scholarly journals IMMUNODIAGNOSIS INFEKSI Aeromonas hydrophila PADA IKAN

2018 ◽  
Vol 36 (1) ◽  
pp. 80
Author(s):  
Yuli Purwandari Kristianingrum ◽  
Sitarina Widyarini ◽  
Kurniasih Kurniasih ◽  
Bambang Sutrisno ◽  
Charles Rangga Tabbu ◽  
...  
Keyword(s):  
Blood Serum ◽  
Polyclonal Antibody ◽  
Aeromonas Hydrophila ◽  
Intraperitoneal Injection ◽  
Staining Method ◽  
Polyclonal Antibodies ◽  
Ulcer Disease ◽  
Tube Test ◽  
Immunohistochemistry Staining ◽  
Motile Aeromonas Septicaemia

Aeromonas hydrophila causes a disease that often infects fish and is known as Motile Aeromonas Septicaemia (MAS), Hemorrhagi Septisemia, Ulcer disease or Red-Sore disease. The   aims of this study were to develop polyclonal antibody of  Aeromonas hydrophila in the rabbits to   confirm the diagnosis of Aeromonas hydrophila  in the fish by immunohistochemistry staining method. Preparation of polyclonal antibodies was performed on the rabbits used to Aeromonas hydrophila bacteria that have been tested biochemically by intravenous and intraperitoneal injection. Doses of Aeromonas hydrophila  bacteria were 109 CPU/ml  of 0.5 ml at first injection, 1 ml at second injection, 2 ml at thirth injection and 3 ml at fourth injection. Blood serum collection was performed at week 5 after injection from  an  ear and intracardial vein. The result of antibody titer was 28 = 1024 which measured by   tube test. Furthermore, polyclonal   antibody was used to immunohistochemistry  staining with 400x dilution. The results of the staining showed that an immunopositive reaction in the liver, skin,lien,  gill, kidney, and heart of fish to Aeromonas hydrophila antibody. The research conclution was polyclonal antibody from rabbit can be used to accurately confirm the diagnosis of Aeromonas hydrophila  based on antigen and antibody reaction. 

Evaluation of germ plasm for resistant to Soybean mosaic virus (SMV)

2020 ◽  
Vol 21 (1) ◽  
pp. 6-9
Author(s):  
Wuye Ria Andayanie
Keyword(s):  
Abiotic Stress ◽  
Environmental Factors ◽  
Mosaic Virus ◽  
Polyclonal Antibody ◽  
Symptom Severity ◽  
Germ Plasm ◽  
Polyclonal Antibodies ◽  
Soybean Mosaic Virus ◽  
High Yield ◽  
High Yields

Soybean superior varieties with high yields and are resistant to abiotic stress have been largely released, although some varieties grown in the field are not resistant to SMV. In addition, the opportunity to obtain lines of hope as prospective varieties with high yield and resistance to SMV is very small. The method for evaluating soybean germplasm is based on serological observations of 98 accessions of leaf samples from SMV inoculation with T isolate. The evaluation results of 98 accessions based on visual observations showed 31 genotypes reacting very resistant or healthy to mild resistant category to SMV T isolate  with a percentage of symptom severity of 0 −30 %. Among 31 genotypes there are 2 genotypes (PI 200485; M8Grb 44; Mlg 3288) with the category of visually very resistant and resistant, respectively and  Mlg 3288  with the category of mild resistant.  They have a good agronomic appearance with a weight of 100 seeds (˃10 g) and react negatively with polyclonal antibodies to SMV, except Mlg 3288 reaction is not consistent, despite the weight of 100 seeds (˃ 10 g). Leaf samples from 98 accessions revealed various symptoms of SMV infection in the field. This diversity of symptoms is caused by susceptibility to accession, when infection occurs, and environmental factors. Keywords—: soybean; genotipe; Soybean mosaic virus (SMV); disease severity; polyclonal  antibody

