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Authors: Sade Aisha Folashade John, Patrick E. Akpaka, Chandrashekhar Unakal, Arvind Kurhade, Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Enterobacteriaceae the leading cause of gastroenteritis. These gram- negative bacteria from species like Escherichia, Shigella, Salmonella, Vibrio and Helicobacter etc are among those that cause severe disease in consumers, especially those that indulge in uncooked seafood. Oysters have been shown to harbour these pathogenic organisms, which pose a major health risk. Numerous outbreaks of gastroenteritis due to oyster consumption have been reported worldwide because of the enteric bacteria as well as viral pathogen contaminationThis research was conducted to identify the Enterobacteriaceae present in oysters sold by vendors in Trinidad. During a five-month period (the rainy season, May - September), a total of 156 oyster samples (comprising 104 oysters samples and 52 water samples of the prepared, unprepared oyster cocktails plus the water used by vendors) were analysed. These samples were collected from roadside vendors operating near to the coastal line of the Western part of Trinidad These were processed at the microbiology unit of the Department of Para-clinical Sciences of the University of the West Indies, St. Augustine using standard microbiology techniques. The antibiotic susceptibility profiles of the recovered organisms were performed using the Kirby Bauer method for the following antibiotics- Tetracycline, Ampicillin, Trimethoprim-Sulfamethoxazole, Cefuroxime and Ceftazidime. The SPSS 21 programme was used to analyse the biometric data and a chi-square test was used to determine if there was any significant difference between the kinds of preparation cocktails with respect to the coliform and organisms found.Results of the 104 oyster samples tested, 112 isolates were obtained, 13.4% (15/112) were E.coli of which 0.1% (1/15) was the 0157:H7 strain; 72.3% (81/112) were Shigella species of which 27.2% (22/81) were Shigella dysenteriae; 14.3% (16/112) were Salmonella species of which 25% (4/16) were Salmonella cubana. The susceptibility profiles of the organisms revealed that 86.5% of Shigella, 100% of E.coli and 93.7% of the Salmonella were resistant to multiple antibiotics. With the recovery of these organisms from these samples, the health of the Trinidadian and foreign consumers of mangrove oysters is at risk particularly from Shigella, E.coli and Salmonella species. As a result, there should be an increased need for public awareness as well as regulations by the Ministry of Health to be made so that illness from these hazardous Enterobacteriaceae can be prevented.
Proper citation: Sade Aisha Folashade John, Patrick E. Akpaka, Chandrashekhar Unakal, Arvind Kurhade, Angel Justiz-Vaillant 2020. Isolation and Characterization of Enteric Bacteria from Oysters. protocols.io dx.doi.org/10.17504/protocols.io.bj6ekrbe Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Binding properties of immunoglobulin-binding protein by double immunodiffusion (Ouchterlony) technique.. protocols.io dx.doi.org/10.17504/protocols.io.bjsjkncn Copy
Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Summary: IFN-γ is a key player in driving cellular immunity. It is capable of orchestrating numerous and vital protective functions to enhance immune responses in cancers and infectious diseases. It exhibits its immunomodulatory effects by enhancing antigen processing and presentation,, inducing an anti-viral state, increasing leukocyte trafficking, boosting the anti-microbial properties of immune cells and by affecting cellular proliferation and apoptosis. [1]ReferenceKak G, Raza M, Tiwari BK. Interferon-gamma (IFN-γ): Exploring its implications in infectious diseases.Biomol Concepts. 2018;9(1):64-79. Published 2018 May 30. doi:10.1515/bmc-2018-0007
Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of human immunoglobulin M (IgM) in serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bj7rkrm6 Copy
Authors: Angel Justiz-Vaillant, Monica F. Smikle
Group: University of the West Indies, [email protected]
Summary: Streptococcal protein G is an immunoglobulin-binding protein that interacts with the Fc region of many mammalian immunoglobulins [1].References1.Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175
Proper citation: Angel Justiz-Vaillant, Monica F. Smikle 2020. Direct ELISA for investigating the binding of Protein-G to immunoglobulins.. protocols.io dx.doi.org/10.17504/protocols.io.bjxrkpm6 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. NK cells and CD4+ T lymphocytes make IL-21. It acts on various immune cells of innate and the adaptive immune systems. IL-21 promotes B and T lymphocyte proliferation and differentiation. It enhances NK cell activity. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-21 in human serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bketkten Copy
