Sterol Carrier Protein-2: Binding Protein for Endocannabinoids

The endocannabinoid (eCB) system, consisting of eCB ligands and the type 1 cannabinoid receptor (CB1R), subserves retrograde, activity-dependent synaptic plasticity in the brain. eCB signaling occurs “on-demand,” thus the processes regulating synthesis, mobilization and degradation of eCBs are also primary mechanisms for the regulation of CB1R activity. The eCBs, N-arachidonylethanolamine (AEA) and 2-arachidonoylglycerol (2-AG), are poorly soluble in water. We hypothesize that their aqueous solubility, and, therefore, their intracellular and transcellular distribution, are facilitated by protein binding. Using in silico docking studies, we have identified the nonspecific lipid binding protein, sterol carrier protein 2 (SCP-2), as a potential AEA binding protein. The docking studies predict that AEA and AM404 associate with SCP-2 at a putative cholesterol binding pocket with ?G values of ?3.6 and ?4.6 kcal/mol, respectively. These values are considerably higher than cholesterol (?6.62 kcal/mol) but consistent with a favorable binding interaction. In support of the docking studies, SCP-2-mediated transfer of cholesterol in vitro is inhibited by micromolar concentrations of AEA; and heterologous expression of SCP-2 in HEK 293 cells increases time-related accumulation of AEA in a temperature-dependent fashion. These results suggest that SCP-2 facilitates cellular uptake of AEA. However, there is no effect of SCP-2 transfection on the cellular accumulation of AEA determined at equilibrium or the IC50 values for AEA, AM404 or 2-AG to inhibit steady state accumulation of radiolabelled AEA. We conclude that SCP-2 is a low affinity binding protein for AEA that can facilitate its cellular uptake but does not contribute significantly to intracellular sequestration of AEA.     

<Emphasis Type="Italic">Agaricus bisporus</Emphasis> supplementation reduces high-fat diet-induced body weight gain and fatty liver development

Obesity is a global epidemic characterized not only by excessive fat deposition but also by important complications such as nonalcoholic liver steatosis. Beneficial antiobesogenic effects have been described for some mushrooms. The current study aimed to demonstrate the protective effect of Agaricus bisporus (AB) supplementation against the metabolic alterations induced by high-fat-diet (HFD) feeding. Eight-week-old C57BL/6J mice were fed for 10 weeks with one of the following diets: (1) control diet (n?=?7), (2) HFD (n?=?7), (3) HFD supplemented with 5% AB (n?=?9), and (4) HFD supplemented with 10% AB (n?=?9). A pair-fed group was also included for the 10% AB group (n?=?6). The impact of AB supplementation on food intake, body weight gain, and liver and fat pad weights was examined. Biochemical, histological, and molecular parameters were also analyzed. Dietary supplementation with 10% AB reduced the HFD-induced increase in body, epididymal, and mesenteric fat weights (p?<?0.01, p?<?0.05, and p?<?0.05, respectively). Supplementation with AB also reduced liver damage in a dose-dependent manner (p?<?0.01 and p?<?0.001). This effect was confirmed by histological analysis that showed that liver steatosis was markedly reduced in mice fed with AB. The beneficial properties of 10% AB supplementation appear to be mediated through a decrease in food intake and via stimulation of mesenteric and hepatic free-fatty acid beta-oxidation, along with a decrease in epidydimal and hepatic expression of CD36. In conclusion, supplementation with AB prevents excessive body weight gain and liver steatosis induced by HFD consumption.     

Effects of Probiotics on Survival,Growth and Digestive Enzymes Activities in Freshwater Prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis> (De Man 1879)

A study was conducted to examine the effects of three probiotics, Lactobacillus sporogenes, Bacillus subtilis and Saccharomyces cerevisiae on the survival, growth and digestive enzymes activities of the freshwater prawn Macrobrachium rosenbergii post larvae (PL). The probiotics, L. sporogenes (4 %), B. subtilis (3 %) and S. cerevisiae (4 %) were taken and mixed with basal diet. Diet without probiotics served as control. These probiotics diets were fed to M. rosenbergii PL for a period of 60 days. After the feeding trail, the growth parameters such as survival, weight gain, specific growth rate and protein efficiency rate were found to be significantly (P < 0.05) higher in 4 % S. cerevisiae incorporated diet fed PL when compared with control. In the case of feed conversion rate just the reverse was seen (P < 0.05) at this concentration. This indicates its superior quality among different concentrations of probiotics tested. Activities of digestive enzymes, such as protease, amylase and lipase were significantly (P < 0.05) higher at this concentration (4 % S. cerevisiae). Some of essential and non-essential amino acids also significantly elevated in probiotics supplemented diet fed prawns. This study indicated that probiotics, S. cerevisiae incorporated diets were beneficial for M. rosenbergii in terms of increasing growth, enzyme and amino acid production.     

