scholarly journals Vertical transmissibility of small ruminant lentivirus

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0239916
Author(s):  
Juscilânia Furtado Araújo ◽  
Alice Andrioli ◽  
Raymundo Rizaldo Pinheiro ◽  
Lucia Helena Sider ◽  
Ana Lídia Madeira de Sousa ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Diagnostic Tests ◽  
Small Ruminant ◽  
Serological Test ◽  
Blood Collection ◽  
Viral Dna ◽  
Viral Isolation ◽  
Source Of Infection ◽  
Pcr Products ◽  
Polymerase Chain

This study aimed to evaluate by means of Nested Polymerase Chain Reaction (nPCR), co-cultivation and sequencing, with genetic comparison between strains (mother/newborn), the occurrence of vertical transmission of Small Ruminant Lentiviruses (SRLV) from naturally occurring nannies infected for their offspring. For the detection of SRLV seropositive progenitors, blood was collected from 42 nannies in the final third of gestation in tubes with and without anticoagulant. The diagnostic tests used were Western Blot (WB) and nPCR. During the period of birth, the same blood collection procedure was performed on 73 newborns at zero hours of birth, with the same diagnostic tests. Seventeen blood samples from seven-day-old kids, proven positive for SRLV by nPCR, chosen at random, were subjected to coculture in goat synovial membrane (GSM) cells for 105 days. The pro-viral DNA extracted from the cell supernatant from the coculture was subjected to nPCR. For DNA sequencing from the nPCR products, nine positive samples were chosen at random, four nannies with their respective offspring, also positive. Each sample was performed in triplicate, thus generating 27 nPCR products of which only 19 were suitable for analysis. Among the 42 pregnant goats, in 50% (21/42) pro-viral DNA was detected by nPCR, while in the WB, only 7.14% (3/42) presented antibodies against SRLV. Regarding neonates, of the 73 kids, 34 (46.57%) were positive for the virus, using the nPCR technique, while in the serological test (WB), three positive animals (4.10%) were observed. The coculture of the 17 samples with a positive result in the nPCR was confirmed in viral isolation by amplification of the SRLV pro-viral DNA. When aligned, the pro-viral DNA sequences (nannies and their respective offspring) presented homology in relation to the standard strain CAEV Co. It was concluded that the transmission of SRLV through intrauterine route was potentially the source of infection in the newborn goats.

Molecular biology can differentiate morphologically indistinguishable myxosporean species: Myxobolus elegans and M. hungaricus (Short communication)

2002 ◽  
Vol 50 (1) ◽  
pp. 59-62 ◽  
Author(s):  
Edit Eszterbauer
Keyword(s):  
Ribosomal Rna ◽  
Dna Sequences ◽  
Valid Species ◽  
Base Pairs ◽  
The Family ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Biological Methods ◽  
Myxosporean Species

Two, morphologically indistinguishable myxosporean species, Myxobolus elegans Kashkovsky, 1966 and M. hungaricus Jaczó, 1940 were differentiated using molecular biological methods. Polymerase chain reaction (PCR) with primers specific for the family Myxobolidae was used to amplify an approximately 1600 base pairs (bp) long fragment of the 18S ribosomal RNA gene. In restriction fragment length polymorphism (RFLP) study with HinfI, MspI and TaqI enzymes, the two parasite species were easily distinguishable. The genetic distinctness was also confirmed by the DNA sequence of their PCR products. Although M. elegans and M. hungaricus are morphologically very similar, based on the results of the PCR-RFLP and the DNA sequences, we concluded that they are valid species.

Spatio-temporal variations and age effect on Toxoplasma gondii seroprevalence in seals from the Canadian Arctic

Parasitology ◽  
2011 ◽  
Vol 138 (11) ◽  
pp. 1362-1368 ◽  
Author(s):  
A. SIMON ◽  
M. CHAMBELLANT ◽  
B. J. WARD ◽  
M. SIMARD ◽  
J. F. PROULX ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Serological Test ◽  
Canadian Arctic ◽  
Temporal Variations ◽  
Test Results ◽  
Source Of Infection ◽  
Food Borne ◽  
Significant Public Health ◽  
Polymerase Chain ◽  
Spatio Temporal

