scholarly journals Molecular characterization and phylogenetic analysis of orf virus isolated from goats in Sokoto metropolis, Nigeria

2021 ◽  
pp. FSO700
Author(s):  
Nafi'u Lawal ◽  
Mubarak Ibrahim ◽  
Dauda Ayomide Onawala ◽  
Muhammad Bashir Bello ◽  
Rabiu Muhammad Aliyu ◽  
...  
Keyword(s):  
Genetic Characterization ◽  
Orf Virus ◽  
Coding Region ◽  
B2l Gene ◽  
Polymerase Chain ◽  
Chicken Eggs ◽  
Unique Amino Acid ◽  
Close Genetic Relationship ◽  
Embryonated Chicken

Aim: The aim of this study was to molecularly characterize orf virus isolated from clinical infections in goats in Sokoto metropolis. Materials & methods: Embryonated chicken eggs were used to isolate orf virus according to the established protocol. Viral DNA was extracted and full coding region of B2L gene was amplified by polymerase chain reaction, sequenced and blasted for identification and phylogenetically analyzed. Results and discussion: The B2L gene sequences of the isolate showed slight variability (96–98.7%) with the reference sequences as it clustered within the same clade with Korean, Zambian and Ethiopian strains, signifying a close genetic relationship. Unique amino acid substitutions were noted. This is the first genetic characterization of B2L gene of orf virus circulating in Nigeria. Conclusion: This study has provided in sight into the genetic diversity of orf virus in the study area.

scholarly journals Molecular Characterization and Phylogenetic Analysis of Orf Virus Isolated From Goats in Sokoto Metropolis, Nigeria

2020 ◽  
Author(s):  
Nafiu Lawal
Keyword(s):  
Molecular Characterization ◽  
Disease Outbreaks ◽  
Coding Region ◽  
Korean Strain ◽  
B2l Gene ◽  
Chicken Eggs ◽  
Unique Amino Acid ◽  
Close Genetic Relationship ◽  
Embryonated Chicken

Aim: Despite the endemic nature of contagious ecthyma in Nigeria, there is limited report on the molecular characterization of the isolates responsible for disease outbreaks. The aim of this study was to molecularly characterize ORFV isolated from clinical infections in goats in Sokoto metropolis. Materials and Methods: Seronegative embryonated chicken eggs were used to isolate ORFV via the chorio allantoic membrane (CAM) route according to the established protocol. Viral DNA was extracted from infected CAM and the full coding region of B2L gene was amplified by PCR and subsequently sequenced by Sanger’s method. The nucleotide sequence results were blasted for identification and phylogenetically analyzed using MEGA and Bioedit softwares. Results and Discussion: The results showed that B2L gene sequences of the ORFV UDUS/01/19/More strain showed slight variability (96- 98.7%) with the reference sequences. Our isolate clustered within the same clade with Korean strain signifying a close genetic relationship. Unique amino acid substitutions were noted in our isolate when compared with other references. This is arguably the first genetic characterisation of B2L gene of ORFV circulating in Nigeria. Conclusion: Our study has provided in sight into the genetic diversity of ORFV in the study area. This is crucial for the design of effective vaccines against the disease which are currently lacking in the country.

Genetic characterization of a mumps virus isolate during passaging in the amniotic cavity of embryonated chicken eggs

2004 ◽  
Vol 99 (2) ◽  
pp. 121-129 ◽  
Author(s):  
Jelena Ivancic ◽  
Dubravko Forcic ◽  
Tanja Kosutic Gulija ◽  
Renata Zgorelec ◽  
Leonida Repalust ◽  
...  
Keyword(s):  
Virus Isolate ◽  
Genetic Characterization ◽  
Mumps Virus ◽  
Amniotic Cavity ◽  
Chicken Eggs ◽  
Embryonated Chicken

scholarly journals Molecular characterization of Orf virus isolates from Kodai hills, Tamil Nadu, India

2019 ◽  
Vol 12 (7) ◽  
pp. 1022-1027
Author(s):  
G. Nagarajan ◽  
R. Pourouchottamane ◽  
G. B. Manjunatha Reddy ◽  
R. Yogisharadhya ◽  
K. Sumana ◽  
...  
Keyword(s):  
Tamil Nadu ◽  
Skin Lesions ◽  
Amino Acid Identity ◽  
Orf Virus ◽  
Research Centre ◽  
B2l Gene ◽  
Bovine Papular Stomatitis Virus ◽  
Polymerase Chain ◽  
Reference Strains

