Oral SessionProceedings

The 17th Asian-Australasian Association of Animal Production Societies Animal Science Congress

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Quality of Liquid Organic Fertilizer from Rabbit fs Urine with The Addition of Nitrifying Bacteria, Urea, and Leucaena leucocephala

Theresia Galuh Wandita, Setyo Rahmat Darmawan, Suharjono Triatmojo, Nanung Agus Fitriyanto

Faculty of Animal Science and Industry, Universitas Gadjah Mada

INTRODUCTIONRabbit (Oryctolagus cuniculus) is herbivore which is very good in feed conversion (Lebas et al. , 1997). Since they barely drink water and only consume forage, rabbit’s urine contains high levels of nitrogen. For a day, a rabbit can produce 100 ml of urine a bunny can produce 25 ml. Urine contains ammonia (NH3)--a colorless gas which is lighter than air and possesses strong odor. Ammonia and other nitrogenous gases results from the digestion of protein, part of which is lost in manure and urine (Atia et al. , 2005). Solid and liquid animal waste is a type of waste that can be used as liquid organic fertilizer through a fermentation process (Salisbury and Ross, 1995). Additional treatment of microbial decomposers can intensively improve the form of organic fertilizer. Pseudomonas is a group of bacteria that is most important in denitrification (Tortora et al. , 2001). Candida is yeast that is commonly found in soil with high organic contents (Prasad, 2005). Urea is inorganic fertilizers, shaped as white crystalline solid, which is highly soluble in water and contents 46% of nitrogen (Boswell et al., 1997). The aim of Leucaena leucocephala addition is to increase the nitrogen content in the organic fertilizer. Therefore, it can be used as nutrient growth for decomposers.

MATERIALS AND METHODSGrowth in liquid conditionThe organisms, Pseudomonas sp. LS3K and Candida sp. LS3T, were grown in basal salt medium (g/L): meat extract, 0.1 biological peptone, 0.1 and NaCl, 0.05. The cultivation was performed at 28℃ at 120 rpm on a shaking incubator for 48 h. Every 3 h, 1 ml sample was read by spectrophotometer (600nm).

Organic fertilizer productionThe fertilizer was made by fermented aerobic process. Each substance (microbial, urea, L. Leucocephala , and control) was used at 1% (v/v) concentration which is put into 500 ml of rabbit’s urine. Fermented process was conducted for a week. Boric acid was also provided to determine ammonia concentration in liquid organic fertilizer.

Ammonia concentrationAmmonia was caught by boric acid which analyzed using Nessler method. Every day, 1 ml sample was read by spectrophotometer (425nm).

Physical, chemical and microbiology parametersPhysical parameters were tested based on physical condition of liquid organic fertilizer, such as pH, color, odor, volume, and temperature. Nitrogen, phosphor, organic carbon, and potassium were measured as chemical parameters of liquid organic fertilizer. Nitrogen phosphor organic carbon was measured by spectrophotometer method (636 889 591 nm). For microbiology parameters, the organisms were grown in basal salt medium (g/L): meat extract, 1.0 biological peptone, 1.0 NaCl, 0.5 and agar powder, 1.5. Isolated microbial (Pseudomonas sp. LS3K and Candida sp. LS3T) was grown by spread plate method. After 3 d incubation, isolated microbial was able to shape colonies.

RESULTS AND DISCUSSIONThe ability of all strains in growing in liquid medium was shown in Fig. 1A it showed different profiles from Pseudomonas sp. LS3K and Candida sp. LS3T. Based on the Fig. 1A, Pseudomonas sp. LS3K directly grew into log phase, while lag phase of the growth of Candida sp. LS3T was occurred at 0 h until 3 h. Log phase of Pseudomonas sp. LS3K confirmed faster compared with the log phase of Candida sp. LS3T log phase of Psedomonas sp. LS3K

