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

2007 ◽  
Vol 55 (4) ◽  
pp. 425-434 ◽  
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.

scholarly journals Molecular Fingerprinting of Pasteurella Multocida Associated with Progressive Atrophic Rhinitis in Swine Herds

1994 ◽  
Vol 6 (4) ◽  
pp. 442-447 ◽  
Author(s):  
Ian A. Gardner ◽  
Rick Kasten ◽  
Graeme J. Eamens ◽  
Kurt P. Snipes ◽  
Randall J. Anderson
Keyword(s):  
Pasteurella Multocida ◽  
Uv Light ◽  
Type A ◽  
Atrophic Rhinitis ◽  
Capsular Type ◽  
Molecular Fingerprinting ◽  
Whole Cell ◽  
Type D ◽  
Progressive Atrophic Rhinitis ◽  
Swine Herds

Ninety-six nasal isolates of Pasteurella multocida from swine herds with progressive atrophic rhinitis were characterized by restriction endonuclease analysis (REA) of whole-cell DNA, ribotyping, and plasmid analysis. For REA, bacterial DNA was digested with SmaI and electrophoresed in 0.7% agarose, and fragments were visualized with UV light. For ribotyping, EcoRI-digested and electrophoresed restriction fragments of whole-cell DNA were transferred to nitrocellulose membranes, hybridized with γ-32P-labeled Escherichia coli ribosomal RNA, and visualized by autoradiography. Phenotypes of isolates were toxigenic capsular type D ( n = 51), nontoxigenic type D ( n = 28), nontoxigenic type A ( n = 16), and toxigenic type A ( n = 1). Plasmids of various sizes were evident in 92.2% and 17.9% of toxigenic and nontoxigenic D strains, respectively, but were absent from all type A strains. Among the 4 phenotypes, there were 17 REA profiles and 6 ribotypes. For 3 of 17 REA patterns, multiple ribotypes were evident, and several REA types were evident in 5 of 6 ribotypes. Thirty-seven isolates of toxigenic capsular type D from Australian herds were either SmaI type B or C and ribotype 2, whereas 14 toxigenic D isolates from the USA and other countries were more heterogeneous (7 REA types and 6 ribotypes). The fingerprinting results provided evidence in support of the hypothesis of a single source infection in Australia associated with the introduction of breeding pigs from overseas.

scholarly journals Comparison of Phenotypic and Genotypic Identification Methods of Pasteurella multocida Serotypes Isolated from Pigs

2018 ◽  
Vol 44 (1) ◽  
pp. 5
Author(s):  
Amanda Figueiredo Amaral ◽  
Raquel Rebelatto ◽  
Catia Silene Klein ◽  
Karine Ludwig Takeuti ◽  
David Emilio Santos Neves de Barcellos
Keyword(s):  
Staphylococcus Aureus ◽  
Multiplex Pcr ◽  
Pasteurella Multocida ◽  
Weighted Kappa ◽  
Type A ◽  
Kappa Coefficient ◽  
Identification Methods ◽  
Type D ◽  
Weighted Kappa Coefficient ◽  
Capsular Typing

Background: Pasteurella multocida serotypes A and D are commonly associated with pneumonia and pleuritis in pigs.  Different phenotypic techniques, such as hyaluronidase and acriflavine tests, and genotyping techniques, such as PCR, are used to distinguish between these serotypes. The objective of this study was to compare the capsular identification methods of type A and type D P. multocida isolated from pigs using both phenotypic (hyaluronidase and acriflavine tests) and genotypic (multiplex PCR) techniques.Materials, Methods & Results: A total of 44 lyophilized P. multocida isolates, obtained between 1981 and 1997 from pig farms at Rio Grande do Sul State, Brazil, were analyzed. The isolates were reactivated in Brain Heart Infusion (BHI) broth and cultured in BHI broth and blood agar supplemented with 5% sheep blood. Colony identity was further confirmed by evaluating colony morphology in blood agar and confirming the absence of growth on MacConkey agar. Bacteria in Tryptone Soy Agar (TSA) were used for the Triple Sugar Iron (TSI), Sulfide-Indole-Motility (SIM), and nitrate, glucose, lactose, sucrose and mannitol fermentation tests. For hyaluronidase test, P. multocida colonies were streaked transversally across the entire plate, approximately 3-5mm apart, in order to observe their lines of growth. Following this, a hyaluronidase producing strain of Staphylococcus aureus was heavily streaked at right angles to the P. multocida lines and the plates were incubated at 37°C for 24 h. Type A isolates were then identified as those with smaller colonies in the region adjacent to the Staphylococcus aureus streak (negative satellitism). For acriflavine test, the isolates were inoculated into tubes containing 2 mL of BHI, incubated at 37°C for 18-24 h, centrifuged (500 g for 15 min) and 1.5 mL of the supernatant was discarded. A 1:1000 solution of acriflavine neutral (0.5 mL) was then added to the residual broth containing bacteria and kept at room temperature. Solutions of acriflavine were freshly prepared each week and stored protected from light. Type D strains were identified by the appearance of a heavy flocculent precipitate within 5 min. DNA extraction by heat shock was performed prior to multiplex PCR for the detection of capsular genes hyaD-hyaC (capsular typing A) and dcbF (capsular typing D). Test of symmetry and a weighted kappa coefficient were used to evaluate correlations and to assess agreement of the  results between the identification methods, respectively. Phenotypic tests showed that two isolates were type D (4.55%), 40 were type A (90.9%) and two (4.55%) were untypable isolates (4.55%) while PCR showed that 38 isolates were type A (86.36%) and six were type D (13.64%). The correlation analysis between the phenotypic and genotypic tests showed that 90.9% of the strains were identified as belonging to the same serotype by both tests and the weighted kappa coefficient (K = 0.633) indicates a substantial agreement between the two tests.Discussion: There was a disagreement between the phenotypic and genotypic results in four of the isolates (9.09%). The phenotypically untypable isolates were classified as type D by multiplex PCR. Nonetheless, we conclude that PCR testing is a more reliable method to differentiate between P. multocida serotypes A and D.

