Detection ofLawsonia intracellularisin faeces of swine from the main producing regions in Brazil

1999 ◽  
Vol 45 (3) ◽  
pp. 230-234 ◽  
Author(s):  
Adriana E.C. Nascimento Chiriboga ◽  
Walter V Guimarães ◽  
Maria Cristina D. Vanetti ◽  
Elza F Araújo
Keyword(s):  
Polymerase Chain Reaction Method ◽  
Lawsonia Intracellularis ◽  
Specific Primers ◽  
Poor Growth ◽  
Mato Grosso ◽  
Proliferative Enteropathy ◽  
Total Incidence ◽  
Faecal Samples ◽  
Dna Fragment ◽  
Polymerase Chain

Swine proliferative enteropathy is an enteric disease caused by Lawsonia intracellularis which affects animals between 6 and 20 weeks of age, causing diarrhea, anorexia, and poor growth. The presence of L. intracellularis was evaluated in the faecal samples of 636 swine from 75 randomly chosen herds in the main swine-producing regions of Brazil. The pathogen was detected by the polymerase chain reaction method (PCR) using L. intracellularis specific primers. A 319-bp DNA fragment specific for L. intracellularis was produced on amplification of DNA from the faeces of pigs with proliferative enteropathy. Equal amounts of DNA extracted from the faeces of animals from the same herd were pooled together and, once L. intracellularis was detected, the faecal material of each animal was analyzed separately. The incidence of L. intracellularis was 33.4% in the state of Santa Catarina, 29.4% in Paraná, 26.3% in Minas Gerais, 16.7% in Mato Grosso, and 7.1% in São Paulo. The presence of the pathogenic agent was detected in samples from 15 farms, representing a total incidence of 20%. Although 46 animals (7.2%) were shown to be infected, 11% did not present any symptoms of swine proliferative enteropathy. The use of PCR allowed the detection of L. intracellularis in swine farms and the evaluation of the incidence of proliferative enteropathy in different regions of Brazil.Key words: proliferative enteropathy, diagnosis, Lawsonia intracellularis, PCR, incidence.

scholarly journals Using PCR to Distinguish Diaporthe phaseolorum and Phomopsis longicolla from Other Soybean Fungal Pathogens and to Detect Them in Soybean Tissues

Plant Disease ◽  
1997 ◽  
Vol 81 (10) ◽  
pp. 1143-1149 ◽  
Author(s):  
A. W. Zhang ◽  
G. L. Hartman ◽  
L. Riccioni ◽  
W. D. Chen ◽  
R. Z. Ma ◽  
...  
Keyword(s):  
Fungal Pathogens ◽  
Nuclear Ribosomal Dna ◽  
Specific Primers ◽  
Phomopsis Longicolla ◽  
Tissue Samples ◽  
Diaporthe Phaseolorum ◽  
Pcr Products ◽  
Fungal Dna ◽  
Dna Fragment ◽  
Polymerase Chain

Restriction fragment length polymorphism analyses of polymerase chain reaction (PCR) amplified DNA were used to distinguish Diaporthe phaseolorum and Phomopsis longicolla isolates from other soybean fungal pathogens. Primers made to the conserved sequences of nuclear ribosomal DNA amplified the internal transcribed spacer (ITS) regions of D. phaseolorum var. meridionalis and P. longicolla. The PCR products were cloned and then sequenced. Specific-primers, Phom.I and Phom.II, were designed from the polymorphic regions of D. phaseolorum and P. longicolla isolates from soybean to distinguish them from other soybean fungal pathogens. These ITS-derived primers amplified a 337-bp-specific DNA fragment from P. longicolla, D. phaseolorum var. meridionalis, D. phaseolorum var. caulivora, D. phaseolorum var. sojae, and Phomopsis spp. from 20 different hosts. No amplified product was observed using DNA of seven other soybean fungal pathogens or soybean DNA. The detection limit of PCR using primers Phom.I and Phom.II was 2.5 × 10-7 dilution of fungal DNA extracted from samples of 10 pooled seeds and as low as a 1:15 (Phomopsis:soybean) ratio when using 10 ng of DNA per μl from each P. longicolla and soybean. PCR did not produce products using primers Phom.I and Phom.II with DNA extracted from noninfected seeds, but specific bands were observed from samples of 10 pooled seeds and from individually infected seeds. A specific band was observed as well from DNA extracts of tissue samples from symptomless plants inoculated with P. longicolla and D. phaseolorum var. sojae.

