Indonesian Journal of Animal and Veterinary Sciences

148 - 152

Authors: R Widiastuti, Indraningsih ., T.B Murdiarti, R Firmansyah

Trenbolone acetate (TBA) is a growth hormone promoter which is implanted into animal to increase the body weight. The implantation of TBA in animal may cause the occurrence of residues of TBA and its metabolite (17-β-trenbolone). The presence of the residue might threat to human health. The aim of this research work was to study the presence of the residue of trenbolone in male Garut sheep which is implanted by TBA. The sheep were divided into 2 groups, those were D1 which were implanted by 40 mg TBA and D2 which were implanted by 60 mg TBA. One animal each from D1 and D2 were killed in the week 1, 2, 3 and 4 after implantation. And 24 hours before slaughtering the urine of D1 and D2 were collected using the metabolism cages. The samples for residue analysis were the organs such as liver, kidney, and the muscles around the implantation sites i.e inner and outer, upper front leg and upper back leg. The samples were extracted by organic solvents and analysed by HPLC. The residue of trenbolone occured in all samples of D1 and D2 which killed on 1 and 2 weeks after implantation. The residues occured on those which were killed 3 weeks after implantation were only in inner and outer samples. The residues in meat and organs were not detected in D1 nor D2 which were killed 4 weeks after implantation. The residue of trenbolone was also detected in the urine and still were detected in D1 and D2 which implanted by TBA for 4 weeks. Key words : trenbolone acetate, 17-β-trenbolone, residue, sheep

158 - 165

Authors: Lily Natalia

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

172 - 178

Authors: Elizabeth Wina, Toto Toharmat, Widia Astuti

Bark, especially from Acacia mangium is a by-product from wood processing industries that commonly found in Indonesiaand in big amount will cause environmental problems. One of the alternatives to utilize bark is for animal feed. The aims of this experiment are to improve the nutritive value of bark by alkali treatments (urea and sodium hydroxide) and to determine the level of substitution of elephant grass by bark. The experiment consisted of 3 in vitro studies and 1 in sacco study. In vitro studies consisted of 1) the use of urea or NaOH by wetting and incubation-method, 2) the use of different concentration of Na OH (0-4%) by soaking method, 3) determination of substitution level of elephant grass by treated bark. In sacco study was conducted at 0, 6, 12, 24, 48 and 72 hours of incubation to compare the degradation of treated bark to elephant grass. The results show that urea treatment did not improve DM or OM digestibilities of bark. Soaking bark in 4% NaOH solution was more effective than wetting and incubation-method in improving in vitro digestibility. (49.26% vs19.56% for soaking and dry-method, respectively). In sacco studyl shows that treated bark had a very high solubility at 0 hour incubation but the degradation at 72 hours incubation was not significantly different from that of 0 hour incubation. The gas produced at in vitro study of treated bark was very low indicated that there was no degradation of bark at all. The level of substitution of elephant grass by treated bark up to 30% gave a non-significant digestibility value to that of 100% elephant grass. In conclusion, bark after tannin-extraction was a better feedstuff for animal feed. The soaking method in 4% NaOH solution improved the digestibility of bark significantly and the level of substitution of elephant grass by treated bark was 30%. Key words: Bark, acacia mangium, urea, sodium hydroxide, digestibility

189 - 194

Authors: Harry T Uhi, Jajat Jachja, Rita Mutia, Dodi Nandika

Termites (Isoptera) as one of local feed in several places in Indonesia just like Irian jaya, is available, easy to find and having protein in a high level. Termites nest which high about 0,5-2 metres above ground is easy to find at all low area merauke which having land areal about 20.000 ha, otherwise in other distrik, it can easily to find in a forest especialy in a new open land for Agriculture and transmigration. The purpose of this experiment was to study the effects of Glyptotermes montanus Kemner termites suplementation as protein resource in the diet of broiler strain Rokky-301. One hundred eighty DOC Rokky-301 were randomly divided in to five treatment with three replicates of 12 chickens each. The treatment diets were basal diet (R0), basal diet + fresh termites 0,5% (R1), basal diet + fresh termites 1% (R2), basal diet + fresh termites 1,5% (R3), and Comercial diet (R4) as positive control. The chickens were fed the treatment diets over 50 days. The results showed that the chicken fed R3 diet had significantly higher body weight gain than did chicken fed other diets except for chicken fed commercial diet (R4). Feed consumption of chicken fed R0 diet was significantly higher than other treatment diets. However 1,5% fresh termites suplementation (R3) significantly reduced feed consumption. Basal diet (R0) had highest feed conversion compared to other treatment diets. However 1,5% fresh termites suplementation (R3) was significantly improved feed conversion compared to other treatment. Key word: Body weight gain, Glyptotermes montanus termites, broiler strain Rokky-301

