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Economic impact of enterotoxigenic Eschericia coli vaccine used to control piglet neonatal colibacillosis Zainal Arifin; Supar .
Jurnal Ilmu Ternak dan Veteriner Vol 1, No 1 (1995)
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Neonatal diarrhoea associated with enterotoxigenic Eschericia coli (ETEC) infections in piglets is common in most piggeries. Control of this disease is difficult with antimicrobial drugs under field conditions . It is due to the high precentage of ETEC strains resistant against there drugs. Field trials of E. coli vaccine were conducted to investigate the effect of dam vaccination on the reduction of diarrhoea and mortality rates. This study was designed in a factorial completely raralomizul block design . Vaccinated and unvaccinated dams was the first factor, while the second factor was location of the piggeries (Jakarta and Bogor) . Vaccination of sows at late gestation using a local polivalent ETEC vaccine appeared to have a dramatic reduction diarrhoea rate (P <_ 0 .0004) and mortality ride (P < 0.(x1001) in piglets. The reduction diarrhoea rate amongs the vaccinated sows in both piggeries was not significantly different (P <_ 0 .2338) . But, the mortality rate of piglets was significantly different in the vaccinated sows (P <_ 0 .026) . The economic impacts using polyvalent ETEC vaccine in controlling colibacillosis are the reduction of diarrhoea and mortality rates which leads to an increase of eveaning in piglets. These will be discussed.

The development of fowl cholera vaccine: II. Pathogenicity and vaccine protection of Pasteurella multocida local isolates in experimental ducks Supar .; Yudi Setiadi; Djaenuri .; Nina Kurniasih; B Poerwadhikarta; Sjafei .
Jurnal Ilmu Ternak dan Veteriner Vol 6, No 2 (2001): JUNE 2001
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Pasteurellosis or fowl cholera in ducks occurs sporadically along the year in many high duck population areas of Java and other parts of Indonesia. Some isolates of Pasteurella multocida from ducks and chicken are kept at the BALITVET culture collection. The aims of this research were to evaluate the pathogenicity of local isolates and imported strains of P. multocida and to study the pasteurellosis local isolate vaccine and protection assay in ducks. Two imported strains of P. multocida (BCC 1359, BCC 1362) and 6 local isolates (BCC 299, BCC 2331, DY1, DY2, 12TG, 15TG) were used in this study. In the pathogenicity assays the imported strains and local isolates were activated in mice and in duct and then in brain hearth infusion broth containing 5% normal sheep serum. Each of broth culture was diluted, each dilution (102 and 104) of strains or isolates was injected intraperitoneally into a group of normal ducks. Antigen for vaccine, each was produced in sheep blood (5%) agar. Cells were harvested and killed with 0.1% formalin. Monovalent, bivalent, and polyvalent vaccines were prepared, at concentration equal to the Macfarland standard tube No 10, and each was adjuvanted with alhydrogel at final concentration of 1.5%. Each vaccine type was injected into a group of 10 week old ducks (8 animals per group), with 0.2 ml/injection. Four weeks later each animal in group were boostered with the same vaccine, dose, route as the previous injection. Before vaccination each animal was bleed through wing vena, then every two weeks, serum was collected and stored at -20oC. Two weeks after boostered, three days after the last blood sample collection, half animal of each group were challenged intraperitonelly with the BCC 2331 and half with DY2 live broth culture. The pathogenicity assays showed the only BCC 2331 and DY2 killed the experimental ducks, the other did not. The animals vaccinated with either BCC 2331, DY2 or bivalent (BCC 2331+DY2) vaccines were protected with either life bacterial challenged of either BCC 2331 or DY2 local isolates. It is likely, P. multocida BCC 2331 and DY2 isolates can be used for pasteurellosis candidate vaccine used in Indonesia, but it still needs more studies in the immunological of protective antigens. Key words: Pasteurella multocida, fowl cholera, ducks, vaccine, protection

