• Parasites, Hosts and Diseases
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• 제49권4호
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• pp.349-356
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• 2011

The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. coli K12 was also applied. The association ratio of E. coli K1 with A. castellanii was 4.3 cfu per amoeba for 1 hr but E. coli K5 with A. castellanii was 1 cfu per amoeba for 1 hr. By invasion and survival assays, E. coli K5 was recovered less than E. coli K1 but still alive inside A. castellanii. E. coli K1 and K5 survived and multiplied intracellularly in A. castellanii. The survival assay was performed under a favourable condition for 22 hr and 43 hr with the encystment of A. castellanii. Under the favourable condition for the transformation of trophozoites into cysts, E. coli K5 multiplied significantly. Moreover, the pathogenic potential of E. coli K1 from A. castellanii cysts exhibited no changes as compared with E. coli K1 from A. castellanii trophozoites. E. coli K5 was multiplied in A. castellanii trophozoites and survived in A. castellanii cysts. Therefore, this study suggests that E. coli K5 can use A. castellanii as a reservoir host or a vector for the bacterial transmission.

• Parasites, Hosts and Diseases
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• 제37권3호
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• pp.181-188
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• 1999

We investigated the value of mitochondrial small subunit rRNA gene (mt SSU rDNA) PCR-RFLP as a taxonomic tool for Acanthamoeba isolates with close inter-relationships. Twenty-five isolates representing 20 species were included in the analysis. As in nuclear 18s rDNA analysis, two type strains (A. astronyxis and A. tubiashi) of morphological group 1 diverged earliest from the other strains, but the divergence between them was less than in 18s riboprinting. Acanthamoeba griffini of morhological group 2 branched between pathogenic (A. culbertsoni A-1 and A. healyi OC-3A) and nonpathogenic (A.palestinensis Reich, A. pustulosa GE-3a, A. royreba Oak Ridge, and A lenticulata PD2S) strains of morphological group 3. Among the remaining isolates of morphological group 2, the Chang strain had the identical mitochondrial riboprints as the type strain of A. hatchetti. AA2 and AA1, the type strains of A. divionensis and A. paradivionensis, respectively, had the identical riboprints as A. quina Vil3 and A. castellanii Ma. Although the branching orders of A. castellanii Neff, A. polyphaga P23, A. triangularis SH621, and A. lugdunensis L3a were different from those in 18S riboprinting analysis, the results obtained from this study generally coincided well with those from 18S riboprinting. Mitochondrial riboprinting may have an advantage over nuclear 18S rDNA riboprinting beacuse the mt SSU rDNAs do not seem to have introns that are found in the 18S genes of Acanthamoeba and that distort phylogenetic analyses.

• 한국축산식품학회지
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• 제31권5호
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• pp.719-726
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• 2011

Pediococcus pentosaceus JWS 939 (JWS 939) is a nonpathogenic bacteriocin-producing probiotic isolated from the duck intestine. This study assessed the immunomodulatory effects of JWS 939 and compared them with those of Lactobacillus rhamnosus GG (LGG), a well-known immune enhancer. The immune-enhancing effects of JWS 939 were measured by measuring the production of nitric oxide (NO) and cytokines in C57BL/6 mouse peritoneal macrophages. In addition, to assess the immune enhancement abilities of JWS 939, in vivo, a Listeria monocytogenes challenge mice model was used. The results showed that heat-killed JWS 939 induced more NO and interleukin (IL)-$1{\beta}$ production in mouse peritoneal macrophages than in LGG, and that oral administration of viable JWS 939 in mice increased more NO, IL-$1{\beta}$, and tumor necrosis factor (TNF)-${\alpha}$ level than in LGG in serum upon L. monocytogenes challenge. In addition, mice fed with JWS 939 had a longer survival time after lethal challenge with L. monocytogenes, and these effects were stronger than those induced by LGG. Collectively, P. pentosaceus JWS 939 is a remarkable strain that, by releasing bacteriocin and enhancing host immune responses, may have potential as a duck feed additive to suppress pathogens.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.155-157
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• 2001

Bifidobacterium spp. is nonpathogenic, gram-positive and anaerobic bacteria, which inhabit the intestinal tract of humans and animals. In breast-fed infants, bifidobacteria comprise morethan 90％ of the gut bacterial population. Bifidobacteria spp. are used in commericial fermented dairy products and have been suggested to exert health promoting effects on the host by maintaining intestinal microflora balances, improving lactose tolerance, reducing serum cholesterol levels, increasing synthesis of vitamins, and aiding the immune enchancement and anticarcinogenic activity for the host. These beneficial effects of Bifidobacterium are strain-related. Therefore continued efforts to improve strain characteristics are warranted. in these respect, development of vector system for Bifidobacterium is very important not only for the strain improvement but also because Bifidobacterium is most promising in serving as a delivery system for the useful gene products, such as vaccine or anticarcinogenic polypeptides, into human intestinal tract. For developing vector system, we have characterized several bifidobacterial plasmids at genetic level and developed several shuttle vectors between E. coli and Bifidobacterium using them. Also, we have cloned and sequenced several metabolic genes and food grade selection marker. Also we have obtained bifidobacterial surface protein, which will be used as the mediator for surface display of foreign genes. Recently we have succeeded in expressing amylase and GFP in Bifidobacterium using our own expression vector system. Now we are in a very exciting stage for the molecular breeding and safe delivery system using probiotic Bifidobacterium strains.

