolates from cases of fowl cholera by multiplex PCR and comparison with phenotypic methods

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.

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Phenotypic and genotypic characterisation of Pasteurella multocida strains isolated from pigs in Hungary

Acta Veterinaria Hungarica ◽

10.1556/avet.55.2007.4.2 ◽

2007 ◽

Vol 55 (4) ◽

pp. 425-434 ◽

Cited By ~ 8

Author(s):

Zsuzsanna Varga ◽

Boglárka Sellyei ◽

T. Magyar

Keyword(s):

Multiplex Pcr ◽

Ornithine Decarboxylase ◽

Pasteurella Multocida ◽

Regional Distribution ◽

Type A ◽

Capsule Type ◽

Type D ◽

Biochemical Characterisation ◽

River Danube ◽

Swine Herds

A total of 146 Pasteurella multocida strains isolated from swine in Hungary in the last 20 years were examined. Biochemical characterisation and PCR-based techniques were used to determine species, subspecies, biovar, capsule type and presence of the toxA gene. Eighty-seven percent of the isolates belonged to P. multocida ssp. multocida , and 98% of these had biovar 3 or were trehalose-or lactose-fermenting or ornithine decarboxylase negative variants of that. Ten percent of the strains were P. multocida ssp. septica , and within this group 80% of the strains showed sorbitol-negative biovars (5, 6 and 7). The rest of the strains (20%) were lactose positive. Only 3% of the porcine isolates were P. multocida ssp. gallicida and 3 out of the 4 strains belonged to the dulcitol-fermenting biovar 8. Using a capsule-specific multiplex PCR, 60% of the strains belonged to capsule type D, 38% to capsule type A, and only 1 isolate had capsule type F. In contrast with data published in the literature, only 3% of capsule type D isolates carried the toxA gene, while this ratio was 41% for the type A strains. A remarkable regional distribution of toxA gene positive strains was observed. All but two isolates were found in swine herds located in the Transdanubian region, separated from other parts of Hungary by the river Danube.

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Identification of Candida Spp. Isolated From Urine by Phenotypic Methods and Multiplex Pcr in Duhok, Iraq

Science Journal of University of Zakho ◽

10.25271/2017.5.1.292 ◽

2017 ◽

Vol 5 (1) ◽

pp. 11-15

Author(s):

Ahmed Mohammed ◽

Jumma Ali ◽

Samir Abdullah

Keyword(s):

Multiplex Pcr ◽

Candida Spp ◽

Phenotypic Methods

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

Scientific Reports ◽

10.1038/s41598-021-88299-0 ◽

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.

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The Current Status of Veterinary Vaccinology: A Review

Journal of Advances in Medical and Pharmaceutical Sciences ◽

10.9734/jamps/2019/v20i230108 ◽

2019 ◽

pp. 1-10

Author(s):

Teferi Mandado

Keyword(s):

Pasteurella Multocida ◽

Scientific Paper ◽

General Information ◽

Current Status ◽

College Of Agriculture ◽

Fowl Cholera ◽

Veterinary Vaccine ◽

Other Information ◽

The University ◽

The 19Th Century

This paper was done starting from February 2017 to July 2017 in Jimma University College of Agriculture and School of Veterinary Medicine. The suffering of different animal species from multiple infectious agents in and around the university leads us to be conscious and enabled us to write this scientific paper which can be acts as the source of information for Veterinary vaccinology. Louis Pasteur in the 19th century demonstrated the ability to protect chickens against fowl cholera (Pasteurella multocida) and thus demonstrated the benefit of vaccination in animals and paved the way for the development of the array of veterinary vaccines which are in use today. Since Pasteur’s work, vaccination against infectious disease have been used successfully to protect animals from many serious diseases some of which were also significant risks to humans. Veterinary vaccine has a parallel way of development in research and development of vaccines in the human field vaccinology today also. Vaccine is a biological preparation that improves immunity to a particular disease. Vaccine contains an agent that resembles a disease-causing microorganism and is often made from weakened or killed forms of the microbe or its toxins. The general information concerning veterinary vaccination such as common vaccination, common methods of veterinary vaccination, principles of vaccination; standardization of veterinary vaccines, generation of vaccine, vaccine formulation, new approaches to veterinary vaccines and few other information were roughly reviewed from scientific journals, experiment results, proceedings, reference books and manuals. The objectives of this paper are to highlight the general current information of Veterinary Vaccinology and to give specific recommendations based on the facts obtained.