Genomic DNA restriction endonuclease from Pasteurella multocida isolated from Indonesia, katha strain and reference strains and analysed by PFGE Supar .
Jurnal Ilmu Ternak dan Veteriner Vol 8, No 3 (2003): SEPTEMBER 2003
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Pasteurella multocida strains are the causative disease agents of wide range of domestic and wild animals in Indonesia. The most important serotypes are associated with Hemorrhagic septicaemic (HS) diseases in cattle and buffaloes, cholera in ducks and chickens. The HS disease associated with P. multocia in large ruminants in Indonesia is controled by killed whole cell vaccines produced by the use of P. multocida Katha strains. There is no discriminatory data of the molecular biology technique has been applied to investigate P. multocida isolates from different geographic locations in Indonesia. The purpose of this studies were to observe the genetic diversity among P. multocida isolated from various geograpic locations and compared with Katha vaccine strain and other reference strains. A total samples of 38 isolates and strains of P. multocida were analysed by means of pulsed-field gel electrophoresis (PFGE). Each sample was grown in nutrient broth, cells were separeted by centrifugation. Whole cell pellet was mixed with agarose and then prepared agarose plugs. The genomic DNA of each sample was digested in situ (plug) with either restriction endonuclease of ApaI and/or BamHI. The digested genomic DNA of each sample was analysed by PFGE, the genomic DNA restricted profile of each sample was compared with others. The use of ApaI restriction endonuclease digestion and analysed by PFGE, demonstrated that 34 out of 38 P. multocia samples could be differentiated into 16 ApaI types, whereas based on the BamHI digestion of these samples were differentiated into 20 BamHI types. Genomic DNA restriction pattern of Indonesian P. multocida isolates originated from cattle and buffaloes associated with haemorrhagic septicaemic diseases demonstrated different pattern to those of vaccine Katha strain, poultry strains as well as the reference strains currenly kept at Balitvet Culture Collection (BCC) unit. Two P. multocida isolates derived from ducks with cholera disease showed similar pattern support the previous findings which exhibited similar pathogecity and vaccine protection. The majority of HS causing P. multocida isolates from some provinces in Indonesia belong to ApaI type 1 (7 isolates), six of these isolates belong to BamHI type 1. The BamHI restriction endonuclease digestion demonstrated higher discriminatory than that of ApaI. These restrection endonucleases digestion combined with PFGE analysis were effective for molecullar defferentiation of P. multocida strains and could be applied to other bacterial veterinary pathogens as well as for local isolate vaccine sellection. Key words: Pasteurella multocida, restriction endonuclease analysis, genomic DNA, PFGE

Development of fowl cholera vaccine: I. Protection of Pasteurella multocida local isolate vaccine against challenge of homologous and heterologous strains. Supar .; Yudi Setiadi; Djaenuri .; Nina Kurniasih; Bhakti Poerwadhikarta; Sjafei .
Jurnal Ilmu Ternak dan Veteriner Vol 6, No 1 (2001): MARCH 2001
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Pasteurella multocida locally isolated from chicken and ducks (BCC 299, BCC 2331, DY1, DY2, 12TG, 15TG) andimported strains (BCC 1359, 1362; HEDDLESTON group 1 and 6 respectively) had been tested for its pathogenicity in theprevious study. The aims of this experiment were to study the preparation of local isolate pasteurellosis vaccines and to determine the protective effect of that vaccines in chicken against the highly pathogenic local isolates of P. multocida. Killed monovalent, bivalent and polyvalent pasteurellosis vaccines were prepared and each was adjunvanted with aluminum hydroxide gel at a final concentration of 1.5% and the cell concentration was equal to the No 10 of MacFarland tube standard. Each of the vaccine prepared was used to vaccinated on a group of six week old of layer chicken (8 per group). Each chicken was subcutaneously injected with 0.2 ml of vaccine, four weeks later each was boostered with similar vaccine with the same dose. Two weeks after giving the boostered vaccine each group of chicken were challenged, half with life bacterium of P. Multocida BCC 2331 and other with DY2. Any chick which survive after challenge was designated as protected by vaccination. Before vaccination 1 ml of blood was drawn from each of chicken and then two weeks apart up to challenge. Serum from each sample was separated and kept in deep freeze until tested by enzyme-linked immunosorbent assay (ELISA). Chicken vaccinated with killed whole cell P. multocida vaccines of monovalent (BCC 2331 or DY2) and bivalent (BCC 2331 + DY2) were protected against challenge with live bacterium of either BCC 2331 or DY2 at rate 67-100%. There was no protection in chicken vaccinated with either BCC 299, DY1, 12TG, 15TG, BCC 1359, or 1362 killed vaccine. Similarly no protection of chicken vaccinated with either DY1 + BCC299, 12TG + 15TG or BCC 1359 + BCC 1362 bivalent vaccines. The protection rate of the polyvalent local isolate vaccine was at average 50-75%. All vaccinated chicken showed the presence of antibody responses againsted the extract cell and whole cell antigens of either P. multocida BCC 2331 or DY2 local isolate as detected by ELISA. The antibody responses from vaccinated chicken against extra cellular antigens prepared from broth cultures of BCC 2331 and DY2 were detected only from vaccinated chicken with vaccine containing killed antigen of BCC 2331 and/or DY2 isolate. It is likely, the local isolate of P. multocida BCC 2331 and DY2 would be benefit for producing inactive fowl cholera vaccine use in Indonesia, but the protective antigen that enhances immune protection should be determined by means of immunoblotting techniques. Key words: Pasteurella multocida, fowl cholera, vaccine, protection