• Parasites, Hosts and Diseases
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• 제37권2호
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• pp.85-92
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• 1999

To determine the pathogenicity of Acanthamoeba spp. isolated in Korea and to develop a isoenzymatic maker, the mortality rate of infected mice, in vitro cytotoxicity against target cells and isoenzyme band pattern were observed. Five isolates of Acanthamoeba spp. (YM-2, YM-3, YM-4, YM-5 and YM-7) were used in this study as well as three reference Acanthamoeba spp. (A. culbertsoni, A. hatchetti, and A. royreba). According to the mortality rate of infected mice, Korean isolated could be categorized into three groups: high virulent (YM-4), low virulent (YM-2, YM-5, YM-7) and the nonpathogenic group (YM-4), In addition, the virulence of Acanthamoeba spp. was enhanced by brain passage in mice. In the cytotoxicity assay against chinese hamster ovary cells, especially, the cytotoxicity of brain-passaged amoebae was relatively higher than the long-term cultivated ones. The zymodeme patterns of glucose-6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), hexokinase (HK), glutamate oxaloacetate transaminase (GOT) and malic enzyme (ME)of Acanthamoeba spp. were different among each isolate, and also between long-term cultrued amoebae and brain passaged ones. In spites of the polymorphic zymodemes, a slow band of G6PD and K, and an intermediate band of MDH were only observed in pathogenic Acanthamoeba spp., which should be used as isoenzymatic makers.

• 식물병연구
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• 제11권2호
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• pp.162-166
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• 2005

차나무(Camellia sinensis O. Kuntze) 겹둥근무늬병의 발생소장을 2004년도 전남 보성군 보성읍 소재 전남농업기술원 보성차시험장에 식재되어 있는 야부기타 품종에서 조사하였다. 차나무 겹둥근무늬병은 6월말부터 발생하기 시작하여 7월말에 최대발병률을 나타내었다. 차나무의 엽권으로부터 차나무 겹둥근무늬병균에 대한 길항미생물을 분리하였으며, 차나무 겹둥근무늬병균에 대하여 가장 길항능력이 우수하고 차나무에는 병원성을 나타내지 않는 BD0310 균주를 겹둥근무늬병 방제용 길항미생물로 선발하였다. 선발된 균주는 배양적, 형태적, 생화학적 특성 조사와 16S rDNA염기서열 분석에 의하여 Bacillus subtilis 로 동정되었다.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.1004-1014
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• 2013

Six nonpathogenic fungal strains isolated from alkaline soils of Buenos Aires Province, Argentina (Acremonium murorum, Aspergillus sidowii, Cladosporium cladosporoides, Neurospora tetrasperma, Purpureocillium lilacinum (formerly Paecilomyces lilacinus), and Westerdikella dispersa) were tested for their ability to produce keratinolytic enzymes. Strains were grown on feather meal agar as well as in solid-state and submerged cultures, using a basal mineral medium and "hair waste" as sole sources of carbon and nitrogen. All the tested fungi grew on feather meal agar, but only three of them were capable of hydrolyzing keratin, producing clear zones. Among these strains, P. lilacinum produced the highest proteolytic and keratinolytic activities, both in solid-state and submerged fermentations. The medium composition and culture conditions for the keratinases production by P. lilacinum were optimized. Addition of glucose (5 g/l) and yeast extract (2.23 g/l) to the basal hair medium increased keratinases production. The optimum temperature and initial pH for the enzyme production were $28^{\circ}C$ and 6.0, respectively. A beneficial effect was observed when the original concentration of four metal ions, present in the basal mineral medium, was reduced up to 1:10. The maximum yield of the enzyme was 15.96 $U_c/ml$ in the optimal hair medium; this value was about 6.5-fold higher than the yield in the basal hair medium. These results suggest that keratinases from P. lilacinum can be useful for biotechnological purposes such as biodegradation (or bioconversion) of hair waste, leading to a reduction of the environmental pollution caused by leather technology with the concomitant production of proteolytic enzymes and protein hydrolyzates.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.459-466
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• 2013