scholarly journals Development of Sequence-specific Primers that Amplify a 1.5-kb DNA Marker for Race 1 Fusarium Wilt Resistance in Cucumis melo L.

HortScience ◽  
1998 ◽  
Vol 33 (2) ◽  
pp. 291-292 ◽  
Author(s):  
W. Patrick Wechter ◽  
Ralph A. Dean ◽  
Claude E. Thomas
Keyword(s):  
Fusarium Wilt ◽  
Cucumis Melo ◽  
Specific Primers ◽  
Breeding Programs ◽  
Breeding Lines ◽  
Race 1 ◽  
Dna Fragment ◽  
Polymerase Chain ◽  
Cucumis Melo L

Two 24-mer primers, MUSKFOM I and MUSKFOM II, were developed that amplify a 1.5-kb DNA fragment in race 1 Fusarium wilt resistant muskmelon (Cucumis melo L.), but not in race 1 susceptible germplasm tested. Three race 1 resistant cultivars and two race 1 resistant breeding lines as well as eight race 1 susceptible lines were analyzed using the two sequence-specific primers in the polymerase chain reaction. These primers should prove valuable for nondestructive determination of Fom 2 gene introgression in breeding programs.

scholarly journals Polymerase Chain Reaction Method to Detect Canis Materials by Amplification of Species-Specific DNA Fragment

2004 ◽  
Vol 87 (5) ◽  
pp. 1195-1199 ◽  
Author(s):  
Hong-Wei Gao ◽  
Bao-Liang Xu ◽  
Cheng-Zhu Liang ◽  
Yi-Bing Zhang ◽  
Lai-Hua Zhu
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Method ◽  
Degraded Dna ◽  
Effective Control ◽  
Heat Denaturation ◽  
Spongiform Encephalopathy ◽  
Chain Reaction ◽  
Animal Blood ◽  
Dna Fragment ◽  
Polymerase Chain

Abstract Rapid identification of mammal materials in feeding stuffs and food is essential for effective control of a potential source of pathogens, such as those that cause bovine spongiform encephalopathy. A convenient polymerase chain reaction (PCR)-based assay was developed for detection and identification of a canis-specific mitochondrial DNA sequence in foodstuffs and food. The amplified canis-specific PCR product was a 213 base pair band from the D-loop DNA fragment of mitochondria, a high copy gene which should improve the possibility of amplifying template molecules of adequate size among the degraded DNA fragments brought about by heat denaturation. The specificity of this method was confirmed by 8 canis blood DNA samples (from different breeds of dog) and 9 noncanis animal blood DNA samples (bovine, sheep, porcine, chicken, fish, donkey, rabbit, deer, horse). This method was able to detect the presence of canis material in foodstuffs and in food mixtures even when the concentration of canis-derived meat was reduced to 0.05%. Furthermore, it did not appear to be affected by prolonged heat treatment. This method was developed for detection of canis materials in feeding stuffs, and occasionally for medical jurisprudence detection of canis-derived materials.

Utilizing a polymerase chain reaction method for the detection of Toxocara canis and T. cati eggs in soil

2007 ◽  
Vol 81 (1) ◽  
pp. 75-78 ◽  
Author(s):  
R. Fogt-Wyrwas ◽  
W. Jarosz ◽  
H. Mizgajska-Wiktor
Keyword(s):  
Polymerase Chain Reaction ◽  
Toxocara Canis ◽  
Polymerase Chain Reaction Method ◽  
Soil Samples ◽  
Chain Reaction ◽  
Egg Development ◽  
Specific Primers ◽  
Reaction Method ◽  
Large Numbers ◽  
Polymerase Chain