Aim: The present study was carried out to find out the causative agent of exanthematous skin lesions in sheep maintained by Southern Regional Research Centre, Mannavanur, Kodai hills, Tamil Nadu. Materials and Methods: Polymerase chain reaction (PCR) with Orf virus (ORFV) B2L gene-specific primers was carried out by employing the total genomic DNA isolated from the scabs as the template. The ORFV isolates from Kodai hills were characterized by the use of bioinformatics tools. Results: The amino acid identity of ORFV isolate 1 from Kodai hills is having 98.14%, 96.29%, and 83.59% identity with reference strains of ORFV, Pseudocowpox virus, and bovine papular stomatitis virus, respectively. Phylogenetic analysis revealed that ORFV isolates from Kodai hills clustered with the other ORFV isolates from different geographical areas of India. Conclusion: The etiological agent of exanthematous skin lesion among sheep of Kodai hills is ORFV.

Characterization of the Original Christmas Disease Mutation (Cysteine 206 → Serine): From Clinical Recognition to Molecular Pathogenesis

1992 ◽  
Vol 67 (01) ◽  
pp. 063-065 ◽  
Author(s):  
Sherryl A M Taylor ◽  
Jacalyn Duffin ◽  
Cherie Cameron ◽  
Jerome Teitel ◽  
Bernadette Garvey ◽  
...  
Keyword(s):  
Nucleotide Substitution ◽  
Novel Mutation ◽  
Factor Ix ◽  
Causative Mutation ◽  
Coding Region ◽  
Body Of Knowledge ◽  
Polymerase Chain ◽  
Splice Junctions

SummaryChristmas disease was first reported as a distinct clinical entity in two manuscripts published in 1952 (1, 2). The eponym associated with this disorder, is the surname of the first patient examined in detail and reported by Biggs and colleagues in a paper describing the clinical and laboratory features of seven affected individuals (3). This patient has severe factor IX coagulant deficiency (less than 0.01 units/ml) and no detectable circulating factor IX antigen (less than 0.01 units/ml). Coding sequence and splice junctions of the factor IX gene from this patient have been amplified in vitro through the polymerase chain reaction (PCR). One nucleotide substitution was identified at nucleotide 30,070 where a guanine was replaced by a cytosine. This mutation alters the amino acid encoded at position 206 in the factor IX protein from cysteine to serine. The non conservative nature of this substitution, the absence of this change in more than 200 previously sequenced factor IX genes and the fact that the remainder of the coding region of this gene was normal, all provide strong circumstantial evidence in favour of this change being the causative mutation in this patient. The molecular characterization of this novel mutation in the index case of Christmas disease, contributes to the rapidly expanding body of knowledge pertaining to Christmas disease pathogenesis.

scholarly journals Characterization of a New Alloantigen (SH) on the Human Neutrophil Fcγreceptor IIIb

Blood ◽  
1997 ◽  
Vol 89 (3) ◽  
pp. 1027-1034 ◽  
Author(s):  
Juergen Bux ◽  
Ernst-Ludwig Stein ◽  
Philippe Bierling ◽  
Patricia Fromont ◽  
Mary Clay ◽  
...  
Keyword(s):  
Nucleotide Sequence Analysis ◽  
Polymorphism Analysis ◽  
Mutational Event ◽  
Coding Region ◽  
Specific Pcr ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Antigen Frequency ◽  
Neonatal Neutropenia

Abstract Polymorphic structures of the neutrophil Fcγreceptor IIIb (FcγRIIIb) result in alloantibody formation that causes alloimmune neonatal neutropenia and transfusion reactions. Alloantigens located on FcγRIIIb include the antigens NA1 and NA2. In four cases of alloimmune neonatal neutropenia, granulocyte-specific alloantibodies directed against a thus far unknown antigen were detected by granulocyte agglutination and immunofluorescence tests in the maternal sera. By the use of the monoclonal antibody–specific immobilization of granulocyte antigens (MAIGA) assay, the new antigen, termed SH, was located on the FcγRIIIb. Nucleotide sequence analysis of the FcγRIIIb coding region from a SH(+) individual showed a single-base C→A mutation at position 266, which results in an Ala78Asp amino acid substitution. A family study confirmed that this nucleotide difference is inherited, and corresponds to the SH phenotype. Serologic typing of 309 randomly selected individuals showed an antigen frequency of 5% in the white population. The same frequency was found by genotyping, for which a technique based on polymerase chain reaction (PCR) using sequence-specific primers (PCR-SSP) was developed. Typing of all SH(+) individuals for NA1 and NA2, and PCR-restriction fragment length polymorphism analysis of the NA-specific PCR products from five SH(+) individuals using the SH-specific endonuclease SfaN I showed that SH antigen is very probably the result of an additional mutational event in the NA2 form of the FcγRIIIB gene. Immunochemical studies also demonstrated that the SH determinants reside on the 65- to 80-kD NA2 isoform of the FcγRIIIb. Our findings show the existence of an additional polymorphism of the FcγRIIIb, which can result in alloantibody formation causing alloimmune neonatal neutropenia.