Authors: Angel Justiz-Vaillant, Norma McFarlane-Anderson
Group: University of the West Indies, [email protected]
Summary: This SpL ELISA can be used to detect specific antibodies in various animal species including human, mouse, rat, dog, rabbit, chicken, monkey, pig and hamster [1].1. De Chateau M, Nilson BH, Erntell M, Myhre E, Magnusson CG, Akerstrom B et al. On the interaction between protein L and immunoglobulins of various mammalian species. Scand J Immunol 1993; 37: 399−405
Proper citation: Angel Justiz-Vaillant, Norma McFarlane-Anderson 2020. Enzyme linked immunosorbent assay for investigating the binding of protein-LA (SpLA) to immunoglobulins. . protocols.io dx.doi.org/10.17504/protocols.io.bjpjkmkn Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. Monocytes and fibroblasts make IL-8. Its principal targets are neutrophils, basophils, mast cells, macrophages, and keratinocytes. It causes neutrophil chemotaxis, angiogenesis, superoxide release, and granule release.[1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-8 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj27kqhn Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Peroxidase-labeled-protein-LAG-anti-IgY conjugate is chemically made. It has unique binding properties. It binds to both avian and mammalian immunoglobulins. It can be used as a reagent in ELISA, Western blotting and dot blot for immunodetection of immunoglobulins and antigens. It may be used to make the immunodiagnosis of infectious diseases involving laboratory, wild, zoo, and farm animals [1].References1.Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175
Proper citation: Angel Justiz-Vaillant 2020. Direct ELISA for investigating the binding of chemically-made Protein-LAG-anti-IgY-peroxidase to both avian and mammalian immunoglobulins.. protocols.io dx.doi.org/10.17504/protocols.io.bjxzkpp6 Copy
Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. [1]The immunoregulatory cytokine IL-41 (also known as meteorin-like protein) is expressed at high levels in the synovium of patients with psoriatic arthritis (PsA).[2]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.2.Onuora S. Novel cytokine, IL-41, linked with PsA.Nat Rev Rheumatol. 2019;15(11):636. doi:10.1038/s41584-019-0314-7
Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of granulocyte macrophage-colony stimulating factor (GM-CSF) in human serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bktbkwin Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Immunoglobulin Y (IgY) is the mayormolecule present in the avian egg yolk. This protein functions as antibody and fulfils important functions in the protection of the embryo against challenging stimuli [1]. IgY has been isolated from the egg yolk of many birds [1]. These antibodies are important therapeutically, but also they have been used as immunological tool in many immunodiagnostic assays.Much time IgY concentration is calculated from the water soluble fraction (WSF), which means that there is not need of treating the preparation with alcohol to separate the protein pellet. Reference1. Justiz Vaillant AA, Ramirez N, Cadiz A, Ferrer B, Akpaka P, et al. (2013) Separation and Reactivity of Avian Immunoglobulin Y. J Chromat Separation Techniq 4: 173. doi:10.4172/2157-7064.1000173
Proper citation: Angel Justiz-Vaillant 2020. Polson method for IgY isolation with modification. . protocols.io dx.doi.org/10.17504/protocols.io.bjiykkfw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Binding properties of Staphylococcal protein A (SpA) to purified immunoglobulins tested by double immunodiffusion (Ouchterlony) technique.. protocols.io dx.doi.org/10.17504/protocols.io.bjsnknde Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. Monocytes mainly produce IL-30 in response to TLR agonists including bacterial LPS. It acts on monocytes, macrophages, dendritic cells, T and B lymphocytes, natural killer cells, mast cells, and endothelial cells. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-30 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj3zkqp6 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of macrophage-colony stimulating factor (M-CSF) in human serum or plasma.. protocols.io dx.doi.org/10.17504/protocols.io.bks7kwhn Copy
Authors: Angel Justiz-Vaillant, Monica F. Smikle
Group: University of the West Indies, [email protected]
Summary: Peptostreptococcal protein L is an immunoglobulin-binding protein that interacts with the Fab region of many mammalian immunoglobulins [1].References1.Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175
