scholarly journals Molecular characterization and Immunodiagnostic potential of various antigenic proteins of Fasciola gigantica species isolated from sheep of North West Himalayan Region

2019 ◽  
Vol 56 (2) ◽  
pp. 93-107
Author(s):  
J. S. Dar ◽  
B. A. Ganai ◽  
R. A. Shahardar ◽  
U. R. Zargar
Keyword(s):  
Small Ruminants ◽  
Fasciola Gigantica ◽  
Ovis Aries ◽  
Digenetic Trematode ◽  
Diagnostic Specificity ◽  
Immunoblotting Analysis ◽  
Accurate Identification ◽  
North West ◽  
Specificity And Sensitivity ◽  
Sds Page

SummaryThe control of the digenetic trematode Fasciola gigantica has been the major challenge in both cattle and small ruminants as there is a paucity of an effective and commercial vaccine. Thus, the accurate identification and prepatent diagnosis of F. gigantica is an essential prerequisite for its successful prevention and control. In the present study, the morphologically identified specimens isolated from the liver and bile ducts of sheep (Ovis aries) were validated through molecular data. The sequence analysis of ITS-2 of our isolates showed high degree of similarity with F. gigantica and F. hepatica using BLAST function of NCBI. The phylogenetic analysis of our isolates showed a close relationship with previously described F. gigantica and F. hepatica isolates from different countries. The antigenic profile of somatic and E/S antigens of F. gigantica were revealed by SDS–PAGE and immunoblotting using sera from sheep naturally infected with F. gigantica. By SDS-PAGE, 20 distinct bands were revealed from crude somatic fraction. Immunoblotting analysis of these proteins with positive sera exhibited 8 sero-reactive bands ranging from 14 to 97 kDa. Among these 38 and 44 kDa bands were quite specific with high diagnostic specificity and sensitivity. The E/S fraction comprised 7 distinct bands, as revealed by SDS-PAGE analysis. Immunoblotting analysis of these proteins with positive sera exhibited 6 antigenic bands ranging from 23 – 54 kDa. Among these 27 and 33 kDa were found to be quite specific with high diagnostic specificity and sensitivity. The present study concludes that the protein bands of 38 and 44 kDa in somatic fraction and 27 and 33 kDa in E/S fraction can be used for the immunodiagnostic purpose for this economically important parasite, which may also entice further studies regarding their vaccine potential.

Molecular characterization and immunodiagnostics of Dicrocoelium dendriticum species isolated from sheep of north-west Himalayan region

2020 ◽  
Vol 94 ◽  
Author(s):  
J.S. Dar ◽  
U. Shabir ◽  
S.A. Dar ◽  
B.A. Ganai
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Ovis Aries ◽  
Diagnostic Specificity ◽  
Immunoblotting Analysis ◽  
Accurate Identification ◽  
North West ◽  
Dicrocoelium Dendriticum ◽  
Specificity And Sensitivity ◽  
Sds Page

Abstract Despite its extensive presence among grazing ruminants, dicrocoeliosis, also known as ‘small liver fluke’ disease, is poorly known and often underestimated by researchers and practitioners in many countries. The accurate identification and prepatent diagnosis of Dicrocoelium dendriticum infection is an essential prerequisite for its prevention and control. In the present study, the morphologically identified specimens isolated from the bile ducts of sheep (Ovis aries) were validated through molecular data. The sequence analysis of the second internal transcribed spacer (ITS-2) of our isolates showed a high degree of similarity with D. dendriticum using the BLAST function of the National Center for Biotechnology Information (NCBI). The phylogenetic analysis of our isolates showed a close relationship with previously described D. dendriticum isolates from different countries. The antigenic profiles of somatic and excretory/secretory (E/S) antigens of D. dendriticum were revealed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and immunoblotting using sera from sheep naturally infected with D. dendriticum. By SDS–PAGE, 16 distinct bands were revealed from crude somatic fraction. Immunoblotting analysis of these proteins with positive sera exhibited six seroreactive bands ranging from 27 to 130 kDa. Among these, the 84 and 130 kDa bands were quite specific, with high diagnostic specificity and sensitivity. The E/S fraction comprised nine distinct bands, as revealed by SDS–PAGE analysis. Immunoblotting analysis of these proteins with positive sera exhibited five antigenic bands ranging from 27 to 130 kDa. Among these, the 130 kDa band was found to be quite specific, with high diagnostic specificity and sensitivity. The present study concludes that the protein bands of 84 and 130 kDa in somatic fraction and 130 kDa in E/S fraction can be used for the immunodiagnostic purpose for this economically important parasite, which may also encourage further studies regarding their vaccine potential.