The effect of vaccination in controling bacterial disease caused by Aeromonas hydrophila in nile tilapia (Oreochromis niloticus)

2014 ◽  
pp. 27
Author(s):  
Arne A Ratulangi ◽  
Reiny Tumbol ◽  
Hengky Manoppo ◽  
Henneke Pangkey
Keyword(s):  
Survival Rate ◽  
Aeromonas Hydrophila ◽  
Oreochromis Niloticus ◽  
Nile Tilapia ◽  
Bacterial Disease ◽  
Immersion Method ◽  
Nile Tilapia Oreochromis Niloticus ◽  
Different Ages ◽  
Motile Aeromonas Septicaemia

This study aims to apply vaccination against bacterial disease. The purpose of vaccination is to trigger the immune respone both non-specific and specific of fish against bacteria Motile Aeromonas Septicaemia (MAS) caused by Aeromonas hydrophila. The vaccination for fish with different ages: 2-3 weeks and 5 weeks were done using immersion method. The fish were re-vaccinated (booster) after two weeks of the first vaccination. The survival rate was < 50 % for juveniles 2-3 weeks and > 50% for juveniles 5 weeks. Survival rate for juveniles 5 weeks was higher than juveniles of 2-3 weeks. This shows that organs of juveniles of 5 weeks were more complete than the 2-3 weeks juveniles. The age of fish is one of the important factors for successfully vaccination. Penelitian ini bertujuan untuk melakukan penerapan vaksinasi terhadap penyakit bakterial. Vaksinasi ditujukan untuk merangsang respon kekebalan non- spesifik dan spesifik pada tubuh ikan terhadap penyakit Motile Aeromonas Septicaemia (MAS) yang disebabkan oleh bakteri Aeromonas hydrophila. Vaksin diberikan pada benih ikan nila yang berbeda umur yaitu 2-3 minggu dan 5 minggu dengan menggunakan metode perendaman. Ikan divaksinasi ulang (booster) setelah 2 (dua) minggu dari vaksinasi yang pertama. Prosentase kelangsungan hidup < 50 % untuk benih umur 2-3 minggu dan > 50% untuk benih umur 5 minggu. Jumlah kematian benih umur 2-3 minggu lebih tinggi dari 5 minggu. Hal ini menunjukkan bahwa fungsi organ benih umur 5 minggu telah lebih lengkap dari pada benih umur 2-3 minggu. Umur ikan merupakan salah satu faktor penting penentu keberhasilan suatu kegiatan vaksinasi.

The use of β-glucan extracted from baker’s yeast (Saccharomyces cerevisiae) to increase non-specific immune system and resistence of tilapia (Oreochromis niloticus) to Aeromonas hydrophila

2013 ◽  
pp. 92
Author(s):  
Ida N Jamal ◽  
Reiny A Tumbol ◽  
Remy E.P Mangindaan
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Immune System ◽  
Aeromonas Hydrophila ◽  
Phagocytic Activity ◽  
Baker’S Yeast ◽  
Baker's Yeast ◽  
Yeast Saccharomyces Cerevisiae ◽  
Randomized Design ◽  
The Difference ◽  
Motile Aeromonas Septicaemia