scholarly journals Typing of isolates of Clostridium perfringensfrom healthy and diseased sheep by multiplex PCR

2012 ◽  
Vol 50 (No. 10) ◽  
pp. 439-442 ◽  
Author(s):  
H. Kalender ◽  
Ertas HB ◽  
B. Cetinkaya ◽  
A. Muz ◽  
N. Arslan ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Type A ◽  
The Other ◽  
Enterotoxin Gene ◽  
Type D ◽  
Enterotoxin Genes ◽  
Healthy Sheep ◽  
Type C ◽  
Alpha Beta

In this study, C. perfringens strains isolated from healthy and diseased sheep were analysed by multiplex PCR in order to to detect the presence of the alpha, beta, epsilon, iota and enterotoxin genes. C. perfringens was isolated from 52 of 104 sheep with enterotoxemia signs and from 61 of 194 clinically healthy sheep. Genotyping of 52 strains from diseased sheep indicated that 33 (64%) were type A, 11 (21%) type D and 8 (15%) type C. Of 61 strains from healthy sheep, 58 (95%) were type A and 3 (5%) type D. The other types of C. perfringens were not detected, and none of the isolates contained the enterotoxin gene. This result indicates that the enterotoxin of C. perfringens does not play the important role in the occurence of enterotoxemia in sheep.

scholarly journals Phenotypic and Genotypic Characterisation of Pasteurella Multocida Strains Isolated from Pigs in Poland

2013 ◽  
Vol 57 (1) ◽  
pp. 29-34 ◽  
Author(s):  
Katarzyna Stępniwska ◽  
Iwona Markowska-Daniel
Keyword(s):  
Pasteurella Multocida ◽  
Type A ◽  
High Susceptibility ◽  
Biochemical Tests ◽  
Internal Organs ◽  
Type D ◽  
Phenotypic Characterisation ◽  
Nasal Swabs ◽  
Typing Methods ◽  
Capsular Typing

AbstractA total of 319Pasteurella multocida(Pm) strains isolated from pigs in Poland were examined. Phenotypic characterisation included: biochemical tests (to determine species, subspecies, and biovar), capsular typing, and antimicrobial susceptibility. Genotypic characterisation included detection of thetoxAgene by PCR. All testedPmstrains were classified asPmsubsp.multocida: 87.2% biovar 3, 10.7%-2 and 0.9%-12. One strain was classified as biovar 1. Three strains ofPmdid not suit any of the biovars. Using capsular typing methods, 77% ofPmstrains isolated from nasal swabs belonged to type D and 33% to type A. AmongPmstrains isolated from internal organs, 59.5% belonged to type A and 40.5% to type D. All the isolates showed a high susceptibility to β-lactams: ampicillin and amoxicilin with clavulonic acid (97.8%), penicillin (86.7%), doxicilline (100%), oxytetracycline (97.8%), and tetracycline (93.2%). It was found that all strains were susceptible to norfloxacin, 97.8% to enrofloxacin, and 95.6% to SxT. 24.4% and 15.6% of the strains were resistant to linco-spectin and tiamulin, respectively. The presence oftoxAgene was confirmed by PCR in 20.8% of the strains isolated from nasal swabs and 29.1% of isolates from internal organs.

scholarly journals CTNI-03. IMAGING FEATURES OF LEPTOMENINGEAL METASTASES OF NON-SMALL CELL LUNG CANCER: A SINGLE-CENTER REAL-WORLD STUDY

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii41-ii41
Author(s):  
Junjie Zhen ◽  
Lei Wen ◽  
Shaoqun Li ◽  
Mingyao Lai ◽  
Changguo Shan ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Type A ◽  
Brain Parenchyma ◽  
Imaging Features ◽  
Type B ◽  
Type D ◽  
Mri Examination ◽  
Mri Features ◽  
Brain And Spinal Cord ◽  
The Brain