205 - 212

Authors: Sudarisman

The aim of this study is to investigate the post challenge clinical responses of Bali cattle vaccinated with an inactivated vaccine of a field isolate Bovine Herpesvirus-1 after being challenged with the same virus. Twenty eight Bali cattle were divided into two groups of ten animals, one of which was vaccinated with a single dose and the other one was vaccinated with a booster dose of vaccination, and the rest of the animals were remained untreated as a control group. One month after the last vaccination, a half number of animals in each group were challenged intravenously with 5 x 108 TCID 50 pathogenic strain of BHV-1 local isolate. Clinical signs were observed from day 0 to day 14 after challenge. These observed included body temperature, mucosal membrane changes of nostril, vagina and palpebrae, hyperlacrimation and hypersecretion of nose and eyes. General clinical changes and mucous membrane lesions of nostril were scored from 0 to 3. The results showed that the non-vaccinated animals: elevated temperature (up to 40.1°C), hyperlacrimation, nasal discharge and mucosal hyperaemia of nostril and vagina after challenge. The duration of illness ranged from 3 to 7 days. Symptoms appeared after an incubation periode of 3 days and disappeared after 5 days (day 9). These clinical symptoms didn’t appear on the vaccinated animals with single or boostered vaccination. One month after vaccination, the antibody titre reached 211 (2048) based on serum neutralisation test (SNT). These vaccinated animals showed good immune respons to BHV-1. Key words: Clinical responses, inactivated vaccine, field isolate, BHV-1, Bali cattle

76 - 82

Authors: Sori B Siregar

Increasing milk production ability of lactating cows through improvement of feeds and feeding frequency was conducted at Tanjungsari Village Cooperative. Fifteen lactating cows of 3-4 months of lactation period were used. The cows were divided into five groups based on their average daily milk production and lactation period. The treatment consisted of RO (control=commonly practiced by farmers), R1 (2.5 kg. concentrate, given three times per head per day). R2 (2 kg. Concentrate plus 13.5 kg roughage given three times per head per day). The experimental design was completely randomized block design. The quality of concentrate given to the lactating cows was a better than that given by the local farmers. Improving feed supplementation and feeding frequency significantly (P<0.01) increased the consumption of dry matter, crude protein and energy of the feeds, and thus increased daily milk production, solid non fat (SNF) and total solid (TS) of the milk (P<0.05). Specific gravity of the milk was not affected by the treatments (P>0.05). Substitution of 0.5 kg concentrate with 13.5 kg roughage resulted in non significant difference on the daily milk production as well as the quality of the milk, thus it gave no economic impact. The increase of daily milk production was followed by the increase of the incomes of the farmers by Rp 905 to Rp 1,425 per head per day. The result of the experiment leads to the assumption that the present feeding practices of the lactating cows should be improved accordingly. Key words: Lactating cows, feeding, management

