Metabolomics Reveals a Key Role for Fumarate in Mediating the Effects of NADPH Oxidase 4 in Diabetic Kidney Disease.

The NADPH oxidase (NOX) isoform NOX4 has been linked with diabetic kidney disease (DKD). However, a mechanistic understanding of the downstream effects of NOX4 remains to be established. We report that podocyte-specific induction of NOX4 in vivo was sufficient to recapitulate the characteristic glomerular changes noted with DKD, including glomerular hypertrophy, mesangial matrix accumulation, glomerular basement membrane thickening, albuminuria, and podocyte dropout. Intervention with a NOX1/NOX4 inhibitor reduced albuminuria, glomerular hypertrophy, and mesangial matrix accumulation in the F1 Akita model of DKD. Metabolomic analyses from these mouse studies revealed that tricarboxylic acid (TCA) cycle-related urinary metabolites were increased in DKD, but fumarate levels were uniquely reduced by the NOX1/NOX4 inhibitor. Expression of fumarate hydratase (FH), which regulates urine fumarate accumulation, was reduced in the diabetic kidney (in mouse and human tissue), and administration of the NOX1/NOX4 inhibitor increased glomerular FH levels in diabetic mice. Induction of Nox4 in vitro and in the podocyte-specific NOX4 transgenic mouse led to reduced FH levels. In vitro, fumarate stimulated endoplasmic reticulum stress, matrix gene expression, and expression of hypoxia-inducible factor-1α (HIF-1α) and TGF-ß. Similar upregulation of renal HIF-1α and TGF-ß expression was observed in NOX4 transgenic mice and diabetic mice and was attenuated by NOX1/NOX4 inhibition in diabetic mice. In conclusion, NOX4 is a major mediator of diabetes-associated glomerular dysfunction through targeting of renal FH, which increases fumarate levels. Fumarate is therefore a key link connecting metabolic pathways to DKD pathogenesis, and measuring urinary fumarate levels may have application for monitoring renal NOX4 activity.

The effect of fermented buckwheat on producing l-carnitine- and γ-aminobutyric acid (GABA)-enriched designer eggs.

BACKGROUND: The potential of fermented buckwheat as a feed additive was studied to increase l-carnitine and γ-aminobutyric acid (GABA) in designer eggs. Buckwheat contains high levels of lysine, methionine and glutamate, which are precursors for the synthesis of l-carnitine and GABA. Rhizopus oligosporus was used for the fermentation of buckwheat to produce l-carnitine and GABA that exert positive effects such as enhanced metabolism, antioxidant activities, immunity and blood pressure control. RESULTS: A novel analytical method for simultaneously detecting l-carnitine and GABA was developed using liquid chromatography/mass spectrometry (LC/MS) and LC/MS/MS. The fermented buckwheat extract contained 4 and 34 times more l-carnitine and GABA respectively compared with normal buckwheat. Compared with the control, the fermented buckwheat extract-fed group showed enriched l-carnitine (13.6%) and GABA (8.4%) in the yolk, though only l-carnitine was significantly different (P < 0.05). Egg production (9.4%), albumen weight (2.1%) and shell weight (5.8%) were significantly increased (P < 0.05). There was no significant difference in yolk weight, and total cholesterol (1.9%) and triglyceride (4.9%) in the yolk were lowered (P < 0.05). CONCLUSION: Fermented buckwheat as a feed additive has the potential to produce l-carnitine- and GABA-enriched designer eggs with enhanced nutrition and homeostasis. These designer eggs pose significant potential to be utilized in superfood production and supplement industries. © 2016 Society of Chemical Industry.

Plant growth promoting effect of Bacillus amyloliquefaciens H-2-5 on crop plants and influence on physiological changes in soybean under soil salinity.

