Numerical Simulation of Deformation in Hot Runner Manifold.

This study simulated the deformation of a hot runner manifold and nozzle assembly during operation, aiming to address potential leaks and premature failure. Both thermal and mechanical models were used simultaneously to accurately capture system behavior. A simplified set of boundary conditions was proposed for efficient problem-solving. Analysis of the simulation results revealed that thermal deformation posed a risk of catastrophic failures and leaks. Deformation from melt pressure was relatively small compared to thermal loading, not exceeding 12%. The study provided design recommendations based on the simulation findings, guiding the development of hot runner systems for improved reliability.

Identification of strA-strB Genes in Streptomycin-Resistant Pseudomonas syringae pv. actinidiae Biovar 2 Strains Isolated in Korea.

Bacterial canker is a devastating disease of kiwifruit caused by the bacterium Pseudomonas syringe pv. actinidiae. Canker disease of kiwifruit in Korea has been controlled using streptomycin for more than two decades. Four streptomycin-resistant strains, belonging to biovar 2, which are found only in Korea, were collected between 2013 and 2014 from different orchards located in Jeju, Korea. The genetic background for streptomycin resistance among P. syringe pv. actinidiae strains were determined by examining the presence of strA-strB or aadA, which are genes frequently found in streptomycin-resistant bacteria, and a point mutation at codon 43 in the rpsL gene. All four streptomycin-resistant strains of P. syringe pv. actinidiae investigated in this study contained strA-strB as a resistant determinant. The presence of the aadA gene and a mutation in codon 43 of the rpsL gene was not identified.

Rapid Numerical Estimation of Pressure Drop in Hot Runner System.

To determine dimensions in the hot runner systems, given a material, it is necessary to predict the pressure drop according to them. Although modern injection molding simulators are able to evaluate such pressure drops, they are expensive and demanding to be employed as a design utility. This work develops a computer tool that can calculate a pressure drop from the sprue to the gate assuming a steady flow of a generalized Newtonian fluid. For a four drop hot runner system, the accuracy has been verified by comparing the obtained results with those by a commercial simulator. This paper presents how to utilize the proposed method in the hot runner design process.

Genomic analyses of Pseudomonas syringae pv. actinidiae isolated in Korea suggest the transfer of the bacterial pathogen via kiwifruit pollen.

Introduction. The bacterial pathogen, Pseudomonas syringae pv. actinidiae (Psa), has emerged as a major threat to kiwifruit cultivation throughout the world. One pandemic strain (from the Psa3 group) has occurred in various geographical regions. It is important to understand how this pathogen is being transmitted.Aim. Although Psa has been found in Korea since 1992, the isolates were until recently of a distinct type (Psa2). Recently, the more virulent Psa3 type has been detected. The purpose of this study was to describe the variety of Psa3 now found in Korea.Methodology. Strains were isolated from kiwifruit plants in Korea and from pollen imported into Korea from New Zealand. The genomes of 10 isolates were sequenced using the Illumina platform and compared to the completely assembled genomes of pandemic Psa3 strains from New Zealand and China. Comparisons were also made with pandemic strains from Chile and non-pandemic Psa3 isolates from China.Results. Six of the 10 Psa3 isolates from Korea show a clear relationship with New Zealand isolates. Two isolates show a distinct relationship to isolates from Chile; one further isolate has a sequence that is highly similar to that of M228, a strain previously isolated in China; and the last isolate belongs to the Psa3 group, but is not a member of the pandemic lineage.Conclusion. This analysis establishes that there have been multiple routes of transmission of the Psa3 pandemic strain into Korea. One route has involved the importation of pollen from New Zealand. A second route probably involves importation from Chile.

Genetic Diversity of the Pear Scab Fungus Venturia nashicola in Korea.

Scab disease caused by Venturia nashicola is of agroeconomic importance in cultivation of Asian pear. However, little is known about the degree of genetic diversity in the populations of this pathogen. In this study, we collected 55 isolates from pear scab lesions in 13 major cultivation areas in Korea and examined the diversity using sequences of internal transcribed spacer (ITS) region, ß-tubulin (TUB2), and translation elongation factor-1α (TEF-1α) genes as molecular markers. Despite a low level of overall sequence variation, we found three distinctive subgroups from phylogenetic analysis of combined ITS, TUB2, and TEF-1α sequences. Among the three subgroups, subgroup 1 (60% of isolates collected) was predominant compared to subgroup 2 (23.6%) or subgroup 3 (16.4%) and was distributed throughout Korea. To understand the genetic diversity among the subgroups, RAPD analysis was performed. The isolates yielded highly diverse amplicon patterns and none of the defined subgroups within the dendrogram were supported by bootstrap values greater than 30%. Moreover, there is no significant correlation between the geographical distribution and the subgroups defined by molecular phylogeny. Our data suggest a low level of genetic diversification among the populations of V. nashicola in Korea.