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Molecular identification by multiplex polymerase chain reaction of Pasteurella multocida in cattle and buffaloes in Baghdad

The Iraqi Journal of Veterinary Medicine ◽

10.30539/iraqijvm.v38i1.261 ◽

2014 ◽

Vol 38 (1) ◽

pp. 99-106

Author(s):

Ihab G. M. AL-Shemmari

Keyword(s):

Polymerase Chain Reaction ◽

Multiplex Pcr ◽

Multiplex Polymerase Chain Reaction ◽

Pasteurella Multocida ◽

Type B ◽

Chain Reaction ◽

Blood Samples ◽

Nasal Swabs ◽

Polymerase Chain ◽

B 31

The aim of this study was to identify pasteurella multocida and their types by PCR in cattle’s and buffaloesi bagdad from March to August 2012 on 204 animals , including 102 cattle and 102 buffaloes at slaughter houses from Baghdad .Blood samples and nasal swaps were collected , before slaughtering and lung tissues of slaughtered animal , and from 54 clinically suspected cases of pasteurellosis , including 27 bovines ,and 27 buffaloes the samples taken included blood and nasal swabs . Pasteurellamultocida were isolated from 94 animals include 49 cattle 45 buffaloes. The typing of the isolates by multiplex PCR for genotyping Pasteuerllamultocida revealed 93 isolates of type B , 31 from cattle and 62 from buffaloes ,and 81 isolates of type A , 55 from cattle and 26 from buffaloes .

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EPIDEMIOLOGICAL INVESTIGATION OF PASTEURELLA MULTOCIDA INFECTION IN POULTRY IN GAZIPUR DISTRICT OF BANGLADESH

Bangladesh Journal of Veterinary Medicine ◽

10.3329/bjvm.v15i2.35517 ◽

2018 ◽

Vol 15 (2) ◽

pp. 91-95

Author(s):

M. R. Hossain ◽

M. M. Meher ◽

M. Afrin

Keyword(s):

Body Weight ◽

Treatment Group ◽

Present Status ◽

Recovery Rate ◽

Production Cost ◽

Pasteurella Multocida ◽

The Body ◽

Post Mortem ◽

Epidemiological Investigation ◽

Fowl Cholera

The present study was undertaken to determine the present status of fowl cholera (FC) infection of poultry in Gazipur district during September 2016 to February 2017. A total of 282 either dead or sick birds (Layer and Sonali) were taken as sample in which the sick birds were subjected to record clinical data and consequently the dead birds were for post mortem examination. Hence, overall 12.41% prevalence was found for FC and about 87.59% was for mixed other diseases. All the birds were grouped into six named as A1, A2 and up to A6 accordingly their age. The highest incidence (29.79 %) was found in A1 (8-20weeks) group and the lowest was 4.26% in A5 (51-60 weeks) group. Furthermore, six FC infected farms were categorized to treat with six specific antibiotics individually. Thereafter, the highest recovery rate (89.6% and 87.5%) was recorded in two farms using Enrofloxacin and Gentamicin particularly. The lowest recovery rate (60%) was in Erythromycin treatment group. However, the layer birds of six farms lost their body weight almost 11.84%, specifically the highest percentages (16.49%) was in treated with Ampicillin and lowest percentages (5.79%) found in a farm treated with Gentamicin. In brief, FC commonly appears just near to laying age and therapeutic management trivially decreases the body weight that ultimately shoots up the production cost.

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Isolation and Identification of Pasteurella multocida from Chicken for the Preparation of Oil Adjuvanted Vaccine

Microbes and Health ◽

10.3329/mh.v2i1.17253 ◽

2013 ◽

Vol 2 (1) ◽

pp. 1-4 ◽

Cited By ~ 2

Author(s):

Samina Ievy ◽

Mohammad Ferdousur Rahman Khan ◽

Md Ariful Islam ◽

Md Bahanur Rahman

Keyword(s):

