​Dermal Response to Experimental Orfvirus (ORFV) Infection in Goats, Mice and Rabbit

2021 ◽  
Author(s):  
Nawab Nashiruddullah ◽  
Debesh Chandra Pathak ◽  
Jafrin Ara Ahmed ◽  
Safeeda Sultana Begum ◽  
Nagendra Nath Barman
Keyword(s):  
Laboratory Animals ◽  
Immunoperoxidase Staining ◽  
Cd4 Cells ◽  
Viral Nucleic Acid ◽  
Virus Latency ◽  
Field Samples ◽  
Intracytoplasmic Inclusions ◽  
Skin Biopsies ◽  
Injured Skin ◽  
Infective Potential

Background: During a study on the outbreak of orf in goats, it was intended to study the disease transmissibility in different hosts from field samples and ascertain the infective potential of the agent in laboratory animals compared to goats. Methods: Cutaneous clinical materials from orf virus (ORFV) infected goats was used to experimentally infect naive goats, rabbits and mice and ascertain its infective potential and transmissibility in different hosts. The processed inoculum was applied topically to mimic a natural transmission through injured skin. Regular skin biopsies were taken that revealed characteristic macroscopic and microscopic lesions typical of orf. Result: Virus inoculum applied on abraded skin in goats successfully established the lesions of orf. A parallel inoculation in rabbit and mice could not successfully reproduce the disease in these unnatural hosts beyond a subtle vesicular stage on 3 dpi with subsequent healing by 7 dpi. The lesions in goats regressed spontaneously by 28 days post-infection (dpi). Intracytoplasmic inclusions were associated only in the vesicular stage. Immunopathological progression was observed by immunoperoxidase staining of CD4+ and CD8+ T-cells which were found to appear by day 5 in the dermis and became more abundant and distributed by day 8, but subsequently reduced in number by 15 dpi. CD4+ cells were found to be more numerous and widespread. Viral antigen in tissues could be demonstrated by 4 dpi by immunohistological methods that increased in signal intensity progressively and disappear by 28 dpi. Similarly, viral nucleic acid in the skin could be detected on day 8 dpi but not on 28 dpi by PCR. The present experiment depicts the ease of disease transmissibility through traumatized skin in the primary hosts, but establishment in unnatural hosts may not be readily achieved. The infection was self-limiting with possibly no virus latency as indicated by immunofluorescence and PCR studies.

A novel tissue preservation medium for immunoperoxidase staining of skin biopsies

1992 ◽  
Vol 127 (6) ◽  
pp. 580-584 ◽  
Author(s):  
A. CARMICHAEL ◽  
G.R. COGHILL ◽  
I.A. CREE ◽  
S. GEORGE ◽  
J.G. LOWE
Keyword(s):  
Immunoperoxidase Staining ◽  
Tissue Preservation ◽  
Skin Biopsies ◽  
Preservation Medium

SURVEILLANCE METHODS FOR VIRUSES IN FOODS1

1969 ◽  
Vol 32 (11) ◽  
pp. 421-425 ◽  
Author(s):  
D. O. Cliver ◽  
J. Grindrod
Keyword(s):  
Food Samples ◽  
Test Method ◽  
Laboratory Animals ◽  
Good Sensitivity ◽  
Tissue Cultures ◽  
Virus Contamination ◽  
Routine Surveillance ◽  
Field Samples ◽  
Moderate Cost ◽  
Further Development

Viruses contaminate foods and sometimes cause illnesses in consumers. Methods have been needed for detection of foodborne viruses both in routine field samples and in samples associated with outbreaks of disease. Viruses are detected by inoculating living hosts such as tissue cultures or laboratory animals. Food samples, made fluid if necessary, can be inoculated directly into the test host. This approach has resulted in several isolations of viruses from field samples of foods. If greater sensitivities are desired, larger samples must be tested. This usually requires that the sample be concentrated before inoculation into the test host, and concentration can be performed only if food solids in the suspension are at a minimum. A family of methods has been developed for extraction and detection of enteroviruses from food samples. More recently, a procedure for dislodging enteroviruses from food surfaces has been devised. These procedures do not possess all of the desired properties of an ideal test method. In particular, they cannot detect all of the viruses known to be transmitted in foods. They do offer the possibility that several samples could be tested per day, with good sensitivity, at a moderate cost per sample. The means for routine surveillance of foods for virus contamination are at hand. Growing points for further development of surveillance methods are discussed.