This study was aimed to identify plant growth-promoting bacterial isolates from soil samples and to investigate their ability to improve plant growth and salt tolerance by analysing phytohormones production and phosphate solubilisation. Among the four tested bacterial isolates (I-2-1, H-1-4, H-2-3, and H-2-5), H-2-5 was able to enhance the growth of Chinese cabbage, radish, tomato, and mustard plants. The isolated bacterium H-2-5 was identified as Bacillus amyloliquefaciens H-2-5 based on 16S rDNA sequence and phylogenetic analysis. The secretion of gibberellins (GA4, GA8, GA9, GA19, and GA20) from B. amyloliquefaciens H-2-5 and their phosphate solubilisation ability may contribute to enhance plant growth. In addition, the H-2-5-mediated mitigation of short term salt stress was tested on soybean plants that were affected by sodium chloride. Abscisic acid (ABA) produced by the H-2-5 bacterium suppressed the NaCl-induced stress effects in soybean by enhancing plant growth and GA4 content, and by lowering the concentration of ABA, salicylic acid, jasmonic acid, and proline. These results suggest that GAs, ABA production, and the phosphate solubilisation capacity of B. amyloliquefaciens H-2-5 are important stimulators that promote plant growth through their interaction and also to improve plant growth by physiological changes in soybean at saline soil.

Specific rhizobacterial resources: characterization and comparative analysis from contrasting coastal environments of Korea.

This study analyzed the rhizobacterial distribution from two coasts, which show contrasting climates and geographical and geological characteristics, to secure specific microbial resources. Furthermore, rhizobacteria were characterized and the results were comparatively analyzed with reference to the characteristics of two coastal environments. For this purpose, three representative halophyte species communities native to the Dokdo Islands and the East Sea coast of Korea were selected. Partial identification of rhizobacteria showed a clear difference between each sampling site and halophyte. Furthermore, isolates were characterized by their growth properties under NaCl or pH gradients related with previous geographical, geological, and climatic studies of the Dokdo Islands and the East Sea coast. A high proportion of the East Sea isolates showed halotolerance, but a high proportion of Dokdo isolates shared halophilic traits. Meanwhile, a higher proportion of East Sea isolates grew at a wider range of pH values than those of the Dokdo Islands. The results of our study suggest that unique rhizobacterial resources developed under specific rhizospheric conditions derived from halophytes interacting with their specific environment, even within the same coastal halophytic species. Therefore, this study proposes the necessity of securing characterized and unique microbial resources to apply to specific environments for the purpose of recovering and restoring sand dunes or salt-damaged agricultural lands.

AMP-activated protein kinase (AMPK) activation inhibits nuclear translocation of Smad4 in mesangial cells and diabetic kidneys.

Diabetic nephropathy is characterized by diffuse mesangial matrix expansion and is largely dependent on the TGF-ß/Smad signaling pathway. Smad4 is required for TGF-ß signaling; however, its regulation has not been well characterized in diabetic kidney disease. Here, we report that high glucose is sufficient to stimulate nuclear translocation of Smad4 in mesangial cells and that stimulation of the major energy sensor AMP-activated protein kinase (AMPK) has a potent effect to block Smad4 nuclear translocation. Activation of AMPK by 5-aminoimidazole-4-carboxamide-1-ß-d-ribofuranoside (AICAR) inhibited high glucose-induced and TGF-ß stimulation of nuclear Smad4. To identify which of the catalytic α-subunits may be involved, small interfering (si) RNA-based inhibition of AMPK α1- or α2-subunit was employed. Inhibition of either subunit reduced overall AMPK activity and contributed to Smad4 nuclear accumulation. In an animal model of early diabetic kidney disease, induction of diabetes was found to markedly stimulate Smad4 protein levels and enhance nuclear accumulation. AMPK activation with AICAR completely prevented the upregulation of Smad4 and reduced mesangial matrix accumulation. We conclude that stimulation of Smad4 in cell culture and in in vivo models of early diabetic kidney disease is dependent on AMPK.

Metabolomics reveals signature of mitochondrial dysfunction in diabetic kidney disease.

