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

Septicemic Infection Suspect In Jenkins' Whipray Pateobatis jenkinsii (Annandale, 1909): A Case Report

A wild-captive male Jenkins' whipray was found dead in a quarantine tank with a clinical sign before death in the form of decreased appetite for a week. The treatment history was oral administration of enrofloxacin antibiotic tablets. The therapy period lasts for ten days. The last treatment was the administration of Hepavit® (liver extract) and intramuscular injection of enrofloxacin antibiotic. One day before the death, blood was collected and then examined for the hematocrit and some parameters of chemical blood. The results of blood examination were found a decrease in blood urea nitrogen (BUN), alkaline phosphatase (ALP), and alanine aminotransferase (ALT) levels, increased glucose level, decreased total protein and albumin levels, and increased globulin level. Anatomical pathology examination was found lesions on the tail, around the eyes, and claspers. Hemorrhagic lesions were found in the mucous layer of the esophagus, stomach, and spiral colon. The blood clot was found under the tunica layer of testicular organs. The liver is damaged by showing a non-homogeneous coloration, organ thickening, congestion, and fragile consistency. Based on the results of the blood examination and was supported by the results of the anatomical pathology examination after death, it is suspected that the fish died due to the condition of septicemia infection during the previous few weeks

Korelasi Virulen gelE dan Pembentukan Biofilm pada Isolat Enterococcus faecalis yang Diisolasi dari Ayam Pedaging

Enterococcus faecalis merupakan patogen oportunistik yang membentuk biofilm dengan menghasilkan gen virulen seperti gelE. Penelitian ini bertujuan untuk mengeksplorasi keberadaan gelE dan mengkaji korelasinya terhadap pembentukan biofilm pada isolat E. faecalis asal ayam. Sampel yang digunakan sebanyak 60 sampel arsip usap kloaka ayam. Isolasi dan identifikasi bakteri menggunakan media agar selektif diferensial KF Streptococcus. Konfirmasi molekuler menggunakan gen spesifik Efac untuk bakteri E. faecalis dan gen gelE untuk deteksi gen virulen gelatinase. Uji biofilm menggunakan teknik spektrofotometer pada densitas optik 630 nm. Data dianalisis secara kuantitatif menggunakan uji chi-square dengan nilai P < 0,05 dianggap signifikan secara statistik. Hasil isolasi dan identifikasi bakteri terhadap 60 sampel arsip asal ayam pedaging yang digunakan dalam studi sebanyak 21 isolat positif terkonfirmasi secara molekuler sebagai bakteri E. faecalis dan memiliki gen virulen gelE. Pada uji biofilm terdapat sebanyak 20 isolat (95,23%) positif kuat (OD630 > 0,130) dan 1 (4,76%) positif lemah (0,065 < OD630 ≤ 0,130) dengan nilai P < 0,05 atau memiliki korelasi secara statistik antara keberadaan gen gelE dengan pembentukan biofilm. Berdasarkan hasil penelitian dapat disimpulkan bahwa gen virulen gelE telah ditemukan pada 21 isolat E. faecalis dan 95,23% mampu membentuk biofilm dengan intensitas positif kuat

Experimental infection of Enterococcus faecalis in red tilapia (Oreochromis hybrid) revealed low pathogenicity to cause streptococcosis

Background: Streptococcosis, as a bacterial disease with broad tropism in fish and one of the causes of septicemia. Enterococcus faecalis is one of the causative agents of streptococcosis that can be isolated in tilapia.Aim: This study was undertaken to complete the reporting gap on the pathogenicity profile and clinical symptoms of E. faecalis bacterial infection in red tilapia (Oreochromis hybrid). The study is expected to provide enriching information regarding recognizable clinical signs in the field that can lead to the diagnosis of streptococcosis caused by E. faecalis, especially in the Indonesian aquaculture environment.Methods: The method used in this artificial infection study using red tilapia, which were divided into two types of route groups infection, namely intraperitoneal (IP) and peroral (PO) with bacterial concentrations given for each route of infection to be 2.1 × 108 CFU ml−1; 2.1 × 107 CFU ml−1; and 2.1 × 106 CFU ml−1. One group was given brain heart infusion broth media sterile as a non-infectious control. Clinical symptoms, changes in swimming habits and consuming feed, external and internal organ lesion, and leukocytes profile changes were observed during the observation period along 14 days to evaluate the infectious effect of each treated fish group. The lethal dose 50 (LD50) was estimated with the Spearman–Kärber method. The evaluation of the leukocyte profile was performed to find leukocytosis as the clinical sign of infection.Results: The results showed variations in clinical symptoms inflicted on fish through death or the moribund stage. The highest mortality occurred in the treatment group of 2.1 × 108 CFU ml−1 with the PO route. The bacterial concentration of 2.1 × 107 CFU ml−1 given either as PO or IP can cause mild infection symptoms but did not cause mortality. The LD50 of the PO and IP route was obtained at 1.99 × 108 CFU ml−1 and 0.79 × 108 CFU ml−1, respectively. The total leukocytes in the infected fish group increased significantly (p < 0.05) by twofold when compared with the non-infectious group. The bacteria’s discovery on the blood smear examination was taken from fresh dead fish or moribund fish in the treatment group of 2.1 × 108 CFU ml−1, for both PO and IP.Conclusion: Enterococcus faecalis with low pathogenicity can lead to septicemia, characterized by a total increase in leukocytes, bacteria’s discovery on the blood smear examination, and various clinical symptoms systemically found in the treated fish. &nbsp