scholarly journals Development and evaluation of recombinant GRA8 protein for the serodiagnosis of Toxoplasma gondii infection in goats

2020 ◽  
Author(s):  
Charoonluk Jirapattharasate ◽  
Ruenruetai Udonsom ◽  
Apichai Prachasuphap ◽  
Kodcharad Jongpitisub ◽  
Panadda Dhepakson
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Field Samples ◽  
Specific Igg

Abstract Background The development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT).Results Western blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT.Conclusion Our findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera

scholarly journals Development and Evaluation of Recombinant GRA8 Protein for the Serodiagnosis of Toxoplasma Gondii Infection in Goats

2020 ◽  
Author(s):  
Charoonluk Jirapattharasate ◽  
Ruenruetai Udonsom ◽  
Apichai Prachasuphap ◽  
Kodcharad Jongpitisub ◽  
Panadda Dhepakson
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Field Samples ◽  
Specific Igg

Abstract Background The development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT).Results Western blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT.Conclusion Our findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera.

scholarly journals Development and evaluation of recombinant GRA8 protein for the serodiagnosis of Toxoplasma gondii infection in goats

2020 ◽  
Author(s):  
Charoonluk Jirapattharasate ◽  
Ruenruetai Udonsom ◽  
Apichai Prachasuphap ◽  
Kodcharad Jongpitisub ◽  
Panadda Dhepakson
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Field Samples ◽  
Specific Igg

Abstract BackgroundThe development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT).ResultsWestern blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT.ConclusionOur findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera

scholarly journals Development and evaluation of recombinant GRA8 protein for the serodiagnosis of Toxoplasma gondii infection in goats

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Charoonluk Jirapattharasate ◽  
Ruenruetai Udonsom ◽  
Apichai Prachasuphap ◽  
Kodcharad Jongpitisub ◽  
Panadda Dhepakson
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Field Samples ◽  
Specific Igg

Abstract Background The development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT). Results Western blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT. Conclusion Our findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera.

The diagnosis of metabolic storage disease in skin biopsies (a light-, electronmicroscopic, and analytical study)

1978 ◽  
Vol 36 (2) ◽  
pp. 648-649
Author(s):  
W. Jurecka ◽  
W. Gebhart ◽  
H. Lassmann
Keyword(s):  
Storage Disease ◽  
Hunter Syndrome ◽  
Histochemical Demonstration ◽  
Sweat Glands ◽  
Type I ◽  
Storage Material ◽  
Scheie Syndrome ◽  
Storage Products ◽  
Skin Biopsies ◽  
Type V

Diagnosis of metabolic storage disease can be established by the determination of enzymes or storage material in blood, urine, or several tissues or by clinical parameters. Identification of the accumulated storage products is possible by biochemical analysis of isolated material, by histochemical demonstration in sections, or by ultrastructural demonstration of typical inclusion bodies. In order to determine the significance of such inclusions in human skin biopsies several types of metabolic storage disease were investigated. The following results were obtained.In MPS type I (Pfaundler-Hurler-Syndrome), type II (Hunter-Syndrome), and type V (Ullrich-Scheie-Syndrome) mainly “empty” vacuoles were found in skin fibroblasts, in Schwann cells, keratinocytes and macrophages (Dorfmann and Matalon 1972). In addition, prominent vacuolisation was found in eccrine sweat glands. The storage material could be preserved in part by fixation with cetylpyridiniumchloride and was also present within fibroblasts grown in tissue culture.