Diabetic kidney disease is the leading cause of ESRD, but few biomarkers of diabetic kidney disease are available. This study used gas chromatography-mass spectrometry to quantify 94 urine metabolites in screening and validation cohorts of patients with diabetes mellitus (DM) and CKD(DM+CKD), in patients with DM without CKD (DM-CKD), and in healthy controls. Compared with levels in healthy controls, 13 metabolites were significantly reduced in the DM+CKD cohorts (P≤0.001), and 12 of the 13 remained significant when compared with the DM-CKD cohort. Many of the differentially expressed metabolites were water-soluble organic anions. Notably, organic anion transporter-1 (OAT1) knockout mice expressed a similar pattern of reduced levels of urinary organic acids, and human kidney tissue from patients with diabetic nephropathy demonstrated lower gene expression of OAT1 and OAT3. Analysis of bioinformatics data indicated that 12 of the 13 differentially expressed metabolites are linked to mitochondrial metabolism and suggested global suppression of mitochondrial activity in diabetic kidney disease. Supporting this analysis, human diabetic kidney sections expressed less mitochondrial protein, urine exosomes from patients with diabetes and CKD had less mitochondrial DNA, and kidney tissues from patients with diabetic kidney disease had lower gene expression of PGC1α (a master regulator of mitochondrial biogenesis). We conclude that urine metabolomics is a reliable source for biomarkers of diabetic complications, and our data suggest that renal organic ion transport and mitochondrial function are dysregulated in diabetic kidney disease.

Phosphate Solubilizing Bacillus megaterium mj1212 Regulates Endogenous Plant Carbohydrates and Amino Acids Contents to Promote Mustard Plant Growth.

The current study was conducted to explore the potential of a phosphate solubilizing soil bacterium, Bacillus megaterium mj1212 for enhancing the growth of mustard plants. The newly isolated bacterial strain mj1212 was identified as B. megaterium using phylogenetic analysis and, its phosphate solubilization ability was shown by the clear zone formation on National Botanical Research Institute's Phosphate medium. Moreover, the phosphate solubilization ability of B. megaterium mj1212 was enhanced by optimal culture conditions at pH 7.0 and 35 °C which might be due to the presence of malic and quinic acid in the culture medium. The beneficial effect of B. megaterium mj1212 in mustard plants was determined by an increasing shoot length, root length and fresh weight of plants. In the biochemical analysis revealed that chlorophyll, sucrose, glucose, fructose and amino acids (Asp, Thr, Ser, Glu, Gly, Ala, Cys, Val, Met, Ilu, Leu, Tyr, Phe, Lys, His, Arg and Pro) were higher in B. megaterium mj1212 treated plants, when compared to their control. The result of present study suggests that B. megaterium mj1212 treatment could be act as phosphate biofertilizer to improve the plant growth.

Fungal Microbiome of Alive and Dead Korean Fir in its Native Habitats.

A rapid decline of Abies koreana has been reported in most of the natural alpine habitats in Korea. It is generally accepted that this phenomenon is due to climate change even though no clear conclusions have been drawn. Most research has focused on abiotic environmental factors, but studies on the relationships between A. koreana and soil fungal microbiomes are scarce. In this study, the rhizoplane and rhizosphere fungal communities in the alive and dead Korean fir trees from its three major natural habitats including Mt. Deogyu, Mt. Halla, and Mt. Jiri in Korea were investigated to identify specific soil fungal groups closely associated with A. koreana. Soil fungal diversity in each study site was significantly different from another based on the beta diversity calculations. Heat tree analysis at the genus level showed that Clavulina, Beauveria, and Tomentella were most abundant in the healthy trees probably by forming ectomycorrhizae with Korean fir growth and controlling pests and diseases. However, Calocera, Dacrymyces, Gyoerffyella, Hydnotrya, Microdochium, Hyaloscypha, Mycosymbioces, and Podospora were abundant in the dead trees. Our findings suggested that Clavulina, Beauveria, and Tomentella are the major players that could be considered in future reforestation programs to establish ectomycorrhizal networks and promote growth. These genera may have played a significant role in the survival and growth of A. koreana in its natural habitats. In particular, the genus Gyoerffyella may account for the death of the seedlings. Our work presented exploratory research on the specific fungal taxa associated with the status of A. koreana.