Motile Aeromonas Septicaemia disease (MAS) attacking tilapia has increased in recent years as a consequence of intensive aquaculture activities, which led to losses in aquaculture industry. The agent causing MAS disease is Aeromonas hydrophila. The disease can be controlled with the β-glucan. As immunostimulants, β-glucans can also increase resistance in farmed tilapia. Studies on the use of β-glucan extracted from baker's yeast Saccharomyces cerevisiae was intended to evaluate the non-specific immune system of tilapia that were challenged with Aeromonas hydrophila. The method used was an experimental method with a completely randomized design consisting of four treatments with three replicats. The dose of β-glucan used as treatments were 0 mg.kg-1 fish (Control), 5 mg.kg-1 fish (B), 10 mg.kg-1 fish (C) and 20 mg.kg-1 fish (D), each treatment as injected three times at intervals of 3 days, the injection volume of 0.5 ml/fish for nine days and resistance surveillance for seven days. The results showed that the difference in the amount of β-glucan and the frequency of the injected real influence on total leukocytes, phagocytic activity and resistance. Total leukocytes, phagocytic activity and resistance to treatment was best achieved by the administration of C a dose of  10 mg.kg-1 of the fish© Penyakit Motil Aeromonas Septicaemia (MAS) yang menyerang ikan nila mengalami peningkatan selama beberapa tahun terakhir sebagai konsekuensi dari kegiatan akuakultur intensif, yang menyebabkan kerugian dalam industri budidaya. Agen utama penyebab penyakit MAS adalah Aeromonas hydrophila. Untuk mengendalikan penyakit tersebut dapat dilakukan dengan pemberian β-glukan. Sebagai imunostimulan, β-glukan juga dapat  meningkatkan resistensi pada ikan nila yang dibudidayakan. Pengkajian mengenai pemanfaatan β-glukan yang diekstrak dari ragi roti Saccharomyces cerevisiae dimaksudkan untuk menguji sistem imun non spesifik ikan nila yang diuji tantang dengan bakteri Aeromonas hydrophila. Metode yang digunakan yaitu metode eksperimen dengan rancangan acak lengkap yang terdiri dari empat perlakuan dan tiga ulangan. Dosis β-glukan  yang digunakan sebagai perlakuan sebesar 0 mg.kg-1 ikan (Kontrol), 5 mg.kg-1 ikan (B), 10 mg.kg-1 ikan (C) dan 20 mg.kg-1 ikan (D), masing-masing perlakuan diinjeksi sebanyak 3 kali dengan interval waktu 3 hari selama 9 hari, volume injeksi 0,5 mL/ekor ikan dan pengamatan resistensi selama tujuh hari. Hasil penelitian menunjukkan perbedaan jumlah β-glukan dan frekuensi pemberian yang diinjeksikan memberikan pengaruh nyata terhadap total leukosit, aktivitas fagositosis dan resistensi. Total leukosit, aktivitas fagositosis dan resistensi terbaik dicapai pada perlakuan C dengan dosis 10 mg.kg-1 ikan©

Immunofluorescence Assay Using Monoclonal and Polyclonal Antibodies for Detection of Staphylococcal Enterotoxins A in Milk

2019 ◽  
Vol 13 (1) ◽  
pp. 137-145 ◽  
Author(s):  
Tzonka Godjevargova ◽  
Zlatina Becheva ◽  
Yavor Ivanov ◽  
Andrey Tchorbanov
Keyword(s):  
Monoclonal Antibody ◽  
Magnetic Nanoparticles ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Food Poisoning ◽  
Staphylococcal Enterotoxin ◽  
Staphylococcal Enterotoxin A ◽  
Fluorescence Immunoassay ◽  
Magnetic Separator ◽  
Milk Samples

