scholarly journals Prevalence of Echinococcus Species in Wild Foxes and Stray Dogs in Qinghai Province, China

2021 ◽  
Author(s):  
Huixia Cai ◽  
Jing Zhang ◽  
Xuefei Zhang ◽  
Yayi Guan ◽  
Xiao Ma ◽  
...  
Keyword(s):  
Alveolar Echinococcosis ◽  
Sequencing Analysis ◽  
Limited Data ◽  
Stray Dogs ◽  
Qinghai Province ◽  
Causal Pathogen ◽  
Polymerase Chain ◽  
Wild Dogs ◽  
Prevalent Species ◽  
Human Echinococcosis

Echinococcosis is a zoonotic parasitic disease that is highly endemic to the Qinghai province of China. Limited data are available on the prevalence of the causal pathogen, Echinococcus spp., in definitive hosts in this region. Thus, the aim of this study was to evaluate the prevalence of Echinococcus spp. in wild foxes and stray dogs in Qinghai province. Five hundred and twenty-eight feces from wild foxes and 277 from stray dogs were collected from 11 counties in the Golog, Yushu, and Haixi prefectures and screened for Echinococcus spp. using copro-DNA polymerase chain reaction (PCR). In total, 5.5% of wild foxes and 15.2% of stray dogs tested positive for Echinococcus spp. The prevalence rates of Echinococcus spp. in wild foxes in Golog, Yushu, and Haixi were 7.3%, 5.2%, and 1.9%, respectively. In stray dogs, these rates were 13.3%, 17.3%, and 0%, respectively. Sequencing analysis determined that Echinococcus multilocularis was the most prevalent species, occurring in 4.0% and 12.6% of wild foxes and stray dogs, respectively. Echinococcus shiquicus was observed in 1.5% of wild foxes and 0.7% of stray dogs. Echinococcus granulosus was observed only in wild dogs, with a prevalence rate of 1.8%. To our knowledge, this is the first report on the prevalence of E. shiquicus in dogs in Qinghai province. The current results improve our understanding of the transmission and dissemination of human echinococcosis and suggest that exposure to the eggs of E. multilocularis harbored by wild foxes and stray dogs may pose a great risk of alveolar echinococcosis to humans in Qinghai province.

scholarly journals Biotinylation of Deoxyguanosine at the Abasic Site in Double-Stranded Oligodeoxynucleotides

2016 ◽  
Vol 2016 ◽  
pp. 1-4
Author(s):  
Chun Wu
Keyword(s):  
Dna Polymerase ◽  
Click Reaction ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Sequencing Analysis ◽  
Abasic Site ◽  
Nylon Membrane ◽  
Abasic Sites ◽  
Polymerase Chain ◽  
Dna Sequencing Analysis ◽  
Endonuclease Digestion

Biotinylation of deoxyguanosine at an abasic site in double-stranded oligodeoxynucleotides was studied. The biotinylation of deoxyguanosine is achieved by copper-catalyzed click reaction after the conjugation of the oligodeoxynucleotide with 2-oxohex-5-ynal. The biotinylation enables visualization of the biotinylated oligodeoxynucleotides by chemiluminescence on a nylon membrane. In order to investigate the biotinylated site, the biotinylated oligodeoxynucleotides were amplified by the DNA polymerase chain reaction. Replacement of guanine opposing the abasic site with adenine generated by the activity of the terminal deoxynucleotidyl transferase of DNA polymerase was detected by DNA sequencing analysis and restriction endonuclease digestion. This study suggests that 2-oxohex-5-ynal may be useful for the detection of the unpaired deoxyguanosine endogenously generated at abasic sites in genomic DNA.

scholarly journals Prevalence and Molecular Epidemiological Data onDirofilaria immitisin Dogs from Northeastern States of India

