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Isolation and Screening of Microbes for The microorganism used as probiotics must be bio-safety, could be cheaply and easily Purwadaria, Tresnawati; Kompiang, I Putu; Darma, Jinadasa; ., Supriati; Sudjatmika, Emi
Indonesian Journal of Animal and Veterinary Sciences Vol 8, No 2 (2003)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (244.605 KB) | DOI: 10.14334/jitv.v8i2.376

Abstract

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
The comparison of cellulase activities from bacteria and molds isolated from termites Purwadaria, Tresnawati; Marbun, Pesta A; Sinurat, Arnold P; Ketaren, Pius P
Indonesian Journal of Animal and Veterinary Sciences Vol 8, No 4 (2003)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (149.599 KB) | DOI: 10.14334/jitv.v8i4.393

Abstract

Screening for choosing the best bacterium and mold producing cellulase was carried out from eight xylanolytic bacteria and three cellulolytic molds isolated from termites. Each bacterium and mold was inoculated on the agar medium containing minerals and carboxymethylcellulose. The diameters of colony and clearing zones were measured after stained with Congo red. Four bacteria: Bacillus larvae XB 1-1, B. larvae XU 2-2, Bacillus PU 2-2, and B. pumilus PU 4-2 showing higher clearing zone ratio were further submerged cultured in PM medium containing 2% wheat pollard in the shaker incubator at room temperature, 150 rpm for 36 and 48 hours. The three molds: Aspergillus flavus S 13, MS 21, and Penicillium nalgiovense S 11 were also cultured in the same way as bacteria, but in Mandels medium containing 2% wheat pollard for three and five days incubation time. B. pumilus PU 4-2 and A. flavus S 13 produced the highest CMCase in each group respectively. All enzyme assays of A. flavus S 13 had higher activities than that of B. pumilus PU 4-2. The specific activity of CMCase, avicelase, FPase, and cellobiohidrolase were the highest on the enzyme of A. flavus S 13 produced from 3 days incubation time, while its β- glucosidase was the best for five days incubation. In term of activities, A. flavus S 13 had the highest ability to produce cellulase. The ability of B. pumilus PU 4-2 to produce cellulase and xylanase at the same time was an additional value.   Key words: Bacillus pumilus, Aspergillus flavus, cellulases
Substitutions of soybean meal with enriched palm kernel meal in laying hens diet Sinurat, Arnold P.; Purwadaria, Tresnawati; Ketaren, Pius P.; Pasaribu, Tiurma
Indonesian Journal of Animal and Veterinary Sciences Vol 19, No 3 (2014)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14334/jitv.v19i3.1081

Abstract

A series of experiment was conducted in order to substitute soybean meal (SBM) with palm kernel cake (PKC) as a protein source in laying hens diet. First experiment was to increase its protein and amino acids content by fermentation process and followed by enzymes supplementation to improve nutrient digestibilities. Second experiment was conducted to evaluate the effectiveness of enzyme- supplemented fermented palm kernel cake (EFPKC) to replace SBM in laying hens diet. The energy (AME) of the PKC, the fermented PKC (FPKC) and the EFPKC was measured by ileal amino acids digestibility (IAAD) in broilers. The AME and the IAAD values of the EFPKC were used for diet formulation in the feeding trial. A feeding trial was performed in laying hens, aged 51 weeks for 8 weeks egg production. Five diets with different levels of substitution of SBM with EFPKC but similar nutrient contents were formulated, i.e.: 1. Control (without EFPKC), 2. 25% of SBM in control diet substituted with EFPKC, 3. 50% of SBM in control diet substituted with EFPKC, 4. 75% of SBM in control diet substituted with EFPKC and 5. 100% of SBM in control diet substituted with EFPKC. Each diet was fed to 24 hens (6 replicates of 4 birds/ replicate). Results of the experiment showed that the fermentation of PKC increased the crude protein and most of the amino acids contents except the threonine and arginine, but decreased its AME.  Supplementation of enzymes (BS4 or CE) improved the ileal amino acid digestibilities of the fermented PKC. However, only BS4 enzymes increased the AME of the EFPKC. About 25% to 50% of the SBM in the diet could be substituted with the EFPKC without any detrimental effect on the performances of laying hens. Key Words: Soybean Meal, Palm Kernel Cake, Fermentation, Enzyme, Egg Production
Effect of enzyme supplementation on nutritive values of fermented palm kernel cake used to substitute soybean meal in broiler diet Sinurat, Arnold P.; Purwadaria, Tresnawati; Purba, Maijon
Indonesian Journal of Animal and Veterinary Sciences Vol 20, No 3 (2015): SEPTEMBER 2015
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (257.23 KB) | DOI: 10.14334/jitv.v20i3.1185