Distribution and diet of demersal Arctic Cod, <Emphasis Type="Italic">Boreogadus saida</Emphasis>, in relation to habitat characteristics in the Canadian Beaufort Sea

Arctic Cod (Boreogadus saida) occur throughout the circumpolar north; however, their distributions at localized scales are not well understood. The seasonal habitat associations and diet preferences across life-history stages of this keystone species are also poorly known, thereby impeding effective regulatory efforts in support of conservation objectives. The distribution of Arctic Cod in the Canadian Beaufort Sea was assessed using bottom trawling in shelf and slope habitats between 20 and 1000 m depths. Highest catch biomasses occurred at 350 and 500 m depth slope stations, coinciding with >0 °C temperatures in the Pacific–Atlantic thermohalocline and Atlantic water mass. Calanus glacialis, Calanus hyperboreus, Themisto libellula, and Themisto abyssorum were identified as key prey species in the diet of Arctic Cod, comprising approximately 86 % of total biomass in guts. Hierarchical cluster analysis with a SIMPROF test identified five statistically significant (p < 0.05) diet groups among gut samples. Arctic Cod shifted from a primarily Calanus diet at shelf stations (<200 m depth) to a Themisto diet in slope habitats (>200 m depth) coinciding with an associated increase in fish standard length with depth. Smaller Arctic Cod fed primarily on Calanus copepods and larger Arctic Cod fed primarily on the larger Themisto species. The habitat and diet associations presented here will inform knowledge of structural and functional relationships in Arctic marine ecosystems, aid in mitigation and conservation efforts, and will enhance our ability to predict the effects of climate change on the local spatial and depth associations of this pivotal marine fish.     

Low 1,5-anhydroglucitol levels are associated with long-term cardiac mortality in acute coronary syndrome patients with hemoglobin A1c levels less than 7.0%

Background

Diabetes mellitus is considered an important risk factor for cardiovascular diseases. High hemoglobin A1c (HbA1c) levels, which indicate poor glycemic control, have been associated with occurrence of cardiovascular diseases. There are few parameters which can predict cardiovascular risk in patients with well-controlled diabetes. Low 1,5-anhydroglucitol (1,5-AG) levels are considered a clinical marker of postprandial hyperglycemia. We hypothesized that low 1,5-AG levels could predict long-term mortality in acute coronary syndrome (ACS) patients with relatively low HbA1c levels.

Methods

The present study followed a retrospective observational study design. We enrolled 388 consecutive patients with ACS admitted to the cardiac intensive care unit at the Juntendo University Hospital from January 2011 to December 2013. Levels of 1,5-AG were measured immediately before emergency coronary angiography. Patients with early stent thrombosis, no significant coronary artery stenosis, malignancy, liver cirrhosis, a history of gastrectomy, current steroid treatment, moderately to severely reduced kidney function (estimated glomerular filtration rate < 45 ml/min/1.73 m²; chronic kidney disease stage 3B, 4, and 5), HbA1c levels ≥ 7.0%, and those who received sodium glucose co-transporter 2 inhibitor therapy were excluded.

Results

During the 46.9-month mean follow-up period, nine patients (4.5%) died of cardiovascular disease. The 1,5-AG level was significantly lower in the cardiac death group compared with that in the survivor group (12.3 ± 5.3 vs. 19.2 ± 7.7 µg/ml, p < 0.01). Kaplan–Meier survival analysis showed that low 1,5-AG levels were associated with cardiac mortality (p = 0.02). Multivariable Cox regression analysis showed that 1,5-AG levels were an independent predictor of cardiac mortality (hazard ratio 0.76; 95% confidence interval 0.41–0.98; p = 0.03).

Conclusion

Low 1,5-AG levels, which indicate postprandial hyperglycemia, predict long-term cardiac mortality even in ACS patients with HbA1c levels < 7.0%.