Proper citation: Angel Justiz-Vaillant, Monica F. Smikle 2020. Direct ELISA for investigating the binding of Peptostreptococcal Protein-L to immunoglobulins.. protocols.io dx.doi.org/10.17504/protocols.io.bjxtkpnn Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. NK cells and CD4+ T lymphocytes make IL-21. It acts on various immune cells of innate and the adaptive immune systems. IL-21 promotes B and T lymphocyte proliferation and differentiation. It enhances NK cell activity. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-21 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj3pkqmn Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Enzyme linked immunosorbent assay (ELISA) for determining the plasma or serum concentration of IL-33 in humans.. protocols.io dx.doi.org/10.17504/protocols.io.bkexktfn Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Streptococcal protein G, type III bacterial Fc receptor, is a globular protein produced by Streptococcal species and is composed of three nearly identical domains, each of 55 amino acids. SpG binds the Fc regions of antibodies IgG from many mammalian species [1-4]. The amino acid sequence of the IgG-binding regions (C1, C2 and C3) of SpG are not homologous to those of the regions (E, D, A, B and C) of SpA. A prerequisite of SpG for successful binding affinity to many IgG is the possession of at least two domains [3].Streptococcal protein G has been shown to have high binding affinity to sera from various mammalian species including human, , cow, pig, rabbit, goat and sheep by using competitive RIA [5] and to cervids, giraffes and peccaries by direct ELISA [2]. More recently using direct ELISA, some new interactions of SpG with free-ranging nondomestic hoofstock (order Artiodactyla) such as addax, bontebok, antelope, bison, elk, impala, kudu/nyala, oryx, muntjac, sheep, and white-tailed deer have been reported [1,2]. SpG does not bind to the Fc region of the immunoglobulin Y (IgY) of avian species [6]. This reagent can be used in ELISA, Western blotting oand Dot blot to detect antigens and antibodies [1]. It is important in the immunodiagnosis of infectious diseases and other problems. Protein L binds to kappa light chains of immunoglobulins from many animal species including human, mouse, rat, chicken, hamster and pig [1].1. Justiz-Vaillant AA, Akpaka PE, McFarlane-Anderson N, Smikle MF. Comparison of techniques of detecting immunoglobulin-binding protein reactivity to immunoglobulin produced by different avian and mammalian species.West Indian Med J. 2013;62(1):12-20.2. Stöbel K, Schönberg A, Staak C. A new non-species dependent ELISA for detection of antibodies to Borrelia burgdorferi s. l. in zoo animals.Int J Med Microbiol. 2002;291 Suppl 33:88-99. doi:10.1016/s1438-4221(02)80018-2.3. Kronvall G. A surface component in group A, C, and G streptococci with non-immune reactivity for immunoglobulin G. J Immunol 1973, 111(5): 1401-6.4. Justiz-Vaillant AA, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 2013, 4: 175.5. Reis KJ, Ayoub EM, Boyle MD. Streptococcal Fc receptors. II. Comparison of the reactivity of a receptor from a group C streptococcus with staphylococcal protein A. J Immunol 1984, 132(6): 3098-102.6. Serhir B, Dubreuil D, Higgins R, Jacques M. Purification and characterization of a 52-kilodalton immunoglobulin G-binding protein from Streptococcus suis capsular type 2. J Bacteriol 1995, 177(13): 3830-6.
Proper citation: Angel Justiz-Vaillant 2020. Preparation of horseradish peroxidase (HRP) conjugated Streptococcal protein-G by the periodate method.. protocols.io dx.doi.org/10.17504/protocols.io.bjkukkww Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Purification of avian egg yolk immunoglobulins using the water dilution method, precipitation with isopropanol and using HiTrap™ Columns.. protocols.io dx.doi.org/10.17504/protocols.io.bkbnksme Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: A protein that combines the binding capacity of SpA, SpG and SpL is not comercially available. It was created in my laboratory by combining these 3 immunoglobulin-binding proteins to horseradish peroxidase by the periodate method [1]. However, a mixture of SpA, SpG and SpL which are comercially available can have the same effect as universal reagent in immunodetection. 1. Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175
Proper citation: Angel Justiz-Vaillant 2020. Universal Immunoblot analysis for investigating Protein-LAG (SpLAG)-binding to mammalian and avian immunoglobulins.. protocols.io dx.doi.org/10.17504/protocols.io.bjsdkna6 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. They have both paracrine and autocrine function. Interleukins are also used in animal studies to investigate aspect related to clinical medicine.[1]T cells produce IL-2. The principal targets are T cells. Its primary effects are T-cell proliferation and differentiation, increased cytokine synthesis, potentiating Fas-mediated apoptosis, and promoting regulatory T cell development. It causes proliferation and activation of NK cells and B-cell proliferation and antibody synthesis. Also, it stimulates the activation of cytotoxic lymphocytes and macrophages. .[1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.
Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-2 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj22kqge Copy
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