(EB) Virus: Latency and Expression in Transplanted and Cultured Human Cells

1971 ◽  
Vol 29 ◽  
pp. 368-369
Author(s):  
B. G. Uzman ◽  
M. M. Kasac ◽  
H. Saito ◽  
A. Krishan
Keyword(s):  
Cell Lines ◽  
Primary Cultures ◽  
Virus Particles ◽  
Barr Virus ◽  
Virus Latency ◽  
Virus Surveillance ◽  
Cultured Human Cells ◽  
Epstein Barr ◽  
Serial Transplantation ◽  
Tubular Arrays

In conjunction with the cultivation and transplantation of cells from human tumors by the Programs of Microbiology and Immunogenetics, virus surveillance by electron microscopy has been routinely employed. Of particular interest in this regard have been 3 cell lines cultured from lymph nodes or spleen of 2 patients with Hodgkin's disease and 1 patient with Letterer-Siwe's disease. Each of these cell lines when transplanted in Syrian hamster neonates conditioned with anti-lymphocyte serum grew as serially transplantable tumors; from such transplants of the 3 cell lines cell cultures were retrieved.Herpes type virus particles (Figs. 1, 2, 3) were found in the primary cultures of all three lines, in frozen thawed aliquots of same, and in cultures retrieved from their tumors growing by serial transplantation in hamsters. No virus was detected in sections of 25 of the serially transplanted tumors. However, in 10 such tumors there were repeated instances of tubular arrays in the cisternae of the endoplasmic reticulum (Fig. 4). On serologic examination the herpes virus was shown to be the Epstein-Barr virus.

Morphogenesis of a New Human Herpes-Type Virus Isolate (Sasha Virus)

1973 ◽  
Vol 31 ◽  
pp. 592-593
Author(s):  
R. F. Zeigel ◽  
W. Munyon
Keyword(s):  
Virus Isolate ◽  
Calf Serum ◽  
Uranyl Acetate ◽  
Human Herpes ◽  
Human Embryonic Lung ◽  
Human Herpes Virus ◽  
Intracytoplasmic Inclusions ◽  
Hel Cells ◽  
And Control

In continuing studies on the role of viruses in biochemical transformation, Dr. Munyon has succeeded in isolating a highly infectious human herpes virus. Fluids of buccal pustular lesions from Sasha Munyon (10 mo. old) uiere introduced into monolayer sheets of human embryonic lung (HEL) cell cultures propagated in Eagles’ medium containing 5% calf serum. After 18 hours the cells exhibited a dramatic C.P.E. (intranuclear vacuoles, peripheral patching of chromatin, intracytoplasmic inclusions). Control HEL cells failed to reflect similar changes. Infected and control HEL cells were scraped from plastic flasks at 18 hrs. of incubation and centrifuged at 1200 × g for 15 min. Resultant cell packs uiere fixed in Dalton's chrome osmium, and post-fixed in aqueous uranyl acetate. Figure 1 illustrates typical hexagonal herpes-type nucleocapsids within the intranuclear virogenic regions. The nucleocapsids are approximately 100 nm in diameter. Nuclear membrane “translocation” (budding) uias observed.

Changes in corneocyte element concentrations occur in skin inner stratum corneum

1990 ◽  
Vol 48 (2) ◽  
pp. 146-147
Author(s):  
R. R. Warner
Keyword(s):  
Stratum Corneum ◽  
Human Skin ◽  
Element Concentration ◽  
Skin Barrier ◽  
Barrier Properties ◽  
Brick Wall ◽  
Inorganic Elements ◽  
Dry Skin ◽  
Skin Biopsies ◽  
Intercellular Lipid

Keratinocytes undergo maturation during their transit through the viable layers of skin, and then abruptly transform into flattened, anuclear corneocytes that constitute the cellular component of the skin barrier, the stratum corneum (SC). The SC is generally considered to be homogeneous in its structure and barrier properties, and is often shown schematically as a featureless brick wall, the “bricks” being the corneocytes, the “mortar” being intercellular lipid. Previously we showed the outer SC was not homogeneous in its composition, but contained steep gradients of the physiological inorganic elements Na, K and Cl, likely originating from sweat salts. Here we show the innermost corneocytes in human skin are also heterogeneous in composition, undergoing systematic changes in intracellular element concentration during transit into the interior of the SC.Human skin biopsies were taken from the lower leg of individuals with both “good” and “dry” skin and plunge-frozen in a stirred, cooled isopentane/propane mixture.

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