Abstract

Two experiments was designed to improve nutritional values of palm kernel cake (PKC) by biofermentation process, followed by enzyme supplementation to substitute soybean meal (SBM) in broilers diet. A factorial of 2 x 2 design was applied in the first experiment, i.e. fermentation process (non fermented PKC and fermented PKC) and enzyme supplementation (no enzyme and +BS4 enzyme). Dry matter (DM) digestibility, AME and amino acids ileal digestibility (IAAD) of the treatment ingredients were measured in broiler chickens. Seven replications were applied for the DM and AME assays and 3 replications for IAAD assay. Second experiment was designed to study the effect of SBM substitution with enzyme supplemented FPKC (EFPKC). Four diets were formulated, i.e., control diet without EFPKC, 10%, 20 and 40% SBM substituted with EFPKC. All diets were formulated to meet the nutrient requirements of broilers. Each diet was fed to broilers from 1 to 35 d. Body weight, feed consumption, FCR and mortalities were measured. Carcass yield, abdominal fat and weight of liver and gizzard were measured at the end of experiment. Results showed that fermentation of PKC increased the DM digestibility, the AME was also increased but not significant. Enzyme supplementation did not affect the DM digestibility and AME of PKC. Fermentation process significantly (P<0.05) decreased IAAD of some indispensable amino acids. However, supplementation of enzyme did not affect the IAAD of indispensable amino acids. Substitution of soybean meal with EFPKC reduced the feed intake and growth rate of broilers. Key Words: Palm Kernel Cake, Fermentation, Enzyme, Broilers, Soybean Meal
Effect of enzyme supplementation on nutritive values of fermented palm kernel cake used to substitute soybean meal in broiler diet Arnold P. Sinurat; Tresnawati Purwadaria; Maijon Purba
Jurnal Ilmu Ternak dan Veteriner Vol 20, No 3 (2015): SEPTEMBER 2015
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (257.23 KB) | DOI: 10.14334/jitv.v20i3.1185

Abstract

Two experiments was designed to improve nutritional values of palm kernel cake (PKC) by biofermentation process, followed by enzyme supplementation to substitute soybean meal (SBM) in broilers diet. A factorial of 2 x 2 design was applied in the first experiment, i.e. fermentation process (non fermented PKC and fermented PKC) and enzyme supplementation (no enzyme and +BS4 enzyme). Dry matter (DM) digestibility, AME and amino acids ileal digestibility (IAAD) of the treatment ingredients were measured in broiler chickens. Seven replications were applied for the DM and AME assays and 3 replications for IAAD assay. Second experiment was designed to study the effect of SBM substitution with enzyme supplemented FPKC (EFPKC). Four diets were formulated, i.e., control diet without EFPKC, 10%, 20 and 40% SBM substituted with EFPKC. All diets were formulated to meet the nutrient requirements of broilers. Each diet was fed to broilers from 1 to 35 d. Body weight, feed consumption, FCR and mortalities were measured. Carcass yield, abdominal fat and weight of liver and gizzard were measured at the end of experiment. Results showed that fermentation of PKC increased the DM digestibility, the AME was also increased but not significant. Enzyme supplementation did not affect the DM digestibility and AME of PKC. Fermentation process significantly (P<0.05) decreased IAAD of some indispensable amino acids. However, supplementation of enzyme did not affect the IAAD of indispensable amino acids. Substitution of soybean meal with EFPKC reduced the feed intake and growth rate of broilers.
Isolation and Screening of Microbes for Poultry Probiotics and Their Growth on Different Sugar Resources Tresnawati Purwadaria; I Putu Kompiang; Jinadasa Darma; Supriati .; Emi Sudjatmika
Jurnal Ilmu Ternak dan Veteriner Vol 8, No 2 (2003): JUNE 2003
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (244.605 KB) | DOI: 10.14334/jitv.v8i2.376