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Sonjoy Kumar Borthakur ◽  
Dilip Kumar Deka ◽  
Saidul Islam ◽  
Dilip Kumar Sarma ◽  
Prabhat Chandra Sarmah
Keyword(s):  
Epidemiological Data ◽  
Elisa Test ◽  
Molecular Techniques ◽  
Diagnostic Methods ◽  
Molecular Tools ◽  
Stray Dogs ◽  
Concentration Technique ◽  
Working Dogs ◽  
Polymerase Chain

The aim of the present study was to determine the prevalence ofDirofilaria immitisin stray, pet, and working dogs (n=413, 266, and 103, resp.) from Guwahati (Assam) and Aizawl (Mizoram), areas located in two Northeastern States of India. Diagnostic methods applied were microscopy (wet film and Knott’s concentration technique), immunological test (Ag ELISA by SNAP 4Dx ELISA kit), and molecular tools (polymerase chain reaction and sequencing), which evidenced 11.38, 18.03, and 13.93% of positive animals, respectively. No significant differences were observed by area (18.23% versus 17.68%) nor by sex (18.1% versus 17.9%), whereas stray dogs proved more infected than other groups (P<0.05). ELISA test evidenced an overall 22.69% of occult infections, mainly in working dogs (60%), and molecular techniques detectedDirofilaria (Nochtiella) repensin 4 stray dogs from Guwahati. Characterization ofD. immitisisolates for ITS-2 region showed close identity with South Asian isolates.

Direct sequencing analysis of transmembrane region of human neu gene by polymerase chain reaction

2006 ◽  
Vol 3 (4) ◽  
pp. 198-201 ◽  
Author(s):  
Hideyuki Saya ◽  
Seiji Ara ◽  
Polly S. Y. Lee ◽  
Jungsil Ro ◽  
Mien-Chie Hung
Keyword(s):  
Polymerase Chain Reaction ◽  
Direct Sequencing ◽  
Sequencing Analysis ◽  
Transmembrane Region ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Transmission of Xylella fastidiosa, Causal Agent of Citrus Variegated Chlorosis, by the Glassy-Winged Sharpshooter, Homalodisca coagulata

Plant Disease ◽  
2006 ◽  
Vol 90 (5) ◽  
pp. 567-570 ◽  
Author(s):  
V. D. Damsteegt ◽  
R. H. Brlansky ◽  
P. A. Phillips ◽  
Avijit Roy
Keyword(s):  
Xylella Fastidiosa ◽  
The United States ◽  
Citrus Variegated Chlorosis ◽  
Homalodisca Coagulata ◽  
Causal Pathogen ◽  
Polymerase Chain ◽  
Species Specific ◽  
Transmission Studies ◽  
Test Plants ◽  
Important Disease

Citrus variegated chlorosis (CVC) is an economically important disease of citrus in Brazil and Argentina. The causal pathogen is a strain of Xylella fastidiosa transmitted by several sharpshooter species. The glassy-winged sharpshooter (GWSS), Homalodisca coagulata, has become an important new pest of citrus and grapevines in California, where it transmits X. fastidiosa strains to several crops including grapes, oleander, and almonds. Transmission studies over a 3-year period at the USDA BSL3-P containment facility at Fort Detrick, MD, utilizing California field-collected GWSS, a Brazilian strain of CVC, and Madam Vinous sweet orange seedlings, have shown a consistent although low level of transmission of CVC. Test plants were observed for CVC symptoms, analyzed by polymerase chain reaction using species-specific primers for X. fastidiosa, membrane entrapment immunofluorescence, and scanning electron microscopy. X. fastidiosa was not detected in field-collected GWSS but was detected in GWSS following feeding on CVC-infected source plants. Transmission of the CVC strain of X. fastidiosa by GWSS increases the risk of establishment of CVC in the United States if it were introduced.

