THE BIOLOGICAL FUNCTIONS OF IMMUNOGLOBULIN Y (IgY) MOLECULES IN AGAINST INFECTION OF Enterococcus faecalis ORIGIN OF RED TILAPIA

Red tilapia (Oreochromis hybrid) is Indonesia's leading freshwater fishery commodity susceptible to streptococcal bacterial infection. Many studies have been conducted on various efforts to prevent and treat this disease, one of which uses the immunoglobulin Y (IgY) molecule from chicken egg yolk. This study aimed to observe the biological function of IgY against Enterococcus faecalis as a cause of streptococcal-like infection. The agglutinin function was conducted by observing the growth of Enterococcus faecalis in brain heart infusion (BHI) broth media which was added with IgY suspension. The function of inhibin was performed using a spectrophotometric method to measure the level of turbidity of the bacterial suspension inoculated with IgY suspension. The bactericidal potential through the complementary activation pathway for red tilapia serum was carried out using a scanning electron microscope (SEM) method to evaluate the topography of the bacterial cell wall. The results of the study can be concluded that IgY anti-Enterococcus faecalis has the potential as an agglutinin, inhibin, and bactericidal agent through its putative potential in complement activation in streptococcal bacterial infections in red tilapia commodities

THE POTENTIAL OF ADJUVANT AGAINST PRODUCTION OF ANTISTREPTOCOCCAL IMMUNOGLOBULIN Y (IGY) IN AQUACULTURE

This study was conducted to explore the potential of adjuvant for the production of immunoglobulin Y (IgY) as antistreptococcosis in layer chicken with mass production orientation. Enterococcus faecalis which causes streptococcosis in the red tilapia was selected as a candidateantigen. The production of immunoglobulin Y (IgY) was carried out on Isa Brown layer chickens and aged around 20 weeks. Furthermore, thechickens were grouped into four groups (A, B, C, and D groups), each consisting of three chickens based on the type of adjuvant, while twochickens were used as a control group. Each group was treated by giving MONTANIDE™ ISA 71R VG adjuvant (A), Freund's adjuvant (B), aluminum potassium sulphate adjuvant (KAl(SO4)2∙12H2O) concentration of 50 ppm in pH 7 (C), and only antigens without adjuvant (D). Chickens were kept for 35 days and each week was checked for presence the IgY antigen in the serum and egg yolk. Booster was conducted on 14th and 28th days of maintenance. The results showed that IgY in treatment group A was detected on day 28 in the serum and day 35 in the yolk. Whereas the treatment group B could be detected on day 35 in the serum. However, the IgY was not detected in the serum and yolk in C, D, and control groups until the end of the maintenance. Based on the results, it can be concluded that the appearance of IgY in serum and yolk in a relatively fast time is obtained in the combination of Enterococcus faecalis antigen with the emulsion of water-in-oil adjuvant (SEPPICMONTANIDE™ ISA 71R VG) compared to the other types of adjuvant that use in this study

Chicken Enterococcus faecalis-induced immunoglobulin Y as a prophylactic and therapeutic agent against streptococcosis in red tilapia (Oreochromis hybrid)

Background and Aim: Streptococcosis is a common bacterial disease in red tilapia, in which Enterococcus faecalis infection has not been widely reported. This study aimed to evaluate the efficacy of pellets that contain chicken E. faecalis-induced immunoglobulin Y (IgY) to treat and prevent streptococcosis in red tilapia. Materials and Methods: We conducted a 28-day study for immunoprophylaxis and immunotherapy, each using four groups with two replications: Healthy control fish (KS), non-IgY pellets (PA and TA), pellets with 25% egg yolk containing E. faecalis-induced IgY (PB and TB), and pellets with 50% egg yolk containing E. faecalis-induced IgY(PC and TC). Indirect enzyme-linked immunosorbent assay was performed on prototype pellets produced with an IgY suspension at 1.63 mg/mL as the standard optical density curve. For the immunoprophylaxis study, pellets of 3% of the average body weight of the experimental fish (0.50 g per fish per day) were given daily until day 14 before the challenge test with E. faecalis (2.1 × 109 Colony-forming unit/mL peroral) on day 15. The data from the observation period on days 15–28 were analyzed. For the immunotherapy study, pellets of 3% of the average body weight (0.50 g per fish per day) were given daily for 21 days (days 8–28) 7 day spost-infection. The data from the immunotherapy study were collected during the observation period on days 8–28. Statistical analysis was performed on non-specific immune variables: Total leukocytes, monocytes, lymphocytes, neutrophils, phagocytic activity, and macrophage capacity; and the semi-quantitative distribution of melanomacrophage centers (MMCs) in the lymphoid organs, such as spleen and liver. Photomacrographic data were analyzed descriptively and qualitatively by comparing the healing process and clinical signs found between experiments in the immunotherapy study. Results: The pellet with 50% egg yolk with an IgY at 2.43 mg/g pellet, 3% of body weight once daily, was the best formula on experimental fish. The administration of this formulation can also increase non-specific immunity and the distribution of MMCs in the spleen and liver with a survival rate of 55% for 14 days of challenge period in the immunoprophylaxis study and 70% for 21 days of therapy period in the immunotherapy study. Conclusion: Immunoglobulin Y can be a prophylactic and therapeutic agent against streptococcal infections caused E. faecalis in red tilapia with an optimum dosage of 2.43 mg/g pellet