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started from 0 h until 18 h and Candida sp. LS3T started from 3 h until 36 h. The differences in the growth profiles affected by genetic potential of microorganisms, medium, and the growth conditions (Prescott et al ., 1999). In the stationary phase, microorganism only actives in metabolic activity, and the cell division is stopped (Prescott et al. , 1999 Tortora et al ., 2001). After 48 h incubation, the color of liquid medium was changed from clear medium into a murky yellow medium.The principle of Nessler method is reaction between Nessler reagent with ammonia in alkali condition that will form a murky brown colloidal dispersion. Based on Fig. 1B, the addition of microbial showed a lower pattern of liquid organic fertilizer it means that the microbial is the best treatment to reduce ammonium concentration in rabbit’s urine compared with the others treatment and control. The data were then analyzed using completely randomized design of statistical analysis. Based on the results, different treatment given on liquid organic fertilizer affected the ammonia concentration (ppm). However, there was no an effect on ammonia concentration when L. Leucocephala was added. Ammonia concentration in the microbial medium was significantly different (P<0.05) with urea medium, meanwhile it was not significantly different with L. leucocephala medium and control. The average of ammonia released per day (ppm) by each treatment was 5936.5 (microbial) 7789.63 (control) 7972.5 (L. leucocephala ) and 30529.02 (urea). It is because of 46% nitrogen contained within the urea that caused ammonia production was higher than other treatments. Ammonia used by microbial as an-organic nitrogen source to cell growth through enzymatic reaction (McCrory and Hobbs, 2001 Satoh et al., 2004).Liquid organic fertilizer from rabbit’s urine was fermented by aerobic process for 8 days. It must be examined for physical analysis every day. The result of physical analysis can be seen in Table 1. The explanation as follow: 1) The volume of liquid organic fertilizer was decreased because the oxygen supply of aerator caused evaporation liquid organic fertilizer with additional treatments has a different temperature and pH. 2) The temperature of liquid organic fertilizer with the addition of microbial and L. leucocephala was showed higher than control and urea. 3) The highest pH found in in the medium in which the microbial was added it reached 10.75. However, pH contained within the medium was the lowest when Leucaena leucocephala is added. Organic compounds degraded by microbial that will produce organic acids. Hence, pH will be decreased. In further step, the microbial will degrade the organic acids. Therefore, pH will be increased and make alkali conditions.From the Table 1, we can see that the medium with the addition of microbial has no odor because microbes play an important role in both production and reduction of malodors (Zhu, 2000). Microbial treatments have been extensively used in municipal livestock waste to degrade organic matter (Low and Chase, 1999) and microbial treatments are emerging to treat livestock waste, since degradation of organic matter in livestock waste relies on microorganisms (Sund et al. , 2001). Some of bacteria have the ability to reduce nitrate in aerobic condition, such as genus Paracoccus, Pseudomonas, Bacillus , and Alcaligenes (Wu et al. , 2013). On the contrary, the medium with the addition of urea had odor because the organic compounds that contained in rabbit’s urine were not degraded.Besides the physical parameters, the liquid organic fertilizer also examined for chemical parameters. Based on the Fig. 2A, the nitrogen concentration of urea that contained within the medium had highest percentage (0.88%) it caused by the nitrogen within the urea was not degraded well and the nitrogen itself was solubility in rabbit’s urine. On the other hand, the lowest nitrogen concentration is in control and microbial medium (0.05%). The result of statistical analysis showed that the nitrogen concentration in liquid organic compounds with the additional treatments was significantly different (P<0.05) with control medium. Organic carbon in liquid organic fertilizer of rabbit’s urine with the additional treatment was shown in Fig. 2B. From the percentage, we can see that the addition of Leucaena leucocephala was the highest organic carbon (0.014%) contained within the medium. Conversely, urea contained within the medium was the lowest organic carbon (0.01%). Based on statistical analysis, organic carbon concentration in liquid organic compounds with the additional treatments was not significantly different with the control.Based on the Fig. 2C, the medium with the addition of L. leucocephala showed the highest phosphor (0.025%) and the lowest phosphor concentration is the medium which contained urea (0.02%) it caused by urea that only contained nitrogen. The result of statistical analysis showed that phosphor concentration in liquid organic compounds with the additional treatments was not significantly different with control medium. Microorganisms used organic carbon as carbon source to produce energy, and when the microorganisms were died, the organic carbon will be released as carbon dioxide/CO2 (Sholikah et al. , 2013). Potassium in liquid organic fertilizer from rabbit’s urine with the additional treatment was shown in Fig. 2D. The highest potassium (0.82%) found in the medium that contained of microbial while the lowest potassium found in control medium (0.63%). Based on statistical analysis, potassium concentration in liquid organic compounds with additional treatments was

The 17th Asian-Australasian Association of Animal Production Societies Animal Science Congress

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significantly different (P<0.05) with control. Hidayati et al. (2011) stated that potassium in substrate substances used by microbial as catalyst. Potassium was bonded and stored within cell by bacteria or fungi and when it was re-degraded, the potassium will be reappeared.Microbiology parameter was analyzed by colony growth in solid medium. Isolated bacteria were grown and maintained by spread plate method. After 3 days incubation, the bacteria were able to shape, and then the colonies were count. The result of colonies computation can be seen in Table 2. The medium with the addition of microbial absolutely showed the best result which was 17.3 x 104 CFU/ml and medium that contained urea showed the worst result, 0.2 x 104 CFU/ml. It definitely caused by Pseudomonas sp. LS3K and Candida sp. LS3T contained within the liquid organic fertilizer. Pseudomonas sp. LS3K and Candida sp. LS3T can grow in the medium with ammonia concentrated. Jenie and Rahayu (1993) stated that microbial growth was influenced by multiple factors, such as the source of energy, protein, mineral, pH, and temperature.

CONCLUSIONSPseudomonas sp. LS3K can grow faster than Candida sp. LS3T. However, the growth of those bacteria was not significantly different. The best treatment to reduce ammonia was using microbial addition. Furthermore, the addition of microbial was significantly different in potassium concentration and microbiology parameter. Then, the addition of urea was significantly different in nitrogen concentration, but it was not significantly different in other parameters. It was unfortunate that the addition of Leucaena leucocephala was not significant in any parameters.

KEYWORD：Animal waste, Liquid organic fertilizer, Nitrifying bacterium, Urea, Leucaena leucocephala

Figure 1. Comparison of microbial growth in liquid medium

The 17th Asian-Australasian Association of Animal Production Societies Animal Science Congress

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Figure 2. Ammonia concentration in liquid organic fertilizer

Figure 3. Profile of liquid organic fertilizer chemical analysis =

(A) Nitrogen, (B) Organic Carbon, (C) Phosphor, (D) Potassium

The 17th Asian-Australasian Association of Animal Production Societies Animal Science Congress

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The 17th Asian-Australasian Association of Animal Production Societies Animal Science Congress

684

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