Molecular Detection of Leptospira Species Serotypes in Iranian Stray Dogs

2019 ◽  
Author(s):  
Saam Torkan ◽  
Hassan Momtaz
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Serum Samples ◽  
Blood Samples ◽  
Stray Dogs ◽  
Wide Range ◽  
Leptospira Spp ◽  
Polymerase Chain ◽  
Leptospira Species

Background and Aims: Leptospirosis is a spirochetal disease with public health importance globally. This disease affects a wide range of domestic and wild animals. Dogs are one of the species most sensitive to Leptospira canicola and Leptospira icterohaemorrhagiae. The present study was concluded to evaluate the prevalence rate of Leptospira species and L. canicola and L. icterohaemorrhagiae serovars in Iranian stray dogs. Materials and Methods: One-hundred and twenty blood samples were first taken from stray dogs. Then the samples were transferred to the laboratory. Sera were extracted from blood samples and genomic DNA was extracted. DNA samples were subjected to conventional polymerase chain reaction. Positive samples for Leptospira spp. were analyzed for presence of L. canicola and L. icterohaemorrhagiaeserovars using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: Nine samples out of 120 serum samples (7.5%) were positive for the flagella gene of the Leptospira spp. Prevalence of Leptospira spp. in serum samples of male and female dogs were 5.4% and 10.86%, respectively. Prevalence of L. canicola and L. icterohaemorrhagiae serovars were 55.55% and 33.33%, respectively. We found that 11.11% of samples were positive for both serovars. Two to three and 3-4 year old dogs had the highest prevalence of Leptospira spp. Conclusions: The considerable prevalence of leptospirta spp. and also their zoonotic serovars among Iranian stray dogs represented an important public health issue regarding the contact of healthy human with these dogs. Identification of infected dogs and their vaccination can inhibit the distribution of Leptospira spp.

scholarly journals The Zoonotic Risk ofAncylostoma ceylanicumIsolated from Stray Dogs and Cats in Guangzhou, South China

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Yuanjia Liu ◽  
Guochao Zheng ◽  
Muhamd Alsarakibi ◽  
Xinheng Zhang ◽  
Wei Hu ◽  
...  
Keyword(s):  
South China ◽  
Phylogenetic Trees ◽  
Sequence Similarity ◽  
Zoonotic Transmission ◽  
Health Concern ◽  
Limited Data ◽  
Stray Dogs ◽  
Significant Public Health Concern ◽  
Zoonotic Risk ◽  
Significant Public Health

Canine and feline hookworm infection is endemic in many countries with zoonotic transmission representing a potentially significant public health concern. However, there is limited data available on the zoonotic transmission of canine and feline hookworms in China. This study was conducted to evaluate the zoonotic risk ofAncylostoma ceylanicumisolated from stray dogs and cats in Guangzhou, south China. Primer pairs CAF/CAR were designed to amplify complete ITS sequences of obtainedA. ceylanicum. The results were compared with fourteen ITS reference sequences of human-derivedA. ceylanicumregistered in GenBank, and phylogenetic trees were established by using NJ and ML methods. The sequence similarity of three dog-derived and five cat-derivedA. ceylanicumwith fourteen human-derivedA. ceylanicumwere 96.8%~100% and 97.8%~100%, respectively. Phylogenetic analysis placedA. ceylanicumisolated from dogs and cats in the same group withA. ceylanicumhuman isolates. Due to the ability ofA. ceylanicumto cause a patent infection in humans, the zoonotic risk arising from dog and cat reservoirs to communities in this region should be determined.

scholarly journals Cell-based culture of SARS-CoV-2 informs infectivity and safe de-isolation assessments during COVID-19

2020 ◽  
Author(s):  
Kerri Basile ◽  
Kenneth McPhie ◽  
Ian Carter ◽  
Susan Alderson ◽  
Hossinur Rahman ◽  
...  
Keyword(s):  
Clinical Sample ◽  
Surrogate Marker ◽  
Upper Respiratory Tract ◽  
Limited Data ◽  
Icu Patients ◽  
Culture Positivity ◽  
The Mean ◽  
Upper Respiratory ◽  
Polymerase Chain ◽  
Virus Replication Cycle