The detection of K88, K99 fimbrial antigen and enterotoxin genes of Escherichia coli isolated from piglets and calves with diarrhoea in Indonesia Supar .
Jurnal Ilmu Ternak dan Veteriner Vol 2, No 1 (1996)
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Enterotoxigenic Escherichia coli (ETEC) strains cause diarrhoeal disease in piglets and calves in Indonesia. These strains possess two virulence factors namely attachment and enterotoxin antigens . These factors could be detected phenotypically and genetically. Haemolytic Escherichia coli (E coli) isolates possessing K88 fimbrial antigen associated with 0-group 108 and 149. They were positive for K88 gene and demonstrated their ability to produce heat labile enterotoxin (LT) and genetically were all positive for LT gene . Seventeen isolates ofE coli K88 which associated with 0-group 149 were positive forSTb gene, other O-serotypes were negative . Ten isolates of Ecoli K88 which associated with 0-group 108 possessed K88, K99, LT and STa genes, but negative for STb gene . However, phenotypically the K99 antigen and STa toxin were not expressed under laboratory conditions, the reason was not well understood . E. coli K99 strains isolated from calves wit h diarrhoea were all associated with 0-group 9 and produced STa toxin when tested by suckling mousse bioassay. The E. coli K99 calf isolates were all hybridized with K99 and STa gene only . It is likely that K99 gene is associated with STa gene . The DNA hybridization technique is more convenience to be used for confirmation diagnosis of colibacillosis, however, not all veterinary laboratories could perform these tests . Keywords: Excherichia coli, probe, gene detection, virulence determinant