Prevalence of Oestrus ovis (Diptera: Oestridae) in sheep from Ituiutaba, south-east region of Brazil

2021 ◽  
Vol 22 ◽  
Author(s):  
Henrique Inhauser Riceti Magalhães ◽  
Ana Caroline Romão da Silva ◽  
Fabiano Braz Romão ◽  
Nadia Grandi Bombonato ◽  
Guilherme Nascimento Cunha
Keyword(s):  
Small Ruminants ◽  
Minas Gerais ◽  
The State ◽  
Ovis Aries ◽  
Oestrus Ovis ◽  
Larval Size ◽  
Stage Of Development ◽  
East Region ◽  
Significant Difference ◽  
Complete Study

Abstract Among the diseases which can afflict the nasal cavities of small ruminants, oestrosis stands out. In Brazil, more specifically in its South-East region, the reports are limited only to the State of São Paulo and to the municipality of Araxá, Minas Gerais. Therefore, it has been sought to assess the parasitic prevalence of Oestrus ovis in sheep farmed in the municipality of Ituiutaba, Minas Gerais-Brazil, while correlating the larval size and stage, and its anatomical localization. Eighty-eight hemiheads of healthy Santa Inês/Dorper crossbreds Ovis aries have been used at random. The larvae in view were then collected and fixated to be quantified and analyzed in regard of size and stage of development. It is concluded that the oestrosis is an existing problem in the municipality of Ituiutaba, this being the first complete study on the prevalence of this parasite in the State of Minas Gerais. By anatomical distribution, only the differences of total larval averages between the frontal sinus and the ventral nasal meatus, the common nasal meatus and the nasopharynx have been significant. In size, the significant difference has been there only upon comparison between the size and the larval stage, information that is crucial for a better understanding of the cyclic progression, of the clinical symptomatology, and animal prophylaxis.

Purification and Characterization of Factor VII Inhibitor Found in a Patient with Life Threatening Bleeding

2000 ◽  
Vol 83 (01) ◽  
pp. 60-64 ◽  
Author(s):  
Seiji Miyamoto ◽  
Atsushi Iwasa ◽  
Masao Ishii ◽  
Kenji Okajima ◽  
Yu-ichi Kamikubo
Keyword(s):  
Light Chain ◽  
Calcium Ion ◽  
Polyacrylamide Gel Electrophoresis ◽  
Factor Vii ◽  
Immunoblotting Analysis ◽  
Calcium Dependent ◽  
Sds Page ◽  
Carboxyglutamic Acid ◽  
Life Threatening ◽  
Gla Domain

