Development of SarS-Cov-2 antigen Detection Kit Based on Immunoglobulin Y (Igy) Using Surface Plasmon resonance (SPr)

SARS disease reappeared at the end of 2019 with a new name as Coronavirus Disease 2019 (COVID-19) caused by a new virus called SARS-CoV-2. This virus has spread throughout the world until recently and caused massive deaths and losses. The nucleic acid test in the form of real-time reverse transcriptase-polymerase chain reaction (RT-PCR) is very important to diagnose COVID-19 in patients, but this method has several drawbacks such as operators who have to be trained, the diagnosis results appear in a relatively long time, and the examination price relatively expensive. This research was conducted to produce immunoglobulin Y (IgY) extracted from chicken egg yolk targeting the S-protein receptor-binding domain (RBD) on SARS-CoV-2 as a component of the surface plasmon resonance (SPR) SARS-CoV-2 antigen detection kit. This research was started by extracting IgY from hyperimmune chicken egg yolks with the polyethylene glycol (PEG) precipitation method and continued with dialysis. The extracted IgY was further purified using thiophilic adsorption chromatography and concentrated by using Amicon® Ultra-15 ultrafiltration. The IgY activity against SARS-CoV-2 RBD was tested qualitatively using the agar gel precipitation test (AGPT) technique and the total protein content was determined using the Lowry method. IgY was tested for its affinity against SARS-CoV-2 RBD using SPR. The IgY concentration obtained was 11 mg/mL. The AGPT result showed the presence of IgY activity against SARS-CoV-2 RBD isolated from egg yolk and chicken serum after 8 weeks after the first vaccination of chickens. The SDS-PAGE results showed a very clear band of IgY characters. The obtained IgY showed adequate interaction with commercial SARS-CoV-2 RBD on an SPR device. The purified IgY was able to bind with protein-S RBD and showed a fairly good affinity for the SARS-CoV-2 antigen sample. The results of these observations indicate that IgY anti-S-protein SARS-CoV-2 can be produced and purified from chicken egg yolk and can be used as a diagnostic component to detect SARS-CoV-2 antigen, especially on SPR.

Detection of Chicken Egg Embryos using BW Image Segmentation and Edge Detection Methods

This study aims to identify chicken egg embryos with the concept of image processing. This concept uses input and output in images. Thus the identification process, which was originally carried out using manual observation, was developed by computerization. Digital images are applied in identification by various image preprocessing, image segmentation, and edge detection methods. Based on these three methods, image processing has three processes: image grayscaling (convert to a grayscale image), image adjustment, and image enhancement. Image adjustment aims to clarify the image based on color correction. Meanwhile, image enhancement improves image quality, using histogram equalization (HE) and Contrast Limited Adaptive Histogram Equalization methods (CLAHE). Specifically for the image enhancement method, the CLAHE-HE combination is used for the improvement process. At the end of the process, the method used is edge detection. In this method, there is a comparison of various edge detection operators such as Roberts, Prewitt, Sobel, and canny. The results of edge detection using these four methods have the SSIM value respectively 0.9403; 0.9392; 0.9394; 0.9402. These results indicate that the SSIM values ​​of the four operators have the same or nearly the same value. Thus, the edge detection method can provide good edge detection results and be implemented because the SSIM value is close to 1.00 (more than 0.93). Image segmentation detected object (egg and embryo), and the continued process by edge detection showed clearly edge of egg and embryo.

Morphfological changes in the membranes of the trachea of guinea pigs in experimental ovalbumin-induced allergic inflammation of the airways.

