scholarly journals Molecular characterization of Pasteurella multocida isolates from swine lungs by Randomly Amplified Polymorphic DNA

2015 ◽  
Vol 46 (1) ◽  
pp. 119-125
Author(s):  
Cristiane Silva Chitarra ◽  
Mayara Inácio Vincenzi da Silva ◽  
Laila Natasha Santos Brandão ◽  
Francielle Cristina Kagueyama ◽  
Stefhano Luis Candido ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Pasteurella Multocida ◽  
Economic Losses ◽  
Clinical Samples ◽  
Atrophic Rhinitis ◽  
Randomly Amplified Polymorphic Dna ◽  
Swine Industry ◽  
Mato Grosso ◽  
Swine Herds

ABSTRACT: Swine respiratory diseases such as atrophic rhinitis and bronchopneumonia caused by Pasteurella (P.) multocida cause important economic losses to the modern swine industry. The purpose of this study was to characterize P. multocida strains isolated from swine lungs by RAPD (Randomly Amplified Polymorphic DNA) to demonstrate their genetic diversity. Ninety-four samples of fragments from lungs with pneumonia and sixty one samples without pneumonia were collected in slaughterhouses in Mato Grosso during the period from December 2009 to March 2010. Clinical cases in 2012 and 2013 were also included in this study. Among the lung fragments with macroscopic lesions, without macroscopic lesions and clinical samples, 40.42%, 4.49% and 100% were positive for P. multocida, respectively. Bacterial identification culturing was confirmed by PCR (polymerase chain reaction) by means of the amplification of the gene kmt1. RAPD technique was performed for 46 isolates, and in every isolate, a total of 7 to 11 amplification bands were detected, composed of 8 clusters based on genetic similarity. Thus, treatment, control and preventive measures should consider the genetic diversity of P. multocida populations in swine herds in order to improve the development of new protocols to produce antimicrobials and vaccines.

scholarly journals Polymorphic microsatellites of analysis in cultivars of taro

2012 ◽  
Vol 30 (1) ◽  
pp. 106-111 ◽  
Author(s):  
Raquel SC Nunes ◽  
Fernanda R Pinhati ◽  
Luciana P Golinelli ◽  
Tiyoko Nair H Rebouças ◽  
Vânia Margaret F Paschoalin ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Local Varieties ◽  
Mato Grosso ◽  
Espírito Santo ◽  
Polymorphic Microsatellites ◽  
Espirito Santo ◽  
Microsatellite Alleles ◽  
Mato Grosso Do Sul ◽  
Simple Sequence

Taro (Colocasia esculenta) is a tuberous plant belonging to the Araceae family whose tuber is the 14th most consumed food crop in the world. Characterized as an unconventional vegetable, taro is grown in Brazil as a subsistence crop, but in recent years began to gain commercial importance, especially in the states of Espirito Santo, Minas Gerais and Rio de Janeiro. To avoid loss of genetic diversity of the local varieties traditionally grown in Brazil a core collection for taro germplasm has been developed by the Instituto Capixaba de Pesquisa, Assistência Técnica e Extensão Rural do estado do Espirito Santo (Incaper). The aim of this study was to perform a molecular characterization of the seven regional core collections. Genetic diversity of the cultivars was investigated by using SSR (Simple Sequence Repeats) polymorphisms, in seven loci (Xuqtem55, Xuqtem73, Xuqtem84, Xuqtem88, Xuqtem91, Xuqtem97 and Xuqtem110). Genetic diversity of the cultivars, based on the seven microsatellite alleles, was evaluated by using the software GelCompar II, showed that the loci Xuqtem73, Xuqtem88 and Xuqtem110 were the most informative, featuring 7, 10 and 8 alleles, respectively, a percentage of cultivars with polymorphic alleles of 85, 57 and 100% and identical PIC of 0.91. Based on Xuqtem110 locus analysis, the seven cultivars were grouped in two clusters. Chinês Regional Incaper cultivar was originated from Chinês cultivar which originated the São Bento cultivar, corroborating previous results. Macaquinho and Chinês cultivars were shown to be the primitive ones originating the allelic collections found in the states of Mato Grosso do Sul and Espirito Santo.