Objectives: Staphylococcus aureus is a Gram-positive microorganism. S. aureus can grow in various foods and cause food poisoning by secreting enterotoxins. The most common enterotoxins involved in food poisoning are staphylococcal enterotoxin A and staphylococcal enterotoxin B, but Staphylococcal Enterotoxin A (SEA) is predominant. The main types of food contaminated with SEs are meat and meat products, poultry and eggs, milk and dairy products. The aim of this study was to develop a rapid and sensitive fluorescence immunoassay for detection of staphylococcal enterotoxin A in milk. Methods: Monoclonal and polyclonal antibodies for SEA were produced and characterized. Competitive fluorescence immunoassay based on Magnetic Nanoparticles (MNPs) was performed and optimized. MNPs were used as a solid carrier of the antibodies. The first step of the assay was immunoreaction between the immobilized antibody onto MNPs and SEA in milk sample. Then the fluorescein-SEA conjugate was added to the sample. Thus, competitive immunoreaction between MNP-mAb/MNP-pAb with SEA and SEA-FITC was performed. These immuno-complexes were separated by a magnetic separator and the obtained supernatants were analyzed. The fluorescent signal from the excess of conjugated SEA was proportional to the SEA contained in the milk. The assay duration was only 30 min. Results: The fluorescence immunoassays performed with polyclonal antibody had linear ranges from 5 pg/mL to 100 ng/mL SEA in a buffer, and from 50 pg/mL to 50 ng/mL SEA in spiked milk samples. While the same assays performed with monoclonal antibody had linear ranges from 1 pg/mL to 20 ng/mL SEA in buffer, and from 10 pg/mL to 10 ng/mL SEA in spiked milk samples. The detection limits of the developed immunoassays performed in milk were: 48 pg/mL with polyclonal antibody and 9 pg/mL with monoclonal antibody. Conclusion: A rapid and sensitive fluorescence immunoassay based on magnetic nanoparticles with a polyclonal and monoclonal antibody for determination of staphylococcal enterotoxin A in milk was developed.

scholarly journals Differential expression of TgMIC1 in isolates of Chinese 1 Toxoplasma with different virulence

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yang Wang ◽  
Chengjian Han ◽  
Rongsheng zhou ◽  
Jinjin Zhu ◽  
Famin Zhang ◽  
...  
Keyword(s):  
Differential Expression ◽  
Rna Sequencing ◽  
Polyclonal Antibody ◽  
Protein Level ◽  
Polyclonal Antibodies ◽  
Mrna Levels ◽  
Quantitative Real Time Pcr ◽  
Sequencing Data ◽  
Predominant Genotype ◽  
High Virulence

Abstract Background The predominant genotype of Toxoplasma in China is the Chinese 1 (ToxoDB#9) lineage. TgCtwh3 and TgCtwh6 are two representative strains of Chinese 1, exhibiting high and low virulence to mice, respectively. Little is known regarding the virulence mechanism of this non-classical genotype. Our previous RNA sequencing data revealed differential mRNA levels of TgMIC1 in TgCtwh3 and TgCtwh6. We aim to further confirm the differential expression of TgMIC1 and its significance in this atypical genotype. Methods Quantitative real-time PCR was used to verify the RNA sequencing data; then, polyclonal antibodies against TgMIC1 were prepared and identified. Moreover, the invasion and proliferation of the parasite in HFF cells were observed after treatment with TgMIC1 polyclonal antibody or not. Results The data showed that the protein level of TgMIC1 was significantly higher in high-virulence strain TgCtwh3 than in low-virulence strain TgCtwh6 and that the invasion and proliferation of TgCtwh3 were inhibited by TgMIC1 polyclonal antibody. Conclusion Differential expression of TgMIC1 in TgCtwh3 and TgCtwh6 may explain, at least partly, the virulence mechanism of this atypical genotype.

scholarly journals Examination of the Efficacy and Cross-Reactivity of a Novel Polyclonal Antibody Targeting the Disintegrin Domain in SVMPs to Neutralize Snake Venom

Toxins ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 254
Author(s):  
Shelby S. Szteiter ◽  
Ilse N. Diego ◽  
Jonathan Ortegon ◽  
Eliana Salinas ◽  
Abcde Cirilo ◽  
...  
Keyword(s):  
Snake Venom ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Cross Reactivity ◽  
Snake Envenomation ◽  
Disintegrin Domain ◽  
The Common ◽  
Neutralization Ability ◽  
New Strategies

