Indonesian Journal of Animal and Veterinary Sciences

76 - 83

Authors: Tresnawati Purwadaria, I Putu Kompiang, Jinadasa Darma, Supriati ., Emi Sudjatmika

produced, and suitable with the environment of the digestive track. Isolation was carried out from commercial culture (containing mixture of Bacillus spp.), digestive tract (proventriculus, small intestine and large intestine) of local and broiler chickens, and commercial yoghurts (fermented milk). Neutral and acidic nutrient agars (NA) were used as the media at room temperature and in the aerobic or anaerobic conditions. Separate colonies were isolated, stained with Gram and spore staining and observed under the microscope. The bacteria which were Gram positive bacillus and can form spores were further identified. Eight different species of Bacillus spp.: B. laterosporus, B. coagulans, B. alvei, B. circulans, B. brevis B. bodius, B. pasteurii, and B. macroides were isolated from the commercial mixture. From the digestive tracts of local and broiler chickens, 13 bacteria and 2 yeasts were isolated, while 5 yeast were obtained from two commercial yoghurts. Those bacteria were facultative aerobic and only grew in neutral condition and not in acidic condition (pH 4.5), while the yeast were either facultative anaerobic also can grow in pH neutral and 4.5. The ability of each isolates to grow in the media containing mixture of minerals and glucose, sucrose or molasses were evaluated. Incubation was carried out in the shaker incubator at 40°C, 150 rpm for 48 hours. Dry matter of the biomass was determined as the growth parameter. All isolates of bacteria and yeast can grow in the substrate containing glucose, sucrose and molasses. Variance analyses show that there were interactions between kind of bacteria and carbon source or between kind of yeast and carbon source (P<0.05). Three isolates of bacteria that had highest production of biomass were B. coagulans on sucrose (the third for molasses), Sp. 9 on glucose, and B. apiarius on sucrose (the sixth on molasses). B. apiarius isolated from digestive tract of local chicken will be useful to be developed as probiotics. The highest production of yeast was the control Saccharomyces cerevisiae on three sugars. Between the isolates from digestive tract of local chicken, Toluraspora delbrueckii AL-15 produced 13.8x10-1mg/ml biomassa on molasses. Key words: Poultry probiotics, bacterial and yeast isolates, glucose, sucrose, molasses

91 - 97

Authors: Endang Triwulaninngsih, P Situmorang, T Sugiarti, R.G Sianturi, D.A Kusumaningrum

This study has been conducted at the Laboratory of Physiology Reproduction, Research Institute for Animal Production (RIAP), Ciawi-Bogor, West Java. Sperms were collected from FH bulls with body weight 613 kg (FH-1) and 480 kg (FH-2) twice a week. Briefly after quality evaluation, semen was diluted in Tris-Citrate buffer medium, containing egg yolk (20% v/v) and (4% v/v) glycerol to get spermatozoa concentration of 50 x 106 per ml. Sperm diluents were added with glutathione (GSH) with doses of 0.0; 0.5; 1.0 and 1.5 mM as treatments A, B, C and D respectively. The diluted semen was then cooled from 35 to 5°C using a cooling machine for 60 minutes then stored in the refrigerator (5°C). Recorded parameters were the survivability of spermatozoa by evaluating the percentage of motile and live, the condition of acrosome and plasma membrane. Data were analysed by completely randomised design with the general linear model (GLM) procedure. The characteristics of collected semen were normal. Viability of spermatozoa stored at 5°C for 0, 1, 4 and 8 days shown by intact acrosomal were 74.42; 69.27; 57.80 and 42.58% for A, B, C and D respectively. Those data were significantly different (P<0.01). Motility, live and intact plasma membrane were 46.72; 52.34; 53.44 and 51.09%; 63.59; 69.11; 68.64; and 66.89%, and 66.01; 69.75; 68.38 and 68.44% for treatment A, B, C and D respectively. Additional 0.5 mM GSH gave the highest (P<0.01) motility, live and intact plasma membrane of sperm. Therefore, it is concluded that the effect of addition 0.5 mM of GSH to the sperm diluents can improve the viability of spermatozoa and possibly protect the spermatozoa from free radical damage. Key words: Glutathione, viability, spermatozoa

