Native Virus Challenge Test Against Vaccinated Chickens With Commercial Active and Inactive IBD Vaccines

Vaccination trial were conducted on two groups of broiler day-old-chick (DOC) using active and inactive IBD vaccines. First Group was further divided into two groups: one received active IBD vaccination at 7 days of age, and the other received oral vaccination at 7 and 21 days of age, respectively. Both subgroups were then challenged against native IBD isolate, K-5, at 21 days of age. The second Group was arranged similar to the first group, but the second Group received inactive IBD vaccination subcutaneously, at 21 days of age. At 35 days of age, all chickens were challenged against native IBD isolate, K-5. The group which received active vaccine showed pathological change of the bursa, correlated to the bursa/body weight indices less than 0.70 (20 %) and the bursa lesion score (BLS) was 1.56. This pathological change was more obvious after second application of the vaccine. The group which received active and inactive vaccines revealed immune responses with mild lesion in the bursa. This im-munity could not protect the chickens after challenged with K-5 isolate, correlated to the bursa/body weight indices less than 0.70 (100 %) and BLS was 4.0. This results show that K-5 isolate belong to different subtype or variant

Enhancement of transient erythropoietin protein expression by valproic acid in CHO‐K1 suspension adapted cells

Erythropoietin (EPO) is a therapeutic protein that is widely used to increase red blood cell production in chronic kidney failure. EPO protein can be produced quickly with a transient gene expression system (TGE). However, the titer produced using TGE is usually lower than the stable gene expression system (SGE). It has been known that TGE can be improved by histone deacetylase inhibitors (iHDACs) such as valproic acid (VPA). This study was conducted to examine the VPA effect on EPO protein expression in CHO‐K1 suspension adapted cells and to find the optimum concentration of VPA on transient EPO protein production. EPO proteins was quantified using the enzyme‐linked immunosorbent assay (ELISA) method. The optimization of VPA concentrations showed that VPA increased the EPO protein yield by up to 2‐fold in transient EPO production, and the optimum concentration of VPA was 4 mM. VPA optimization was very helpful to obtain the maximum increase in the transiently expressed protein. Furthermore, this study can be used as a model to produce EPO proteins or other recombinant proteins rapidly with TGE of CHO‐K1 suspension adapted cells

Deteksi Spesies Brucella pada Kambing di Rumah Potong Hewan Jakarta

Brucellosis is a zoonosis and occupational diseases transmision. The diseases caused by bacterial and attack multiple species of animals. Common species that infects goats as the most pathogenic species (zoonotic) is Brucella melitensis; however, the species B. abortus could also infect goats. The study purposed to find out the brucellosis seropositive in goat in Jakarta slaughterhouse and to detect caused agent of brucellosis. Sampling was done through slaughtered goats that come from brucellosis endemic area. The samples were collected fromslaughtered mature female goats i.e serum, goat milk, vaginal swab, mamary gland, limphoglandula supramamary, limph, and uterus. The detection method was used i.e patological lession, serological, culture and PolymeraseChain Reaction (PCR) technique. The serological detection of brucellosis in goats was done parallelly between Rose Bengal Test (RBT), Complement Fixation Test (CFT) and Enzyme Linked Immunosorbent Assay (ELISA). The results of this study demonstrated that out of the 119 serum samples serologically tested, negative for RBT, one was positive for CFT and none were positive with ELISA. Patological observation in the Brucella predilection organs, there were 5 goat carcases showed pathological lession (vagina discharge, hemoragy at limphand limphoglandula, crumbly limph and there were pus in uterus). The serum samples that had reacted positively and the organs with pathological lesion were confirmed further with PCR, bacterial isolation and identification.The PCR test results and the culture of milk samples, vaginal swabs and organs did not reveal any Brucella spp bacteria (B. abortus, B. melitensis, B. ovis dan B. suis) and also vaccine strains of RB51. Based on these results, it was concluded that brucellosis in goats on Java Island was a 0.84% seropositive (confidence interval 95%; 0.00826 - 0.00854) (1/119), although the species of Brucella that had infected them remains unknown

