1.Detection of Helicobacter spp. in gastric, fecal and saliva samples from swine affected by gastric ulceration.
Patrizia Casagrande PROIETTI ; Annalisa BIETTA ; Chiara BRACHELENTE ; Elvio LEPRI ; Irit DAVIDSON ; Maria Pia FRANCIOSINI
Journal of Veterinary Science 2010;11(3):221-225
The aim of this study was to evaluate the presence of Helicobacter (H.) spp. in swine affected by gastric ulceration. Stomachs from 400 regularly slaughtered swine were subjected to gross pathological examination to evaluate the presence of gastric ulcers. Sixty-five samples collected from ulcerated pars esophagea and 15 samples from non-ulcerated pyloric portions were submitted to histopathological and molecular analyses, to detect Helicobacter spp., H. suis and H. pylori by PCR. Feces and saliva swabs were also collected from 25 animals in order to detect in vivo the presence of Helicobacter spp.. Gastric ulcers were detected in 373 cases (93%). The presence of ulcers in association with inflammatory processes was further confirmed by histological examination. Forty-nine percent (32/65) of the ulcerated esophageal portions as well as 53% (8/15) of the non-ulcerated pyloric portions were positive for Helicobacter spp. by PCR. The Helicobacter spp. positive samples were also positive for H. suis, while H. pylori was not detected. These results were confirmed by restriction enzyme analysis. With regard to feces and saliva samples, 15/25 (60%) and 16/25 (64%) were positive for Helicobacter spp. PCR, respectively but all were negative in H. suis and H. pylori specific PCR.
Animals
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Feces/*microbiology
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Helicobacter/*isolation & purification
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Polymerase Chain Reaction/veterinary
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Restriction Mapping/veterinary
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Saliva/*microbiology
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Stomach/*microbiology
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Stomach Ulcer/microbiology/pathology/*veterinary
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Swine
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Swine Diseases/*microbiology/pathology
2.Differential diagnosis of Salmonella gallinarum and S. pullorum using PCR-RELP.
Myeong Kyu PARK ; Kyoung Seong CHOI ; Myeong Chul KIM ; Joon Seok CHAE
Journal of Veterinary Science 2001;2(3):213-219
Salmonellosis in poultry of Korea is a significant health problem, which causes substantial economic losses. The most common causative agents of chicken salmonellosis ar S. gallinarum and S. pullorum. Traditional methods used to detect Salmoenella spp. In chicken are tedious, time consuming and confer little guarantee of sensitivity and species specificity. Therefore, a rapid and sensitive method for the differentiation of Salmonella serogroup D was assessed. We first amplified the rfbS genes by PCR and characterized the amplified product by nucleotide sequence analysis. The homology of nucleotide sequence was 99.7% between S. gallinarum and S. pullorum rfbS genes. Further comparisons of the sequences of S. gallinarum, S. gallinarum fied strain, S. pullorum and S. typhi(GenBank Accession No.M29682) showed a homology of 98.3%. The predicted amino acid sequence homology was 97.1%, 97.1% and 97.5%, respectively. Based on this comparison of these nucleotide sequences, we found unique restriction enzyme sites within the rfbS genes of S. gallinarum and S. pullorum. Thus, the PCR products could be further digested with restriction enzymes TfiI and PleI for use in a restriction fragment length polymorphism (RELP) technique. This method can be applied in the differential diagnosis between S. gallinarum and S. pullorum.
Animals
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Base Sequence
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*Chickens
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Diagnosis, Differential
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Polymerase Chain Reaction/veterinary
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Polymorphism, Restriction Fragment Length
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Poultry Diseases/*diagnosis/microbiology
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Restriction Mapping/veterinary
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Salmonella/*classification/genetics/isolation&purification
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Salmonella Infections, Animal/*diagnosis/microbiology
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Sensitivity and Specificity
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Sequence Homology, Amino Acid
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Sequence Homology, Nucleic Acid
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Species Specificity
3.Genetic characterization of porcine circovirus-2 field isolates from PMWS pigs.
Journal of Veterinary Science 2002;3(1):31-39
PMWS is a new emerging disease in swine herds worldwide. Field isolates of PCV-2, a putative major causative agent of PMWS, were isolated and genetically characterized. Viral genome of two field isolates (PC201DJ and PC201SS) from pigs showing typical PMWS was sequenced. The nucleotide sequence homology with other PCV-2 isolates was ranging from 95% to 99% in complete viral genomic sequence. The highly conserved nonanucleotide motif of replication origin was identical to that of other PCV-2 isolates. To determine the genetic heterogeneity of PCV-2 isolates, the phylogenetic tree based on the complete genome of PCV-2 isolates were constructed. Two PCV-2 field isolates were closely related to Canadian isolates of PCV-2. PCV-2 isolated from field may have an origin of North America and is possibly originated from importation of breeding stocks. The result indicates that although the genome of PCV-2 is relatively stable in general, minor genetic variations exist among PCV-2 isolates from the different geographic locations. These differences of viral genome might have an important implication for genetic characteristics of PCV-2 infection. Three major immunorelevant epitopes of capsid protein showed variations in amino acid sequences. Also, the variance of amino acid sequence in antigenic epitope existed between two Korean PCV-2 isolates.
Animals
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Base Sequence
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Circoviridae Infections/*veterinary
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Circovirus/classification/genetics/*isolation & purification
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Cloning, Molecular
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Conserved Sequence
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DNA Primers
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Genome, Viral
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Korea
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Molecular Sequence Data
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Phylogeny
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Polymerase Chain Reaction
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Restriction Mapping
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Sequence Alignment
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Sequence Homology, Nucleic Acid
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Swine
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Swine Diseases/*virology
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Wasting Syndrome/*veterinary/virology
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Weaning