Cross Reaction of Serum in Salmonella enteritidis- Vaccinated Chicken to Some Salmonella enterica Serotypes (REAKSI SILANG SERUM AYAM YANG DIVAKSIN DENGAN SALMONELLA ENTERITIDIS TERHADAP BEBERAPA SEROTIPE SALMONELLA ENTERICA)

Salmonella spp. has been recognized as the major cause of food-borne illness in humans worldwidecausing remain relevant to public health. Poultry vaccination is one promising strategy to mitigateSalmonella infection in poultry and, in turn, in humans as well. The objective of this study was to assessthe potential of cross-reaction of serum in Salmonella enteritidis-vaccinated chicken to some serotype ofSalmonella enterica. Four female, Isa Brown layer chickens (20 weeks old), were vaccinated with S. enteritidisstrain Sm24/Rif12/Ssq (intra vena) to induced the production of specific antibodies in serum. Crossreactionof serum in S. enteritidis-vaccinated chicken were assess with agar gel immunodiffusion test(AGID) with S. enteritidis, S. pullorum, S. typhimurium, S. typhi, and Escherichia coli antigens. Serumcould react with S. enteritidis and all types of S enterica used in this study (S. pullorum, S. typhimurium,S. typhi), but could not react with E. coli. The potential of cross-reaction of serum in S. enteritidis-vaccinatedchicken to some serotypes of S. enterica may play a role in reducing the infection caused by that serotype

DEVELOPMENT OF A RAPID DIAGNOSTIC KIT FOR DETECTION OF SALMONELLA ENTERITIDIS IN FOOD USING INDIRECT COAGGLUTINATION TECHNIQUE

The objective of this study was to develop a rapid, simple, cheap, sensitive, and specific assay for detection of Salmonella Enteritidis in food. The kit-prototype was developed by using indirect coagglutination technique with three main components, namely Staphylococcus aureus Cowan I, rabbit IgG anti-chicken Fc IgY and chicken IgY anti-S. Enteritidis. Isa Brown layer chickens were used to produce specific antibodies against S. Enteritidis. Monospecific antisera were prepared by absorption method. Staphylococcus aureus Cowan I was coupled with rabbit IgG anti-chicken Fc IgY and monospecific antisera anti-S. Enteritidis. Kit-prototype was compared with multiplex polymerase chain reaction to determine sensitivity and specificity of kit-prototype. Artificially inoculated food sample was used to determine the limit of detection of kit-prototype in a food sample. Indirect coagglutination kit-prototype was able to differentiate positive control from negative control without self-agglutination reaction. This assay has a high specificity to S. Enteritidis without significant cross-reactivity towards other bacteria. Kit-prototype was able to detect 108 CFU/mL of S. Enteritidis in the buffer and 1 CFU/mL of S. Enteritidis in a food sample after selective enrichment procedure. The application of this kit was able to give a fast result (reaction can be observed in 10 sec), to be applied in a sample without extraction in the preparation of antigen and to reduce detection time of S. Enteritidis in food until 4 days