Abstract BACKGROUND According to EANO-ESMO clinical practice guidelines, the MRI findings of LM are divided into 4 types, namely linear enhancement (type A), nodular enhancement (type B), linear combined with nodular enhancement (type C), and sign of hydrocephalus (type D). METHODS The MRI features of brain and spinal cord in patients diagnosed with NSCLC-LM in Guangdong Sanjiu Brain Hospital from 2010 until 2019 were investigated, and then were classified into 4 types. The imaging features were analyzed. RESULTS A total of 80 patients were enrolled in the study. The median age of the patients was 53.5 years old, and the median time from the initial diagnosis to the confirmed diagnosis of LM was 11.6 months. The results of enhanced MRI examination of the brain in 79 cases showed that the number of cases with enhancements of type A, B, C and D were 50 (63.3%), 0, 26 (32.9%) and 3 (3.8%), respectively, and that LM with metastases to the brain parenchyma was found in 42 cases (53.2%). The results of enhanced MRI examination of spinal cord in 59 cases showed that there were only enhancements of type A and C in 40 cases (67.8%) and 3 cases (5.0%), and no enhancement sign in the other 16 cases (27.2%). CONCLUSION MRI examination of brain and spinal cord will improve the detection rate of LM. The MRI features of NSCLC-LM in real world are mainly characterized by the linear enhancements of brain and spinal cord, followed by linear combined with nodular enhancement. The enhancements of type B and type D are rare in clinic. Almost half of the patients have LM and metastases to the brain parenchyma. Therefore, the differentiation of tumor metastases is needed to be paid attention to for the early diagnosis and the formulation of reasonable treatment plans.

MRI analysis and surgical treatment of Wassel Type IV duplicated thumbs

2021 ◽  
pp. 175319342098321
Author(s):  
Anyuan Wang ◽  
Jian Ding ◽  
Long Wang ◽  
Tinggang Chu ◽  
Zhipeng Wu ◽  
...  
Keyword(s):  
Type A ◽  
Surgical Reconstruction ◽  
Mri Findings ◽  
Level Of Evidence ◽  
Type D ◽  
Type Iv ◽  
Secondary Operations ◽  
Type C

We present the MRI findings for 39 Wassel Type IV duplicated thumbs in 38 patients. We found that MRI revealed the morphology of the cartilaginous connection between the thumb anlages and the location of the deviation corresponding to the classification of Horii, which allowed precise preoperative planning of corrective osteotomies. All 39 thumbs were available for follow-up after surgical reconstruction at a mean of 29 months (range 25 to 39). Four out of nine Horii Type A cases and all 12 Type B, as well as the six Type C and the six Type D cases, achieved good results according to the Tada scoring system. Five Type A cases achieved fair results with residual stiffness of the interphalangeal joint. No secondary operations were needed. We conclude that MRI proved useful in subclassifying Wassel Type IV duplicated thumbs and may aid in planning the osteotomies needed for their reconstruction. Level of evidence: IV

scholarly journals Virulence Determinants and Antimicrobial Profiles of Pasteurella multocida Isolated from Cattle and Humans in Egypt

Antibiotics ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 480
Author(s):  
Mohamed Sabry Abd Elraheam Elsayed ◽  
Samah Mahmoud Eldsouky ◽  
Tamer Roshdy ◽  
Lamia Said ◽  
Nahed Thabet ◽  
...  
Keyword(s):  
Pasteurella Multocida ◽  
Efflux Pump ◽  
Virulence Gene ◽  
Molecular Classification ◽  
Multiple Drug ◽  
Virulence Determinants ◽  
Type D ◽  
Multiple Drug Resistant ◽  
Class 1 ◽  
Methylase Gene

Pasteurella multocida is a Gram-negative bacterium that causes drastic infections in cattle and humans. In this study, 55 isolates were recovered from 115 nasal swabs from apparently healthy and diseased cattle and humans in Minufiya and Qalyubia, Egypt. These isolates were confirmed by kmt1 existence, and molecular classification of the capsular types showed that types B, D, and E represented 23/55 (41.8%), 21/55 (38.1%), and 11/55 (20.0%), respectively. The isolates were screened for five virulence genes with hgbA, hgbB, and ptfA detected in 28/55 (50.9%), 30/55 (54.5%), and 25/55 (45.5%), respectively. We detected 17 capsular and virulence gene combinations with a discriminatory power (DI) of 0.9286; the most prevalent profiles were dcbF type D and dcbF type D, hgbA, hgbB, and ptfA, which represented 8/55 (14.5%) each. These strains exhibited high ranges of multiple antimicrobial resistance indices; the lowest resistances were against chloramphenicol, ciprofloxacin, amoxicillin/clavulanic acid, and levofloxacin. The macrolide–lincosamide–streptogramin B methylase gene erm(Q), with erm(42) encoding MLSB monomethyltransferase, mph(E) encoding a macrolide efflux pump, and msr(E) encoding macrolide-inactivating phosphotransferase were present. The class 1 and 2 integrons and extended-spectrum β-lactamase genes intl1, intl2, blaCTX-M, blaCTX-M-1, and blaTEM were detected. It is obvious to state that co-occurrence of resistance genes resulted in multiple drug-resistant phenotypes. The identified isolates were virulent, genetically diverse, and resistant to antimicrobials, highlighting the potential risk to livestock and humans.

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