KR19990007756A - Novel Livestock Wastewater Microbial Strains and Their Biological Uses - Google Patents

Abstract

The present invention relates to a novel livestock wastewater treatment strain and a biological use of the treated water, wherein the strain is isolated and cultured from the strains to purify livestock wastewater flowing out from pigs or barn and treated livestock wastewater using these strains Provides the use of water The treated water produced provides for use as deodorant and fertilizer.

Description

New Livestock Wastewater Treatment Microbial Strains and Biological Uses of Treated Water

The present invention relates to novel microbial strains suitable for livestock wastewater treatment and the biological use of treated water remaining after treatment using the same.

More specifically, the present invention, unlike the conventional activated sludge method, is a technique for separating mixed microorganism strains suitable for livestock wastewater and injecting the new strain into the livestock wastewater to purify it, and as a deodorant or liquid ratio of the purified treated water. It relates to a new use.

Conventional wastewater treatment methods are mainly to reduce the biological oxygen demand (BOD) and chemical oxygen demand (COD) of the wastewater by culturing activated sludge in the aeration tank of the wastewater treatment tank, and most of the wastewater to be treated is factory wastewater or domestic wastewater. .

Therefore, until now, there are no methods or facilities for treating livestock wastewater flowing from pigs or barn, so that they have been polluted in the environment by causing natural decay or discharge into rivers.

The present invention has been made in view of the above points to provide a useful and novel livestock wastewater treatment microorganism strains related to the method of collecting the wastewater generated in the barn in the collection tank first treatment and then biological treatment again. For that purpose.

Another object of the present invention is to provide a use of the treated water as a deodorant and liquid ratio in relation to a method for recycling livestock wastewater (hereinafter referred to as treated water) treated by the wastewater treated microorganism strain of the present invention.

The object of the present invention is to isolate the wastewater treatment strain from the soil, identify it, and then treat the strain with livestock wastewater to derive the test results, purify the wastewater, and analyze the water quality of the remaining treated water. It was achieved by investigating the deodorizing effect by spraying on and examining the growth process by spraying on horticultural plants as liquid fertilizer.

Hereinafter, the specific configuration and operation of the present invention.

1 to 5 is a photograph showing each microbial strain of the present invention.

The present invention comprises the steps of separating and identifying wastewater treatment strains; Treating livestock wastewater using the strain; Spraying the water to be applied to the barn or the bathroom to investigate the deodorizing effect; The dilution of the water to be treated of the present invention is used for horticultural crops to evaluate the value as liquid ratio.

Hereinafter, the specific configuration and operation of the present invention will be described with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example 1 Isolation Identification of Wastewater Treatment Strains

The wastewater treatment strain of the present invention selects a total of five mixed microorganism strains of bacteria and actinomycetes predominant species collected from pigs, and then cultures them in livestock wastewater or artificial media, isolates them, and identifies them as JSB 98.0. It was deposited on September 28, 1998 with the accession number KCTC 0524BP to the Gene Bank of the Institute of Biotechnology. Five dominant strains were identified as Micrococcus kristinae HO778 (FIG. 1), corynebacterium glutamicum HO780 (FIG. 2), corynebacterium glutamicum HO777 (FIG. 3), paracocus denitrificans HO779 (FIG. 4), and Bacillus mycoides HO781 (FIG. 5).

Experimental Example 1: Total bacteria

The total number of aerobic bacteria that survived in 1 mL of the well-mixed samples was counted by serial dilution in Humic acid medium. Actinomycetes were counted by continuous dilution in the culture medium, 1.12 × 10 ⁸ cfu / mL, and the number of fungi were 7.1 × 10 ⁵ cfu / mL was identified.

Experimental Example 2: Isolation of Dominant Fungi

The number of microorganisms observed in the plate count agar medium was 13 or more when visually classified, and each of them was counted to select five bacteria having a population of 10% or more relative to the total number of bacteria. Occupied 10-20%.

Strain 1: microcolony with cream colony, 20%

Strain 2: white and sticky colony, 18%

Strain 3: yellow colony, 14%

Strain 4: plane brownish colony, 11%

Strain 5: rhizoid from colony, 10%

Experimental Example 3: Identification of Dominant Bacteria

Strain 1 (HO 778):

(1) Morphology and Culture Characteristics

Granular colonies are round, intimate, convex and smooth, pale creamy, globular, 0.5-1.1 μm in diameter, mostly tetramolecular or irregular, forming gram-like, non-motile, spore-free .

(2) Physiological biochemical and chemical classification

* Catalase positive and oxidase positive were shown.

Assimilation test: L-arabinose, D-ribose (D-ribose), D-xylose, acetic acid, methyl pyruvate, mono- Methyl-succinate, propionic acid, putrescine (from Biolog GP).