107 - 113

Authors: N.L.P Indi Dharmayanti, Risa Indriani, Darminto

Infectious Bronchitis is a contagious and acute respiratory disease in chickens caused by infectious bronchitis virus (IBV).Antigenic differences in IBV are associated with changes in the sequence of the spike glycoprotein (S). The subunit S1 which demonstrates more sequence variability than S-2 have been identified as hypervariable region (HVR-1 and 2). There were several IB virus field isolates included I-37 have been identified in Indonesia by serum neutralization method. However, gene sequence variation in HVR subunit S-1 had not yet been identified. Isolate I-37 was close to the serotype Connecticut 46 (Conn 46). The aim of this study is to identify sequence variation of HVR subunit S-1 gene of isolate I-37 produced by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and sequencing. Several procedures were carried out in the study including virus titration, propagation and was concentrated from the allantoic fluid infected with IBV. Then, RNA was extracted for RTPCR. urther the product was sequnced and its homology with IBV references from GenBank was compared by GenMac version 8.0. Result showed that isolate I-37 produced 515 bp of amplification product. Isolate I-37 and Conn 46 are same serotype, yet their HVR subunit S-1 nucleotides and amino acids (protein) differ by 6.9% and 15.6% respectively. It might be concluded that isolate I-37 was variant of Conn 46. Key words: Sequences variation, IBV, I-37 field isolate, HVR subunit S-1 gene

122 - 133

Authors: Muharam Saepulloh, Hope G Rovira

Infectious laryngotracheitis (ILT) is an acute, highly contagious respiratory disease of poultry characterized by respiratory disorder such as coughing with blood exudate from the trachea. The disease is caused by Herpesvirus of the family Herpesviridae and subfamily of Alphaherpesvirus. ILT is worldwide distribution and has been reported to be present in the Philippines since 1980. Since then, confirmation of subsequent outbreaks were not reported. Isolation was conducted from nine commercial layer chicken farms located at Lipa City, Batangas from May to July 2002. Tracheal and lung extracts were processed and inoculated into embryonated chicken eggs by chorio-allantoic membrane (CAM) inoculation. Five samples produced typical pock lesions in CAM after the second passage. Lesions observed were yellowish pocks with opaque edges, distributed throughout the CAM. A vaccine strain of the virus used as the positive control also produced similar pock lesions. Serological confirmation using the Agar Gel Immunodiffusion (AGID) test showed sharp precipitation lines reacting to a standard reference ILTV antisera (anti-NS175). All five isolates produced lines of precipitate identity among themselves and the positive control. This study confirms that the 2002 disease outbreak in the commercial layer chicken farms in Lipa City, Batangas was due to the ILTV. Key words: Infectious laryngotracheitis virus, agar gel immunodiffusion, isolation, pocks lesion, identification, CAM

139 - 145

Authors: A.P Sinurat, T Purwadaria, M.H Togatorop, T Pasaribu

Feed additives are commonly added in poultry feed as a growth promotant or to improve feed efficiency. The most common feed additive used is antibiotic at sub-therapheutic doses, although there is a controversy on its impact on human health. Previous results showed that Aloe vera gel could improve feed efficiency in broilers and an in vitro study showed that the extract have an antibacterial effect. Therefore, a further experiment was designed to study the response of broilers to Aloe vera gel or its extract as feed additives. Aloe vera was prepared in dry gel or chloroform-extract and included in the diet at levels of 0.25; 0.50 and 1.00 g/kg (equal to dry gel). Standard diets with or without antibiotic were also formulated as control and a commercial diet was included for comparison. The diets were fed to broilers from day old to 5 weeks. Each treatment has 9 replicates and 6chicks/replicate. Parameters observed were: feed consumption, weight gain and feed convertion ratios. Carcass yield, abdominal fat levels, relative weight of liver, gizzard, tractus digestivus and length of tractus digestivus were also measured at the end of feeding trial. The results showed that Aloe gel and its extract did not influence body weight gain and feed consumption of broilers significantly (P>0.05), but improved feed convertion slightly (3.50%). The response in this trial was similar as thosecommercial diet and diet added with antibiotic. There was no significant (P>0.05) effect of Aloe vera bioactives on carcass yield, abdominal fat level and relative weight of liver. However, Aloe vera gel and its extract tend to increase gizzard weight, gastro intestinal weight and length. The Aloe vera gel and its extract also reduced the total count of aerobic bacteria in the digesta of tractus digestivus. It is concluded that the Aloe vera gel improve feed efficiency in broilers by increasing the size of tractus digestivus and reducing the total count of aerobic bacteria in the gastro intestinal tract. Key words: Aloe vera, feed additives, broilers, anti-bacteria