Correction to: Survey of coastal inland pollution sources and their influence on seawater quality in Doam bay, Korea.

The original version of this article unfortunately contained an error in the affiliation section.

Survey of coastal inland pollution sources and their influence on seawater quality in Doam bay, Korea.

Inland pollution sources of Doam bay were investigated from August to October in 2013. A total of 210 sources including rivers, streams, domestic, agricultural and industrial discharge points were identified along the coast, including 32 sources that had outflow. Agricultural sources were the largest inland pollution sources (139, 66.2%). Fecal coliform concentrations were measured. These data were combined with water discharge data to determine daily loads of pollutants discharged from each source into the bay. Fecal coliform concentrations were the highest in domestic discharges. However, they only had slight influence because their discharge volume was small. The most significant pollution source was Tamjin River (St.85) due to large amount of discharge volume. The influence of St.85 reached almost half of Doam bay. Fecal coliform levels of streams increased after rainfall, but decreased overtime. Domestic pollution sources were not affected upon rain event.

Occurrence and Epidemics of Bacterial Canker of Kiwifruit in Korea.

Bacterial canker is the largest limiting factor in the cultivation and production of kiwifruit worldwide. Typical symptoms comprise necrotic spots on leaves, canker and dieback on canes and trunks, twig wilting, and blossom necrosis. Pseudomonas syringae pv. actinidiae (Psa), which is the causal agent of kiwifruit bacterial canker, is divided into four biovars based on multilocus sequence analysis of different genes, additional PCR testing of pathogenic genes (argKtox cluster, cfl, and various effector genes), and biochemical and physiological characterization. Bacterial canker caused by Psa biovar 2 designated Psa2 was detected for the first time on the green-fleshed kiwifruit cultivar Hayward in 1988 and the yellow-fleshed kiwifruit cultivar Hort16A in 2006 in Korea. Psa biovar 3 designated Psa3, responsible for the current global pandemics of kiwifruit bacterial canker, began to appear in Korea in 2011 and caused tremendous economic losses by destroying many vines or orchards of yellow-fleshed kiwifruit cultivars in one or several growing seasons. Bacterial canker epidemics caused by both Psa2 and Psa3 are prevalent in Korea in recent years. In this review, we summarize the symptomatology, etiology, disease cycle, diagnosis, and epidemiology of kiwifruit bacterial canker in Korea.

Incidence Rates of Major Diseases of Kiwiberry in 2015 and 2016.

Incidence rates of diseases in kiwiberry orchards were investigated monthly from late June to late September in Gwangyang and Boseong in 2015 and 2016. The impact of postharvest fruit rot was investigated during ripening after harvest. Bacterial canker was only observed on one single tree in 2015, but black rot, powdery mildew, leaf spot and blight, and postharvest fruit rot diseases were problematic throughout the study period in both 2015 and 2016. Incidence rates of the diseases varied with kiwiberry cultivar, region and sampling time. Incidence rates of powdery mildew, leaf spot and blight diseases increased significantly during the late growing stages near fruit harvest, while black rot peaked in late August. Incidence rate of postharvest fruit rot on fruit without fruit stalks was less than half of fruit with fruit stalks, regardless of kiwiberry cultivars. Among the four cultivars, Mansu was relatively resistant to black rot and postharvest fruit rot diseases. In our knowledge, this is the first report of various potential pathogens of kiwiberry in Korea.

Development of Carrot Medium Suitable for Conidia Production of Venturia nashicola.

The causal fungus of pear scab, Venturia nashicola, grows slowly and rarely produces conidia on artificial media in the laboratory, but it produced conidia on the Cheongah medium containing Cheongah powder. V. nashicola grew too slow to produce conidia until 15 days after cultivation but produced conidia with 4 × 104 conidia/plate 30 days after cultivation on the Cheongah medium containing 1% Cheongah powder. V. nashicola showed a peak production of conidia with 4.5 × 105 conidia/plate 60 days after cultivation on the carrot medium containing 2% carrot powder, one of the constituents of Cheongah powder. The carrot medium is considered to be the best medium to obtain conidia of V. nashicola in the laboratory until now. This is the first report on the development of a suitable medium for conidia production of V. nashicola, as far as we know.