94 - 99

Authors: I.P Kompiang, Supriyati ., M.H Togatorop, S.N Jarmani

An experiment was conducted to study the effect of free choice feeding on the performance of native chicken and to determine its protein and energy requirements for growth. Eight hundreds and seventy 2-weeks old native chicken were used, and divided into three treatment groups, with 5 replicates: (I) Complete feed (II) Choice feed A (two levels of protein, similar in energy content) and (III) Choice feed B (energy and protein sources). Feed and water were given ad libitum during the 10 weeks trial. Feed intake (protein and energy) and body weight were recorded biweekly. Body weight gain (BWG) of treatment I was 852 + 33 grams/head, similar to treatment II, 858 + 28 grams/head, and both significantly (P<0.05) better than treatment III (800 + 42 grams/head). Energy consumption and its energy efficiency ratio (EER) value were similar for all treatments, 9226 + 149; 9203 + 739, and 8706 + 383 kcal/head and 10.84 + 0.34, 10.75 + 1.03, and 10.89 + 0.27 kcal/gram for treatment I, II and III, respectively. There was a significant (P<0.05) difference between treatment on protein intake, 556 + 8, 506 + 15, and 454 + 25 grams/head for treatment I, II, and III, respectively. Protein energy ratio (PER) value of treatment III (0.57 ± 0.02) was similar to treatment II (0.57 ± 0.02) and both were significantly better (P<0.05) than treatment I (0.65 ± 0.02). It was concluded that the native chicken, given a correct choice of feed, has an ability to determine its energy and protein requirements. During the growth periods, 2-12 weeks old, the bird require feed with 16% protein and 2900 kcal metabolized energy/kg. Key word: Native chicken, feeding system, free choice

107 - 112

Authors: A.P Sinurat, Tatty Purwadaria, T Pasaribu, J Darma, I.A.K Bintang, M.H Togatorop

Drying proces, especially with heating often affects the nutritive values of feed ingredients. Therefore, this experiment was designed to study the responses of broiler chickens when fed with ration containing fresh or dried fermented palm oil sludge(FPOS). Experimental diets with different levels of fresh or dried FPOS (5, 10, and 15% equally to dried FPOS) were formulated with similar nutrient contents. A control diet with no FPOS was also included. Each diet was fed to 30 broiler chickens (5 replicates of 6 birds) for 5 weeks. Performances (body weight, feed consumption, feed conversion, and mortalities) of chickens were recorded. Carcass percentage and abdominal fat content was also measured at the end of feeding trial. Data obtained were subjected to analysis of variance in a completely randomized design and different between means were tested by orthogonal contrast procedures. Results of the experiment showed that body weight gain (BWG) of control birds was not significantlydifferent with BWG of birds fed with FPOS. Birds fed with dried FLS gain more weight than those fed with fresh FPOS (1048 vs 981 g/bird). Increasing of dietary FPOS levels decreased BWG, but 10% inclusion was still tolerable. Feed consumption of control diet was significantly (P<0.05) less than feed consumption of diet with FPOS. Increasing of dried FPOS to 15% did not affect feed consumption, but increasing of fresh FPOS significantly increased feed consumption. Feed conversion (FCR) of control diet was significantly better than the FPOS diet (2.07 vs 2.13). Increasing levels of dried FPOS from 5 to 10% did not affect the FCR, but further increasing to 15% significantly worsen the FCR. Increasing of fresh FPOS from 5 to 10 or 15 significantly worsen the FCR. Dry matter intake, mortalities, carcass percentage, liver weight, and abdominal fat levels of broilers were not significantly affected by dietary treatments (P>0.05). Conversion of feed dry matter to body weight gain of control diet was not significantly (P>0.05) different with those diets with FPOS. However, dry matter conversion of dried FPOS was significantly better than the fresh FPOS. Increasing levels of FLS from 5 to 15 significantly worsen the feed dry matter conversion but not with inclusion of 10% FPOS. Therefore, it is concluded that there is no superior advantage of feeding fresh FPOS as compared with dried FPOS. Inclusion of 10% dried or fresh FPOS in the diets did not affect growth performances of broiler chickens. Key words: Palm oil sludge, dried, fermented, broilers

120 - 125

Authors: Supar ., Yudi Setiadi, Djaenuri ., Nina Kurniasih, B Poerwadhikarta, Sjafei