Snake envenomation can result in hemorrhage, local necrosis, swelling, and if not treated properly can lead to adverse systemic effects such as coagulopathy, nephrotoxicity, neurotoxicity, and cardiotoxicity, which can result in death. As such, snake venom metalloproteinases (SVMPs) and disintegrins are two toxic components that contribute to hemorrhage and interfere with the hemostatic system. Administration of a commercial antivenom is the common antidote to treat snake envenomation, but the high-cost, lack of efficacy, side effects, and limited availability, necessitates the development of new strategies and approaches for therapeutic treatments. Herein, we describe the neutralization ability of anti-disintegrin polyclonal antibody on the activities of isolated disintegrins, P-II/P-III SVMPs, and crude venoms. Our results show disintegrin activity on platelet aggregation in whole blood and the migration of the SK-Mel-28 cells that can be neutralized with anti-disintegrin polyclonal antibody. We characterized a SVMP and found that anti-disintegrin was also able to inhibit its activity in an in vitro proteolytic assay. Moreover, we found that anti-disintegrin could neutralize the proteolytic and hemorrhagic activities from crude Crotalus atrox venom. Our results suggest that anti-disintegrin polyclonal antibodies have the potential for a targeted approach to neutralize SVMPs in the treatment of snakebite envenomations.

scholarly journals The Ability of Zika virus Intravenous Immunoglobulin to Protect From or Enhance Zika Virus Disease

2021 ◽  
Vol 12 ◽  
Author(s):  
Amelia K. Pinto ◽  
Mariah Hassert ◽  
Xiaobing Han ◽  
Douglas Barker ◽  
Trevor Carnelley ◽  
...  
Keyword(s):  
Virus Infection ◽  
Polyclonal Antibody ◽  
Zika Virus ◽  
Virus Disease ◽  
Polyclonal Antibodies ◽  
Antibody Therapeutics ◽  
The World ◽  
Flavivirus Infection

The closely related flaviviruses, dengue and Zika, cause significant human disease throughout the world. While cross-reactive antibodies have been demonstrated to have the capacity to potentiate disease or mediate protection during flavivirus infection, the mechanisms responsible for this dichotomy are still poorly understood. To understand how the human polyclonal antibody response can protect against, and potentiate the disease in the context of dengue and Zika virus infection we used intravenous hyperimmunoglobulin (IVIG) preparations in a mouse model of the disease. Three IVIGs (ZIKV-IG, Control-Ig and Gamunex®) were evaluated for their ability to neutralize and/or enhance Zika, dengue 2 and 3 viruses in vitro. The balance between virus neutralization and enhancement provided by the in vitro neutralization data was used to predict the IVIG concentrations which could protect or enhance Zika, and dengue 2 disease in vivo. Using this approach, we were able to define the unique in vivo dynamics of complex polyclonal antibodies, allowing for both enhancement and protection from flavivirus infection. Our results provide a novel understanding of how polyclonal antibodies interact with viruses with implications for the use of polyclonal antibody therapeutics and the development and evaluation of the next generation flavivirus vaccines.

scholarly journals The Isolation DNA Chromosomal DNA from Local Isolate of Aeromonas hydrophila Bacteria from East Java

2018 ◽  
Vol 6 (1) ◽  
Author(s):  
Retno Sri Wahjuni ◽  
M. Gandul Atik Yuliani
Keyword(s):  
Aeromonas Hydrophila ◽  
Economic Loss ◽  
Dna Purification ◽  
Bacterial Disease ◽  
Bacterial Cells ◽  
Chromosomal Dna ◽  
Fish Farms ◽  
Antigenic Protein ◽  
Ulcer Disease ◽  
Improved Sanitation

Aeromoniasis is a bacterial disease caused by Aeromonas hydrophila that affects fish and shrimps in ponds and aquariums. This is an ulcer disease that causes petechiae in scales of fish and may be fatal. It can cause economic loss if it is not treated with medication accompanied with improved sanitation. It affects numerous freshwater fish farms in East Java. A previous study has characterized an antigenic protein derived from the outer membrane protein of A. hydrophila. In order to perform sequencing of the DNA coding for this protein, we first conducted a study for isolating DNA fragments of A. hydrophila via four stages: cultivation and harvesting of bacterial cells, cell lysis, DNA purification, and the concentration of chromosomal DNA. The results will be used as a predictive immunogenic determinant by the method by Kolaskar and Tongaonkar. Keywords: Aeromonas hydrophila, chromosomal DNA, immunogenic determinant

scholarly journals Subclass-restricted IgG polyclonal antibody production in mice injected with lipid A-rich lipopolysaccharides.