Development of alpha hemolytic verotoxigenic Escherichia coli vaccine: Verotoxic antibody responses in experimental animals, mice, rabbits and dairy cows Supar .; B Peorwadikarta; Nina Kurniasih; Djaenuri .
Jurnal Ilmu Ternak dan Veteriner Vol 4, No 1 (1999): MARCH 1999
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Escherichia coli isolates showing alpha hemolytic on blood agar were found in calves with bloody diarrhoea from Bandung, Sukabumi and Bogor. Three E. coli isolates designated as (B34c, B909, B910) were pathogenic for mice and selected for vaccine candidates. Crude supernatant of tryptic soy broth (TSB) cultures precipitated with saturated ammoniumsulphate caused cytopathic effect on vero cell line. For vaccine preparation the isolates were grown on tryptic soy agar (TSA). Whole cell inactive vaccine containing equal proportion of each isolate was prepared and emulsified in aluminum hydroxide gel at a final concentration of 1.5% and cell concentration equal to the tube number 10 of MacFarland standard. Mice and rabbits were injected subcutaneously of monovalent vaccine with the dose of 0.2 ml and 1 ml respectively. Four weeks later mice or rabbits were boostered with the same dose. Before injection each animal was bleed, subsequently every two weeks period up to 4 weeks after the second injection. The sera were separated and kept at -20oC up to serological assay with an enzyme linked immunoassay (ELISA) was done. Vaccinated mice with VTEC were challenged with homologous isolate did not die, where as 60% of unvaccinated mice died within 2 days post challenged. Pregnant cows were given 2 doses of three valent vaccine 6 weeks and again 2 weeks before expected date of calving. Calves born were given colostrum and milk of the own mother. The three day old calf was challenged with three isolates of verotoxigenic E. coli studied. Calves born from vaccinated cows did not show diarrhoea at post challenged, whereas calf from unvaccinated cow demonstrated bloody diarrhoea at post challenged. From the experimental animals (mice and calves) demonstrated the presence of protection against challenged. The antiverotoxic antibody probably involved in important role of immunoprotection. These were supported by the presence of high titer anti verotoxic antibody in serum of experimental animals (mice and cows), as well as in experimental rabbits, but none in the unvaccinated animals. Key words: E. coli, alpha hemolytic, verotoxic, dysentery, vaccine

Antigenicity and Immunogenicity of Salmonella enteritidis: Its Implication for Diagnosis and Development of Local Isolate Vaccine for Poultry Tati Ariyanti; Supar .
WARTAZOA, Indonesian Bulletin of Animal and Veterinary Sciences Vol 18, No 4 (2008): DECEMBER 2008
Publisher : Indonesian Center for Animal Research and Development

Genus Salmonella consists of more than 2,400 serovars, which can be identified by means of serological method based on the variation of their somatic (O), flagellar (H) and capsular antigens (Vi). Salmonella serovars which are able to cause disease in animal or domestic animal are limited, such as: S. pullorum and S. gallinarum which are well adapted to poultry, cause fowl typhoid, S. cholerasuis causes disease in swine. S. typhimurium and S. enteritidis can infect all animals and humans. S. typhimurium and S. enteritidis could be isolated from salmonellosis of poultry, meat, milk and eggs. The prevalence of those isolates within the last two decades tends to increase. Pathogenic Salmonella serovars can infect both animals and humans, colonize the intestinal epithelial cells lead to diarrhoea. Salmonella spp. may enter the lower layer of epithelial cells and the lymphoid vascular system. Humoral antibody and cell mediated immunity responses may develop. Extraintestinal shedding or dissemination of Salmonella spp. may occur and multiply, this may cause latent infections and spread to the environment. Serologic diagnosis of infected animals can be done by means of serum or whole blood agglutination tests with whole cell antigen or ELISA with LPS coated tray, might demonstrate cross reactions among serovars within the one group. ELISA antibody by using fimbrial SEF14 antigen demonstrated specific diagnosis of S. enteritidis infection. The use of S. enteritidis inactive vaccines stimulates high humoral antibody response and protection against challenged homologous serovar within one group (D). The secretory antibody in mucosal surface of intestine and cell mediated immunity were not stimulated after vaccination with inactive Salmonella vaccine. Inactive vaccines (local isolate of S. enteritidis) which was developed and evaluated on experimental layer chicken produced protection against challenged homologous and may be used to control vertical transmission salmonellosis through eggs and can be used to improve the safety of animal food products for human consumption. Key words: Salmonella enteritidis, antigenicity, immunogenicity, diagnosis, vaccines for poultry

Animal and Human Leptospirosis in Indonesia Kusmiyati .; Susan M Noor; Supar .
WARTAZOA, Indonesian Bulletin of Animal and Veterinary Sciences Vol 15, No 4 (2005): DECEMBER 2005
Publisher : Indonesian Center for Animal Research and Development