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

134 - 140

Authors: Risa Indriani, Darminto

Infectious bronchitis (IB) is an acute highly contagious viral respiratory disease of poultry caused by Coronavirus. IBV infection consists of many serotypes and can only be controlled by vaccination. An effective IB vaccine should be prepared from local isolates, due to the antigenic variation among serotypes. The aims of this research were to develop inactivated IB vaccine derived from IBV local isolate and to determine the efficacy of that vaccine in layer flocks. Five layer chicken groups were used in this experiments, group I was vaccinated with commercial IBV live vaccine thrice, group II was vaccinated with commercial IBV live vaccine once and repeated with inactivated local IBV isolate twice, group III was vaccinated with commercial IBV live vaccine once and repeated with commercial inactivated twice, group IV was vaccinated with IBV live vaccine once, and group V was not vaccinated. After the chickens reached at a stable egg production they were challenged with IBV local isolates. Antibody responses were examined by means of haemagglutination hibitition (HI) test and HI titres were expressed as log2 of the reciprocal of the highest dilution of serum causing inhibition of a log2 HA titre of 2. The mean titres of antibody responses of chicken in group I, II, III, IV, and V was 4.9 ± 0.87, 6.8 ± 0.97, 7.7 ± 0.46, 2.9 ± 0.94, and 2.0 ± 1.67 respectively. The levels of protection against challenges were determined by viral isolation, this in group I, II, III, IV, and V was 63, 73, 60, 50, and 0% respectively. Clinical symptom of egg quality was slightly reduced in group I, IV, and V and it were unchanged in group II and III. Group II gave better in number of egg production than the other groups. The results indicated that the IBV inactivated localisolate vaccine gave high titres of antibody and higher protection rates than that of commercial IBV inactivated vaccine. Inaddition, IBV local isolate vaccinated group prevented from declining egg production after challenged with IBV local isolate. Key words: Infectious bronchitis, layer, antibody titre, vaccine, challenge virus

7 - 13

Authors: I-W Mathius, Dwi Yulistiani, E Wina, B Haryanto, A Wilson, A Thalib

High producing ewes required high amounts of energy, which can not be met through feeding conventional ration. Therefore, supplementing energy should be done, especially protected-energy. The purpose of this study was to determine the effects of additional of protected fat/energy on the consumption of ration and the performance of ewes and lambs. Thirty-four local ewes were randomly allotted to four levels of by-pass energy groups, in which parts of energy content in the concentrate was replaced with rumen-protected fatty acids in the proportion of 0 (R1), 5 (R2), 10 (R3), and 20% (R4). Consumption, nutrient digestibility and animal performance (ewes and lambs) were determined by standard procedures. Results showed that the daily dry matter intake during pregnancy (g/kg BW0.75) did not differ among diet (72.1 + 2.56). The crude fibre intake varied slightly between treatment (P>0.05), those being slightly lower intake on the highest replacement of energy content in concentrate diet (R4). Dry matter digestibility were 52.4 + 0.24, 52.1 + 0.30; 51.8 +0.28; and 51.3 + 0.81 in groups R1, R2, R3, and R4, respectively, which were non-significantly different from each other. These results confirmed that the by-pass energy sources replacements did not affect rumen fermentation. The same tendency was also found for crude protein digestibility (P>0.05) i.e. 60.1; 61.8; 61.2; and 60.2 for R1, R2, R3, and R4 respectively. The average daily gain of ewes during gestation period was affected (P<0.05) by by-pass energy replacement and the effect was linear with increasing by-pass energy replacement. Significant difference was not found in the total birth weight of lambs and among dietary treatments. Lambs growth rate during the first 8 week of lactation was the highest on ewes fed diet containing the highest amount of by-pass energy sources (R4). It can be concluded that by-pass fatty acids can be used in ewes’ ration without affecting rumen fermentation and gave a positive respons to animal performance. Key words: By-pass energy, ewes ration, gestation and lactating phase