1981 ◽  
Vol 153 (2) ◽  
pp. 324-338 ◽  
Author(s):  
S Izui ◽  
R A Eisenberg ◽  
F J Dixon
Keyword(s):  
Nude Mice ◽  
Polyclonal Antibody ◽  
Lipid A ◽  
Polyclonal Antibodies ◽  
Serum Levels ◽  
Igg Subclass ◽  
Igg Antibodies ◽  
Athymic Nude Mice ◽  
Bacterial Lipopolysaccharides ◽  
Autoantibody Formation

The effects of five distinct bacterial lipopolysaccharides (LPS) on the induction of polyclonal IgM and IgG antibodies, including polyclonal autoantibody formation, were investigated in several strains of mice. Injections of most LPS preparations that contained polysaccharide transiently induced only IgM polyclonal antibodies. However, LPS from Salmonella minnesota R595 (R595 LPS), which had a particularly high content of lipid A but lacked O-antigen polysaccharide, induced a markedly prolonged IgM and IgG polyclonal antibody response in mice, including athymic nude mice, but not in LPS-unresponsive C3H/HeJ mice. Polyclonal IgM and IgG production peaked in sera on day 8 and day 15, respectively, and remained higher than control values 2 mo after the injection. The IgG induced by R595 LPS was strictly restricted to IgG2b and Igg3 subclasses in normal mice. In contrast, in athymic nude mice which have normally lower levels of IgG1 and IgG2a than normal mice, R595 LPS stimulated the production of all the IgG subclasses and reconstituted serum levels of IgG1 and IgG2a up to, but not higher than, control values of normal mice. These findings suggest that different mechanisms regulate production of each IgG subclass after stimulation with LPS.

scholarly journals The third member of the Tetrahymena CCT subunit gene family, TpCCTα, encodes a component of the hetero-oligomeric chaperonin complex

1997 ◽  
Vol 326 (1) ◽  
pp. 21-29 ◽  
Author(s):  
Helena SOARES ◽  
Luisa CYRNE ◽  
Cristina CASALOU ◽  
Bruno EHMANN ◽  
Claudina RODRIGUES-POUSADA
Keyword(s):  
Gene Expression ◽  
Gene Family ◽  
Polyclonal Antibody ◽  
Similar Pattern ◽  
Synthetic Peptides ◽  
Polyclonal Antibodies ◽  
Recombinant Yeast ◽  
Subunit Gene ◽  
The Third ◽  
Tubulin Genes

The sequence of a third member of the Tetrahymenapyriformis chaperonin CCT (‘chaperonin containing TCP1’) subunit gene family is presented. This gene, designated TpCCTα, is the orthologue of the mouse chaperonin gene TCP1/CCTα. To characterize the CCT complex in this ciliate, we have produced polyclonal antibodies against synthetic peptides based on C-terminal sequences deduced from the primary sequences of the TpCCTα, TpCCTγ and TpCCTη subunits. We have also used polyclonal antibodies produced against recombinant yeast CCTα and CCTβ subunits. Using these antibodies, we show that Tetrahymena cells contain a hetero-oligomeric CCT chaperonin comprising at least seven distinct subunits. Three of these were assigned to specific TpCCT genes, whereas a fourth was recognized by the polyclonal antibody against yeast CCTβ, suggesting that this gene is also present in the ciliate. The CCT complex also contains other unidentified proteins that were recognized by the polyclonal antibody UM-1, raised against the putative ATP binding domain of the chaperonin proteins. TpCCTα gene expression was shown in exponentially growing cells and cells regenerating their cilia for different periods to have a similar pattern to the previously identified genes TpCCTγ and TpCCTη, and also to tubulin genes.

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