SummaryWe recently observed a patient with acquired inhibitor-induced F.VII deficiency whose plasma level of F.VII was < 1.0%. However, the biochemical nature of the inhibitor has not yet been clarified. In the present study, we purified the F.VII inhibitor from the patient’s plasma by using activated F.VII (F.VIIa)-conjugated gel and characterized the inhibitor. The results showed that the inhibitor comprised two kinds of antibodies: one was eluted with EDTA (antibody 1) and the other with glycine-HCl buffer (pH 2.3) (antibody 2) from the F.VIIa affinity gel. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting analysis of these inhibitors demonstrated that both antibodies had features of immunoglobulin G1 (IgG1) with κ and λ-light chains. Antibody 1 bound to the immobilized F.VIIa with a high affinity in the presence of calcium ion, while antibody 2 bound to the F.VIIa very weakly and the binding was independent of calcium ion. Immunoblotting analysis demonstrated that antibody 1 bound to the light chain of F.VIIa after reduction with 2-mercaptoethanol, while it did not react with either the γ carboxyglutamic acid (Gla)-domainless light chain of F.VIIa or the heavy chain with the protease domain. Antibody 1 markedly inhibited the activity of tissue factor-F.VIIa complex. Based on these observations, it is suggested that F.VIIa autoantibody (antibody 1) recognizes the calcium-dependent conformation within or near the Gla domain and inhibits F.VIIa activity by interacting with the light chain.

Disease and mortality in small ruminants in the North West Province of Cameroon

1989 ◽  
Vol 21 (3) ◽  
pp. 191-196 ◽  
Author(s):  
K. J. N. Ndamukong ◽  
M. M. H. Sewell ◽  
M. F. Asanji
Keyword(s):  
Small Ruminants ◽  
North West ◽  
The North ◽  
North West Province

scholarly journals Digenetic Trematode Parasites in Two Fresh Water Fishes Ophiocephalus Punctatus and Xenentodon Cancila From North West Himalayas

2021 ◽  
Author(s):  
Sukanya Rajput ◽  
Seema Langer
Keyword(s):  
Fresh Water ◽  
Digenetic Trematode ◽  
Host Fish ◽  
Mean Intensity ◽  
North West ◽  
Trematode Infection ◽  
Xenentodon Cancila ◽  
The Mean ◽  
Union Territory ◽  
Trematode Parasites

Abstract The study was conducted during September 2018-August 2019 to study the digenetic trematode infection in fresh water fishes of some of the water bodies viz. Gho-manhasan, Chakrali and Chadwal of Jammu region of J&K union territory A total of 220 fishes comprising Ophiocephalus punctatus and Xenentodon cancila belonging to families Channidae and Belonidae respectively were examined. A total of 4 digenetic trematode parasites belonging to 4 different families i.e., Euclinostomum heterostomum (Clinostomidae Luhe, 1901); Phyllodistomum tripathi (Gorgoderidae Looss, 1901); Genarchopsis piscicola (Hemiuridae, Luhe, 1901), and Bucephalopsis karvei (Bucephalidae Poche, 1907) were detected. The overall prevalence of digenetic trematode infection was 65.90% and the mean intensity was 3.58. Among these Genarchopsis piscicola showed the highest prevalence (40.38%) with mean intensity 2.95 in the host fish Xenentodon cancila, while in other species the prevalence ranged between 26.23% and 34.62%. Present study authenticates the presence of several species of digenetic trematode parasites in the fishes inhabiting freshwater of J&K union territory.

scholarly journals Microsphere Suspension Array Assays for Detection and Differentiation of Hendra and Nipah Viruses

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Adam J. Foord ◽  
John R. White ◽  
Axel Colling ◽  
Hans G. Heine
Keyword(s):  
Human Health ◽  
Virus Type ◽  
Nipah Virus ◽  
Hendra Virus ◽  
Diagnostic Specificity ◽  
Suspension Array ◽  
Specificity And Sensitivity ◽  
Nucleoprotein N ◽  
Encoding Genes ◽  
Single Reaction

Microsphere suspension array systems enable the simultaneous fluorescent identification of multiple separate nucleotide targets in a single reaction. We have utilized commercially available oligo-tagged microspheres (Luminex MagPlex-TAG) to construct and evaluate multiplexed assays for the detection and differentiation of Hendra virus (HeV) and Nipah virus (NiV). Both these agents are bat-borne zoonotic paramyxoviruses of increasing concern for veterinary and human health. Assays were developed targeting multiple sites within the nucleoprotein (N) and phosphoprotein (P) encoding genes. The relative specificities and sensitivities of the assays were determined using reference isolates of each virus type, samples from experimentally infected horses, and archival veterinary diagnostic submissions. Results were assessed in direct comparison with an established qPCR. The microsphere array assays achieved unequivocal differentiation of HeV and NiV and the sensitivity of HeV detection was comparable to qPCR, indicating high analytical and diagnostic specificity and sensitivity.