21 - 27

Authors: Benny Gunawan, Tike Sartika

The experiment aimed to produce native chicken with faster growth rate that is to achieve body weight of more than 1 kg at 3 months of age. Three hundreds and thirty three crossbreeds (PK) day old chicken from the results of artificial insemination between male Pelung and second generation of selection of female native chicken, were used in this experiment. One hundred and eighty purebreds native chickens were also used as control population. They were put in grower cages with density of 10 birds per cage and each cage was treated as a single unit of replication. Feeds during experiment were given and divided into 3 phases that is Starter Feeds I (Protein 21% and Energy 3000 kcal/kg) for chicken between 0-21 days of age, Starter Feeds II (Protein 19% and Energy 2900 kcal/kg) for chicken between 22-42 days and Grower Feeds (Protein 17% and Energy 2900 kcal/kg) for chicken between 43-84 days. Variables which were recorded were weekly body weight until 12 weeks of age, feed consumption, feed conversion, mortality, carcass weight and simple economic ratio (B/C ratio). The results of the experiment showed that body weight at 12 weeks of crossbred (PK) was significantly higher than the pure native chicken (1000 vs 923 g) (P<0.05). Feed consumption was not significantly different (3037 vs 3036 g/bird/12 weeks), but the feed conversion of the crossbred was significantly better than the purebred (3.09 vs 3.4) (P<0.05). Carcass weight and carcass components were not significantly different between the two breeds, but they were different between sexes (P<0.05). Simple economic calculation showed that crossbreeds produced higher profit than purebreds with the benefit cost ratio were 1.31 for crossbred (PK) and 1.2 for purebred native chicken. The mortality rate during the experiment was low, that is 5.5% for crossbred and 6.36% for purebred. Key words: Crossbreeding, male Pelung, female native chicken

34 - 37

Authors: Cece Sumantri, T Morzumi, N Hamashima

The study was done to detect the incident of deletion and mutation in exon 14th of Mx1 gene in pig. Six hundred base pairs at the position (1937 to 2537) of the 14th exon of the pig Mx1 gene was amplified by polymerase chain reaction (PCR) from 15 breed of pig DNA sample. The amplified PCR products were digested by Nar I enzyme that called restriction fragment length polymorphism (PCR-RFLP) technique. The results show genetic polymorphism at the 14th exon of pig Mx1 gene. The Nar I digested revealed three phenotyphic variation (C/C, C/N and N/N, designated for Nar I cut homozygote, heterozygote and for Nar I no cut homozygote, respectively). The Nar I (N/N or C/N) type is corresponding to (1) the deletion 11 bp at the position 2064 to 2075. This type was observed in Landrace breed. (2) the incidence of two point mutation at the position 2065 Guanine (G) change to Thymine (T) and at the position 2124 Guanine (G) change to Adenine (A).This type was observed in Chinese native pig (Meishan) and Vietnamese native pig. Key words: Deletion, mutation, pig Mx1 gene and PCR-RFLP

45 - 50

Authors: Abubakar ., Triyantini ., R Sunarlim, H Setiyanto, Nurjannah

A study on the effect of temperature and time of pasteurization on the milk quality during storage was carried out using fresh milk. The aim of the experiment was to asses the storage time of pasteurized milk for consumption without nutrient losses. A completely randomized factorial design, 2 x 8 was used, with pasteurization temperature (T), consisted of 2 levels, the low temperature long time (LTLT), i.e. fresh milk was warmed at 65oC for 30 minutes (T1) and the high temperature short time (HTST), i.e. fresh milk was warmed at 71oC for 15 seconds (T2); and storage time (S), consisted of 8 levels, i.e. 0, 3, 6, 9, 12, 15, 18, and 21 hours respectively, as the factors, with 3 replicates. Parameters measured were alcohol test, water, fat, and protein concentrations, and microbial population of pasteurized milk during storage. Data were analyzed using analysis of variance and simple linear regression. The result showed that water and fat concentrations and microbial population was not significantly different (P>0.05) in pasteurization temperature treatment, but was significantly different (P<0.05) due to storage time treatment. Meanwhile, the protein concentration was significantly different (P<0.05) either in pasteurization temperature or storage time. It was concluded that pasteurized milk was still suitable for consumption at 15-21 hours storage, while protein concentration tended to be better when was pasteurized at 65oC. Key words: Pasteurized milk, storage quality

59 - 67

Authors: Supar ., Yudi Setiadi, Djaenuri ., Nina Kurniasih, Bhakti Poerwadhikarta, Sjafei