Outbreak and Spread of Bacterial Canker of Kiwifruit Caused by Pseudomonas syringae pv. actinidiae Biovar 3 in Korea.

A bacterial pathogen, Pseudomonas syringae pv. actinidiae (Psa), is a causal agent of kiwifruit bacterial canker worldwide. Psa biovar 3 (Psa3) was first detected in 2011 at an orchard in Dodeok-myeon, Goheunggun, Jeonnam Province in Korea. In this study, we present the results of an epidemiological study regarding Psa3 occurrence on kiwifruit orchards in Korea for the period of 2013 to 2015. Since the first detection of Psa3 in 2011, there was no further case reported by 2013. However, Psa3 was rapidly spreading to 33 orchards in 2014; except for three orchards in Sacheonsi, Gyeongnam Province, most cases were reported in Jeju Island. Entering 2015, bacterial canker by Psa3 became a pandemic in Korea, spreading to 72 orchards in Jeju Island, Jeonnam, and Gyeongnam Provinces. Our epidemiological study indicated that the first Psa3 incidence in 2011 might result from an introduction of Psa3 through imported seedlings from China in 2006. Apart from this, it was estimated that most Psa3 outbreaks from 2014 to 2015 were caused by pollens imported from New Zealand and China for artificial pollination. Most kiwifruit cultivars growing in Korea were infected with Psa3; yellow-fleshed cultivars (Yellow-king, Hort16A, Enza-gold, Zecy-gold, and Haegeum), red-fleshed cultivars (Hongyang and Enza-Red), green-fleshed cultivars (Hayward and Daeheung), and even a kiwiberry (Skinny-green). However, susceptibility to canker differed among cultivars; yellow- and red-fleshed cultivars showed much more severe symptoms compared to the green-fleshed cultivars of kiwifruit and a kiwiberry.

Development of Specific Markers for Identification of Biovars 1 and 2 Strains of Pseudomonas syringae pv. actinidiae.

Pseudomonas syringae pv. actinidiae, the causal agent of canker in kiwifruit, can be divided into three biovars (biovars 1, 2, and 3). Strains belonging to biovar 1 produce phaseolotoxin and were isolated in Japan and Italy before 2008. Strains of biovar 2 produce coronatine instead of phaseolotoxin and have been isolated only in Korea. Strains belonging to biovar 3 produce neither phaseolotoxin nor coronatine and are responsible for the global outbreak of bacterial canker of kiwifruit in recent years. The biovar 3-specific primer set was developed in a previous work. In this study, two sets of PCR primers specific to strains of biovars 1 and 2, respectively, were developed based on random amplified polymorphic DNA analyses. Primers PsaJ-F and PsaJ-R produced a 481-bp region with genomic DNA of biovar 1 strains, whereas primers PsaK-F and PsaK-R amplified a 413-bp region present only in the genome of biovar 2 strains.

Specific and Sensitive Detection of the Pear Scab Fungus Venturia nashicola by SYBR Green Real-Time PCR.

A new improved PCR method has been developed for the rapid, reliable, and sensitive detection of Venturia nashicola, a destructive pathogen of scab disease in Japanese pear. The translation elongation factor-1 alpha gene-derived PCR primers specifically amplified a 257-bp-sized DNA band of the target gene from the genomic DNA of V. nashicola. No amplicon was produced from the genomic DNA of other Venturia spp. and reference fungal species tested. With the high detection limit of 10 fg DNA content, our real-time method could be used for the quarantine inspection and field monitoring of V. nashicola.

Molecular Characteristics of Pseudomonas syringae pv. actinidiae Strains Isolated in Korea and a Multiplex PCR Assay for Haplotype Differentiation.

The molecular features of Pseudomonas syringae pv. actinidiae strains isolated in Korea were compared with strains isolated in Japan and Italy. Sequencing of eight P. syringae pv. actinidiae and three P. syringae pv. theae strains revealed a total of 44 single nucleotide polymorphisms across 4,818 bp of the concatenated alignment of nine genes. A multiplex PCR assay was developed for the detection of P. syringae pv. actinidiae and for the specific detection of recent haplotype strains other than strains isolated since the 1980s in Korea. The primer pair, designated as TacF and TacR, specifically amplified a 545-bp fragment with the genomic DNA of new haplotype of P. syringae pv. actinidiae strains. A multiplex PCR conducted with the TacF/TacR primer pair and the universal primer pair for all P. syringae pv. actinidiae strains can be simultaneously applied for the detection of P. syringae pv. actinidiae and for the differentiation of new haplotype strains.

