• Title/Summary/Keyword: fecal DNA

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Prevalence and Phylogenetic Analyses of Trichuris suis in Pigs in Hunan Province, Subtropical China

  • Tan, Lei;Wang, Aibing;Yi, Jing;Liu, Yisong;Li, Jiayu;Liu, Wei
    • Parasites, Hosts and Diseases
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    • v.56 no.5
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    • pp.495-500
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    • 2018
  • Trichuris suis infection in pigs is ubiquitous in intensive and extensive farms, which causes potential threat to human health. The objective of this research was to investigate the prevalence of T. suis in pigs in Hunan province. Total 2,267 fresh fecal samples distributed in 28 pig farms from 7 different administrative regions (Hunan province) were evaluated for the existence of T. suis eggs using saturated NaCl floating method. The average infection rate of T. suis in pigs was 8.91% in Hunan province. To determine genetic variation of the gained T. suis isolates in the present study, the internal transcribed spacer (ITS) regions from nuclear ribosomal DNA (rDNA) of 7 T. suis isolates were cloned and analyzed. Nucleotide diversities were 1.0-3.5% and 0-3.8% for ITS-1 and ITS-2, respectively. Phylogenetic analyses indicated that all isolates collected in the present study and T. suis available in Genbank generated a monophyletic clade. The present investigation revealed high infection rates of T. suis in pigs in Hunan province, which shed light on making effective measures to prevent and control T. suis infection in pigs in Hunan province.

Effects of feed intake on the diversity and population density of homoacetogens in the large intestine of pigs

  • Matsui, Hiroki;Mimura, Ayumi;Maekawa, Sakiko;Ban-Tokuda, Tomomi
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.12
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    • pp.1907-1913
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    • 2019
  • Objective: Homoacetogens play important roles in the production of acetate in the large intestine of monogastric mammals. However, their diversity in the porcine large intestine is still unknown. Marker gene analysis was performed to assess the effects of energy level on the diversity and population densities of homoacetogens in porcine feces. Methods: Crossbred pigs were fed high or low energy-level diets. The high-intake (HI) diet was sufficient to allow a daily gain of 1.2 kg. The low-intake (LI) diet provided 0.6 times the amount of energy as the HI diet. Genetic diversity was analyzed using formyltetrahydrofolate synthetase gene (FHS) clone libraries derived from fecal DNA samples. FHS DNA copy numbers were quantified using real-time polymerase chain reaction. Results: A wide variety of FHS sequences was recovered from animals in both treatments. No differences in FHS clone libraries between the HI and LI groups were found. During the experimental period, no significant differences in the proportion of FHS copy numbers were observed between the two treatment groups. Conclusion: This is the first reported molecular diversity analysis using specific homoacetogen marker genes from the large intestines of pigs. There was no observable effect of feed intake on acetogen diversity.

Development of oligonucleotide microarray system for differential diagnosis of enteric viruses in diarrheic fecal samples in pigs

  • Park, Nam-Yong;Kim, Yong-Hwan;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.489-496
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    • 2007
  • An oligonucleotide microarray system was developed for the simultaneous detection of porcine epidemic diarrhea virus, transmissible gastroenteritis virus, porcine enteric calicivirus, porcine group A and C rotavirus. RNAs of the reference viruses and porcine diarrhea samples were extracted and amplified using one-step multiplex RT-PCR in the presence of cyanine 5-dCTP and hybridized on the microarray chip that spotted the virus-specific oligonucleotides. This system were approximately 10-to 100-fold higher in sensitivity than conventional RT-PCR, and the assay time was less than 3 hours. The relative sensitivity and specificity were 92% and 72.2%, respectively, based on 102 porcine diarrhea samples using RT-PCR as gold standard. These results suggested that the oligonucleotide microarray system in this study be probably more reliable and reproducible means for detecting porcine enteric viruses and that it could be of substantial use in routine diagnostic laboratories.

Axenic cultivation and characterization of Giardia lamblia isolated from humans in Korea

  • Park, Soon-Jung;Yong, Tai-Soon;Yang, Hye-Won;Lee, Du-Ho;Lee, Kyung-Won
    • Parasites, Hosts and Diseases
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    • v.37 no.2
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    • pp.121-125
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    • 1999
  • Inoculation of human fecal cysts to suckling Mongolian gerbils, two Giardia lamblia isolates, Kl and K2, were established as axenic cultures. Using this in vitro culture, both two Giardia isolates were grouped by using two genetic analysis. With genetic analysis of SSS-rDNA sequences, both K1 and K2 were found as members of hopkins'group 1, despite some nucleotide differences noticed in K1 (5 differences/292 bases.). The other genetic study used PCI-RFLP of the tim (triose phosphate isomerase) Nash's group 2 can bot be a separate group, but a part of Hopkins' group 1.

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Molecular Identification of Oesophagostomum and Trichuris Eggs Isolated from Wild Japanese Macaques

  • Arizono, Naoki;Yamada, Minoru;Tegoshi, Tatsuya;Onishi, Kotaro
    • Parasites, Hosts and Diseases
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    • v.50 no.3
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    • pp.253-257
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    • 2012
  • Natural habitat fragmentation and reducing habitat quality have resulted in an increased appearance of Japanese macaques, Macaca fuscata (Gray, 1870), in suburban areas in Japan. To investigate the risk of zoonotic infections, a coprological survey of helminth eggs passed by wild Japanese macaques was carried out in 2009 and 2010 in Shiga Prefecture, Japan. Microscopic examination found helminth eggs in high prevalence, and nucleotide sequencing of DNA extracted from the eggs identified Oesophagostomum cf. aculeatum and Trichuris trichiura. A fecal culture also detected infective larvae of Strongyloides fuelleborni. These zoonotic nematodes pose a potential health issue to local people in areas frequented by Japanese macaques.