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

210 - 214

Authors: L.H Prasetyo, T Susanti

alternative to increasing productivity and production efficiency. Crossbreeding has been used widely for increasing productivity, and in this experiment Alabio and Mojosari ducks were crossed reciprocally in order to evaluate their egg production and egg quality, when compared to their parental breeds. Four genotypes AA, AM, MA and MM were compared and each consisted of 50 layer ducks. Each animal was kept in individual cage, and individual egg production and egg quality were recorded. They were given layer feed containing 20% crude protein and 3000 kcal/kg metabolisable energy. Results showed that the crossbred ducks (AM and MA) laid eggs earlier than their parental breeds (AA and MM), and MA laid earlier than AM. The weight of first eggs of the crossbreeds did not differ significantly, AM was the same as MM and MA the same as AA. For the body weight at first lay, MA and AM weighed in between their parental breeds, AA was the heaviest and then followed by MA, AM and MM. For the 3-month egg production, MA ducks laid significantly more egg than the other 3 genotypes, and this is related to the age of first lay being the earliest. In terms of egg quality such as egg weight, yolk color, weight and thickness of shell, weight of the white and HU value, the crossbred ducks showed quality in between AA and MM. The AM ducks tended to be closer to MM and MA closer to AA, and this shows a strong influence of maternal effects. The average level of heterosis for weight of first egg is 2.41% and for 3-month egg production is 2.1%, but for MA cross the heterosis level is 11.69% for egg production. The overall results showed that crossbreeds between Alabio and Mojosari have the potential in improving production traits in the effort to increase productivity and production efficiency. Key words : Duck, crossbred, production egg

219 - 223

Authors: Kuswandi ., Muchji Martawidjaja, Zulbardi Muhammad, Bambang Setiadi, Didi Budi Wiyono

A complete block design was applied to employ 40 weaner goats, initial weighing average of 14.6 kg, which were grouped into 5 supplementary treatments, i.e. 125, 250, 300, 350 dan 400 g of skim milk/d. Each animal received.1.25 kg gliricidia leaves. Mixed pasture was consumed freely. The research was conducted in Tanjungrejo village, Tongas district, Probolinggo. A digestibility study was conducted for 7 days. Measurements included nutrient intakes and digestibilities, liveweight gain, and feed conversion ratio. Data were analyzed using the GLM procedure of SAS (SAS, 1986). The results showed that the medium skim milk level (300 g/d) gave the highest dry component digestibility. Cell wall digestibility from the group given 125-300 g skim milk/d was higher (P<0.05) than those given 350-400 g/d. No significant (P>0.05) difference in organic matter or protein digestibility was found among treatments. Supplementation with 350 g skim milk/d gave higher protein or digestible protein consumption (P<0.05), while supplementation with 125 g/d gave the best feed utilization efficiency. Levels at 125-250 g skim milk/d gave weight gain of about 65 g/d, while at more than 250 g gave 57 g/d or less. The best feed conversion ratio (7.8) was obtained in the group fed 125 g skim milk/d. In order to optimize microbial activity in the rumen, provision of slowly rumen degradable protein sources such as legumes, or rumen undegradable protein is of great importance. Key words: Rumen degradable N, weaner goats, digestibility, average daily gain

229 - 233

Authors: E.T Mergawati, E.M Kaiin, K Eriani, N.D Yanthi, Indriawati

The purpose of this study was to examine the effect of medium combination of IVM and IVC on the in vitro development of bovine embryos. The study involved 4 groups in a 2 (IVM medium) x 2 (IVC medium) factorial in a randomized block design. Each group was replicated for 5 times. The treatments were as follows: TCM-199/CR1aa (T1); TCM-199/SOF (T2); B- 199/CR1aa (T3) and B-199/SOF (T4). Oocytes were aspirated from ovaries collected at local abattoirs using aspiration medium of PBS supplemented with 3% FCS and 0.1% Penicillin and Streptomycin. The oocytes were matured in medium of TCM-199 or B-199 supplemented with 10% FCS, hormones: 10μg/ml FSH+ 10μg/ml hCG+ 1μg/ml Estradiol. Maturation was maintained at 37oC for 22 hours in 5% CO2 incubator with high humidity. A method of BRACKETT & Oliphant (BO) was used to fertilize the matured oocytes. The fertilization was incubated for 7 hours in the 5% CO2 incubator. Two culture media of CR1aa or SOF/AA/BSA were used to develop the fertilized oocytes undergo to morula and blastocyst embryos. The findings showed that the proportion of oocytes cleaved and formation of blastocysts were affected significantly by a combination of IVM and IVC media (P<0.05). A combination of B-199/SOF (T4) resulted in a higher blastocyst rate (32%) than others (T3= 29%; T2=T1= 23%). This study suggests that either SOF/AA/BSA or CR1aa has similar competence in development of bovine embryos in vitro. Key words: Bovine embryo, IVF, maturation medium, culture medium