     

Ribosome engineering and fermentation optimization leads to overproduction of tiancimycin A,a new enediyne natural product from <Emphasis Type="Italic">Streptomyces</Emphasis> sp. CB03234

Tiancimycin (TNM) A, a recently discovered enediyne natural product from Streptomyces sp. CB03234, showed rapid and complete killing of cancer cells and could be used as a payload in antibody drug conjugates. The low yield of TNM A in the wild-type strain promoted us to use ribosome engineering and fermentation optimization for its yield improvement. The Streptomyces sp. CB03234-R-16 mutant strain with a L422P mutation in RpoB, the RNA polymerase β-subunit, was obtained from the rifamycin-resistant screening. After fermentation optimization, the titers of TNM A in Streptomyces sp. CB03234-R-16 reached to 22.5 ± 3.1 mg L^?1 in shaking flasks, and 13 ± 1 mg L^?1 in 15 L fermentors, which were at least 40-fold higher than that in the wild-type strain (~ 0.3 mg L^?1). Quantitative real-time RT-PCR revealed markedly enhanced expression of key genes encoding TNM A biosynthetic enzymes and regulators in Streptomyces sp. CB03234-R-16. Our study should greatly facilitate the future efforts to develop TNM A into a clinical anticancer drug.     

Combined or Individual Effects of Dietary Probiotic <Emphasis Type="Italic">Pedicoccus acidilactici</Emphasis> and Nucleotide on Growth Performance,Intestinal Microbiota,Hemato-biochemical Parameters,and Innate Immune Response in Goldfish (<Emphasis Type="Italic">Carassius auratus</Emphasis>)

This study investigated the effect of dietary supplementation of probiotic Pedicoccus acidilactici and nucleotide (separately or combined) on growth performance, intestinal microbiota, hemato-immunological parameters, and immunity response in goldfish (Carassius auratus). Fish (average weight 5–6 g) were acclimatized and divided into eight experimental diets supplemented with P. acidilactici of different concentrations (0.1, 0.2, and 0.3% diet) and nucleotides (0 and 0.5% diet) for 6 months. Fish fed with experimental diets showed significant differences in terms of final weight, weight gain, feed conversion ratio, daily growth rate, and condition factor when compared to control diet (P < 0.05). Fish fed with probiotic (0.3%) separately and combined with nucleotide (0.5%) had highest RBC and WBC when compared to other diets (P < 0.05), while the highest values for Hb and Hct as well as total protein, glucose, albumin, and globulin were observed in probiotic (0.2%) and nucleotide (0.5%) combined diet. Serum lysozyme and anti-protease activities were significantly higher in probiotic (0.1 and 0.2%) and nucleotide (0.5%) combined diets. Similarly, these two diets combined showed the highest colonization of P. acidilactici when compared to other diets. In conclusion, combined dietary probiotic and nucleotide improve the growth performance, hemato-biochemical parameters, and intestine growth in C. auratus.     

Maternal high-fat diet during pregnancy and lactation affects hepatic lipid metabolism in early life of offspring rat

We investigated whether maternal over-nutrition during pregnancy and lactation affects the offspring’s lipid metabolism at weaning by assessing liver lipid metabolic gene expressions and analysing its mechanisms on the development of metabolic abnormalities. Female Sprague–Dawley rats were fed with standard chow diet (CON) or high-fat diet (HFD) for 8 weeks, and then continued feeding during gestation and lactation. The offspring whose dams were fed with HFD had a lower birth weight but an increased body weight with impaired glucose tolerance, higher serum cholesterol, and hepatic steatosis at weaning. Microarray analyses showed that there were 120 genes differently expressed between the two groups. We further verified the results by qRT-PCR. Significant increase of the lipogenesis (Me1, Scd1) gene expression was found in HFD (P<0.05), and up-regulated expression of genes (PPAR-α, Cpt1α, Ehhadh) involved in β-oxidation was also observed (P<0.05), but the Acsl3 gene was down-regulated (P<0.05). Maternal over-nutrition could not only primarily induce lipogenesis, but also promote lipolysis through an oxidation pathway as compensation, eventually leading to an increased body weight, impaired glucose tolerance, elevated serum cholesterol and hepatic steatosis at weaning. This finding may provide some evidence for a healthy maternal diet in order to reduce the risk of metabolic diseases in the early life of the offspring.     