The lactic acid bacterium Streptococcus thermophilus is widely used as a starter culture for the production of dairy products. Whole-genome sequencing is expected to utilize the genetic basis behind the metabolic functioning of lactic acid bacterium (LAB), for development of their use in biotechnological and probiotic applications. We sequenced the whole genome of Streptococcus thermophilus MTCC 5461, the strain isolated from a curd source, by 454 GS-FLX titanium and Ion Torrent PGM. We performed comparative genome analysis using the local BLAST and RDP for 16S rDNA comparison and by the RAST server for functional comparison against the published genome sequence of Streptococcus thermophilus CNRZ 1066. The whole genome size of S. thermophilus MTCC 5461 is of 1.73Mb size with a GC content of 39.3%. Streptococcal virulence-related genes are either inactivated or absent in the strain. The genome possesses coding sequences for features important for a probiotic organism such as adhesion, acid tolerance, bacteriocin production, and lactose utilization, which was found to be conserved among the strains MTCC 5461 and CNRZ 1066. Biochemical analysis revealed the utilization of 17 sugars by the bacterium, where the presence of genes encoding enzymes involved in metabolism for 16 of these 17 sugars were confirmed in the genome. This study supports the facts that the strain MTCC 5461 is nonpathogenic and harbors essential features that can be exploited for its probiotic potential.

• 대한의생명과학회지
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• 제17권1호
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• pp.7-12
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• 2011

Free-living amoebae ingest several kinds of bacteria. In other words, the bacteria can survive within free-living amoeba. To determine how Escherichia coli K1 isolate causing neonatal encephalitis and non-pathogenic K12 interact with free-living amoebae, e.g., Acanthamoeba castellanii (T1), A. astronyxis (T7), Naegleria fowleri, association, invasion and survival assays were performed. To understand pathogenicity of free-living amoebae, in vitro cytotoxicity assay were performed using murine macrophages. T1 destroyed macrophages about 64% but T7 did very few target cells. On the other hand, N. fowleri which needed other growth conditions rather than Acanthamoeba destroyed more than T1 as shown by lactate dehydrogenase (LDH) release assay. In association assays for E. coli binding to amoebae, the T7 exhibited significantly higher association with E. coli, compared with the T1 isolates (P<0.01). Interestingly, N. fowleri exhibited similar percentages of association as T1. Once E. coli bacteria attach or associate with free-living amoeba, they can penetrate into the amoebae. In invasion assays, the K1 (0.67%) within T1 was observed compared with K12 (0%). E. coli K1 and K12 exhibited high association with N. fowleri and bacterial CFU. To determine the fate of E. coli in long-term survival within free-living amoebae, intracellular survival assays were performed by incubating E. coli with free-living amoebae in PBS for 24 h. Intracellular E. coli K1 within T1 (2.5%) and T7 (1.8%) were recovered and grown, while K12 were not found. N. fowleri was not invaded and here it was not recovered.

• 동의생리병리학회지
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• 제20권4호
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• pp.799-806
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• 2006

Fungi cause a number of plant and animal diseases. Because fungi are more chemically and genetically similar to animals than other organisms, this makes fungal diseases very difficult to treat. Human fungal infections are uncommon in normally healthy persons, being confined to conditions such as candidiasis (thrush) and dermatophyte skin infections such as athlete's foot. However, in the immunocompromised host, a variety of normally mild or nonpathogenic fungi can cause potentially fatal infections. Furthermore, the relative ease with which people can now visit 'exotic' countries provides the means for unusual fungal infections to be imported into this country. Fungal infections or mycoses are classified depending on the degree of tissue involvement and mode of entry into the host. These are Cutaneous, Subcutaneous, Systemic, and Opportunistic. As listed above, in superficial mycoses infection is localised to the skin, the hair, and the nails. An example is 'ringworm' or 'tinea', an infection of the skin by a dermatophyte. Ringworm refers to the characteristic central clearing that often occurs in dermatophyte infections of the skin. Dermatophyte members of the genera Trycophyton, Microsporum and Epidermophyton are responsible for the disease. Tinea can infect various sites of the body, including the scalp (tinea capitis), the beard (tinea barbae) the foot (tinea pedis: 'athlete's foot') and the groin (tinea cruris). All occur in the United Kingdom although tinea infections, other than pedis, are now rare. Candids albicans is a yeast causing candidiasis or 'thrush' in humans. As a superficial mycoses, candidiasis typically infects the mouth or vagina. C. albicans is part of the normal flora of the vagina and gastrointestinal tract and is termed a 'commensal' However, during times of ill health or impaired immunity the balance can alter and the organism multiplies to cause disease. Antibiotic treatment can also alter the normal bacterial flora allowing C. albicans to flourish. If we study mycoses of the orient medicine, we can improve the medical skills about mycoses.