Cell Fatty Acid Composition: C15: 0 anteiso 58.35%

C17: 0 anteiso 26.04%

C16: 0 iso 13.08%

C16: 0 2.53%

(3) judgment

Micrococcus kristinae (98% confidence)

Strain 2 (HO780):

(1) Morphology and Culture Characteristics

The nutritious colonies were smooth, finished, round, slightly pale, and generally pale yellow in white. According to the broth culture, the turbidity caused by the flocculant sediment was controlled, and short Gram-positive bacilli or ellipsoids were 0.7-1.0 × 1.0-3.0 μm, and they were formed singly or in pairs or irregular lumps. In the late cell culture stage, it was very short and mostly ellipsoid. Metachromatic granules formed prior to staining, lacking automatic force, and did not form ecdospore.

(2) Physiological biochemical and chemical classification

* Catalase positive, urease positive, nitrate reduction, caseinase negative, and gelatin solubilizer negative were shown.

Assimilation test: D-ribose, D-xylose, acetic acid, propionic acid (Biolog GP)

Cellular fatty acid composition: C18: 1 w9c 65.55%

C16: 0 26.61%

C17: 0 6.32%

C18: 0 1.52%

＊ Main menaquinone is MK-9 (H ₂ )

(3) judgment

Corynebacterium glutamicum (85% confidence)

Strain 3 (HO777):

(1) Morphology and Culture Characteristics

The nutritive colonies were smooth, finished, round, slightly pale, slightly dark yellow, and turbidity was controlled by flocculant sediment by broth culture.

Short Gram-positive bacillus or 0.7-1.0 × 1.0-3.0 size ellipsoid, isolated, paired or irregular lumps, very short in late stage of cell culture, and ellipsoid was mostly cocci. Pre-staining formed granules, no automatic force and no endospoa.

(2) Physiological biochemical and chemical classification

* Catalase positive, urease positive, nitrate reduction, caseinase negative, and gelatin solubilizer negative were shown.

Assimilation test: β-cyclodextrin, cellobiose, D-fructose, α-D-glucose, malcoat Maltotriose, D-mannose, 3-methyl glucose, palatinose, D-psicose, D-ribose, Sucrose, turanose, D-xylose, acetic acid, L-malic acid, L-asparagine, L-glutamic acid acid), uridine (from Biolog GP)

＊ Cell Fatty Acid Composition: C18: 1 w9c 57.46%

C16: 0 38.01%

C18: 0 1.45%

C15: 0 1.06%

＊ Main menaquinone is MK-9 (H ₂ )

(3) judgment

Corynebacterium glutamicum (99% confidence)

Strain 4 (HO779):

(1) Morphology and Culture Characteristics

The nutritious colonies were 2-3 mm in diameter, mostly round, smooth, finished, shiny, with a faint brownish white color. It showed globular cells (0.5-0.9 μm in diameter) or short rods (0.9-1.2 μm in length) and developed singly, paired or crusted, without resting periods, without gram-negative and automatic force.

(2) Physiological biochemical and chemical classification

* Catalase positive, oxidase positive, and nitric acid were reduced to nitrogen oxides and nitrogen molecules under anaerobic conditions, and escurin, starch, gelatin and Tween 80 hydrolysis were negative, phosphatase and urease negative.

* Assistance test: L-ribose, D-ribose, D-xylose, β-hydroxybutyric acid, D D-lactic acid methyl ester, L-lactic acid, methyl pyruvate, mono-methyl succinate

Cellular fatty acid composition: C18: 1w7c / w9t / w12t 82.98%

C10: 0 3OH 5.93%

C16: 1 iso I / C14: 0 3OH 3.29%

C18: 0 2.50%

C20: 1 w9t 1.65%

C17: 0 1.30%

C19: 0 1.27%

(3) judgment

Paracoccus denitrificans (85% confidence)

Strain 5 (HO781):

(1) Morphology and Culture Characteristics

Unique root colonies formed on agar were formed and chain-shaped bacilli were formed. Colonies were monotonous or frosted glass. Gram-positive, non-motile, spores ellipsoid, located in the center of the cell.

(2) Physiological biochemical and chemical classification

Assimilation test: L-arabinose (L-arabinose), D-arabitol (D-arabitol), D-ribose (D-ribose), D-alanine (D-alanine), adenosine Adenosine-5'-monophosphate (Biolog GP).

Cellular fatty acid composition: C15: 0 iso 20.71%

C15: 0 iso 2OH / C16: 1 w7c 14.98%

C13: 0 iso 11.25%

C17: 0 iso 8.64%

C12: 0 iso 6.24%

C14: 0 6.02%

Iso C17: 1 w5c 5.16%

Biochemical properties Characteristics result Catalase + Anaerobic growth + Boggs-Pro Scout Exam - Acid origin D-glucose + L-Arabinose - D-xylose - D-mannitol - Casein hydrolysis + Starch hydrolysis + Propionic acid - Citric Acid + Nitrate reduced to nitrite + Indole formation - Growth pH 6.8 Nutritional Broth + 5.7, SDA + NaCl growth 2% + 5% + 7% - 10% - Growth temperature 30 ℃ + 37 ℃ + 50 ℃ - 55 ℃ -

(3) judgment

Bacillus mycoides (90% confidence)

Experimental Example 4

Photographs of the dominant bacteria in Differential Interference Microscopy (× 1000) is shown in Figures 1 to 5.