Functional characterization of the sucrose isomerase responsible for trehalulose production in plant-associated Pectobacterium species.

Fifty-three plant-associated microorganisms were investigated for their ability to convert sucrose to its isomers. These microorganisms included one Dickeya zeae isolate and 7 Enterobacter, 3 Pantoea, and 43 Pectobacterium species. Eleven out of the 53 strains (21%) showed the ability to transform sucrose to isomaltulose and trehalulose. Among those, Pectobacterium carotovorum KKH 3-1 showed the highest bioconversion yield (97.4%) from sucrose to its isomers. In this strain, the addition of up to 14% sucrose in the medium enhanced sucrose isomerase (SIase) production. The SIase activity at 14% sucrose (47.6 U/mg dcw) was about 3.6-fold higher than that of the negative control (13.3 U/mg dcw at 0% sucrose). The gene encoding SIase, which is comprised a 1776 bp open reading frame (ORF) encoding 591 amino acids, was cloned from P. carotovorum KKH 3-1 and expressed in Escherichia coli. The recombinant SIase (PCSI) was shown to have optimum activity at pH 6.0 and 40 °C. The reaction temperature significantly affected the ratio of sucrose isomers produced by PCSI. The amount of trehalulose increased from 47.5% to 79.1% as temperature was lowered from 50 °C to 30 °C, implying that SIase activity can be controlled by reaction temperature.

Biruloquinone, an acetylcholinesterase inhibitor produced by lichen-forming fungus Cladonia macilenta.

At present, acetylcholinesterase (AChE) inhibitors are the first group of drugs to treat mild to moderate Alzheimer's disease (AD). Although beneficial in improving cognitive and behavioral symptoms, the effectiveness of AChE inhibitors has been questioned since they do not delay or prevent neurodegeneration in AD patients. Therefore, in the present study, in order to develop new and effective anti-AD agents from lichen products, both the AChE inhibitory and the neuroprotective effects were evaluated. The AChE inhibitory assay was performed based on Ellman's reaction, and the neuroprotective effect was evaluated by using the MTT method on injured PC12 cells. One AChE inhibitor (IC50 = 27.1 microg/ml) was isolated by means of bioactivity-guided isolation from the extract of lichen-forming fungus Cladonia macilenta, which showed the most potent AChE inhibitory activity in previous screening experiment. It was then identified as biruloquinone by MS, and 1H- and 13C-NMR analyses. The inhibitory kinetic assay suggested that biruloquinone is a mixed-II inhibitor on AChE. Meanwhile, biruloquinone improved the viability of the H2O2- and beta-amyloid-injured PC12 cells at 1 to 25 microg/ml. The protective effects are proposed to be related to the potent antioxidant activities of biruloquinone. These results imply that biruloquinone has the potential to be developed as a multifunctional anti- AD agent.

Marinivirga aestuarii gen. nov., sp. nov., a member of the family Flavobacteriaceae, isolated from marine environments, and emended descriptions of the genera Hyunsoonleella, Jejuia and Pontirhabdus and the species Hyunsoonleella jejuensis, Jejuia pallidilutea and Pontirhabdus pectinivorans.

Two orange, rod-shaped, Gram-reaction-negative, aerobic bacterial strains devoid of flagella and gliding motility, designated strains KYW371(T) and KS18 were isolated from a seawater sample and a shellfish Ruditapes philippinarum, respectively, collected from Gwangyang Bay, Republic of Korea. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the two strains belonged to the family Flavobacteriaceae; and that strain KYW371(T) was most closely related to Algibacter mikhailovii LMG 23988(T) (96.7 % sequence similarity), Pontirhabdus pectinivorans JC2675(T) (96.3 %), Postechiella marina M091(T) (95.6 %) and Hyunsoonleella jejuensis CNU004(T) (95.3 %). The 16S rRNA gene sequence similarity (98.8 %) and DNA-DNA relatedness (78.1 %) between strains KYW371(T) and KS18 indicated that these two strains represented a single species. The predominant cellular fatty acids of strain KYW371(T) were iso-C15 : 1 G, iso-C15 : 0, iso-C15 : 0 3-OH and iso-C17 : 0 3-OH. Flexirubin-type pigments were absent. MK-6 was the only isoprenoid quinone and the DNA G+C content was 34.8-36.6 mol%. Data from this taxonomic study employing a polyphasic approach suggested that the isolates represent a novel species in a new genus in the family Flavobacteriaceae, for which the name Marinivirga aestuarii gen. nov., sp. nov. is proposed. The type strain is KYW371(T) ( = KCTC 23449(T) = JCM 17452(T)), and an additional strain of the species is KS18 ( = KCTC 23128 = JCM 16845). Emended descriptions of the genera Hyunsoonleella, Jejuia and Pontirhabdus and the species Hyunsoonleella jejuensis, Jejuia pallidilutea and Pontirhabdus pectinivorans are also proposed.