scholarly journals Use of ELISA to Detect Toxigenic Pasteurella Multocida in Atrophic Rhinitis in Swine

1992 ◽  
Vol 4 (4) ◽  
pp. 419-422 ◽  
Author(s):  
Terry L. Bowersock ◽  
Tom Hooper ◽  
Ronald Pottenger
Keyword(s):  
Tissue Culture ◽  
Pasteurella Multocida ◽  
Atrophic Rhinitis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cytotoxicity Assays ◽  
Highly Sensitive ◽  
Nasal Secretions ◽  
Toxigenic Strains ◽  
Swine Herds ◽  
Toxigenic Pasteurella Multocida

The use of an enzyme-linked immunosorbent assay (ELISA) as a means of detecting dermone-crotoxin-producing strains of Pasteurella multocida was investigated. The assay was evaluated as a means to identify toxigenic P. multocida isolates recovered from nasal secretions of swine with atrophic rhinitis. The sensitivity and specificity of the ELISA for detecting dermonecrotoxin-producing P. multocida strains were compared to those of mouse-inoculation and cytotoxicity assays. The ELISA was highly sensitive and more specific than animal inoculation or tissue culture assay and is thus a more effective method for screening swine herds for the presence of toxigenic strains of P. multocida. The ELISA is a rapid, effective, economical way to identify toxigenic P. multocida isolates.

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 Characterization of Pasteurella multocida isolated from dead rabbits with respiratory disease in Fujian, China

2019 ◽  
Author(s):  
Jinxiang Wang ◽  
Lei Sang ◽  
Shikun Sun ◽  
Yanfeng Chen ◽  
Dongjin Chen ◽  
...  
Keyword(s):  
Respiratory Disease ◽  
Pasteurella Multocida ◽  
Sequence Type ◽  
Economic Losses ◽  
Pcr Screening ◽  
Resistance Rates ◽  
And Control ◽  
First Time ◽  
Sequence Types

Abstract Background: Pasteurella multocida is one of the important pathogens that infect rabbits, causing major economic losses in commercial rabbit farming. In this study, 205 P. multocida isolates recovered from lungs of dead rabbits with respiratory disease were defined by capsular serogroups, lipopolysaccharide (LPS) genotypes and multi-locus sequence types, screened virulence factors and antimicrobial susceptibility. Results: The 205 isolates were assigned into 2 capsular types, A and D, and 2 LPS genotypes, L3 and L6. When combining capsular types with LPS genotypes, 4 serotypes were detected. A:L3 (51.22%, 105/205) was the most predominant serotype, followed by A:L6 (24.88%, 51/205), D:L6 (19.02%, 39/205) and D:L3 (4.88%, 10/205). The 205 isolates were grouped into 3 sequence types, ST10, ST11 and ST12. ST12 (56.10%, 115/205) was the most prevalent sequence type, followed by ST10 (24.88%, 51/205) and ST11 (19.02%, 39/205). In the 205 isolates, virulence associated genes ptfA , fur , hgbB , ompA , ompH and oma87 were positive in the PCR screening, whereas the toxA and tbpA genes were negative. Notably, the 156 capsular serogroup A isolates carried the pmHAS gene. All the 205 isolates were susceptible to most of the used antibiotics, except for streptomycin, gentamycin, kanamycin and ceftriaxone, and the resistance rates of which were 27.80%, 15.61%, 9.27% and 2.44%, respectively. Conclusions: This study, for the first time, described the prevalence and characteristics of P. multocida causing respiratory disease in rabbits in Fujian Province, which might be useful for tracking the epidemic strains and development of efficient vaccines and methods to prevent and control the pathogen.

scholarly journals Porcine Epidemic Diarrhea Virus 3C-Like Protease-Mediated Nucleocapsid Processing: Possible Link to Viral Cell Culture Adaptability