69 Invitro production of ovine embryos using either fresh or frozen-thawed semen

2021 ◽  
Vol 33 (2) ◽  
pp. 141
Author(s):  
G. Márquez-Márquez ◽  
A. Velázquez-Roque ◽  
F. Villaseñor-González ◽  
M. Kjelland ◽  
H. Álvarez-Gallardo ◽  
...  
Keyword(s):  
Semen Quality ◽  
Small Ruminants ◽  
Physiological Saline ◽  
Ovis Aries ◽  
Penicillin G ◽  
Domestic Sheep ◽  
Assisted Reproductive Technique ◽  
Physiological Saline Solution ◽  
Α Level ◽  
Fresh Semen

Invitro embryo production (IVP) is an important tool for genetic improvement in small ruminants. Semen quality is one of the most important aspects to consider for the success of this assisted reproductive technique. With ovine IVP, it is a common practice to use fresh semen for IVF, but this could be a problem because the differences between ejaculates from the same animal are well documented and a source of variation in IVP results. The objective of this research was to compare the effect of fresh and frozen–thawed domestic sheep (Ovis aries) semen on IVF for ovine IVP. The research was carried out in the reproduction laboratory at the Palominos Ranch (Jalisco, México). The IVP was performed with a continuous invitro culture system. Ovaries (n=186) were collected from a slaughterhouse (León, México) and transported to the laboratory within 2h in physiological saline solution (0.9% NaCl) supplemented with penicillin G (100IU mL−1) and streptomycin sulphate (100µg mL−1). For IVP, IVF-Bioscience™ media were used for IVM, IVF, and invitro culture (IVC). For IVM, the cumulus–oocyte complexes (COCs) were selected (only grades 1 and 2) and matured for 24h at 38.5°C in 5% CO2 in air and 100% humidity. Matured oocytes (n=1000) were invitro fertilized using either fresh or frozen–thawed semen (Triladyl™; Minitube) from the same sheep, at a concentration of 2×106 sperm mL−1, for 18h in 38.5°C, 5% CO2 in air, and 100% humidity. The presumptive zygotes were denuded by pipetting and set in IVC until Day 7 at 38.5°C, 5% CO2, 5% O2, and 90% N2 at 100% humidity. The percentages of cleavage, embryos with more than 6 cells, and blastocysts on Day 7 of culture were evaluated, based on the initial number of oocytes entering into IVM. Statistical analyses were carried out with the GLM procedure of SAS software (version 9.3; SAS Institute Inc.) to evaluate the results of fresh versus frozen–thawed (α level=0.05). Rates of cleavage, embryos with more than 6 cells, and blastocysts on Day 7 were similar (P&gt;0.05): fresh 52.3±3.0%, 43.6±2.6%, and 34.3±2.9%, respectively; frozen–thawed: 53.3%±3.0, 41.1±2.6%, and 33.3±2.9%, respectively. In conclusion, under the conditions of this research, the use of fresh and frozen–thawed semen had similar results for ovine IVP.

Management and productivity of small ruminants in the North West Province of Cameroon

1989 ◽  
Vol 21 (2) ◽  
pp. 109-119 ◽  
Author(s):  
K. J. N. Ndamukong ◽  
M. M. H. Sewell ◽  
M. F. Asanji
Keyword(s):  
Small Ruminants ◽  
North West ◽  
The North ◽  
North West Province
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