SUMMARYToxoplasmosis is a significant public health threat for Inuit in the Canadian Arctic. This study aimed to investigate arctic seals as a possible food-borne source of infection. Blood samples collected from 828 seals in 7 Canadian Arctic communities from 1999 to 2006 were tested for Toxoplasma gondii antibodies using a direct agglutination test. Polymerase chain reaction (PCR) was used to detect T. gondii DNA in tissues of a subsample of seals. Associations between seal age, sex, species, diet, community and year of capture, and serological test results were investigated by logistic regression. Overall seroprevalence was 10·4% (86/828). All tissues tested were negative by PCR. In ringed seals, seroprevalence was significantly higher in juveniles than in adults (odds ratio=2·44). Overall, seroprevalence varied amongst communities (P=0·0119) and by capture year (P=0·0001). Our study supports the hypothesis that consumption of raw seal meat is a significant source of infection for Inuit. This work raises many questions about the mechanism of transfer of this terrestrial parasite to the marine environment, the preponderance of infection in younger animals and the natural course of infection in seals. Further studies to address these questions are essential to fully understand the health risks for Inuit communities.

scholarly journals Sequence analysis of the cytochrome c oxidase subunit 1 gene of Sarcoptes scabiei isolated from goats and rabbits in East Java, Indonesia

2019 ◽  
Vol 12 (7) ◽  
pp. 959-964
Author(s):  
Nunuk Dyah Retno Lastuti ◽  
Ali Rohman ◽  
Didik Handiyatno ◽  
Dony Chrismanto ◽  
Kurnia Desiandura
Keyword(s):  
Cytochrome C ◽  
Cytochrome C Oxidase ◽  
Dna Sequences ◽  
Educational Software ◽  
Sarcoptes Scabiei ◽  
Pcr Products ◽  
Reverse Primer ◽  
Polymerase Chain ◽  
Cox 1 ◽  
Forward Primer

Aim: This study aimed to sequence the Cytochrome c oxidase (COX-1) gene sequence from mitochondrial DNA of Sarcoptes scabiei isolated from Lamongan goats and Mojokerto rabbits, align it with DNA isolated from Zi'gong rabbit (GenBank accession No. EU256389.1), and produce a phylogenetic analysis of S. scabiei COX-1 gene. Materials and Methods: S. scabiei mites were obtained from goats and rabbits, and DNA was extracted using QIAamp DNA Mini Kit. The forward and reverse primer sequences were designed based on the DNA sequence of an S. scabiei COX-1 gene isolated from the Zi'gong rabbit (5'-TCTTAGGGGCTGGATTTAGTATG-3' and 5'-AGTTCCTCTACCAGTTCCAC-3', respectively). To confirm sequencing output, the sequence resulting from the reverse primer was inverted and aligned to the sequence from the forward primer using Clone Manager Professional Version 9 for Windows (Scientific & Educational Software; http://www.scied.com). This alignment was subsequently used to build a phylogenetic tree, using the Neighbor- Joining method, in the MEGA6 program (https://www.megasoftware.net/). Results: Polymerase chain reaction (PCR) products from S. scabiei isolates from Lamongan goats and Mojokerto rabbits produced bands of around 290 bp with 2% agarose gel electrophoresis. Comparing the DNA sequences of the S. scabiei COX-1 gene with those isolated from Lamongan goats and Mojokerto rabbits showed 99% homology. Conclusion: PCR products of the S. scabiei COX-1 gene isolated from Lamongan goats and Mojokerto rabbits were around 290 bp long. The sequences had more than 99% homology. The sequences of the COX-1 gene of S. scabiei from Lamongan goats and Mojokerto rabbits were relatively close to the sequence of the gene in S. scabiei obtained from various hosts according to National Center for Biotechnology Information data.

scholarly journals DETERMINATION OF THE INFECTIVE STRAIN OF HYDATID CYST IN IRAQI CATTLE BY USING CO1 GENE

2017 ◽  
Vol 48 (2) ◽  
Author(s):  
M. J. Muhaidi
Keyword(s):  
Hydatid Cyst ◽  
Gene Bank ◽  
Hydatid Cysts ◽  
Chain Reaction ◽  
Source Of Infection ◽  
Pcr Products ◽  
Nucleotide Database ◽  
Polymerase Chain ◽  
Sheep Strain

Hydatid Cysts were obtained  from liver, lungs, spleen, heart, and peritoneal cavity of 15 cows, from different Iraqi regions  between December 2014 and October 2015. Hydatid cysts (protoscoleces) were used for mitochondrial DNA extraction by using mechanical grinder, and the purification of mtDNA was done by (promega kit, USA). "The mitochondrial cytochrome c oxidase subunit 1 (CO1) gene" was used as target for "polymerase chain reaction (PCR)" which successfully amplified the targeted this gene with 450 bp. The PCR products were purified and partial sequences were determine. The obtained sequences were aligned with the corresponding region of co1 gene in the Gene Bank nucleotide database to confirm the infection with hydatid cyst sheep strain (G1) in Iraq. The amplified CO1 targeted region was analyzed to obtain the  phylogenetic tree. G1 genotype was the most common strain and the actual source of infection of  Iraqi's  cattle. All of 15 samples were G1 strain (sheep strain) according to  the partial sequences of (CO1) genes.