Example 2 Treatment of Livestock Wastewater

The wastewater from the barn is collected in a collection tank and sequentially introduced into the mixed microorganism strain treatment tank separated in Example 1, and oxygen is supplied at the lower portion of the treatment tank by a BLOWER so that dissolved oxygen is maintained at 2 ppm or more, and the aeration time is 18 hours / day. Maintained.

This Example was repeated three times at the Health and Environment Institute of Seoul from June 28, 1998 to July 6. 1998, the results are shown in Table 2.

Livestock Wastewater Treatment Performance Inspection items Test result (mg / L) Judgment BOD 24.0 pass SS 52.5 pass Note: Seoul National Institute of Health and Environment

Example 3 Water Quality Analysis of Treated Water Obtained from Treatment of Livestock Wastewater

The amount of treated water produced according to Example 2 of the present invention generally corresponds to the amount of influent. In order to investigate the use of the treated water produced according to the present invention, the microbiological and physicochemical tests were performed to obtain water quality test results as shown in Table 3 below.

Water quality test result of treated water of the present invention (1998. 8. 20. Daesang Livestock Science Research Institute) sample Nitrous acid nitrogen Nitrate Nitrate Total hardness Fe pH Escherichia coli General bacteria Standard value (drinking water) NT Less than 10ppm 300ppm or less 0.3ppm or less 5.8 to 8.0 NT / 500cc 100 or less / 1cc This invention (treated water) 0.07 0.12 129.56 0.14 7.87 3,500 370 Note: NT = Not Detected

As can be seen from the experimental results, the treated water according to the present invention was found to be inferior as drinking water in physicochemical properties, but it was found that bacteria were higher than the reference value in microbiological properties.

On the other hand, in order to confirm the use as an inorganic fertilizer for the sample of the treated water of the present invention, the result of performing quantitative analysis of the inorganic component was evaluated as having the value as shown in Table 4.

Quantitative Analysis of Inorganic Components (Base Science Support Institute) ingredient Content (ppm) ingredient Content (ppm) Na 314 Ca 29.5 K 710 Fe 0.4 Mg 29.9 Cu 0.5 P 27.1 Zn 0.2 (Note) 1: 1998. 10. 19. Analysis result 2: Mn, Al, Cr, Cd, Pb, As, Hg is not detected in the sample

Example 4 Investigation of Use of By-Product Treated Water According to the Present Invention

The use of by-product treated water obtained in accordance with the present invention was investigated.

First, 1L of water to be treated in premature ejaculation was sprayed three times a day to pigs, barns, and toilets, and the smells of pig meal, milk powder, and phosphorus were completely removed. This shows that the deodorizing effect of the by-product treated water according to the present invention is very strong.

On the other hand, in order to confirm the use of the by-product treated water according to the present invention as a liquid ratio, the experimental ground and general groundwater sprayed 50-100 times of the treated liquid with a sprinkler three times a day to the greenhouse and golf course grass of a nearby farmhouse Experimental results of the control spraying bay was obtained as shown in the following Table 5 and Table 6.

Plant cultivation test results (pepper) division Control 50 * liquid 100 * liquid 150 * liquid Remarks Pn 23.7 25.7 24.8 24.2 Chl 2.10 2.78 2.92 2.73 Cs 3.24 4.54 4.72 4.48 Growth 7.9 90.5 88.5 87.7 Products 34.7 48.4 47.2 45.8 Note: Pn: Photosynthesis rate (unit: μmol CO _2m -2S-1) Chl: Chlorophyll content (unit: mg / g) Cs: Pore conductance (unit: cm / s) Growth: Length growth (unit: cm) Deletion amount (unit: number)

As can be seen from the experimental results, it can be seen that the photosynthesis rate, chlorophyll content, pore conductance, length growth, and total loss are significantly improved compared to the control.

Grass growth test results division Control 50 * liquid 100 * liquid 150 * liquid Pn 5.22 4.30 9.28 8.82 Cs 0.30 0.82 0.73 0.68 Tr 4.8 10.8 9.72 8.92 Note: Pn: Photosynthesis rate (unit: μmol CO _2m -2S-1) Cs: Pore conductance (unit: cm / s) Tr: transpiration (unit: mg H ₂ O / m ² / sec)

As can be seen from the above experimental results, the experimental group treated with 50 × liquid of the present invention to be treated with liquid ratio improved the photosynthesis rate and transpiration activity by 50%, respectively, compared to the control group.

As apparent from the above examples and experimental examples, the present invention has the effect of providing a novel mixed strain that can efficiently treat livestock wastewater. In addition, the treated water treated by the novel strain of the present invention has an excellent deodorizing effect and growth promoting effect of the plant, and thus has an effect of providing a deodorant or a liquid fertilizer, which is a very useful invention for environmental conservation and fertilizer industry.

Claims (3)

Novel Microbial Strain JSB-98.0 (KCTC 0524BP) Useful for Livestock Wastewater Treatment The livestock wastewater treatment microorganism strain JSB-98.0 (KCTC 0524BP) of Claim 1 is used for processing the livestock wastewater, and using the remaining untreated water. The method according to claim 2, wherein the use of the water to be treated is a fertilizer and a deodorant.