Hemato-Immunological Responses and Disease Resistance in Siberian Sturgeon <Emphasis Type="Italic">Acipenser baerii</Emphasis> Fed on a Supplemented Diet of <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis>

A feeding trial was conducted to investigate the effects of different levels of dietary Lactobacillus plantarum on hemato-immunological parameters and resistance against Streptococcus iniae infection in juvenile Siberian sturgeon Acipenser baerii. Fish (14.6 ± 2.3 g) were fed three experimental diets prepared by supplementing a basal diet with L. plantarum at different concentrations [1 × 10⁷, 1 × 10⁸ and 1 × 10⁹ colony-forming units (cfu) g^?1] and a control (non-supplemented basal) diet for 8 weeks. Innate immune responses (immunoglobulin (Ig), alternative complement activity (ACH50) and lysozyme activity) were significantly higher in fish fed the 1 × 10⁸ and 1 × 10⁹ cfu g^?1 L. plantarum diet compared to the other groups (P < 0.05). Furthermore, fish fed on various levels of L. plantarum significantly showed higher red blood cell (RBC), hemoglobin (Hb), white blood cell (WBC) and monocyte compared to those of the control group (P < 0.05). At the end of the feeding experiment, some fish were challenged with S. iniae to quantify the level of disease resistance. The mortality after S. iniae challenge was decreased in fish fed a probiotic. These results indicated that dietary supplementation of L. plantarum improved immune response and disease resistance of Siberian sturgeon juvenile.     

Effects of neutrophils peptide-1 transgenic <Emphasis Type="Italic">Chlorella ellipsoidea</Emphasis> on the gut microbiota of male Sprague–Dawley rats,as revealed by high-throughput 16S rRNA sequencing

Rabbit neutrophils peptide-1 (NP-1) is a type of defensin that possesses a broad spectrum of antimicrobial activity. Chlorella ellipsoidea is a new eukaryotic expression system for exogenously producing NP-1. The NP-1 transgenic C. ellipsoidea can be directly added into feed as antimicrobial agent without any purification procedure for the NP-1 peptide. However, the effects of C. ellipsoidea and NP-1 on the host gut microbiota should be explored before application. In this study, diets containing different concentrations (1.25, 2.5, and 5 %) of C. ellipsoidea and NP-1 transgenic C. ellipsoidea were administered to male Sprague–Dawley rats. Compared with the chow diet control group, none of the experimental groups showed any significant differences in their growth indices, and no histopathological damage was observed. The phylotypes of gut microbiota in the control group, the 5 % C. ellipsoidea diet group and the 5 % NP-1 transgenic C. ellipsoidea diet group were determined by 16S rRNA sequencing. The results showed that both 5 % experimental groups had shifted community memberships of gut microbiota. In particular, the 5 % NP-1 transgenic C. ellipsoidea diet exhibited an increased abundance of most Gram-positive bacterial taxa and a reduced abundance of most Gram-negative bacterial taxa, and it promoted the growth of some lactic acid bacterial genera. Lactic acid bacteria, especially the Bifidobacterium and Lactobacillus, have been widely reported to be benefic effects on the host. Thus NP-1 transgenic C. ellipsoidea is promising feed additive and gut regulator, as it have the potential to increase the abundance of Bifidobacterium and Lactobacillus in gut microbiota of animal.     

Milk and Dairy Products Intake Is Associated with Low Levels of Lead (Pb) in Workers highly Exposed to the Metal

Lead (Pb) is a toxic metal, frequently associated with occupational exposure, due to its widespread use in industry and several studies have shown high Pb levels in workers occupationally exposed to the metal. The aim of this study was to evaluate the influence of milk and dairy products (MDP) on Pb levels in blood (B-Pb), plasma (P-Pb), and urine (U-Pb), in workers from automotive battery industries in Brazil. The study included 237 male workers; information concerning diet and lifestyle were gathered through a questionnaire, and B-Pb, P-Pb, and U-Pb were determined by ICP-MS. Mean B-Pb, P-Pb, and U-Pb were 21 ± 12, 0.62 ± 0.73 μg/dL, and 39 ± 47 μg/g creatinine, respectively. Forty three percent of participants declared consuming ≤3 portions/week of MDP (classified as low-MDP intake), while 57% of individuals had >3portions/week of MDP (high-MDP intake). B-Pb and P-Pb were correlated with working time (r _s  = 0.21; r _s  = 0.20; p < 0.010). Multivariable linear regressions showed a significant influence of MDP intake on B-Pb (β = ?0.10; p = 0.012) and P-Pb (β = ?0.16; p < 0.010), while no significance was seen on U-Pb. Our results suggest that MDP consumption may modulate Pb levels in individuals highly exposed to the metal; these findings may be due to the Pb-Ca interactions, since the adverse effects of Pb are partially based on its interference with Ca metabolism and proper Ca supplementation may help to reduce the adverse health effects induced by Pb exposure.     