scholarly journals A Begomovirus Isolated from Chlorotic and Stunted Soybean Plants in Mexico is a New Strain of Rhynchosia golden mosaic virus

Plant Disease ◽  
2006 ◽  
Vol 90 (7) ◽  
pp. 972-972 ◽  
Author(s):  
J. Méndez-Lozano ◽  
L. L. Perea-Araujo ◽  
R. D. Ruelas-Ayala ◽  
N. E. Leyva-López ◽  
J. A. Mauricio-Castillo ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Dna Sequences ◽  
Sequence Similarity ◽  
Pcr Amplification ◽  
Nucleotide Identity ◽  
Degenerate Primers ◽  
Viral Dna ◽  
Field Samples ◽  
Pcr Products ◽  
Soybean Plants

Soybean (Glycine max Merr.) is an alternative crop during the summer in Sinaloa, a northern state of Mexico. During the last 4 years, symptoms of yellowing, curled leaves, and stunting have been observed on soybean plantings, and a scrutiny of field samples collected in 2003 identified a begomovirus related to Pepper golden mosaic virus in symptomatic plants (4). A new survey was conducted during the summer of 2004 when the soybean disease was prevalent in the region. Affected plants appeared as patches displaying symptoms ranging from mild to severe yellow mosaic with leaf deformation and stunted growth in several parcels of commercial fields of northern Sinaloa. More than 100 samples from symptomatic soybean plants and weeds growing within the same fields were collected and analyzed for the presence of begomoviruses using DNA hybridization with the coat protein gene of Pepper huasteco yellow vein virus as a probe. Thirty-eight soybean, 12 Rhynchosia sp., and 14 sunflower hybridization-positive samples were subsequently used for polymerase chain reaction (PCR) amplification with the degenerate primers pRep-DGR and pCP70-Mot (1). PCR products were cloned into pGEM-T Easy vector (Promega, Madison, WI) and sequenced. The amplified viral DNA (915 nt) from two soybean plants, Sb1 and Sb2 (GenBank Accession Nos. AY955101 and AY957561, respectively), one isolate from Rhynchosia minima (GenBank Accession No. AY955102), and one from Heliantus annum (GenBank Accession No. AY957560) were sequenced and compared with DNA sequences available at NCBI database using BLAST. The highest sequence similarity was obtained with the two known isolates of Rhynchosia golden mosaic virus, RhGMV [Honduras] (GenBank Accession No. AF239671), and RhGMV [Chiapas] (GenBank Accession No. AF408199), displaying a nucleotide identity of approximately 89% with the Sinaloa isolates. Sequence comparisons of the latter isolates showed that viruses in the weeds were 97% identical to one of the soybean isolates, RhGMV-Sb1, but differed significantly (88% of nucleotide identity) from the second soybean isolate, RhGMV-Sb2. The complete genome A sequence of RhGMV-Sb1 was determined using PCR amplification of viral DNA with four degenerate primers recently described (2), cloning of overlapping PCR products into pGEM-T Easy vector (Promega) and sequencing. The 2,604-bp DNA-A of RhGMV-Sb1 (GenBank Accession No. DQ347950) was compared with the homologous genome of RhGMV [Chiapas] and RhGMV [Honduras] using the CLUSTAL alignment method (MegAlign, DNASTAR software, London) and an overall nucleotide identity of 89.2 and 88.6%, respectively, was determined. Current taxonomic criteria for begomoviruses establish that a DNA-A sequence identity lower than 93% with other isolates of a virus is indicative of a separate strain (3). Therefore, the virus identified in this study is a new strain of RhGMV that is provisionally named Rhynchosia golden mosaic virus-Soybean [Mexico:Sinaloa:2004]. This is the first soybean-infecting begomovirus from the American continent whose genome A has been completely characterized as of today. References: (1) J. T. Ascencio-Ibañez et al. Plant Dis. 86:692, 2002. (2) R. De La Torre-Almaraz et al. Plant Dis. 90:378, 2006. (3) C. Fauquet et al. Arch. Virol. 150:2151, 2005. (4) J. Mendez-Lozano et al. Plant Dis. 90:109, 2006.

scholarly journals Detection of Canine Parvovirus in Baghdad city by PCR technique

2012 ◽  
Vol 36 (0E) ◽  
pp. 95-98
Author(s):  
Ahmed F. Ahmed
Keyword(s):  
Gel Electrophoresis ◽  
Canine Parvovirus ◽  
Chain Reaction ◽  
Viral Dna ◽  
Fatal Disease ◽  
The Third ◽  
Mutant Strains ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Hemorrhagic Enteritis