Aster × chusanensis Growth and Phenolic Acid Composition under Different Cultivation Temperatures.

Plants of the Asteraceae family have been cultivated worldwide for economic, medicinal, and ornamental purposes, including genera such as Aster, Helianthus, and Cosmos. Numerous studies examined their secondary metabolites; however, those of Aster × chusanensis, which is a natural hybrid species in South Korea, are unclear, and optimized propagation methods should be identified. We analyzed phenolic acid concentrations in each part of Aster × chusanensis through HPLC. Further, we investigated the growth characteristics and secondary metabolite concentrations under various growth temperatures using division propagation, followed by growing at 20, 25, and 30 °C in a growth chamber. Chlorogenic acid was the primary compound, which was particularly high in the leaves. The growth characteristics did not differ significantly between temperatures, and 30 °C was most efficient for phenolic acid biosynthesis. Our results provide valuable information on optimized propagation and secondary metabolite concentrations under different temperatures of Aster × chusanensis.

Role of Nox2 in diabetic kidney disease.

NADPH oxidase (Nox) isoforms have been implicated in contributing to diabetic microvascular complications, but the functional role of individual isoforms in diabetic kidney are unclear. Nox2, in particular, is highly expressed in phagocytes and may play a key inflammatory role in diabetic kidney disease. To determine the role of Nox2, we evaluated kidney function and pathology in wild-type (WT; C57BL/6) and Nox2 knockout (KO) mice with type 1 diabetes. Diabetes was induced in male Nox2 KO and WT mice with a multiple low-dose streptozotocin protocol. Groups were studied for kidney disease after 8 and 20 wk of diabetes. Hyperglycemia and body weights were similar in WT and Nox2 KO diabetic mice. All functional and structural features of early and later stage diabetic kidney disease (albuminuria, mesangial matrix, tubulointerstitial disease, and gene expression of matrix and transforming growth factor-ß) were similar in both diabetic groups compared with their respective nondiabetic groups, except for reduction of macrophage infiltration and monocyte chemoattractant protein-1 in the diabetic Nox2 KO mice. Systolic blood pressure by telemetry was surprisingly increased in Nox2 KO mice; however, the systolic blood pressure was reduced in the diabetic WT and Nox2 KO mice by tail-cuff. Interestingly, diabetic Nox2 KO mice had marked upregulation of renal Nox4 at both the glomerular and cortical levels. The present results demonstrate that lack of Nox2 does not protect against diabetic kidney disease in type 1 diabetes, despite a reduction in macrophage infiltration. The lack of renoprotection may be due to upregulation of renal Nox4.

Cadophora malorum Cs-8-1 as a new fungal strain producing gibberellins isolated from Calystegia soldanella.

Fourteen endophytic fungi with different colony morphologies were isolated from the roots of Calystegia soldanella. Endophytic fungi isolated from C. soldanella were identified by internal transcribed spacer (ITS) region. To verify plant growth promotion (PGP), culture filtrates of isolated endophytic fungi were treated in Waito-c rice (WR) and C. soldanella seedlings. Culture filtrates of Cs-8-1 fungal strain had advanced PGP activity. The presence of physiologically bioactive gibberellins (GA) GA(1) (1.213 ng ml(-1)), GA(3) (1.292 ng ml(-1)), GA(4) (3.6 ng ml(-1)), GA(7) (1.328 ng ml(-1)), other inactive GA(9) (0.796 ng ml(-1)) and GA(12) (0.417 ng ml(-1)), GA(20) (0.302 ng ml(-1)), GA(24) (1.351 ng ml(-1)), GA(34) (0.076 ng ml(-1)), and GA(53) (0.051 ng ml(-1)) in culture filtrates of Cs-8-1 fungal strain was detected. The Cs-8-1 fungal strain was confirmed as a producer of GAs. Molecular analysis of sequences showed high similarity of 99% to Cadophora malorum. Consequentially, the Cs-8-1 fungal strain was identified as a new C. malorum producing GAs.