Production of a Mixture of Fengycins with Surfactant and Antifungal Activities by <Emphasis Type="Italic">Bacillus</Emphasis> sp. MA04, a Versatile PGPR

Bacillus sp. strain MA04 a plant growth-promoting rhizobacteria (PGPR) showed hemolytic activity on blood agar plates, and the supernatant from liquid culture in nutrient broth at 24 h exhibited emulsification activity, suggesting the production of biosurfactants. In antagonist assays, the supernatant showed antifungal activity against phytopathogenic fungi such as Penicillium expansum, Fusarium stilboides, Sclerotium rolfsii y Rhizoctonia solani, finding a reduction of mycelial growth of all fungi tested, ranging from 35 to 69%, this activity was increased with time of culture, accomplishing percentages of inhibition up to 85% with supernatants obtained at 72 h. Then, the crude biorsurfactant (CB) was isolated from the supernatant in order to assay its antagonistic effect on the phytopathogens previously tested, finding an increase in the inhibition up to 97% at 500 mg/L of CB. The composition of CB was determined by infrared spectroscopy, identifying various functional groups related to lipopeptides, which were purified by high-performance liquid chromatography and analyzed by MALDI-TOF/TOF–MS, revealing a mixture of fengycins A and B whose high antifungal activity is been widely recognized. These results show that PGPR Bacillus sp. MA04 could also contribute to plant health status through the production of metabolites with antimicrobial activity.     

Protein malnutrition during fetal programming induces fatty liver in adult male offspring rats

We evaluated the effects of protein malnutrition on liver morphology and physiology in rats subjected to different malnutrition schemes. Pregnant rats were fed with a control diet or a low protein diet (LPD). Male offspring rats received a LPD during gestation, lactation, and until they were 60 days old (MM group), a late LPD that began after weaning (CM), or a LPD administrated only during the gestation-lactation period followed by a control diet (MC). On day 60, blood was collected and the liver was dissected out. We found a decrease in MM rats’ total body (p < 0.001) and liver (p < 0.05) weight. These and CM rats showed obvious liver dysfunction reflected by the increase in serum glutamic pyruvic transaminase (SGOT) (MM p < 0.001) and serum glutamic pyruvic transaminase (SGPT) (MM and CM p < 0.001) enzymes, and liver content of cholesterol (MM and CM p < 0.001) and triglycerides (MM p < 0.01; CM p < 0.001), in addition to what we saw by histology. Liver dysfunction was also shown by the increase in gamma glutamyl transferase (GGT) (MM, MC, and CM p < 0.001) and GST-pi1 (MM and CM p < 0.001, MC p < 0.05) expression levels. MC rats showed the lowest increment in GST-pi1 expression (MC vs. MM; p < 0.001, MC vs. CM; p < 0.01). ROS production (MM, CM, and MC: p < 0.001), lipid peroxidation (MM, CM, and MC p < 0.001), content of carbonyl groups in liver proteins (MM and CM p < 0.001, MC p < 0.01), and total antioxidant capacity (MM, CM, and MC p < 0.001) were increased in the liver of all groups of malnourished animals. However, MM rats showed the highest increment. We found higher TNF-α (MM and CM p < 0.001), and IL-6 (MM and CM p < 0.001) serum levels and TGF-β liver content (MM p < 0.01; CM p < 0.05), in MM and CM groups, while MC rats reverted the values to normal levels. Pro-survival signaling pathways mediated by tyrosine or serine/threonine kinases (pAKT) (MM and CM p < 0.001; MC p < 0.01) and extrasellular signal-regulated kinase (pERKs) (MM p < 0.01; CM p < 0.05) appeared to be activated in the liver of all groups of malnourished rats, suggesting the presence of cells resistant to apoptosis which would become cancerous. In conclusion, a LPD induced liver damage whose magnitude was related to the developmental stage at which malnutrition occurs and to its length.     