2016 ◽  
Vol 91 (2) ◽  
Author(s):  
Peera Jaru-Ampornpan ◽  
Juggragarn Jengarn ◽  
Asawin Wanitchang ◽  
Anan Jongkaewwattana
Keyword(s):  
Cell Culture ◽  
Growth Retardation ◽  
Porcine Epidemic Diarrhea Virus ◽  
Vaccine Development ◽  
Wild Type Virus ◽  
Economic Losses ◽  
Swine Industry ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

ABSTRACT Porcine epidemic diarrhea virus (PEDV) causes severe diarrhea and high mortality rates in newborn piglets, leading to massive losses to the swine industry worldwide during recent epidemics. Intense research efforts are now focusing on defining viral characteristics that confer a growth advantage, pathogenicity, or cell adaptability in order to better understand the PEDV life cycle and identify suitable targets for antiviral or vaccine development. Here, we report a unique phenomenon of PEDV nucleocapsid (N) cleavage by the PEDV-encoded 3C-like protease (3Cpro) during infection. The identification of the 3Cpro cleavage site at the C terminus of N supported previous observations that PEDV 3Cpro showed a substrate requirement slightly different from that of severe acute respiratory syndrome coronavirus (SARS-CoV) 3Cpro and revealed a greater flexibility in its substrate recognition site. This cleavage motif is present in the majority of cell culture-adapted PEDV strains but is missing in emerging field isolates. Remarkably, reverse-genetics-derived cell culture-adapted PEDVAVCT12 harboring uncleavable N displayed growth retardation in Vero E6-APN cells compared to the wild-type virus. These observations altogether shed new light on the investigation and characterization of the PEDV nucleocapsid protein and its possible link to cell culture adaptation. IMPORTANCE Recurrent PEDV outbreaks have resulted in enormous economic losses to swine industries worldwide. To gain the upper hand in combating this disease, it is necessary to understand how this virus replicates and evades host immunity. Characterization of viral proteins provides important clues to mechanisms by which viruses survive and spread. Here, we characterized an intriguing phenomenon in which the nucleocapsids of some PEDV strains are proteolytically processed by the virally encoded main protease. Growth retardation in recombinant PEDV carrying uncleavable N suggests a replication advantage provided by the cleavage event, at least in the cell culture system. These findings may direct us to a more complete understanding of PEDV replication and pathogenicity.

scholarly journals Genetic Characterization of Porcine Epidemic Diarrhea Virus in China Between 2014 and 2018: Emergence of the G1c Subtype

2020 ◽  
Vol 40 (04) ◽  
pp. 474-478
Author(s):  
Wenqiang Jiao
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Phylogenetic Trees ◽  
Genetic Characterization ◽  
Diagnostic Methods ◽  
Clinical Samples ◽  
Swine Industry ◽  
Diarrhea Virus ◽  
Positive Rate ◽  
Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV) has caused substantial economical loss to the Chinese swine industry. To illustrate the genetic characterization of PEDV circulating in China, 205 clinical samples between 2014 and 2018 were collected from 7 provinces in China. 93.17% (191 of 205) of the intestinal and fecal samples were positive for PEDV. 25 S1 amino acid (aa) together with 27 ORF3 genes from 8 provinces were sequenced and analyzed. The phylogenetic trees based on the S1 and ORF3 genes were constructed by the neighbor-joining method using MEGA 7 software. PEDV prevalence was 86.96% (40 of 46) of the swine farms in the 8 provinces and the PEDV positive rate was 93.17% (191 of 205) in the tested samples. Genetic analysis showed CH-JIANGXI-1-2016 CH-JIAGNXI-2-2016, CH-JIANGXI-3-2016 and CH-JIANGXI-2017 had three notable insertions or deletions occurred at aa 59-62, 160, and 139 (140) when compared to all of the strains in this study; moreover, phylogenetic analysis indicated that the four isolates formed a new branch significantly different from G1a, G1b and Indel subtype based on S1 gene: that is the G1c subtype. More research is needed to determine whether the insertions and deletions had biological influence on the virus. The results acquired in the present study showed the genetic diversity of PEDV circulating in 8 provinces, providing information for the development of new diagnostic methods and new vaccines

scholarly journals Molecular Detection and Characterization of Pasteurella multocida Infecting Camels in Marsabit and Turkana Counties, Kenya