Specific and Sensitive Detection of Venturia nashicola, the Scab Fungus of Asian Pears, by Nested PCR.

The fungus Venturia nashicola is the causal agent of scab on Asian pears. For the rapid and reliable identification as well as sensitive detection of V. nashicola, a PCR-based technique was developed. DNA fingerprints of three closely related species, V. nashicola, V. pirina, and V. inaequalis, were obtained by random amplified polymorphic DNA (RAPD) analysis. Two RAPD markers specific to V. nashicola were identified by PCR, after which two pairs of sequence characterized amplified region (SCAR) primers were designed from the nucleotide sequences of the markers. The SCAR primer pairs, designated as D12F/D12R and E11F/E11R, amplified 535-bp and 525-bp DNA fragments, respectively, only from genomic DNA of V. nashicola. The specificity of the primer sets was tested on strains representing three species of Venturia and 20 fungal plant pathogens. The nested PCR primer pair specific to V. nashicola was developed based on the sequence of the species-specific 525-bp DNA fragment amplified by primer set E11F/E11R. The internal primer pair Na11F/Na11R amplified a 235-bp fragment from V. nashicola, but not from any other fungal species tested. The nested PCR assay was sensitive enough to detect the specific fragment in 50 fg of V. nashicola DNA.

Production of anti-Helicobacter pylori metabolite by the lichen-forming fungus Nephromopsis pallescens.

The present study was conducted to evaluate the antibacterial activity of lichen-forming fungi (LFF) against Helicobacter pylori, and to optimize the culture conditions of LFF for maximum production of natural antibiotics against H. pylori. To accomplish this, a screening assay was first conducted among 19 species of LFF. The extract of Nephromopsis pallescens (KOLRI-040516) exhibited the strongest anti-ff. pylori activity. Bioautograghic TLC and HPLC analysis identified usnic acid as the main antibacterial substance produced by JV. pallescens. The growth of JV. pallescens and production of antibacterial substances produced by the fungus were then investigated under several culture conditions including the culture media, initial medium pHs, incubation temperatures, and the degree of aeration. The results indicated that culture in MY medium with an initial pH of 6.0, a temperature of 15°C and a low degree of aeration supported the largest usnic acid production of the fungus (16.4 ug usnic acid/g dry biomass). Especially, aeration was found to be an important factor that affect both growth and usnic acid production of N. pallescens.

Gangjinia marincola gen. nov., sp. nov., a marine bacterium of the family Flavobacteriaceae.

A novel strictly aerobic, orange-pigmented, Gram-stain-negative bacterium, designated strain GJ16(T), was isolated from coastal seawater of Gangjin Bay, the southernmost part of the Korean peninsula, and subjected to a polyphasic taxonomic study. It grew optimally at 25-30 °C, at pH 7.0-8.0 and in the presence of 3 % NaCl. Comparative 16S rRNA gene sequence analysis revealed that strain GJ16(T) formed a distinct lineage within the family Flavobacteriaceae and shared less than 91.2 % 16S rRNA gene sequence similarity with members of the genera Leptobacterium, Zhouia, Winogradskyella, Dokdonia and Krokinobacter. The predominant cellular fatty acids were iso-C(15 : 0) (40.2 %), iso-C(15 : 1) G (12.8 %), iso-C(17 : 0) 3-OH (11.2 %) and C(15 : 0) (6.6 %). The G+C content of the genomic DNA was 39.4 mol% and the major respiratory isoprenoid quinone was MK-6. On the basis of phenotypic and genotypic data, strain GJ16(T) represents a novel species in a new genus in the family Flavobacteriaceae, for which the name Gangjinia marincola gen. nov., sp. nov. is proposed; the type strain is GJ16(T) (=KCTC 22649(T) =JCM 16082(T)).