Four Unrecorded Aspergillus Species from the Rhizosphere Soil in South Korea.

The genus Aspergillus is commonly isolated from various marine and terrestrial environments; however, only a few species have been studied in rhizosphere soil. As part of the Korean indigenous fungal excavation project, we investigated fungal diversity from rhizosphere soil, focusing on Aspergillus species. A total of 13 strains were isolated from the rhizosphere soil of three different plants. Based on phylogenetic analysis of ß-tubulin and calmodulin and morphological characteristics, we identified five Aspergillus species. A. calidoustus and A. pseudodeflectus were commonly isolated from the rhizosphere soil. Four species were confirmed as unrecorded species in Korea: A. calidoustus, A. dimorphicus, A. germanicus, and A. pseudodeflecuts. The detailed morphological descriptions of these unrecorded species are provided.

Biodegradative Activities of Fungal Strains Isolated from Terrestrial Environments in Korea.

Polylactic acid (PLA) and polycaprolactone (PCL) are commercially available bioplastics that are exploited worldwide, and both are biodegradable. The PLA and PCL polymer-degrading activity of 30 fungal strains that were isolated from terrestrial environments were screened based on the formation of a clear zone around fungal colonies on agar plates containing emulsified PLA or PCL. Among them, five strains yielded positive results of biodegradation. Strains Korean Agricultural Culture Collection (KACC) 83034BP and KNUF-20-PPH03 exhibited PCL degradation; two other strains, KACC 83035BP and KNUF-20-PDG05, degraded PLA; and the fifth strain, KACC 83036BP, biodegraded both tested plastics. Based on phylogenetic analyses using various combinations of the sequences of internal transcribed spacer (ITS) regions, RPB2, LSU, CAL, and ß-TUB genes, the above-mentioned strains were identified as Apiotrichum porosum, Penicillium samsonianum, Talaromyces pinophilus, Purpureocillium lilacinum, and Fusicolla acetilerea, respectively. Based on our knowledge, this is the first report on (i) plastic biodegraders among Apiotrichum and Fusicolla species, (ii) the capability of T. pinophilus to degrade biodegradable plastics, (iii) the biodegradative activity of P. samsonianum against PCL, and (iv) the accurate identification of P. lilacinum as a PLA biodegrader. Further studies should be conducted to determine how the fungal species can be utilized in Korea.

Geographical Isolation and Root-Associated Fungi in the Marine Terrains: A Step Toward Establishing a Strategy for Acquiring Unique Microbial Resources.

This study aimed to understand whether the geo-ecological segregation of native plant species affects the root-associated fungal community. Rhizoplane (RP) and rhizosphere (RS) fungal microbiota of Sedum takesimense native to three geographically segregated coastal regions (volcanic ocean islands) were analyzed using culture-independent methods: 568,507 quality sequences, 1399 operational taxonomic units, five phyla, and 181 genera were obtained. Across all regions, significant differences in the phyla distribution and ratio were confirmed. The Chao's richness value was greater for RS than for RP, and this variance coincided with the number of genera. In contrast, the dominance of specific genera in the RS (Simpson value) was lower than the RP at all sites. The taxonomic identity of most fungal species (95%) closely interacting with the common host plant was different. Meanwhile, a considerable number of RP only residing fungal genera were thought to have close interdependency on their host halophyte. Among these, Metarhizium was the sole genus common to all sites. These suggest that the relationship between potential symbiotic fungi and their host halophyte species evolved with a regional dependency, in the same halophyte species, and of the same natural habitat (volcanic islands); further, the fungal community differenced in distinct geographical regions. Importantly, geographical segregation should be accounted for in national culture collections, based on taxonomical uniqueness.

Rhizoplane and Rhizosphere Fungal Communities of Geographically Isolated Korean Bellflower (Campanula takesimana Nakai).

