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Latex agglutination test for field diagnosis of haemorrhagic septicaemia Lily Natalia
Jurnal Ilmu Ternak dan Veteriner Vol 6, No 3 (2001): SEPTEMBER 2001
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Pasteurella multocida is bacterial pathogens that cause haemorrhagic septicaemia (HS) in cattle and buffaloes. Various tests have been used to differentiate types of P. multocida, as well as to diagnose this specific disease. A latex agglutination test has been developed for the detection of P. multocida B:2 which is the causal agent of HS. This test is a rapid and simple test suitable for local laboratorium to diagnose HS cases in the field. A heat stable antigen consisting of mainly lipopolysaccharide (LPS) extract of formalin killed P. multocida 0019 was used to produce specific antibody against P. multocida B:2. The antiboy was then used to sensitise latex particles. Latex agglutination test have been used to screen some P. multocida field isolates and this test have been proven to be specific, simple and easy to be used in detecting P. multocida B:2. The specificity is due to antibodies recognising LPS or LPS protein complexes. Sensitised latex was stable at 4° C for at least12 months. This test should be used as an aid to diagnosis and employed principally to confirm and support clinical and post mortem findings of HS. Keywords: Pasteurella multocida B:2, haemorrhagic septicaemia (HS), diagnosis, latex agglutination test

Prevention of enterotoxacmiu in transported cattle Lily Natalia; Sudarisman .; M Darodjat
Jurnal Ilmu Ternak dan Veteriner Vol 2, No 1 (1996)
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Fatal enterotoxaemia of transported cattle is frequently reported in Indonesiu. Acute enteritis and fatal enterotoxaemia of cattle and buffaloes in Indonesia are associated with toxigenic Clostridium perfringenr type A. The outbreaks could have been caused by some kinds of stress, such as a possible change in nutrition or management as well as transportation . To reduce mortality rate caused by enterotoxaemia, an effective vaccine against the disease was produced . The vaccine was made in an alum precipitated toxoid form, prepared from Clostridlumperfringens type A toxin, which was then tested for safety in mice and for its capacity in generating high immunity in cattle. The vaccine was then used to immunise transported cattle as an attempt to reduce mortality rate and to observe antibody response of cattle following vaocletdon. The results showed that mortality in vaccinated was lower than in non-vaccinated groups of cattle. From field observation, it was obvious that alum precipitated toxoid vaccine could produce good immune response against enterotoxaemia in cattle . It was also evidence that this vaccine could reduce mortality in transported cattle . Keywords: Enterotoxaemia, vaccine, transportation, cattle

Development of ELISA technique for detecting Clostridium novyi alpha toxin Lily Natalia
Jurnal Ilmu Ternak dan Veteriner Vol 2, No 3 (1997)
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14334/jitv.v2i3.69

the investigation of infectious necrotic hepatitis . Modification of the indirect ELISA technique in the system used is an inhibition ELISA for antigen . The test samples were liver fluid from dead animals suspected from infectious necrotic hepatitis. To analyse for alpha toxin, microtiter plates were coated with antigen or alpha toxin . The liver fluid sample thought to contain antigen or toxin was then mixed together with reference antiserum containing specific alpha antitoxin of Cl. novyi. Enzyme labelled antiglobulin was then added, followed by enzyme substrate . The difference in colour change between a reference sample containing no antigen or toxin and the test sample solution indicates the amount of antigen or toxin in the test samples . This is a competitive assay; high toxin concentrations result in less colour at the end of the test . The sandwich ELISA technique was sensitive enough to detect as little as 390 ng/ml toxin in liver fluid sample. These results indicate that the ELISA technique is useful for detecting alpha toxin of Cl. novyi and for diagnosing infectious necrotic hepatitis . Keywords: ELISA technique, Clostridium novyi alpha toxin, infectious necrotic hepatitis

Clostridial necrotic enteritis: field cases and the role of vaccination against coccidiosis to the incidence in broiler chicken Lily Natalia; A Priadi
Jurnal Ilmu Ternak dan Veteriner Vol 8, No 1 (2003): MARCH 2003
Publisher : Indonesian Center for Animal Research and Development (ICARD)

A study of necrotic enteritis in broiler chicken was carried out in West Java. Isolates of highly toxigenic C. perfringens of type C were isolated from field cases of necrotic enteritis which most frequently occured in chicken with vaccine-inducedcoccidial lesions. Experimental study in broiler chicken was conducted to determine the predisporing factor for necroric enteritis. Three possible factors were evaluated, those were feeding with high animal protein ingreadients, vaccination with live attenuated anticoccidial vaccine, and feeding with spores of C. perfringens type A and C. The occurrence of intestinal coocidial leions in chicks vaccinated with life attenuated anticoccidial vaccine was demonstrated. The highest mortality and the significant intestinal lesions of chickens were observed in group treated with live attenuated anticoccidial vaccine and C. perfringens type A and C spores (P<0.01). From the experiment, it was found that vaccine induced-coccidial lesions and C. perfringens type A and C were the predisposing factors of necrotic enteritis. These results suggest that the concurrent infection with coccidia and C. perfringens has a synergistic effect on mortality and intestinal lesions in necrotic enteritis. Key words : Clostridial necrotic enteritis, broiler chicken, coccidiosis