scholarly journals Genetic Differentiation of Clariid Populations using Microsatellite Markers in Kano State Rivers

2021 ◽  
pp. 35-45
Author(s):  
I. O. Suleiman ◽  
R.O. Okeke ◽  
J. M. Madu ◽  
A. U. Umar ◽  
O.M Akinsola ◽  
...  
Keyword(s):  
Genetic Variation ◽  
Genetic Differentiation ◽  
Dna Extraction ◽  
Genetic Characterization ◽  
Caudal Peduncle ◽  
Fta Cards ◽  
Fish Samples ◽  
Polymerase Chain ◽  
Outbred Populations

This study aimed to investigate the genetic characterization of strains of Clariid fish species in some river bodies in Kano State using microsatellite markers.One hundred and seventy seven Clariid fish samples (Clariasgariepinus and Heterobranchuslongifilis) were collected from six rivers (Thomas, Ghari, Tiga dam, Duddurun Gaya, Karaye and Bagwai) in Kano state. Blood sample was taken from each fish sample by severing the caudal peduncle and drained into FTA cards for DNA extraction, Polymerase Chain Reaction and electrophoresis to determine genetic variation between the Clariid fish populations.Genealex 6.4 software package was used to analyse the resolve bands from DNA extraction to determine their base pair and genetic variation. Results showed that the Fst values ranged from 0.00 to 0.66, Fit ranged from -0.04 to 0.12, Fis ranged from -0.35 to -0.26. It indicated a large number of gene flow (exchange) among the populations with a range of 0.46 to 0.87. There was an established magnitude of genetic divergence (91.86%) among the populations as shown by the result of the percentage polymorphism which depends on the number of alleles detected per locus and their frequencies. It can be concluded that since there was no inbreeding as shown in the study, none of the population exhibited genetic uniqueness. The populations had a high genetic differentiation between populations but moderate differentiation within populations. The populations were outbred populations; an indication that relatives avoided mating in the population.

scholarly journals Genetic diversity of Blastocystis in kindergarten children in southern Xinjiang, China

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Meng Qi ◽  
Zilin Wei ◽  
Ying Zhang ◽  
Qiyuan Zhang ◽  
Juanfeng Li ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Genetic Characterization ◽  
Intestinal Parasites ◽  
Small Subunit ◽  
Kindergarten Children ◽  
Chain Reaction ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Southern Xinjiang

Abstract Background Blastocystis is one of the most common intestinal parasites in humans and various animals worldwide. Few studies are available regarding the genetic characterization of Blastocystis infections in humans in China. Methods In the present study, 609 fecal samples were collected from two- to six-year-old kindergarten children in southern Xinjiang and were examined by polymerase chain reaction (PCR). Results The infection rate of Blastocystis was 14.3% (87/609); no significant difference was observed among counties and between sexes. Blastocystis subtypes ST1 (n = 38), ST2 (n = 8), and ST3 (n = 41) were identified by sequence analysis of the small subunit ribosomal RNA gene. Genetic polymorphisms were observed at the intra-subtype level, including seven variations for ST1 (ST1A to ST1G), four for ST2 (ST2A to ST2D), and two for ST3 (ST3A and ST3B); with ST1F and ST2B being new variations. Conclusions ST1 and ST3 are the two common Blastocystis subtypes in the study area. More extensive studies in both humans and animals in different regions are needed to better characterize the transmission of Blastocystis.