Abstract

The microorganism used as probiotics must be bio-safety, could be cheaply and easily 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. macrolides 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. apiaries on sucrose (the sixth on molasses). B. apiaries 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 13.8x10-1 mg/ml biomassa on molasses.
The comparison of cellulase activities from bacteria and molds isolated from termites Tresnawati Purwadaria; Pesta A Marbun; Arnold P Sinurat; Pius P Ketaren
Jurnal Ilmu Ternak dan Veteriner Vol 8, No 4 (2003): DECEMBER 2003
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (149.599 KB) | DOI: 10.14334/jitv.v8i4.393

Abstract

Screening for choosing the best bacterium and mold producing cellulase was carried out from eight xylanolytic bacteria and three cellulolytic molds isolated from termites. Each bacterium and mold was inoculated on the agar medium containing minerals and carboxymethylcellulose. The diameters of colony and clearing zones were measured after stained with Congo red. Four bacteria: Bacillus larvae XB 1-1, B. larvae XU 2-2, Bacillus PU 2-2, and B. pumilus PU 4-2 showing higher clearing zone ratio were further submerged cultured in PM medium containing 2% wheat pollard in the shaker incubator at room temperature, 150 rpm for 36 and 48 hours. The three molds: Aspergillus flavus S 13, MS 21, and Penicillium nalgiovense S 11 were also cultured in the same way as bacteria, but in Mandels medium containing 2% wheat pollard for three and five days incubation time. B. pumilus PU 4-2 and A. flavus S 13 produced the highest CMCase in each group respectively. All enzyme assays of A. flavus S 13 had higher activities than that of B. pumilus PU 4-2. The specific activity of CMCase, avicelase, FPase, and cellobiohidrolase were the highest on the enzyme of A. flavus S 13 produced from 3 days incubation time, while its β- glucosidase was the best for five days incubation. In term of activities, A. flavus S 13 had the highest ability to produce cellulase. The ability of B. pumilus PU 4-2 to produce cellulase and xylanase at the same time was an additional value.   Key words: Bacillus pumilus, Aspergillus flavus, cellulases
Optimization of BS4 Enzyme production with Different Substrate Thickness and Type of Trays Tuti Haryati; Arnold P Sinurat; H Hamid; Tresnawati Purwadaria
Jurnal Ilmu Ternak dan Veteriner Vol 24, No 4 (2019): December 2019
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14334/jitv.v24i4.2055

Abstract

BS4 enzyme that is produced from solid substrate fermentation (SSF) on coconut cake with Eupenicillium javanicum BS4 in tray bioreactor has been applied as a feed additive. It increases the nutritional value of animal feedstuff. The BS4 production on SSF may be influenced by the better aeration through the perforated trays or by the thinner substrate. The aim of this research is to optimize the production of BS4 with different substrate thicknesses and types of trays. The trial was carried out using a factorial randomized design (2x2x3) with 6 replicates. The first factor was the type of trays: i.e., non-perforated and perforated tray. The second factor was the thickness of the substrate: i.e., 1.5 and 3.0 cm, while the third factor was the duration of fermentation: i.e. 5, and 7 days. The variables observed were moisture content, dry matter loss (DML), mannanase and saccharification activities, soluble protein content, their specific activities, and yield. Statistical analysis showed no interactions between the three factors, but there were interactions between types of trays and substrate thicknesses, as well as type of trays and incubation times on the mannanase activity and yield of mannanase. The results showed that DML was observed on day 7 were around 31.43- 36.89. The highest mannanase activity was observed on the non-perforated tray with 3 cm thickness on day 7. The saccharification activity towards palm kernel meal was better in the non-perforated tray on day 7 but not influenced by The yield value of mannanase and saccharification activities on a non-perforated tray with 3.0 cm thickness on day 7 was also the highest. Based on energy efficiency and the cost of production, it can be concluded that the optimum condition to produce the BS4 enzyme was observed in the non-perforated tray with 3 cm thickness and fermented for 7 days.
Substitutions of soybean meal with enriched palm kernel meal in laying hens diet Arnold P. Sinurat; Tresnawati Purwadaria; Pius P. Ketaren; Tiurma Pasaribu
Jurnal Ilmu Ternak dan Veteriner Vol 19, No 3 (2014): SEPTEMBER 2014
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (277.333 KB) | DOI: 10.14334/jitv.v19i3.1081