Phenotypic and Serotypic Characterization of Staphylococcus aureus Strains from Subclinical Mastitis Cattle (KARAKTERISASI SECARA FENOTIPE DAN SEROTIPE STAPHYLOCOCCUS AUREUS YANG BERASAL DARI MASTITIS SUBKLINIK PADA SAPI)

Staphylococcus aureus is known as a major causative agent of mastitis in dairy cattle. In the presentstudy, 104 isolates of Staphylococcus originated from subclinical mastitis cattle characterized for thephenotypic properties and the presence of Staphylococcal protein A (Spa). Some bacteria were resistancesagainst several antibiotics were also studied, such as erythromycin, streptomycin, tetracycline, cefepime,nitrofurantoin, amikacin, chloramphenicol, and ciprofloxacin. About 78% of the isolated were moderatelysensitive to nitrofurantoin, while 89% were highly resistant to cefepime and ciprofloxacin. Using thevarious mammals’ sera, seven isolates out of 104 revealed the presence of Spa

PHENOTYPIC AND GENOTYPIC ANALYSIS OF A TRUEPERELLA BERNARDIAE STRAIN ISOLATED FROM A DOG

The present study was designed to characterize phenotypically and genotypically the Trueperella bernardiae strain (T. bernardiae P5459/15) isolated from the purulent thelitis of a dog. The species identity could be confirmed by phenotypical investigations, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis, and sequencing various molecular targets. Sequencing the 16S rDNA, the β subunit of bacterial RNA polymerase encoding gene rpoBand, and the glyceraldehyde-3-phosphate dehydrogenase encoding gene gap revealed sequence similarities to the type strain T. bernardiae DSM 9152 of 99.6%, 99.3% and 99.1%, respectively. The present study is the first phenotypic and genotypic characterization of the T. bernardiae strain isolated from a dog

PHENOTYPIC AND GENOTYPIC ANALYSIS OF AN ARCANOBACTERIUM PLURANIMALIUM ISOLATED FROM A MUSKOX (OVIBOS MOSCHATUS)

The present study was designed to characterize an <i>Arcanobacterium pluranimalium</i> strain isolated from a muskox (<i>Ovibos moschatus</i>) phenotypically, by MALDI-TOF MS analysis and genotypically using various molecular targets. The phenotypic properties, the MALDI-TOF MS analysis and sequencing the 16S rRNA gene, the β subunit of bacterial RNA polymerase encoding gene <i>rpoB</i>, the glyceraldehyde 3-phosphate dehydrogenase encoding gene <i>gap</i>, the elongation factor tu encoding gene <i>tuf</i> and the pluranimaliumlysin encoding gene <i>pla<i/> allowed a successful identification of the isolated strain as <i>A. pluranimalium</i>. Gene <i>pla</i> could also be detected by a previously described loop-mediated isothermal amplification (LAMP) assay. This is first report on the isolation and characterization of <i>A. pluranimalium</i> originated from a Muskox

Hydroxyapatite-Gold Modified Screen-Printed Carbon Electrode for Selective SARS-CoV‑2 Antibody Immunosensor

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), or coronavirus disease 2019 (COVID-19), is still spreading worldwide; therefore, the need for rapid and accurate detection methods remains relevant to maintain the spread of this infectious disease. Electrochemical immunosensors are an alternative method for the rapid detection of the SARS-CoV-2 virus. Herein, we report the development of a screen-printed carbon electrode immunosensor using a hydroxyapatite-gold nanocomposite (SPCE/HA-Au) directly spray-coated with the immobilization receptor binding domain (RBD) Spike to increase the conductivity and surface electrode area. The HA-Au composite synthesis was optimized using the Box–Behnken method, and the resulting composite was characterized by UV–vis spectrophotometry, TEM-EDX, and XRD analysis. The specific interaction of RBD Spike with immunoglobulin G (IgG) antibodies was evaluated by differential pulse voltammetry and electrochemical impedance spectroscopy methods in a [Fe(CN)6]4–/3– solution redox system. The IgG was detected with a detection limit of 0.0561 pg mL–1, and the immunosensor had selectivity and stability of 103–122% and was stable until week 7 with the influence of storage conditions. Also, the immunosensor was tested using real samples from human serum, where the results were confirmed using the chemiluminescent microparticle immunoassay (CMIA) method and showed satisfactory results. Therefore, the developed electrochemical immunosensor can rapidly and accurately detect SARS-CoV-2 antibodies