Genetic characterization of Spermophagus niger (Coleoptera: Chrysomelidae: Bruchinae: Amblycerini) pest associated to seeds of Sorrel (Hibiscus sabdariffa L.) in Burkina Faso

2016 ◽  
Vol 6 (1) ◽  
pp. 07-14
Author(s):  
Jean Christophe Koussoubé ◽  
Fatimata Mbaye ◽  
Cheikh Abdou Khadre Mbacké Dia ◽  
Mbacké Sembène ◽  
Antoine Sanon
Keyword(s):  
Amino Acid ◽  
Burkina Faso ◽  
Genetic Parameters ◽  
Genetic Characterization ◽  
Mitochondrial Gene ◽  
Hibiscus Sabdariffa ◽  
Polymerase Chain ◽  
The One ◽  
First Time

In Burkina Faso, the seeds of sorrel, Hibiscus sabdariffa L. are attacked by a pest identified morphologically as Spermophagus niger which is maintained all year on seeds and causing considerable damages. In the current study, for the first time, genetic characterization for S. niger was performed to determine its genetic identity and place it in its phyletic group. Mitochondrial gene, the Cytochrome oxidase I (COI) of the pest was partially sequenced after extraction and amplification by Polymerase Chain Reaction (PCR). Then the variability of genetic parameters namely the number of polymorphic and monomorphic sites, the frequencies of the different nucleotides and amino acid composition were determined. The nucleotide sequence of S. niger ob-tained was submitted in Genbank and the accession number is KU710716. Nucleotide sequences of S. niger obtained and those of different species of Spermophagus and Z. subfasciatus available in the GenBank database, we determined the percentage of similarity on the one hand and kinship through Phylogenetics reconstructions on the other hand. The results showed the absence of polymorphic sites for 406 sites obtained with 36.5% of thymine, 17.5% of cytosine, adenine 31% and 15% of guanine. Leucine was the majority amino acid (14.50%); the lysine was minority amino acid (0.76%) and cysteine was absent. The percentage of similarity obtained and phylogenetics reconstructions showed that S. niger is very close to the different species of Spermophagus particularly S. drak and different from Z. sub-fasciatus.

An Epidemiological Study of Sheep and Goat pox Outbreaks in the Sudan

Food Biology ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Ahmed Zein Elabdeen Mahmoud ◽  
Abdelmalik Ibrahim Khalafalla ◽  
Muaz Magzob Abdellatif
Keyword(s):  
Virus Isolation ◽  
Chorioallantoic Membrane ◽  
Vero Cells ◽  
Fibroblast Cells ◽  
Chick Embryo Fibroblast ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Species Specific ◽  
Chicken Eggs ◽  
Embryonated Chicken

Sheep and goat pox Outbreaks occurred in different geographic areas of Sudan and most strikingly, were highly species specific. Two outbreaks in Gedarif State in June. 2013 affected no goats and outbreak in Khartoum state in March. 2015 affected no sheep despite communal herding; affected goats were vaccinated with 0240 strain. Clinically, the disease was characterized by fever, depression and eruption of generalized pox lesions. Mortality rate ranged between 5.2 and 6.7% with a mean of 6.1%. Isolation of viruses succeed on Lamb testes cell culture at passage four, the diseases were diagnosed using virus neutralisation test and polymerase chain reaction. Sheeppox and goatpox isolates grew well in lamb testes and Vero cells. In MDBK however, both viruses induced slight CPE that reached 60% in 9 days. On the other hand, both isolates induced no CPE in chick embryo fibroblast cells. Virus isolation attempts failed on chorioallantoic membrane of embryonated chicken eggs.

Importance of flagella and enterotoxins for Aeromonas virulence in a mouse model

2007 ◽  
Vol 53 (2) ◽  
pp. 261-269 ◽  
Author(s):  
Keya Sen ◽  
Dennis Lye
Keyword(s):  
Drinking Water ◽  
Virulence Factor ◽  
Genetic Characterization ◽  
Aeromonas Veronii ◽  
Aeromonas Caviae ◽  
Polymerase Chain ◽  
Immunocompromised Mice ◽  
Immunocompromised Hosts ◽  
Virulence Factor Genes

A genetic characterization of eight virulence factor genes, elastase, lipase, polar flagella (flaA/flaB, flaG), lateral flagella (lafA), and the enterotoxins alt, act, and ast, was performed using polymerase chain reaction with 55 drinking water and nine clinical isolates. When 16 Aeromonas hydrophila strains, seven Aeromonas veronii strains, and seven Aeromonas caviae strains exhibiting different combinations of virulence factor genes were tested in immunocompromised mice by intraperitoneal injection, only those strains that had one or more of the enterotoxins flaA, flaB, and either flaG or lafA showed signs of being virulent. The correlation was seen in 97% (29/30) of the strains, which included strains from drinking water. Thus, Aeromonas water isolates have the potential to be pathogenic in immunocompromised hosts.

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