Background. In recent years, there has been an increase in the number of allergic diseases of the respiratory organs, especially in children. The predictor of the further development of bronchial asthma is sensitizing at an early age to the allergens of chicken egg. The use of new knowledge about the allergenic components of the chicken egg will predict the risks and clinical features of the disease. Despite the importance, the problem of morphogenesis of allergic inflammation of the wall of the trachea is not sufficiently studied, so far many issues are not found in morphology and require further research. Objective.The aim of the study is to establish morphological changes in the tracheal membranes in experimental ovalbumin-induced allergic inflammation of the airways of guinea pigs. Methods. The thickness of tracheal wall of 48 male guinea pigs was investigated by histological, morphometric, statistical methods on the twenty-third, thirty-sixth, thirty-sixth and forty-fourth days after the initiation of the experimental ovalbumin-induced allergic inflammation of the airways. Results. We have found, that maximum statistically significant thickening is shown in the late period of tracheal mucosa in 2 times on the 44th day of observation and tracheal submucosa in the 3rd experimental group on the 36th day of observation (increasing coefficient 2) compared to the control. However, the thinning of tracheal submucosa is observed in the early period of the inflammatory process on the 23rd and 30th day of observation. It has been proved, that the allergic inflammation of the tissues of the trachea caused by the sensitization and allergization of ovalbumin leads to the change in the thickness of layers of trachea in the chronobiological aspect. Conclusion. On the 23rd and 30th days of the experiment, thinning of tracheal mucosais observed due to damage of epithelial cells. Thickening of tracheal mucosa and submucosa was found in the third and fourth groups of observation (late period of allergic inflammation) compared with animals of the intact group and the control group due to an increase in the area of loose connective tissue, which is a consequence of the continuation of the allergic inflammatory process in the trachea after the end of the experiment.

Effect of pack carburizing with chicken egg shell powder agent and vibrator quenching on the mechanical properties of AISI 9310 steel

The effects of vibrator quenching (QV) on the carbon content, microstructure, and mechanical properties (surface hardness number, wear resistance) in the pack carburizing of AISI 9310 steel were studied. The aim of this research is to increase the surface hardness and improve the wear resistance of AISI 9310 steel. The problem that often occurs in the quenching treatment after pack carburizing is that the thick cooling medium does not evenly wet the surface of the specimen, so that the cooling rate is not uniform, the impact is the distribution of the specimen surface hardness is not the same. Therefore, it is necessary to research the implementation of the vibrator in the quenching treatment. The specimens were treated with pack carburizing at a temperature of 875 °C, soaking time for 3 hours. The carburizing agent consisted of chicken egg shell powder (CESP) and rice husk charcoal (RHC) with various weight ratios of 5 %:95 %, 15 %:85 %, and 30 %:70 %. Followed by quenching treatment using a 10 % cane molasses cooling medium and vibrator. Hardness testing was carried out using a Vickers microhardness tester, wear resistance test using the pin-on-disc method, and a scanning electron microscope (SEM-EDX) was used to observe changes in the microstructure and carbon elemental content on the specimen surface. The results showed that the application of VQ caused the formation of a small martensite microstructure while without VQ it was large martensite and a few of residual ferrite. The highest surface hardness number is 685 kg/mm2, the wear resistance is 0.32 cm/mg for pack carburizing, using carburizing agent 70 % RHC, 30 % CESP and VQ. VQ causes a more even distribution of the thick cane molasses cooling medium so that the cooling rate of the specimens is uniform.

A Pioneer Study on a Non-invasive Method for in Ovo Chicken Egg Sexing.

Background: Day-old male chicks culling is one of the world most inhumane problems in the poultry industry. Every year, seven billions of male chicks are being killed in laying-hen hatcheries, due to their higher feed exchange rate, lower management compare to female ones and production costs. This work reports a novel non-invasive method for the gender identification of chicken eggs. Four electrodes were attached onto each egg during the incubation process for the data gathering period of fourteen days. Standard polymerase chain reaction (PCR) based chicken gender determination protocol was applied to the eggs on the last day of the incubation process to get the gender information. Result: A relationship between the collected data and the gender of the egg was built, and it was found to have a reliable connection, indicating that by measure the impedance data of the eggs from day 9 of incubation with the four electrodes setting and applying the self-normalization technique, we can determine the chicken egg gender.Conclusion: This is a pioneer founding, proving that impedance spectroscopy can be use to sexing of chicken eggs, before its life has been formed, relieving the poultry industry from such an ethical burden.