AbstractA polymerase chain reaction (PCR) technique has been used for the differentiation of T. canis and T. cati eggs isolated from soil and previously identified from microscopical observations. The method, using specific primers for the identification of the two Toxocara species, was assessed in both the field and laboratory. Successful results were obtained when only a single or large numbers of eggs were recovered from 40 g soil samples. The method is sensitive, allows analysis of material independent of the stage of egg development and can be adapted for the recovery of other species of parasites from soil.

scholarly journals Molecular identification of Pentatrichomonas hominis in animals in central and western Thailand

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Aongart Mahittikorn ◽  
Ruenruetai Udonsom ◽  
Khuanchai Koompapong ◽  
Rachatawan Chiabchalard ◽  
Chantira Sutthikornchai ◽  
...  
Keyword(s):  
Polymerase Chain Reaction Method ◽  
Small Subunit ◽  
Reservoir Host ◽  
Molecular Techniques ◽  
Nested Polymerase Chain Reaction ◽  
Faecal Samples ◽  
Zoonotic Parasite ◽  
Polymerase Chain ◽  
Pentatrichomonas Hominis ◽  
Rna Genes

Abstract Background Pentatrichomonas hominis inhabits the digestive tracts of several vertebrates, such as humans, monkeys, pigs, dogs, cats and rats. This protozoan was originally considered a commensal of the digestive tract but has subsequently been identified as a potential zoonotic parasite and a causative agent of diarrhoea. Molecular techniques are considered more sensitive and specific to detect P. hominis. This study aimed to determine the presence and genetic diversity of P. hominis in animals in Thailand. A total of 403 faecal samples were collected from 119 cats, 55 dogs, 73 goats, 35 monkeys, 55 cattle and 66 pigs, and the presence of P. hominis was determined using the nested polymerase chain reaction method. Sequence analysis of small-subunit ribosomal RNA genes was used to determine the genotype of the organism. Results Twenty-six samples (26/403, 6.45%) were positive for P. hominis. The highest prevalence was found in cats (21/119; 17.65%), followed by cattle (3/55; 5.45%) and dogs (2/55; 3.64%). Seven out of 26 nucleotides demonstrated 100% sequence identity with existing sequences; additionally, 16 novel sequence patterns were identified. All nucleotide sequences of P. hominis-positive samples were shown in the same branch with the previously described P. hominis sequences found in humans, dogs and goat. Conclusion This is the first study on P. hominis infections in animals in Thailand. Our findings revealed that the prevalence of P. hominis was significantly higher in cats than in cattle and dogs. Cats were the main reservoir host; however, P. hominis can infect several kinds of animals. Therefore, the proper waste management of animals is necessary to reduce and prevent infection in the community.

scholarly journals Karakterisasi Molekuler Papaya ringspot virus tipe P pada Tanaman Mentimun di Jawa

2018 ◽  
Vol 14 (3) ◽  
pp. 75
Author(s):  
Listihani Listihani ◽  
Tri Asmira Damayanti ◽  
Sri Hendrastuti Hidayat ◽  
Suryo Wiyono
Keyword(s):  
Ringspot Virus ◽  
Mosaic Symptom ◽  
Papaya Ringspot Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Specific Primers ◽  
Central Java ◽  
Dna Fragment ◽  
Polymerase Chain