Leptospirosis is a disease caused by Leptospira spp. infection. It is a zoonotic disease that is world-widely distributed, particularly in the tropics, including Indonesia. Infected animals are the source of leptospirosis for humans, and these animals are secreting pathogens into the environment. The clinical signs off leptospirosis may vary from mild to severe and dead tray occur without a proper treatment. Due to the unspecific clinical sign, laboratory examination is required. However, isolation and identification the organism is time-consuming. Serological test is the most frequent way to confirm the clinical diagnosis, to determine prevalence in the community, and to conduct epidemiological studies. Vaccination with the appropriate antigen is used for controlling leptospirosis in animals. The multivalent Leplospira vaccine in Indonesia is developed according to the different types of serovar found in the field. The use of the appropriate vaccine combined with a good sanitation management could control leptospirosis in animals in the future. Key words: Leptospirosis, zoonosis, diagnosis, control

Fowl Cholera and Its Control Prospect With Locally isolated Pasteurella multocida Bivalent Vaccines Tati Ariyanti; Supar .
WARTAZOA, Indonesian Bulletin of Animal and Veterinary Sciences Vol 18, No 1 (2008): MARCH 2008
Publisher : Indonesian Center for Animal Research and Development

Pasteurellosis or fowl cholera disease which associated with Pasteurella multocida group A and D infections occurred sporadically in many parts of the world, including in Indonesia. The pathogenic activity of P. multocida in chickens were based on lipopolysacharide (LPS) antigens associated with group A and D capsules, and the resistance factor of complement mediated bacteriolysis in animals. In order to reduce common bacterial infections, antibiotics were routinely used as feed additive or by drinking water, but fowl cholera cases still occur. Fowl cholera control by vaccinations have been used more than a hundred years ago by means of inactive vaccine, but imported inactive vaccine was reported not effective due to lack of cross protection against heterologous serotype. At present, many local P. multocida isolates from chicken and ducks from many areas in Indonesia were characterised for their antigenicity, immunogenicity and prepared as monovalent or bivalent vaccine. Only the monovalent vaccine prepared from BCC 2331 or DY2 demonstrated the presence of immunoprotection against homologous and heterologous challenged with live bacteria. The prototype bivalent vaccine consisting of BCC 2331 + DY2 demonstrated high degree of cross protection against challenged individual with or mixed of BCC 2331 + DY2 at average of 60 – 75% and 75 – 100%, respectively. Monovalent and bivalent vaccine prepared from other isolates including imported reference strains of P. multocida demonstrated no protection in experimentally vaccinated ducks and chicken against challenged with live bacteria of neither BCC 2331 nor with DY2. From these retrospective studies, it was concluded that the local isolates P. multocida designated as BCC 2331 and DY2 could be used as candidates of prototype vaccine or master seed vaccine but their effectiveness still need to be evaluated under field conditions. Key words: Pasteurella multocida, Indonesian isolates, inactive vaccines, fowl cholera

The Role of Salmonella Enteritidis in Chicken and its Product Tati Ariyanti; Supar .
WARTAZOA, Indonesian Bulletin of Animal and Veterinary Sciences Vol 15, No 2 (2005): JUNE 2005
Publisher : Indonesian Center for Animal Research and Development

Free pathogenic microorganism of food derived from animals is a prerequisite for human consumption . One of the important pathogenic microorganisms originated from animal product of food is Salmonella. Salmonella enteritidis is frequently found in chicken and spreads vertically as well as horizontally products (eggs, meats and meat products) by direct or indirect contact. Salmonella that contaminated animal product of food can cause foodborne disease in human . Foodborne disease associated with Salmonella occurred in some parts of the world including Indonesia . This problem needs attention from the government, producers and consumers. In the animal production especially chicken, it is demanded to provide animal food and their products free from Salmonella . This is an important indicator of safety food condition . Salmonella control programs in the animal production level begin with raising free-Salmonella day old chick with free Salmonella feed, good farm environmental sanitation . Further more, the monitoring program of Salmonella in farm and post harvest process needs to be conducted . Appropriate handlings of animals and their products are important to obtain food of animal products that are healthy and safe for human consumption . Key word: Salmonella enteritidis, contamination, meat, egg

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