• Journal of Animal Environmental Science
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• v.12 no.1
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• pp.41-44
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• 2006

Broiler manure has much nutrient that can be used as the organic fertilizer to enhance the fertility of soil. However, if it is used directly without biodegradation of organic materials and destruction of weed seed and harmful bacteria, it can produce the generation of weed and disease of plant. Composting of manure is a biodegradation of organic materials into inorganic materials and humus. To proceed biodegradation of manure effectively and enhance the composting performance, optimum environmental condition for microbial growing should be maintained. Environmental variables which can influence the growing activity of microbes are moisture content, pH, porosity, C/N of the composting materials and oxygen supply quantity. Oxygen and porosity are usually supplied by aeration or mixing of materials. This study was intended to investigate the effect of mixing frequency on the composting performance. Mixing of composting materials was performed by turning the bioreactor up and down by hand without any mechanical energy. The broiler manure was captured from the greenhouse type broiler ham as the compounds of broiler manure and rice-hulls, which were used as the base materials. To compost the compounds of broiler manure and rice-hulls, dairy manure was mixed to get appropriate characteristics of composting material. Composting temperature over $55^{\circ}C$ for killing pathogen and weed seed was maintained for longer period with increase of mixing frequency.

• Reproductive and Developmental Biology
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• v.32 no.1
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• pp.45-49
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• 2008

We have previously produced transgenic (TG) mice expressing the human lactoferrin (hLF), interleukin-10 (hIL-10), and thrombopoietin (hTPO) proteins in the milk. In this study, we examined whether simple crossbreeding between two kids of a single transgenic mouse can produce double transgenics co-expressing two human proteins.. The hLF male, and the hIL-10 male were crossbred with the hIL-10 and hTPO females, and the hTPO female, respectively. PCR analysis for genotyping showed 32%, 23% and 24% double transgenic rates for hLF/hIL-10, hLF/hTPO, and hIL-10/hTPO transgenes, respectively. We analyzed the expression levels of the human proteins from double transgenic mice and compared those with their single transgenic siblings. All double transgenic co-expressed two human proteins at comparable levels to singles', unless hTPO was not co-expressed: for hLF, 1.1 mg/ml in hLF/hIL-10, whereas 0.5 mg/ml in hLF/hTPO; for hIL-10, 4.1 mg/ml in hIL-10/hLF, whereas 1.4 mg/ml in hIL-10/hTPO. Ihe downregulation of hTPO to half level of singles' was observed in double transgenic mice. The possible reason why hTPO co-expressed might lead to down-regulation of another human protein was discussed. These results suggested that double transgenic generated by crossbreeding between two singles' could be useful system for bioreactor.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.69-75
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• 2013

Enrofloxacin is a fluoroquinolone antibiotic approved for the treatment of infections in animals. Because of the side effects to consumers of animal products, the maximum residue limits (MRLs) of enrofloxacin in animal tissues for consumption are regulated. In this study, a monoclonal antibody (mAb) against enrofloxacin was prepared and characterized for the development of a direct competitive enzyme-linked immunosorbent assay (ELISA). The obtained mAb, Enro44, was highly specific for enrofloxacin and had a 50% inhibition concentration ($IC_{50}$) of 1.99 ng/ml in a competitive ELISA, and the limit of detection (LOD) was 0.50 ng/ml. The cross-reactivity of the mAb with other quinolones and fluoroquinolones was lower than 0.01%. The subclass of the mAb Enro44 was identified as IgG1. The antigen (Ag)-captured direct competitive ELISA using the mAb Enro44 was tested on different spiked samples, including chicken muscle, cattle milk, and cattle urine, and the assay demonstrated recoveries of 82-112%, 80-125%, and 78-124%, respectively. Furthermore, the quantitation of enrofloxacin obtained from the ELISA and from high-performance liquid chromatography (HPLC) was in good agreement, with the linear regression coefficient between 0.933 and 1.056. The cDNAs encoding a heavy-chain Fd fragment (VH and CH1) and a light chain of the mAb Enro44 were cloned and sequenced. Taken together, the results obtained reveal a potential use of this mAb in an ELISA for the detection of enrofloxacin in food samples. The information of amino acid sequence of this mAb will be useful for further modification and production of the mAb in a bioreactor.