Immune Response ◽

Pasteurella Multocida ◽

Challenge Infection ◽

Cholera Vaccine ◽

Isolation And Identification ◽

Passive Hemagglutination ◽

Fowl Cholera ◽

Adjuvanted Vaccine ◽

Group A ◽

The Mean

The research work was performed for the isolation and identification of Pasteurella multocida from field cases, preparation of oil adjuvanted vaccine from isolated strain and determination of its efficacy. Samples were collected from suspected dead birds of three poultry farms of Bangladesh (Code name: M and R). The P. multocida isolates were Gram negative, non-motile, non- spore forming rod occurring singly or pairs and occasionally as chains or filaments. Biochemically P. multocida ferment basic sugar and consistently produced acid except from maltose and lactose. After isolation formalin killed oil adjuvanted Fowl cholera vaccine was prepared in Laboratory of the Department of Microbiology and Hygiene, BAU and this experimental vaccine (3.2x108 CFU/ml) was administered in nine weeks old White Leg Horn chickens at the different dose rate through intramuscular (IM) route in each selected group A (1ml alum precipitated vaccine), B (0.5ml alum precipitated vaccine), C (1ml oil adjuvanted vaccine) and D (0.5ml oil adjuvanted vaccine). Pre-vaccinated sera were collected from all groups of birds. The mean of Passive Hemagglutination (PHA) titers of post-vaccination were 51±17.8, 76.8±17, 89.6±17, and 115±17.81 in group A, B, C and D respectively which consist of 5 birds in each. The vaccine produced better immune response when boostering with the similar dose and route at 15 days after primary vaccination. The mean PHA titers were higher at group D than other groups after boostering. Challenge infection was conducted on all the vaccinated and control group (n=5) of birds after 15 days of vaccination which protect 93.75% of birds and the PHA titers from different groups analyzed to determine the protective capacity of vaccinated chickens against challenge exposure. It was demonstrated that experimental oil adjuvanted fowl cholera vaccine with 0.5ml dose produce higher immune response against challenge infection and found to be safe. Microbes and Health, June 2013, 2(1): 1-4DOI: http://dx.doi.org/10.3329/mh.v2i1.17253

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Rapid virulence typing of Pasteurella multocida by multiplex PCR

Research in Veterinary Science ◽

10.1016/j.rvsc.2009.04.004 ◽

2009 ◽

Vol 87 (3) ◽

pp. 355-357 ◽

Cited By ~ 21

Author(s):

Sina Atashpaz ◽

Jalal Shayegh ◽

Mohammad Saied Hejazi

Keyword(s):

Multiplex Pcr ◽

Pasteurella Multocida ◽

Virulence Typing

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Natural Selection in the Chicken Host Identifies 3-Deoxy-d-manno- Octulosonic Acid Kinase Residues Essential for Phosphorylation of Pasteurella multocida Lipopolysaccharide

Infection and Immunity ◽

10.1128/iai.00457-10 ◽

2010 ◽

Vol 78 (9) ◽

pp. 3669-3677 ◽

Cited By ~ 7

Author(s):

Marina Harper ◽

Andrew D. Cox ◽

Frank St. Michael ◽

Mark Ford ◽

Ian W. Wilkie ◽

...

Keyword(s):

Defense Mechanisms ◽

Pasteurella Multocida ◽

Inner Core ◽

Single Amino Acid ◽

Sequencing Analysis ◽

Wild Type ◽

Fowl Cholera ◽

Mutant Strains ◽

High Doses

ABSTRACT Pasteurella multocida is the causative agent of a number of diseases in animals, including fowl cholera. P. multocida strains simultaneously express two lipopolysaccharide (LPS) glycoforms (glycoforms A and B) that differ only in their inner core structure. Glycoform A contains a single 3-deoxy-d-manno-octulosonic acid (Kdo) residue that is phosphorylated by the Kdo kinase, KdkA, whereas glycoform B contains two unphosphorylated Kdo residues. We have previously shown that P. multocida mutants lacking the heptosyltransferase, HptA, produce full-length glycoform B LPS and a large amount of truncated glycoform A LPS, as they cannot add heptose to the glycoform A inner core. These hptA mutants were attenuated in chickens because the truncated LPS made them vulnerable to host defense mechanisms, including antimicrobial peptides. However, here we show that birds inoculated with high doses of the hptA mutant developed fowl cholera and the P. multocida isolates recovered from diseased birds no longer expressed truncated LPS. Sequencing analysis revealed that the in vivo-derived isolates had mutations in kdkA, thereby suppressing the production of glycoform A LPS. Interestingly, a number of the spontaneous KdkA mutant strains produced KdkA with a single amino acid substitution (A112V, R123P, H168Y, or D193N). LPS structural analysis showed that complementation of a P. multocida kdkA mutant with wild-type kdkA restored expression of glycoform A to wild-type levels, whereas complementation with any of the mutated kdkA genes did not. We conclude that in P. multocida KdkA, the amino acids A112, R123, H168, and D193 are critical for Kdo kinase function and therefore for glycoform A LPS assembly.

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