Isolation and Characterization of Two Rare Mucoralean Species with Specific Habitats

  • Lee, Seo Hee;Nguyen, Thuong T.T.;Lee, Hyang Burm
    • Mycobiology
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    • v.46 no.3
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    • pp.205-214
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    • 2018
  • The order Mucorales, the largest in number of species within the Mucoromycotina, comprises typically fast-growing saprotrophic fungi. During a study of the fungal diversity of undiscovered taxa in Korea, two novel mucoralean strains, CNUFC-GWD3-9 and CNUFC-EGF1-4, were isolated from specific habitats including freshwater and fecal samples, respectively. On the basis of their morphological characteristics and sequence analyses of internal transcribed spacer and large subunit ribosomal DNA, the CNUFC-GWD3-9 and CNUFC-EGF1-4 isolates were confirmed to be Gilbertella persicaria and Pilobolus crystallinus, respectively. It is ecologically, pathologically, and mycologically significant to find such rare zygomycetous fungi in such specific habitats.

Isolation and Characterization of Three Unrecorded Zygomycete Fungi in Korea: Cunninghamella bertholletiae, Cunninghamella echinulata, and Cunninghamella elegans

  • Nguyen, Thuong T.T.;Choi, Young-Joon;Lee, Hyang Burm
    • Mycobiology
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    • v.45 no.4
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    • pp.318-326
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    • 2017
  • In a survey of undiscovered taxa in Korea, three zygomycete fungal strains-EML-W31, EML-HGD1-1, and EML-RUS1-1-were isolated from freshwater, grasshopper fecal, and soil samples in Korea. On the basis of the morphological characteristics and phylogenetic analysis of internal transcribed spacer and 28S rDNA, the isolates of EML-W31, EML-HGD1-1, and EML-RUS1-1 were confirmed to be Cunninghamella bertholletiae, Cunninghamella echinulata, and Cunninghamella elegans, respectively. These species have not been previously described in Korea.

Biomarker-directed Targeted Therapy in Colorectal Cancer

  • John M. Carethers
    • Journal of Digestive Cancer Research
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    • v.3 no.1
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    • pp.5-10
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    • 2015
  • With advances in the understanding of the biology and genetics of colorectal cancer (CRC), diagnostic biomarkers that may predict the existence or future presence of cancer or a hereditary condition, and prognostic and treatment biomarkers that may direct the approach to therapy have been developed. Biomarkers can be ascertained and assayed from any tissue that may demonstrate the diagnostic or prognostic value, including from blood cells, epithelial cells via buccal swab, fresh or archival cancer tissue, as well as from cells shed into fecal material. For CRC, current examples of biomarkers for screening and surveillance include germline testing for suspected hereditary CRC syndromes, and stool DNA tests for screening average at-risk patients. Molecular biomarkers for CRC that may alter patient care and treatment include the presence or absence of microsatellite instability, the presence or absence of mutant KRAS, BRAF or PIK3CA, and the level of expression of 15-PGDH in the colorectal mucosa. Molecularly targeted therapies and some general therapeutic approaches rely on biomarker information. Additional novel biomarkers are on the horizon that will undoubtedly further the approach to precision or individualized medicine.

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Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces

  • Wu, Yao-Dong;Wang, Qi-Qi;Wang, Meng;Elsheikha, Hany M.;Yang, Xin;Hu, Min;Zhu, Xing-Quan;Xu, Min-Jun
    • Parasites, Hosts and Diseases
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    • v.59 no.2
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    • pp.167-171
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    • 2021
  • Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation.

Development of a Rapid Method for the Screening of Conjugated Linoleic Acid (CLA)-Producing Strains of Bifidobacterium breve

  • Choi, Sun-Hae;Lee, Kyoung-Min;Kim, Kwan-Hu;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.38 no.4
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    • pp.806-815
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    • 2018
  • This study was performed to isolate some strains of Bifidobacterium breve from fecal materials of neonates and to screen them for the biotransformation activity of converting linoleic acid into conjugated linoleic acid (CLA). Fecal samples were collected from twenty healthy neonates between 14 and 100 days old, and four hundred colonies were randomly selected from a Bifidobacterium selective transoligosaccharide medium. A duplex polymerase chain reaction technique was developed for the rapid and accurate molecular characterization of the B. breve strains that have been reported to show the species-specific characteristic of CLA production. They are identified by 16S ribosomal DNA, fructose-6-phosphate phosphoketolase encoding genes (xfp), and rapid pulsed field gel electrophoresis. Thirty-six isolates were identified as B. breve, and just two of the 12 neonates were harboring B. breve strains. Each isolate showed different CLA-producing ability in the spectrophotometric assay. All of the positive strains from the primary spectrophotometric assay were confirmed for their CLA-producing activities using gas-chromatographic analysis, and their conversion rates were different, depending on the strain isolated in this study. Some strains of B. breve were successfully isolated and characterized based on the CLA-producing activity, and further studies are necessary to characterize the enzyme and the gene responsible for the enzyme activity.