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

Rescue with an anti-inflammatory peptide of chickens infected H5N1 avian flu

Chickens suffering from avian flu caused by H5N1 influenza virus are destined to die within 2 days due to a systemic inflammatory response. Since HVJ infection (1,2) and influenza virus infection (3,4) cause infected cells to activate homologous serum complement, the systemic inflammatory response elicited could be attributed to the unlimited generation of C5a anaphylatoxin of the complement system, which is a causative peptide of serious inflammation. In monkeys inoculated with a lethal dose of LPS (4 mg/kg body weight), inhibition of C5a by an inhibitory peptide termed AcPepA (5) rescued these animals from serious septic shock which would have resulted in death within a day (6). Therefore, we tested whether AcPepA could also have a beneficial effect on chickens with bird flu. On another front, enhanced production of endothelin-1 (ET-1) and the activation of mast cells (MCs) have been implicated in granulocyte sequestration (7). An endothelin receptor derived antisense homology box peptide (8) designated ETR-P1/fl was shown to antagonize endothelin A receptor (ET-A receptor) (9) and reduce such inflammatory responses as endotoxin-shock (10) and hemorrhagic shock (11), thereby suppressing histamine release in the circulation (12). Thus, we also administered ETR-P1/fl to bird flu chickens expecting suppression of a systemic inflammatory response

Protection from avian influenza H5N1 virus infection with antibody-impregnated filters

There is worldwide concern over the possibility of a new influenza pandemic originating from the highly pathogenic avian H5N1 influenza viruses. We herein demonstrate that functional air filters impregnated with ostrich antibodies against the hemagglutinin of the H5N1 virus protect chickens from death by H5N1 transmission. These results suggest that the use of ostrich antibody-impregnated filters might be a powerful way to prevent the transmission of H5N1

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

Genetic characterization of S1 gene of infectious bronchitis virus isolated from commercial poultry flocks in West Java, Indonesia

Background and Aim: Infectious bronchitis (IB) is still a major problem among poultry industry in Indonesia, IB outbreaks continue to happen even in vaccinated flocks. The emergence of new IB virus (IBV) variants might lead to mismatching between vaccine virus strain and circulating virus strain, this may be a reason of vaccination failure. Information about circulating IBV in a region is important to decide which IB vaccine should be used. However, information about recent IBV strains which circulated in Indonesia and their genetic characters were limited; therefore, the aim of our research was to determine the genetic characterization of S1 gene of IBV isolated from commercial poultry flocks in West Java, Indonesia. Materials and Methods: A total of 47 viral isolate samples collected from chickens with clinical sign and reduced in egg production. Six IB live vaccines were used as control, the reference vaccines represent IBV strains including H120, H52, 4/91, CR88, 233A, and 1-96. Primers XCe2+ and XCe2- were used to amplify S1 gene partially. Results: Twenty-six of 47 samples showed positive result to S1 gene of IBV by reverse transcription-polymerase chain reaction. Three IBV isolates, Indonesia/K233A31/18, Indonesia/K4A9/17, and Indonesia/P3/17, were selected for nucleotide sequencing. Phylogenetic analysis of 352 nucleotides of the partial S1 gene shows that isolates Indonesia/K4A9/17 and Indonesia/K233A31/18 have 100% homology with IBV vaccine strain 4/91, while isolate Indonesia/P3/17 has 100% homology with IBV vaccine strain 233A. Conclusion: Our result indicates that at least two IBV strains were circulating among poultry in West Java, Indonesia, which is IBV close to vaccine strain 4/91 and 233A. The present study provides updates on the circulating IBV in commercial poultry flocks in West Java, Indonesia, and might use as guidance on selecting a proper IB vaccine strain to improve IB vaccination efficacy in certain region

Deteksi Virus Avian Influenza H5N1 pada Anak Ayam Umur Satu Hari dengan Teknik Imunohistokimia

Avian Influenza (AI) or bird flu caused by virus H5N1 is still present in Indonesia. The Department ofAgriculture of Indonesia has banned poultry distribution from endemic to non endemic area, except fordistribution of day old chick (DOC). The aim of this research was to detect AI virus infection in DOCdistributed from AI endemic to AI non endemic areas. Two hundred and forty DOCs from farms in WestJava and Banten were collected from Soekarno Hatta airport. Their antibody titers were examined againstAI virus by Haemaglutination Inhibition (HI) test. The AI virus detected in tissues (trachea, lung, heart,kidney, liver, and intestine) by immunohistochemistry technique. Detection of AI virus using anti AI H5N1monoclonal antibody was conducted AEC as chromogen. The result showed that 66,2% of DOC were positiveAI and 33,8% were negative AI. The 66,2% of positive samples, 43,3% showing the presence of AI antigenin trachea, lung and intestine, and 22,9% were presence in liver and kidney. DOCs were infected AI viruswith subclinical symptoms and they were potential as the source of rapid AI spread in Indonesia. It istherefore important to take a very cautious measure to prevent the spread of AI via DOC from AI endemicto free area