Spore associated bacteria of arbuscular mycorrhizal fungi improve maize tolerance to salinity by reducing ethylene stress level

The role of spore associated bacteria of arbuscular mycorrhizal fungi (AMF) in improving plant growth and alleviating salt stress is a potential area to explore. In the present study, 22 bacteria isolated from the spore walls of AMF were identified to contain 1-aminocyclopropane-1-carboxylate deaminase. These were tested for their ability to improve seed germination and alleviate salt stress in the early growth of maize. Among the isolates, 19 bacteria that were able to grow at 4?% NaCl were used for germination assay. Two bacteria and seven bacteria significantly improved maize seed germination at 100 mM NaCl and 200 mM NaCl, respectively. Based on the presence of plant growth promoting (PGP) characters and the ability to improve seed germination, five strains were chosen for further experiments. At 0 mM NaCl, all the strains were able to increase maize shoot and root growth significantly. At 25 mM NaCl, except for Bacillus aryabhattai S210B15, all the strains were able to increase shoot and root growth significantly. At 50 mM NaCl, Bacillus aryabhattai S110B3 and B. aryabhattai S210B15 significantly improved shoot length, whereas, Pseudomonas koreensis S2CB35 and B. aryabhattai S210B15 significantly increased root length. Although salinity increased ethylene production in maize, bacterial inoculation significantly reduced the ethylene level at 0, 25 and 50 mM NaCl. Among the five strains, only P. koreensis S2CB35 showed the presence of PGP functional traits of nifH, acdS and nodA genes.     

Effects of Dietary <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> on Growth Performance,Digestive Enzymes and Gut Morphology of <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

A 15-day feeding trial was conducted to investigate the effect of dietary Lactobacillus plantarum on growth performance, digestive enzyme activities and gut morphology of juvenile Pacific white shrimp, Litopenaeus vannamei (initial body weight = 7.96 ± 0.59 g). Four microbound diets were formulated to contain fermentation supernatant (FS), live bacteria (LB), dead bacteria (DB), and cell-free extract (CE) of L. plantarum. Results indicated that final weight was significantly higher in FS, DB, and CE group in comparison to the control group (P < 0.05). The maximum weight gain rate (WGR) and specific growth rate (SGR) of the CE diet group were significantly higher than that of other groups (P < 0.05). The FCR of CE diet group was lower than that of the control, LB, DB, and FS diets groups (P < 0.05). The highest digestive enzyme activities (amylase, lipase, and pepsin activity) in the hepatopancreas and gut of shrimp were observed in the CE diet group. Histological study revealed that dietary CE diet could significantly increase the enterocytes height of shrimp. The administration of cell-free extract of L. plantarum could effectively improve the growth performance of L. vannamei via the improvement of digestive enzyme activities and the enterocytes height of shrimp. The results of this study will be essential to promote application of probiotics in shrimp aquaculture.     

Clinical and Microbiological Investigation of Fungemia from Four Hospitals in China

In this study, fungemia cases from four tertiary hospitals located in Shanghai and Anhui province in China from March 2012 to December 2013 were enrolled to investigate clinical features, species distribution, antifungal susceptibility and strain relatedness. During the study period, 137 non-duplicate cases and their corresponding isolates were collected. Six different genera of fungi were identified, of which Candida spp. were the most common (126/137, 91.97 %), with C. albicans predominating (48/137, 35.03 %). The non-Candida fungi rate reached 8.03 % (11/137), and Pichia spp. was the most common (5/137, 3.65 %). Compared with C. albicans, non-C. albicans fungi-associated fungemia was more likely in younger (p = 0.004) and male patients (χ ² = 6.2618, p = 0.0123) and patients from ICUs (χ ² = 6.3783, p = 0.0116). Overall, the susceptible/WT rates of common Candida spp. to fluconazole, itraconazole, voriconazole, flucytosine, amphotericin B and caspofungin were 74.63, 92.31, 93.16, 96.58, 100 and 95.69 %, respectively. C. tropicalis and C. guilliermondii had a low susceptibility to fluconazole: 79.95 and 77.78 %, respectively. No isolates were resistant/WT to caspofungin, but C. parapsilosis and C. guilliermondii had high MIC₉₀ values; 1 and 2 mg/L, respectively. In terms of genotyping, MLST was taken for C. glabrata and C. tropicalis, while microsatellite marker analysis was used for C. albicans and C. parapsilosis. C. glabrata was predominantly clone ST7, accounting for 75 %, while the other isolates showed genetic diversity. Considering the increased proportion of non-C. albicans fungi and the presence of endemic clones of C. glabrata, it is essential to undertake additional surveillance of fungemia.