Fungal communities in the rhizoplane (RP) and rhizosphere (RS) of geographically isolated C. takesimana habitats in different environments such as oceanic (Seodo, the Dokdo Islands), coastline (Sadong, Ulleungdo Island), and inland (Taeha, Ulleungdo Island) regions were analyzed by MiSeq sequencing. In total, 1279 operational taxonomic units (OTUs) were obtained and they were further classified into 185 genera belonging to five phyla. The total number of fungal taxa in the RP samples was lower than those in the RS samples in all the sampled locations, providing an indication of the existence of a certain level of the selective pressures from the host plant. The richness of the RP in the Dokdo Islands was higher than that of Ulleungdo Island, but the richness of the RS in the Dokdo Islands was lower than that of Ulleungdo Island. These results suggest evidence for strong effects of a harsh geo-climate on the RP and RS fungal diversities in the Dokdo Islands. Additionally, a total of 82 fungal genera were identified in all three RP samples and 63 genera (77%) were uniquely found in each of the geographical regions and 43 genera (52.4%) showed high dependency on the C. takesimana vegetation. It was found that the genus Mortierella was the most dominant taxon in all the samples. The geo-ecological isolation of the Korean bellflower may have caused unique formation of the RP and RS fungal communities in the natural habitats.

TRB3 is stimulated in diabetic kidneys, regulated by the ER stress marker CHOP, and is a suppressor of podocyte MCP-1.

The prevalence of diabetic nephropathy continues to rise, highlighting the importance of investigating and discovering novel treatment strategies. TRB3 is a kinase-like molecule that modifies cellular survival and metabolism and interferes with signal transduction pathways. Herein, we report that TRB3 expression is increased in the kidneys of type 1 and type 2 diabetic mice. TRB3 is expressed in conditionally immortalized podocytes; however, it is not stimulated by elevated glucose. The diabetic milieu is associated with increased oxidative stress and circulating free fatty acids (FFA). We show that reactive oxygen species (ROS) such as H(2)O(2) and superoxide anion (via the xanthine/xanthine oxidase reaction) as well as the FFA palmitate augment TRB3 expression in podocytes. C/EBP homologous protein (CHOP) is a transcription factor that is associated with the endoplasmic reticulum stress response. CHOP expression increases in diabetic mouse kidneys and in podocytes treated with ROS and FFA. In podocytes, transfection of CHOP increases TRB3 expression, and ROS augment recruitment of CHOP to the proximal TRB3 promoter. MCP-1/CCL2 is a chemokine that contributes to the inflammatory injury associated with diabetic nephropathy. In these studies, we demonstrate that TRB3 can inhibit basal and stimulated podocyte production of MCP-1. In summary, enhanced ROS and/or FFA associated with the diabetic milieu induce podocyte CHOP and TRB3 expression. Because TRB3 inhibits MCP-1, manipulation of TRB3 expression could provide a novel therapeutic approach in diabetic kidney disease.

A New Species of Thelonectria and a New Record of Cephalotrichum hinnuleum from Gunwi and Ulleungdo in Korea.

Three fungal strains belonging to the class Sordariomycetes were isolated from soils collected from Gyeongsangbuk-do in Korea. They were identified as Cephalotrichum hinnuleum (UD CT 1-3-3 and KNU-19GWF1) and Thelonectria chlamydospora sp. nov. (UD ST 1-2-1). T. chlamydospora sp. nov. was morphologically identical to T. truncata, but its specific macroconidial dimensions, lower number of septations, and chlamydospore diameter render it distinct from the strains of the genus Thelonectria. The strains UD CT 1-3-3 and KNU-19GWF1 were developed flat, velvety to felty, and golden gray to brown-gray after 14 days of incubation at 25 °C on PDA. These strains were produced polyblastic conidiogenous cells and conidia were pale brown to brown, smooth, thin-walled, subglobose to ellipsoidal, arranged in chains, and the diameters of 6.7-9.0 × 3.7-5.1 µm. The strains were also confirmed by using the multi-locus genes using internal transcribed spacer (ITS) regions, partial large subunit (LSU), translation elongation factor 1α (TEF1-α), ß-tubulin (TUB2), and actin (ACT) genes. This is the discovery of T. chlamydospora sp. nov. and Cephalotrichum hinnuleum, a new record from Korea.