Phage typing and sensitivity test to antibiotics of Salmonella enteritidis isolates from Indonesia Sri Poernomo; Adin Priadi; Lily Natalia
Jurnal Ilmu Ternak dan Veteriner Vol 11, No 2 (2006): JUNE 2006
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Salmonella enteritidis (SE) is frequently implicated in disease outbreaks such as human food poisoning. Phage typing have been proved to be a valuable and sensitive tool in the control of SE infections. The ability of phage to distinguish varieties among apparently identical serotypes led to the development and acceptance of phage typing as a significant epidemiological procedure. To determine the epidemiological pattern of SE, phage typing of 53 SE isolated from various sources in Indonesia during 1991–1999, has been conducted using 16 typing phages of phage typing scheme of SE obtained from the International Collaborating Center for Enteric Phage typing, Central Public Health Laboratory, Colindale, UK. The lyse blood isosensitest was then used to test the sensitivity of the Salmonella isolates to antibiotics. The phage typing results obtained that of 53 Salmonella isolates there were one S. infantis, one S. berta, and 46 SE phage type 4, 2 SE phage type 7 (from chicken and water), 1 SE phage type 6 (from chicken) and 2 SE phage type 1 (from chicken). SE phage type 4 isolates comprised of 2 isolates from human, 19 isolates from chicken (young and adult), 17 isolates from day old chicks, 4 isolates from fluff, 2 isolates from chicken meat, 1 isolate from poultry farm water, 1 isolate from dog organ. These findings indicated that contaminated chicken appeared to be the sources of human and dog for SE infection. The results of sensitivity test of the isolates to antibiotics showed that most of the Salmonella isolates from Indonesia were resistant to the antibiotics tested. Key Words: Salmonella Enteritidis, Phage typing, Sensitivity test, Indonesia, Chicken

Effects of irradiation on the survival of bacterial contaminants in food Lily Natalia; A. Priadi; Z. Irawati
Jurnal Ilmu Ternak dan Veteriner Vol 14, No 1 (2009): MARCH 2009
Publisher : Indonesian Center for Animal Research and Development (ICARD)

The primary concern about microbial safety of irradiated food is the survival of pathogenic spore forming bacteria. Clostridium sporogenes was selected as the spore forming test organism for conducting inoculated pack studies for its similarities to the most toxigenic Cl. botulinum, in radiation resistance. Minimum radiation dose applied (45 kGy under cryogenic condition, in -790C) was determined to eliminate Cl. sporogenes spores and other bacterial contaminants in different kind of Indonesian chicken and beef dishes. In separate studies, irradiation doses of 3 – 7 kGy at cryogenic condition was used to improve the microbiological safety of a number chilled prepared meals. The dishes or ready to eat foods were packaged in air impermeable pouches. Irradiation process was carried out after inoculation on chicken and beef dishes with certain amounts of Cl. sporogenes spores. The evaluation of colony count differences between the irradiated and unirradiated foods revealed the effect of radiation on the survival of bacterial spores or other bacterial contaminants. It was demonstrated that a minimum dose of 45 kGy under cryogenic condition eliminated the spore of Cl. sporogenes, Bacillus spp and Staphylococcus spp. Irradiation at doses 5-7 kGy significantly reduced some potential pathogenic microorganisms in samples without affecting quality up to 3 months of storage at the refrigeration temperature. Key Words: Food Safety, Irradiation, Bacterial Contaminants

Infection of Ornithobacterium rhinotracheale (ORT) in chickens in Indonesia Adin Priadi; Lily Natalia
Jurnal Ilmu Ternak dan Veteriner Vol 11, No 1 (2006): MARCH 2006
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Ornithobacterium rhinotracheale is a bacterium identified as a new species in 1994 and generally associated with respiratory distress in chickens. From 214 of sinus swabs, tracheal swabs, lungs, airsac, liver heart blood samples and yolk sacs of chickens suffered from respiratory distresses, 6 isolates of O. rhinotracheale were isolated. These isolates were obtained from tracheal swabs of broiler chickens aged between 28-35 days old and broiler breeder of 32 weeks old. Upon incubated on blood agar for 48 hours at 37oC in a 5% CO2 atmosphere, round, convect and grey colonies with diametres of 1-2 mm were observed. The bacteria were pleomorphic, Gram negative rods, negative catalase and positive oxidase. Biochemically, the bacteria did not change potassium nitrate, tryptophan, glucose, arginine, urea, esculin, gelatine, arabinose, mannose, mannitol, N-acetyl-glucosamine, maltose, gluconate, caprate, adipate, malate, citrate and phenyl-acetate in API 20 NE system but β –galactosidase was produced. In the API 20 NE system, the isolates were identified as 0020004, 0060004, 0020104 codes. Tracheitis, air sacculitis, pneumonia and cheesy air sacs were pathological changes generally found in chickens infected with O. rhinotracheale. Trachea is the most important organ for the isolation of O. rhinotracheale. Key Words: Ornithobacterium rhinotracheale, Infection, Chicken, Indonesia