241 - 249

Authors: Suhardono ., D.B Copeman

Field of irrigated rice paddy was the most suitable habitat as breeding site of snail Lymnaea rubiginosa, the intermadiate host of Fasciola gigantica and other trematodes. Fluctuations in the population of fresh water snail, L. rubiginosa in irrigated rice fields and their infection with L. rubiginosa and other trematodes were studied in five villages in the subdistrict of Surade, provivine of West Java. Snail were sampled based on time collection (half man hour) each site of collection. The samples of the snails were further counted and examined for the presence of larval trematodes. The result indicated that snails died during the dry season except those in persistent aquatic refuges such as streams and springs. Surviving snails recolonised rice fields near villages by passive transfer with water from refuges early in the wet season. Some recolonosation may also have resulted from haching of snail eggs deposited in habitats which had not been dried for more than a few weeks. Recolonisation with snails of further rice fields from a village occurred during the later period of the wet season. No snail infected with F. gigantica was found in the distance of more than 200 m from a village. Snail with the highest prevalence of infection occurred in rice fields which received effluent from a cattle pen were fertilised with bovine faeces. Each snail was only infected with one species of tramatodes. Infection with echinostone larvae was most common. Kay words: L. rubiginosa, trematode infection, population dynamic

255 - 259

Authors: T.B Murdiati, G Adiwinatai, D Hiladsari

Intestinal parasites such Haemonchus contortus is usually controlled by management improvement and regular administration of anthelmintic. However, there is an indication of H. contortus resistance to several anthelmintic available in the market, which makes medicinal plants as an alternative anthelmintic and mengkudu or noni fruit (Morinda citrifolia) have been reported as an effective anthelmintic. To trace the active compounds responsible for anthelmintic activity against H. contortus, the mengkudu fruit was continuosly extracted into hexane, chloroform, metanol and water, followed by in-vitro study on the anthelmintic activity. The in-vitro anthelmintic activity was base on the ability of the extracts to kill the worm and the ability of the extracts to prevent egg development. The study suggested that chloroform fraction which contains alkaloid and anthraquinon have the highest anthelmintic activity and showed significant different compared to control (P≤ 0.05). Key words: Morinda citrifolia, anthelmintic, Haemonchus contortus

144 - 149

Authors: Sukarsih ., Sujitno Partoutomo, Robert Tozer, Edy Satria, Gene Wijeffels, George Riding

A colony of the Old World Screwworm fly, Chrysomya bezziana, has been established and maintained at Balai Penelitian Veteriner, Bogor, West Java, Indonesia since 1994. Culture on a convenient, defined diet able to sustain long term reproducibility of insect performance is described. The long-term viability of the colony has been further assured with an effective and reliable culturing procedure, with the genetic diversity and robustness of the colony being maintained through regular infusions of wild-type field isolates. The colony has been a reliable source of larval material used for the isolation and identification of protective antigens in the ACIAR funded Screwworm Fly Vaccination Project. Larvae have been used in the development and routine application of assays employed to evaluate vaccine efficacy. The colony continues to be a valuable resource for both screwworm fly vaccine and other new research includi the evaluation of screwworm fly attractants and lures. Key words: Chrysomya bezziana, culture, colony, larvae, vaccination