2021 ◽  
Author(s):  
Justus Kyalo Kasivalu ◽  
George Isanda Omwenga ◽  
Gabriel Oluga Aboge
Keyword(s):  
Genetic Diversity ◽  
Dna Sequences ◽  
Pasteurella Multocida ◽  
Marker Gene ◽  
Climatic Conditions ◽  
Marker Genes ◽  
Northern Kenya ◽  
Pcr Multiplex ◽  
Limited Food

Abstract BackgroundInfection with Pasteurella multocida is abundant in Kenya yet there is scarce information on their genetic diversity. Pasteurella multocida is considered to be one of the normal flora in the respiratory tract of camels and other animals but it becomes pathogenic and causes pasteurellosis when the resistance of the camel body is diminished by harmful environmental influences. Close herding, overwork, limited food supply, and wet climatic conditions are stresses that seem to speed the spread of the infection. Conventional PCR, Multiplex PCR and sequencing were applied to enhance identification of Pasteurella multocida at any level of specificity viz; strain, species, and genus. These molecular tools were applied to confirm the presence and genetic diversity of Pasteurella multocida in 102 blood and 30 nasal swab samples collected from Marsabit and Turkana counties in Kenya. Kmt1 gene was used as the marker gene for Pasteurella multocida and hyaD-hyaC, bcbD, dcbF, ecbJ, and fcbD as marker genes for capsular groups. A study done in northern Kenya noted that in Africa pasteurellosis infections causing death in camels (Camelus dromedarius) have been existing since 1890 though the real cause of this disease remains elusive and needs further study. The study was done to detect Pasteurella multocida and characterize its capsular types by application of molecular biology toolsResultsTwenty one Kenyan isolates were confirmed to be Pasteurella multocida and only capsular group E was detected in both counties. Pasteurella multocida sequences were found to be highly conserved, however isolates detected in Kenya were found to be genetically related to other isolates from African and other parts of the world. ConclusionsThe study confirm that the camels were infected by Pasteurella multocida of capsular type E in Marsabit and Turkana Counties of Kenya. DNA sequences were found to be homologous to Pasteurella multocida thereby confirming that the camels were infected by Pasteurella multocida.

scholarly journals Genetic Diversity and Prevalence of Porcine Circovirus Type 2 in China During 2000-2019

2021 ◽  
Vol 8 ◽  
Author(s):  
Ning Li ◽  
Jing Liu ◽  
Jiali Qi ◽  
Feng Hao ◽  
Lei Xu ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Wide Distribution ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Amino Acid Sequences ◽  
Porcine Circovirus ◽  
Economic Losses ◽  
Swine Industry ◽  
Emerging Virus

As the major pathogen for porcine circovirus-associated disease (PCVAD), porcine circovirus type 2 (PCV2) is no longer treated as an emerging virus anymore. The wide distribution of PCV2 infection in China causes huge economic losses in the swine industry. Currently, it is generally believed that PCV2 has eight genotypes (PCV2a to PCV2h), with PCV2a, PCV2b, and PCV2d being widely distributed. To comprehensively explore the genetic diversity and prevalence of PCV2 in China, PCV-2 sequences submitted from China in the GenBank database were retrieved. With a total of 714 PCV2 strains were retrieved, we found that early-submitted PCV2 sequences were mainly collected from coastal provinces in the southeast part of China, which may indicate PCV2 was initially circulating in those regions. From 2002 to 2008, PCV2b was the dominant prevalent genotype in those retrieved sequences. From 2009, PCV2d became the dominant genotype in those sequences, dropping a hint that a potential shift of PCV2b to PCV2d might occur in 2009, which is similar to the patterns at the global level. In addition to the PCV2a, PCV2b, and PCV2d genotypes, novel strains were also characterized. We further revealed that the amino acid sequences consistency of PCV2a Cap is higher than those in other genotypes. Together, this study provided clues for the possible prevalent genotypes and dynamics of genetic diversity in China from 2000 to 2019.

Sign in / Sign up

Export Citation Format

Share Document