Background The detection of SARS-CoV-2 by real-time polymerase chain reaction (PCR) in respiratory samples collected from persons recovered from COVID-19 does not necessarily indicate shedding of infective virions. By contrast, the isolation of SARS-CoV-2 using cell-based culture likely indicates infectivity, but there are limited data on the correlation between SARS-CoV-2 culture and PCR. Here we review our experience using SARS-CoV-2 culture to determine infectivity and safe de-isolation of COVID-19 patients. Methods 195 patients with diverse severity of COVID-19 were tested (outpatients [n=178]), inpatients [n=12] and ICU [n=5]). SARS-CoV-2 PCR positive samples were cultured in Vero C1008 cells and inspected daily for cytopathic effect (CPE). SARS-CoV-2-induced CPE was confirmed by PCR of culture supernatant. Where no CPE was documented, PCR was performed on day four to confirm absence of virus replication. Cycle threshold (Ct) values of the day four PCR (Ctculture) and the PCR of the original clinical sample (Ctsample) were compared, and positive cultures were defined as a Ctsample - Ctculture value of greater than or equal to 3. Findings Of 234 samples collected, 228 (97%) were from the upper respiratory tract. SARS-CoV-2 was only successfully isolated from samples with Ctsample values <32, including in 28/181 (15%), 19/42 (45%) and 9/11 samples (82%) collected from outpatients, inpatients and ICU patients, respectively. The mean duration from symptom onset to culture positivity was 4.5 days (range 0-18 days). SARS-CoV-2 was significantly more likely to be isolated from samples collected from inpatients (p<0.001) and ICU patients (p<0001) compared with outpatients, and in samples with lower Ctsample values. Conclusion SARS-CoV-2 culture may be used as a surrogate marker for infectivity and inform de-isolation protocols.

scholarly journals RANCANGAN PRIMER SPESIFIK GEN MACROPHAGE MANNOSE RECEPTOR (MMR) UNTUK POLYMERASE CHAIN REACTION (PCR) DAN SEKUENSING DEOXYRIBO NUCLEIC ACID (DNA)

2018 ◽  
Vol 22 (2) ◽  
pp. 158
Author(s):  
Yani Triyani ◽  
Nurizzatun Nafsi ◽  
Lelly Yuniarti ◽  
Nanan Sekarwana ◽  
Endang Sutedja ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequencing ◽  
Mannose Receptor ◽  
Primer Design ◽  
Sequencing Analysis ◽  
Nucleotide Polymorphisms ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Polymerase Chain ◽  
Dna Sequencing Analysis

The order (sequencing) determinationof DeoxyribonucleicAcid (DNA) bases is the gene’s most basic information, using the methodof Polymerase Chain Reaction (PCR) as its stage. A key factor of successful detection by PCR is specific PCR primer design choice. Thedetection of diversity of Mycobacterium Mannose Receptor (MMR) gene, responsible for recognizing mannosylated antigen structureof Mycobacterium tuberculosis (M.tb) by DNA sequencing of exon 7 chromosome 10p12, related to susceptiblity for PulmonaryTuberculosis(TB), was first performed in China in 2012. The purpose of this study was to find specific primerfromboth design originatedfrom the research in China/primer I and my own design/primer IIby using Primer3 software. This study was based on 10 healthy subjectsand was a preliminary study of a research titled. The Relationship of Single Nucleotide Polymorphisms (SNPs) of Macrophage MannoseReceptor Gene to Pulmonary Tuberculosis Cases. The examination materials consist of 3 mL of EDTA blood and DNA extraction from itsbuffy coat. The resulting DNA was processed by PCR to amplify MMR gene with primer I and II. The primer I successfully amplified DNAfragments up to 780bp while primer II only 329 bp. The MMR gene DNA sequencing analysis was performed on the amplification resultof both kinds primers by using DNA Baser and Ensembl−BLAST software. The results were different, DNA sequencing result by using theprimer I was found in several chromosomes and also in several loci. Whereas, by using the primer II, it was only found in chromosome10 and in the same locus. Based on this study, it can be concluded that the specific primer design is one of the most important factorsin the success of DNA sequencing.

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