160 - 169

Authors: Tony Voucolo, Florentina Supriyanti, Sri Muharsini, Gene Wijffe

The construction and use of cDNA libraries for the isolation of genes encoding candidate antigens for use in a recombinant vaccine against Chrysomya bezziana is described. RNA was isolated and mRNA purified from first and third instar larvae of Chrysomya bezziana and used in the synthesis of two cDNA libraries in the bacteriophage vector λ ZAP express®. These libraries were screened using Digoxigenin-labeled DNA probes obtained from two independent approaches. First, a homolog approach used probes designed from previously characterized peritrophic membrane genes identified from the related myiasis fly, Lucilia cuprina. Secondly, a de novo approach used amino-terminal and internal peptide sequence information derived from purified Chrysomya bezziana peritrophic membrane proteins to generate DNA probes. Three peritrophic membrane genes were identified and characterized. Chrysomya bezziana peritrophin-48 was identified using the homolog approach and, Chrysomya bezziana peritrophin-15 and Chrysomya bezziana peritrophin-42 were identified using the de novo approach. The identification of these genes as encoding candidate antigens against Chrysomya bezziana has allowed the production of recombinant proteins for use in vaccination trials. Key words: cDNA library, peritrophin, peritrophic membrane, Chrysomya bezziana, Lucilia cuprina, vaccine

177 - 184

Authors: Sri Muharsini, Tony Voucoloco

Pichia pastoris has been investigated as a means to express recombinant forms of two putative peritrophic membrane antigens from Chrysomya bezziana, Cb-peritrophin-42 (Cb42) and Cb-peritrophin-48 (Cb48). Recombinant Cb48 was expressed as a secreted and glycosylated protein. The yield of recombinant protein was 8 mg per litre of culture. In contrast, recombinant Cb42 was not expressed at detectable levels in Pichia pastoris, probably due to A + T rich sequence which may cause premature transcriptional termination. To expedite Cb42 expression in yeast, Cb42 was divided into two domains: Cb42A and Cb42B. Cb42B was successfully expressed in Pichia pastoris, yielding 0.4 mg per litre of culture. However, Cb42A was not expressed. This work demonstrates that although Pichia pastoris offers considerable benefits as an expression system producing high level of glycosylated protein, success may vary from protein to protein. Key words: Chrysomya bezziana, peritrophin, Pichia pastoris

192 - 196

Authors: Sukarsih ., Sujitno Partoutomo, Gene Wijffels, Tony Voucoloco, Peter Willadsen

Recombinant forms of a number of peritrophic membrane proteins from the screwworm fly Chrysomya bezziana have been assessed in vitro and in vivo for their efficacy as antigens in vaccination against the tissue-invasive, larval form of the parasite. The proteins included Cb15 and Cb42 expressed in Escherichia coli and Cb48 expressed in both Escherichia coli and Pichia pastoris. In all cases, the in vitro assays of larval growth on serum from vaccinated sheep failed to show inhibition of larval weight gain or any detrimental effect on larval survival relative to controls. Chrysomya bezziana Cb48 has a significant degree of sequence identity with the antigen PM48 from Lucilia cuprina. Feeding Lucilia cuprina larvae on antisera to Cb48 induced a small but statistically significant reduction in weight gain, as does feeding on antisera to PM48. In vivo, larvae feeding on sheep vaccinated with Escherichia coli-expressed Cb15 and Cb42 and Pichia pastoris-expressed Cb48 showed marginally greater weight gain and survival which was equal to or greater than that on non-vaccinated sheep. The significance of these observations is discussed. Key words: Chrysomya bezziana, recombinant antigen, peritrophin, vaccination

201 - 203

Authors: Peter Willadseni

Methods for the culture of the Old World Screwworm Fly Chrysomya bezziana on a small laboratory scale, the production of life stages and tissues suitable for vaccine study, and the in vitro and in vivo assessment of vaccine effects have all been developed. These methods would be applicable to further studies on screwworm control by many alternative technologies. It has been shown to be possible to induce dramatic immunological effects on feeding larvae and to isolate and characterise individual antigens. However, a recombinant vaccine remains elusive. Current understanding of the effects of vaccination is discussed and suggestions made for possible future research directions. Key words: Chrysomya bezziana, Old World screwworm fly, vaccine, recombinant antigen, sheep