• Journal of Animal Environmental Science
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• v.16 no.2
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• pp.143-152
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• 2010

Conditions for artificial culture of Lemna Paucicostata and its nutritional values were examined in this study. Lemna P. was cultured using artificial wastewater and a bioreactor (total volume $2,630\;cm^3$, working volume $2,240\;cm^3$) was operated at conditions of 6,250 lux and $28^{\circ}C$. Water flow affected the growth of Lemna P.: growth rate was very high (more than $1.1\;d^{-1}$) at a condition of no-water movement, but it was very low (less than $0.15\;d^{-1}$) when water moved slowly. The growth of Lemna P. was higher in $16h\;d^{-1}$ light cycle than in Sand $24h\;d^{-1}$, and it was also severely affected by the initial $NH_4$-N levels of wastewater. The growth rate of Lemna P. was high in lower $NH_4$-N level, indicating that the growth rate is in inverse proportion to $NH_4$-N concentration in wastewater. However, the contents of crude protein (CP) of Lemna P. were proportional to the initial $NH_4$-N concentration. The CP contents of Lemna P. cultured at 2, 10, 50 and 100 $NH_4$-N mg $L^{-1}$ was 18, 24, 37, 43%, respectively, showing the Lemna P. cultured at 50 and $100\;mg\;L^{-1}$ had similar protein contents to linseed (CP 35%), cottonseed (CP 38%) and soybean (CP 45%). Fat, protein, fiber, NDF and ADF contents of Lemna P. harvested at conditions of $16h\;d^{-1}$ light cycle and less than $2\;mg\;L^{-1}$ of $NH_4$-N level was 2.8, 18, 27, 20, 41 and 65.7%, respectively. Since the growth rate of Lemna P. was very high (more than $1.1\;d^{-1}$) at those conditions, it was convinced that mass production of valuable protein and fiber sources are feasible. In particular, since the Lemna P. has unsaturated fatty acids found mainly in animal fat as well as beneficial fatty acids to health such as C18:ln9c, C18:2n6c, C20:5n3 and C22:2, the Lemna P. biomass would be a highly valuable alternative feed source to grains.

• Journal of Life Science
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• v.26 no.3
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• pp.275-281
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• 2016

Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.87-94
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• 2002

This study was performed during the four seasons for the production of transgenic pigs containing the Cellulase Digest Gene. Purebred Landrace gilts and sows approximately 8∼15 months of age (n=126) were used for the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality fur zygote collection. Synchronization and superovulation procedures were used that cyclic gilts were synchronized with 20mg altrenogest (ALT) per day for 9 days after PG600 administration followed by superovulation with 1000 IU pregnant mares serum gonadotropin (PMSG) and 750IU human chorionic gonadotrophin (hCG). The cellulase digestion gene for microinjection is rat elasterase promoter (rEl) linked to CelD gene. After hormone treatment, 1,422 embryos were collected from 91 donors and 95.6% (1,359/1,422) embryos were in 1-cell stage which can be visualized the pronuclei for DNA microinjection. A total of 725 DNA microinjected embryos transferred into 35 recipients and produced 65 piglets from 13 litters. Pregnancy rate according to the number of transferred embryos to recipients was higher the group which received 21 to 24 embryos (50.0%) than other groups 20.0% in less and 33.3% in more. A tail tissue was collected from 65 piglets for biopsy. PCR screening was performed on each DNA sample using two separate sets of primers specific for the 5'- and 3'-flanking region of the rEl-CelD gene. Five of the 65 piglets (7.69%) were positive for the transgene. This study provide useful information regarding production of transgenic pig for bioreactor research.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.37-37
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• 2005

Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone consisting of non-covalently linked two subunits, the ${\alpha}$ and ${\beta}$ subunit. It has been used as a infertility drug for ovulation to mimic luteinizing hormone $(LH).^{1)}$ A stable cell line was established by transfection of Rc/CMV-i-dhfr-hCG, expression vector containing hCG ${\alpha}-$ and ${\beta}-genes$, into dihydrofolate reductase-deficient CHO cells and subesquent methotrexate-mediated gene amplification. Anchorage-dependent CHO cells were adapted into a serum-free and/or animal component-free suspension medium through gradual serum weaning for the hCG production. The established cell line showed typical morphological characteristics and growth profile of CHO cells, and could produce FSH with passage-to-passage consistency. The high density perfusion culture of the CHO cells was carried out in Celligen Plus bioreactor equipped with a spin-filter as a internal cell retention device. The cell density reached up to $>1x10^{7}$ cells/ml in less than 7 days and a perfusion-control strategy based on cellular consumption rates of glucose was $established.^{2)}$ Biologically active recombinant hCG was purified by a series of chromatographic steps including anion exchange chromatography and hydrophobic interaction chromatography to homogeneity. The highly purified recombinant hCG was characterized for physicochemical, immunological and biological properties.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2006.11a
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• pp.43-58
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• 2006

As a bioreactor, bird has proved to be most efficient system for producing useful therapeutic proteins. More than half of the egg white protein content derives from the ovalbumin gene with four other proteins(lysozyme, ovomucoid, ovomucin and conalbumin) present at levels of 50 milligrams or greater. And the naturally sterile egg also contains egg white protein at high concentration allowing for a long shelf life of recombinant protein without loss in activity. In spite of these advantages, transgenic procedures for the bird have lagged far behind because of its complex process of fertilized egg and developmental differences. Recently, a system to transplant mouse testis cells from a fertile donor male to the seminiferous tubules of an infertile recipient male has been developed. Spermatogenesis is generated from transplanted cells, and recipients are capable of transmitting the donor haplotype to progeny. After transplantation, primitive donor spermatogonia migrate to the basement membrane of recipient seminiferous tubules and begin proliferating. Eventually, these cells establish stable colonies with a characteristic appearance, which expands and produces differentiating germ cells, including mature spermatozoa. Thus, the transplanted cells self-renew and produce progeny that differentiate into fully functional spermatozoa. In this study, to develop an alternative system of germline chimera production that operates via the testes rather than through developing embryos, the spermatogonial stem cell techniques were applied. This system consisted of isolation and in vitro-culture of chicken testicular cells, transfer of in vitro-maintained cells into heterologous testes, production of germline chimeras and confirmation of germline transmission for evaluating production of heterologous, functional spermatozoa.

• KSBB Journal
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• v.13 no.4
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• pp.404-410
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• 1998

Three types of adsorbents were developed by immobilizing synthetic zeolite, Philipsite-Gismonine, in alginate, cellulose acetate and dialysis membrane for the in situ removal of ammonium ion which inhibits growth and productivity of animal cells such as CHO cells producing tPA. Ammonium ion removal efficiency and cell growth promoting effect with various immobilized adsorbents were evaluated and the membrane type was selected as an optimal immobilized adsorbent. The experiments were then simulated by adding 8mM ammonium chloride and immobilized adsorbent in order to validate the removal effect under high density cell cultures. The results showed increase in maximum cell density by three times, in cell viability, and in tPA productivity by 40%. And it was found that the promoting effects were more significant in case of high ammonium ion concentration system. It was also found that the optimum addition time for immobilized adsorbents was 48 hr in the absence of ammonium chloride addition and 72 hr in the presence of ammonium chloride addition.

• Journal of Animal Environmental Science
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• v.1 no.1
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• pp.55-64
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• 1995

A wastewater purifying system using phototrophic bacterium, Rhodopseudomonas capsulata, is currently in operation in several countries, One of them, is a continuously aerated bioreactor(CABR) system, which treats concentrated swin wasterwater using small amounts of phototrophic bacteria as additive bacterial seeding. Using this plant, total biochemical oxygen demand was decreased to 13%, and most of volatile fatty acids were removed. About 40% of the wastewater(Influx) was evaporated during aerobic digestion for 24h, and 60% of that erupted in a decodorized foam(Efflux). The efflux had enough nutrients, N, P and K kor growing plant, as well as organic matters. When the efflux was applied to Italian ryegrass with high dose, fresh shoot and root weights were significantly greater, and $NO_3-N$ contents of the dried shoot were lower than those of control plant (CDU). These results indicate that the CABR plant is useful for reduction and deodorization of swine wastewater and the efflux from CABR can be used for crop production as an organic fertilizer.