Clostridial necrotic enteritis in chicken associated with growth rate depression Adin Priadi; Lily Natalia
Jurnal Ilmu Ternak dan Veteriner Vol 13, No 1 (2008): MARCH 2008
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Clostridium perfringens (C. perfringens) is a normal inhabitant of the intestinal tract of chickens as well as a potential pathogen causing necrotic enteritis. C. perfringens only causes necrotic enteritis when it transforms from non-toxin producing type to toxin producing type. The alpha toxin, (phospholipase C) is believed to be a key to the occurrence of Clostridial necrotic enteritis (CNE). The best known predisposing factor is mucosal damage, caused by coccidiosis that damages the intestinal lining, making the gut susceptible to infections including C. perfringens. The purpose of this study was to observe the chicken performance in experimental CNE and field cases of CNE. Diagnosis of CNE were made by latex agglutination test, isolation and identification of the agent. Pathological and histopathological changes were also observed. Experimentally, NE could be reproduced when Eimeria sp and C. perfringens spores are inoculated in chicken. Signs of an NE are wet litter and diarrhea, and an increase in mortality is not often obvious. The depression of growth rate and feed efficiency of chicken become noticeable by week 5 because of damage to the intestine and the subsequent reduction in digestion and absorption of food. Subclinical form of CNE was also frequently found in the field, leading to significant decreases in performance. Chicken gut samples examinations revealed that subclinical form of CNE causes damage to the intestinal mucosa caused by C. perfringens leads to decreased digestion and absorption, increased feed conversion ratio and reduced weight gain. Dual infection with C. perfringens and Eimeria sp. was frequently found in field. The results of these studies provide evidence for C. perfringens as a causative bacteria for growth depression. Key Words: Clostridial Necrotic Enteritis, Chicken, Growth Depression

Production and purification of Bacillus anthracis protective antigen Simson Tarigan; Rahmat S Adji; Lily Natalia
Jurnal Ilmu Ternak dan Veteriner Vol 10, No 3 (2005): SEPTEMBER 2005
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Protective antigen (PA) plays crucial roles in the pathogenicity and virulence of Bacillus anthracis. Animals or human immunised with the protein acquire a complete protection against the disease. In addition to vaccine, PA can also be developed into a sensitive diagnostic test for anthrax. The purpose of this study was to produce PA using a culture medium easily obtained, and to develop a simple and effective technique for purification of the protein. To produce PA, B. anthracis Sterne 34F2 strain was first grown on blood agar, then bacterial colonies were suspended and incubated for 2 hours in RPMI-1640 supplemented with NaHCO3 and Tris. Protein components in the culture supernatant were separated consecutively with Phenyl sepharose, Qsepharose and Superdex-200 columns. This order was used in order to simplify and speed up the purification process. The PA contained in the fractions was detected by a dot blot or an ELISA using commercial PA specific antibody. The PA was absorbed strongly by the phenyl sepharose whereas other proteins were absorbed weakly or not absorbed at all. When these PA-containing fractions were loaded into Q-sepharose column, PA was absorbed considerably weaker than contaminated proteins. Although the level of purity obtained from the Q-sepharose column was satisfactory, further separation on Superdex produced an even higher purity. However, on SDS-PAGE analysis, the purified PA was seen as a two-band protein (54.7 and 29.2 kDa) because of nicked proteolysis. On an immunoblot assay, only the 54.7 band was recognised by the PA-specific antibody. Despite the nick proteolysis, the PA purified in this study was considered to retain its biological activities. Key Words: Bacillus anthracis, Protective Antigen, Protein Purification

Evaluation of antibody responses of cattle and buffaloes to Clostridium perfringens type A vaccine using ELISA Lily Natalia
Jurnal Ilmu Ternak dan Veteriner Vol 1, No 3 (1995)
Publisher : Indonesian Center for Animal Research and Development (ICARD)

An enzyme-linked immunosorbent assay (ELISA) was developed to detect specific antitoxin Clostridium perfringens type A. It was based on the use of purified alpha toxin of Clostridium perfringens type A as the coating antigen, which was then linked to specific alpha antitoxin . Horse raddish peroxidase labelled IgG was used as the conjugate, and 2,2-azino-bis (3- ethylbenzthiazoline-6-sulfonic acid) (ABTS) as the substrate . The ELISA was used to evaluate vaccination results on cattle and buffaloes against enterotoxaemia caused by Clostridium perfringens type A.

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