Moleculer Characterization of Papaya ringspot virus type P on Cucumber in JavaInfection of Papaya ringspot virus (PRSV) on cucumber plants showing mosaic symptom was detected using specific antibody.  Further investigation was conducted to determine molecular characters and status of PRSV infecting cucumber in Java.  Infection of PRSV was detected from leaf samples collected from the field using dot immunobinding assay (DIBA).  Disease frequency caused by PRSV infection reached 81.11%, 95.86%, 91.66%, and 92.3% in East Java, Central Java, Yogyakarta, and West Java, respectively.  Characterization of PRSV isolates was conducted by reverse transcription polymerase chain reaction (RT-PCR) using specific primers for PRSV-P and PRSV-W, followed by cloning, and DNA sequencing.  DNA fragment of 470 bp was successfully amplified using specific primers for PRSV-P from several samples from Nganjuk, Brebes, Kulon Progo, and Subang; but no amplification was achieved using specific primers for PRSV-W.  Nucleotide and amino acid analysis showed high homology among PRSV-P isolates from Nganjuk, Brebes, Kulon Progo, and Subang, i.e. 98.6%-99.7% and 99.3%-100%, respectively.  This is an indication of a low genetic variation among PRSV-P from Java. Further phylogenetic analysis indicated that PRSV-P isolate cucumber is in the same cluster with PRSV-P isolate papaya from Bali, Indonesia.  This is the first report of PRSV-P infecting cucumber in Indonesia.

scholarly journals Occurrence of Lawsonia intracellularis in horses raised in three regions of the state of Paraná, Brazil

2021 ◽  
Vol 42 (5) ◽  
pp. 2867-2876
Author(s):  
Tatiane Caleffo ◽  
◽  
Vinicius Dahm ◽  
Jéssica Gonçalves dos Santos ◽  
Arthur Colombari Cheng ◽  
...  
Keyword(s):  
Real Time ◽  
Clinical Manifestations ◽  
Epidemiological Studies ◽  
The State ◽  
Serological Testing ◽  
Lawsonia Intracellularis ◽  
Chain Reaction ◽  
The West ◽  
Proliferative Enteropathy ◽  
Polymerase Chain

Lawsonia intracellularis is a bacterium already described in several species and most prevalent in pigs, in which it causes enteric problems. Horses can also be affected, developing a disease known as equine proliferative enteropathy, which results from the proliferation of intestinal crypt cells in response to infection by the bacterium. Despite the existence of reports of the disease in several countries, including Brazil, there are still no reports of the disease or epidemiological studies of its occurrence in symptomatic or asymptomatic horses in the state of Paraná. Thus, the present study was conducted to examine the occurrence of L. intracellularis in asymptomatic horses raised in the west, northwest and north regions of Paraná by means of serological testing and the real-time polymerase chain reaction (qPCR) technique. In the serological approach, the immunoperoxidase monolayer assay (IPMA) technique was employed. Feces were processed and subjected to qPCR. In total, samples were collected from 162 animals from 20 farms. Of these, 9/162 (5.55%) showed specific antibodies against L. intracellularis. Real-time PCR, on the other hand, identified 7/162 (4.32%) fecal samples positive for the presence of the bacterium. When the techniques were compared, none of the samples was positive by both, demonstrating that, for a better diagnosis, they must be performed together. In contrast to most reports in horses, the present study describes higher serological and molecular occurrence in animals older than two years. These results are of great epidemiological relevance, as they indicate that the bacterium is present in the sampled regions of the state of Paraná. Therefore, the disease must be included in the differential diagnosis of diseases with similar clinical manifestations.

scholarly journals Molecular selection of Beta vulgaris L. breeding materials with genes of resistance to abiotic stresses

2019 ◽  
Vol 1 (1) ◽  
pp. 16-20
Author(s):  
A. A. Nalbandyan ◽  
T. P. Fedulova ◽  
A. S. Hussein
Keyword(s):  
Sugar Beet ◽  
Molecular Genetic ◽  
Polymerase Chain Reaction Method ◽  
Root Knot Nematode ◽  
Chain Reaction ◽  
Specific Primers ◽  
Allele Specific ◽  
Molecular Selection ◽  
Polymerase Chain ◽  
Selection Of

In the work, the results of sugar beet breeding materials' molecular-genetic studying for presence of genes of resistance to root-knot nematode, rhizomania and powdery mildew are presented. Testing of plants was conducted using polymerase chain reaction method. The genes R6m-1, Rz1 and Rz2, Pm were identified with the help of 5 one-chain RAPD and 4 allele-specific primers. Aim of the studies is to screen sugar beet varieties for presence of the abovementioned genes of resistance. Domestic and foreign sugar beet hybrids were an object of the studies.

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