Canine parvovirus 2 (CPV2) is a highly contagious and fatal disease of dogs, causingacute hemorrhagic enteritis and myocarditis. In this study different mutant strains of the viruswere characterized by polymerase chain reaction (PCR).The fecal samples from infected dogssuspected for CPV2 infection were collected in a suitable medium. The viral DNA from fecalsamples was extracted using specific kits, PCR were carried out with five different primer,pCPV-2ab and pCPV-2b, to distinguish the strain prevalent in field condition. The primerpCPV-2ab recognized both variant CPV-2a and CPV-2b, whereas the primer pCPV-2brecognized only the variant CPV-2b, using the third primer pCPV to recognize the residualbase pair, enabling the differentiation of CPV-2a variant from CPV-2b in field isolates. Thedifferent PCR products were further analyzed by using gel electrophoresis.

A Study of the Efficiency of Modern Domestic Disinfectants in the System of TB Control Activities

2015 ◽  
Vol 2 (2) ◽  
pp. 26-31 ◽  
Author(s):  
A. Paliy ◽  
A. Zavgorodniy ◽  
B. Stegniy ◽  
A. Gerilovych
Keyword(s):  
Molecular Genetic ◽  
Critical Role ◽  
Bactericidal Effect ◽  
Chain Reaction ◽  
Tb Control ◽  
Source Of Infection ◽  
Polymerase Chain ◽  
And Control ◽  
Chemical Groups ◽  
Test Objects

Due to the absence of elaborated effi cient means for specifi c prevention of bovine tuberculosis, it is ex- tremely important to detect and eliminate the source of infection and to take veterinary and sanitary preven- tive measures. Here the critical role is attributed to disinfection, which breaks the epizootic chain due to the elimination of pathogenic microorganisms in the environment and involves the application of disinfectants of different chemical groups. Aim. To study the tuberculocidal properties of new disinfectants DZPT-2 and FAG against atypical mycobacteria Mycobacterium fortitum and a TB agent Mycobacterium bovis. Methods. The bacteriological and molecular-genetic methods were used. Results. It was determined that DZPT-2 prepara- tion has bactericidal effect on M. fortuitum when used in the concentration of 2.0 % of the active ingredient (AI) when exposed for 5–24 h, while disinfectant FAG has a bactericidal effect in the concentration of 2.0 % when exposed for 24 h. Disinfectant DZPT-2 in the concentration of 2.0 % of the AI, when exposed for 5–24 h, and FAG preparation in the concentration of 2.0 %, when exposed for 24 h, and with the norm of consump- tion rate of 1 cubic decimeter per 1 square meter disinfect the test-objects (batiste, wood, glazed tile, metal, glass), contaminated with the TB agent M. bovis. Conclusions. Disinfecting preparations of DZPT-2 in the concentration of 2.0 % of AI when exposed for 5 h and FAG in the concentration of 2.0 % when exposed for 24 h may be used in the complex of veterinary and sanitary measures to prevent and control TB of farm ani- mals. The possibility of using the polymerase chain reaction as an additional method of estimating tuberculo- cide activity of disinfectants was proven.

scholarly journals DMSO Improves the Ski-Slope Effect in Direct PCR

2021 ◽  
Vol 11 (4) ◽  
pp. 1943
Author(s):  
Joo-Young Kim ◽  
Ju Yeon Jung ◽  
Da-Hye Kim ◽  
Seohyun Moon ◽  
Won-Hae Lee ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Analytical Techniques ◽  
Direct Pcr ◽  
Dna Profile ◽  
Novel Technologies ◽  
Specific Amplification ◽  
Polymerase Chain ◽  
Dna Profiles

Analytical techniques such as DNA profiling are widely used in various fields, including forensic science, and novel technologies such as direct polymerase chain reaction (PCR) amplification are continuously being developed in order to acquire DNA profiles efficiently. However, non-specific amplification may occur depending on the quality of the crime scene evidence and amplification methods employed. In particular, the ski-slope effect observed in direct PCR amplification has led to inaccurate interpretations of the DNA profile results. In this study, we aimed to reduce the ski-slope effect by using dimethyl sulfoxide (DMSO) in direct PCR. We confirmed that DMSO (3.75%, v/v) increased the amplification yield of large-sized DNA sequences more than that of small-sized ones. Using 50 Korean buccal samples, we further demonstrated that DMSO reduced the ski-slope effect in direct PCR. These results suggest that the experimental method developed in this study is suitable for direct PCR and may help to successfully obtain DNA profiles from various types of evidence at crime scenes.

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