Mycoplasma is a vertically transmitted disease causing severe economic losses in poultry industry, so this study aimed to prepare a local inactivated a bivalent Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) vaccine. Vaccine was adjuvanted with Montanide ISA70 then evaluated for safety, sterility, and potency. The vaccine efficacy was evaluated in 50 SPF one day old chicks which divided into 5 groups 10 birds each. Chicks were vaccinated with 1 st dose at 7 days of age and boostered with the 2 nd dose after 3 weeks. Three weeks post vaccination with 2 nd dose birds were challenged, chicks were kept under observations for 10 weeks. During the experiment data of clinical signs, daily mortality, body weight, postmortem, serum samples and nasal swabs were collected. serum samples were collected weekly after first vaccination till the third week after challenge and humoral immune response was estimated against Mycoplasma strains using the enzyme-linked immunosorbent assay (ELISA) test and results revealed that vaccines gave a high protective antibody titer, and the vaccine gave 80% protection against MG and 90% against MS. From our result locally prepared inactivated bivalent vaccine gave a promising result in birds protection against serious avian pathogens and considered as an effective tool for the prevention of mycoplasmosis in layers and breeders chicken farms in Egypt.
Aim:The present work aimed to develop lateral flow immunochromatographic strip (ICS) test for detection of Salmonella Enteritidis (SE) specific antibodies in chicken sera.Materials and Methods:A rapid lateral flow immunochromatographic test (LFIT) has been developed, in which SE Group D antigen labeled with the gold chloride molecules laid on the conjugate pad. Staphylococcus aureus protein A was used as capture antibody at the test line (T) of a nitrocellulose (NC) membrane and anti-SE antigen-specific rabbit antibodies were used as capture antibody at the control line (C) of the NC strip in the lateral flow layout device.Results:Using the developed LFIT, the minimal amount of SE-specific antibodies that can be detected in chicken serum sample was 1427 enzyme-linked immunosorbent assay (ELISA) unit/100 µl that was equal to 0.1 µg (Ab)/100 µl sample. 100 suspected serum samples collected from a poultry flock were tested with the prepared SE-LFIT kits and the locally prepared stained Salmonella antigen, and the results were compared with those obtained from examination of these samples with Salmonella Group D antibody ELISA kit as the gold standard test. The sensitivity, specificity, and accuracy of the prepared SE-LFIT antigen kits were 94.4%, 90%, and 94%, respectively, while those obtained with stained Salmonella antigen were 88.8%, 90%, and 89%, respectively.Conclusion:The developed test is a simple field rapid test of high sensitivity, specificity, and accuracy that can improve and facilitates rapid field surveillance of salmonellosis among chickens.
Aim:This work was conducted to study the phenotypic and genotypic characterization of locally isolated Salmonella strains (Salmonella Pullorum, Salmonella Enteritidis, and Salmonella Typhimurium) from poultry used in the preparation of Salmonella antigens in Egypt.Materials and Methods:The phenotypic characterization of Salmonella strains was done using standard microbiological, biochemical, and serological techniques. Molecular identification was done using different sets of primers on different genes using different polymerase chain reaction (PCR) techniques.Results:The phenotypic characterization of Salmonella strains was confirmed. Molecular identification revealed detection of 284 bp fragment of InvA gene in all studied Salmonella strains. Furthermore, multiplex PCR was used for more confirmation of being Salmonella spp., generally at 429 bp as well as genotyping of Salmonella Typhimurium and Salmonella Enteritidis at 559 and 312 bp, respectively, in one reaction.Conclusion:The locally isolated field Salmonella strains were confirmed phenotypically and genotypically to be Salmonella Enteritidis, and Salmonella Typhimurium and could be used for the preparation of Salmonella antigens.
Hyperimmune serum against peste des petites ruminants virus was successfully prepared in horses where it was found to have specific peste des petites ruminants virus (PPRV) neutralizing antibodies titer of 1024/ml as determined by serum neutralization test (SNT). Quality control testing of such serum revealed that it was free from bacterial, fungal and mycoplasma contaminants as tested on specific media and safe when inoculated in sheep. Passive induced immunity in sheep was persisted with a protective level up to 5 weeks post inoculation as determined by SNT and ELISA. On the other side, inoculation of PPR vaccine with antisera in sheep showed similar findings with slight rise in antibody titer. Depending on the obtained results, it could be concluded that horse anti-PPR serum is of a significant importance and can help to protect and control PPR infections especially in case of outbreaks that need rapid management.
Background and Aim: Brucellosis is a zoonotic disease with a worldwide distribution. It has a serious impact on the health of humans and animals, along with a negative impact on the economy. This study aimed to prepare and evaluate the diagnostic performance of a lateral flow immunochromatographic test (LFIT) nanogold diagnostic kit for detecting brucellosis in sheep.
Materials and Methods: A rapidly developed LFIT, in which lipopolysaccharide conjugates with nanogold molecules, was placed on the conjugate pad. One hundred ovine serum samples were tested to detect Brucella antibodies (Ab) using the prepared lateral flow immunochromatography assay (LFA) kit and Rose Bengal test. The evaluation of specificity, sensitivity, and accuracy for LFIT and Rose Bengal plate test was conducted using the P04310-10 IDEXX brucellosis ovine/ caprine Ab enzyme-linked immunosorbent assay (ELISA) test (gold standard).
Results: The lower amount of Brucella Ab in the ovine serum samples was detected and was 1.58 S/P ratio ELISA titer/100 μL using LFIT and with Rose Bengal to detect 1.86 S/P ratio ELISA. The results showed that the developed LFIT had high specificity with no cross-reactivity with other tested bacteria. The calculated sensitivity, specificity, and accuracy of LFIT and Rose Bengal test using the P04310-10 IDEXX brucellosis ovine/caprine Ab ELISA test (gold standard) were 74% and 89%, 81% and 59%, and 76.9% and 66%, respectively.
Conclusion: The present results showed interesting results implying that the LFIA strip test could be used as a substantial diagnostic tool for field screening ovine Brucella as an essential step in the control of brucellosis. However, further studies for the validation of the present findings are necessary.
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