Fungal Clusters and Their Uniqueness in Geographically Segregated Wetlands: A Step Forward to Marsh Conservation for a Wealth of Future Fungal Resources.

Here, we investigated fungal microbiota in the understory root layer of representative well-conserved geographically segregated natural wetlands in the Korean Peninsula. We obtained 574,143 quality fungal sequences in total from soil samples in three wetlands, which were classified into 563 operational taxonomic units (OTU), 5 phyla, 84 genera. Soil texture, total nitrogen, organic carbon, pH, and electrical conductivity of soil were variable between geographical sites. We found significant differences in fungal phyla distribution and ratio, as well as genera variation and richness between the wetlands. Diversity was greater in the Jangdo islands wetland than in the other sites (Chao richness/Shannon/Simpson's for wetland of the Jangdo islands: 283/6.45/0.97 > wetland of the Mt. Gariwang primeval forest: 169/1.17/0.22 > wetland of the Hanbando geology: 145/4.85/0.91), and this variance corresponded to the confirmed number of fungal genera or OTUs (wetlands of Jangdo islands: 42/283 > of Mt. Gariwang primeval forest: 32/169 > of the Hanbando geology: 25/145). To assess the uniqueness of the understory root layer fungus taxa, we analyzed fungal genera distribution. We found that the percentage of fungal genera common to two or three wetland sites was relatively low at 32.3%, while fungal genera unique to each wetland site was 67.7% of the total number of identified fungal species. The Jangdo island wetland had higher fungal diversity than did the other sites and showed the highest level of uniqueness among fungal genera (Is. Jangdo wetland: 34.5% > wetland of Mt. Gariwang primeval forest: 28.6% > wetland of the Hanbando geology: 16.7%).

Microbial control measures for soft ice cream in franchise brands: A comparative analysis of microbial analysis and manufacturing practices.

The aim of this study was to establish a complementary direction of the franchise food hygiene guideline to control microbial risks. We evaluated current measures of preventing microbial spoilage and ensuring microbiological safety of food in the food franchise industry. Manufacturing practices were assessed using microbiological analyses, third-party food safety audits, and existing hygiene guidelines. Microbial load indicators of food, manufacturing processes, work environment, and workers were also analyzed to track microbial proliferation. We audited manufacturing practices and processes and analyzed the cleaning and sanitation clauses. We found high contamination of heterotrophic bacteria and detected coliforms in some products. There was no direct contamination by food handlers, and the sterility of raw materials was satisfactory. The main issues were structural complexities of equipment, which hindered cleaning and disinfection, and disinfection practices of franchise brands. Store-level microbial control may be compromised due to the industrial nature of small stores operating collectively. We thus issued complementary guidelines. Improved collective microbiological safety may be ensured through implementation of the revised cleaning and sterilization regulations.

Penicillium from Rhizosphere Soil in Terrestrial and Coastal Environments in South Korea.

Penicillium, the most common genus plays an important ecological role in various terrestrial and marine environments. However, only a few species have been reported from rhizosphere soil. As part of a project to excavate Korean indigenous fungi, we investigated rhizosphere soil of six plants in the forest (terrestrial habitat) and sand dunes (coastal habitat) and focused on discovering Penicillium species. A total of 64 strains were isolated and identified as 26 Penicillium species in nine sections based on morphological characteristics and the sequence analysis of ß-tubulin and calmodulin. Although this is a small-scale study in a limited rhizosphere soil, eight unrecorded species and four potential new species have been identified. In addition, most Penicillium species from rhizosphere soil were unique to each plant. Penicillium halotolerans, P. scabrosum, P. samsonianum, P. jejuense, and P. janczewskii were commonly isolated from rhizosphere soil. Eight Penicillium species, P. aurantioviolaceum, P. bissettii, P. cairnsense, P. halotolerans, P. kananaskense, P. ortum, P. radiatolobatum, and P. verhagenii were recorded for the first time in Korea. Here, we provide the detailed morphological description of these unrecorded species.