Abstract

A series of experiment was conducted in order to substitute soybean meal (SBM) with palm kernel cake (PKC) as a protein source in laying hens diet. First experiment was to increase its protein and amino acids content by fermentation process and followed by enzymes supplementation to improve nutrient digestibilities. Second experiment was conducted to evaluate the effectiveness of enzyme- supplemented fermented palm kernel cake (EFPKC) to replace SBM in laying hens diet. The energy (AME) of the PKC, the fermented PKC (FPKC) and the EFPKC was measured by ileal amino acids digestibility (IAAD) in broilers. The AME and the IAAD values of the EFPKC were used for diet formulation in the feeding trial. A feeding trial was performed in laying hens, aged 51 weeks for 8 weeks egg production. Five diets with different levels of substitution of SBM with EFPKC but similar nutrient contents were formulated, i.e.: 1. Control (without EFPKC), 2. 25% of SBM in control diet substituted with EFPKC, 3. 50% of SBM in control diet substituted with EFPKC, 4. 75% of SBM in control diet substituted with EFPKC and 5. 100% of SBM in control diet substituted with EFPKC. Each diet was fed to 24 hens (6 replicates of 4 birds/ replicate). Results of the experiment showed that the fermentation of PKC increased the crude protein and most of the amino acids contents except the threonine and arginine, but decreased its AME.  Supplementation of enzymes (BS4 or CE) improved the ileal amino acid digestibilities of the fermented PKC. However, only BS4 enzymes increased the AME of the EFPKC. About 25% to 50% of the SBM in the diet could be substituted with the EFPKC without any detrimental effect on the performances of laying hens.
Apparent Induction of Xylanase by Bacillus pumilus PU4-2 using Pretreated Substrates SHERLY WIDJAJA; TRESNAWATI PURWADARIA; PIUS PERTUMPUN KETAREN
Microbiology Indonesia Vol. 2 No. 1 (2008): April 2008
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (47.031 KB) | DOI: 10.5454/mi.2.1.9

Abstract

Bacillus pumilus PU4-2 produces xylanase (&acirc;-1,4-D-xylan xylanohydrolase; EC 3.2.1.8) in wheat pollard with high activity. Water and NaOH-soaked pollard were used in this research to enhance the production of assayable enzyme. Enzyme activity was produced in minimal media containing 3% w/v untreated or water or NaOH-soaked pollards in 250 ml flasks incubated on shaker incubator at 30&deg;C and 150 rpm for 36 h. The production was also compared to untreated oatmeal known as an inducer substrate. The highest xylanase activity was obtained by using untreated pollard as a sole carbon source. The enzyme activity was 157 U ml-1 with specific activity at 718 U mg-1. Xylanase production using different soaking time for water pretreated pollard also confirmed that untreated pollard was the best inducer. The production was not influenced by different water soaking times used to remove reducing sugar. Although pretreatment decreased the reducing sugar, the reduction did not enhance assayable enzyme levels. The production was best induced by the soluble oligosaccharides of untreated pollard. We conclude that B. pumilus PU4-2 was able to produce xylanase with reducing sugar content up to 660 ppm present in production medium. With this reducing sugar level, repression of enzyme production was not detected in the production medium.