Isolation of azole-resistant Aspergillus spp. from food products.

The prevalence of azole-resistant Aspergillus fumigatus is increasing worldwide and is speculated to be related to the use of azole pesticides. Aspergillus spp., the causative agent of aspergillosis, could be brought into domestic dwellings through food. However, studies on azole-resistant Aspergillus spp. in food products are limited. Therefore, we aimed to isolate Aspergillus spp. from processed foods and commercial agricultural products and performed drug susceptibility tests for azoles. Among 692 food samples, we isolated 99 strains of Aspergillus spp. from 50 food samples, including vegetables (22.9%), citrus fruits (26.3%), cereals (25.5%), and processed foods (1.8%). The isolates belonged to 18 species across eight sections: Aspergillus, Candidi, Clavati, Flavi, Fumigati, Nidulantes, Nigri, and Terrei. The most frequently isolated section was Fumigati with 39 strains, followed by Nigri with 28 strains. Aspergillus fumigatus and A. welwitschiae were the predominant species. Ten A. fumigatus and four cryptic strains, four A. niger cryptic strains, two A. flavus, and four A. terreus strains exceeded epidemiological cutoff values for azoles. Aspergillus tubingensis, A. pseudoviridinutans, A. lentulus, A. terreus, and N. hiratsukae showed low susceptibility to multi-azoles. Foods containing agricultural products were found to be contaminated with Aspergillus spp., with 65.3% of isolates having minimal inhibitory concentrations below epidemiological cutoff values. Additionally, some samples harbored azole-resistant strains of Aspergillus spp. Our study serves as a basis for elucidating the relationship between food, environment, and clinically important Aspergillus spp.

[Prevalence and Antimicrobial Resistance of Campylobacter jejuni/coli and Analysis of Macrolide Resistance Isolates from Retail Meat in Tokyo, Japan (2010-2019)].

This study aimed to investigate the prevalence and antimicrobial sensitivity of Campylobacter jejuni and Campylobacter coli in retail meat (chicken, beef, pork, venison, wild boar, horse, lamb and mutton) in Tokyo (Japan) from 2010 to 2019. Furthermore, the resistance mechanism of erythromycin (EM)-resistant strains was analysed. C. jejuni had a highly positive rate in domestic chicken meat (53.4%, 334/626 samples), domestic chicken offal (49.3%, 34/69 samples), and domestic beef offal (28.3%, 47/166 samples), while C. coli had a high positivity rate in domestic pork offal (31.7%, 44/139 samples). The positive rate of C. jejuni was significantly higher in offal than that in meat in domestic beef, while the positive rate of C. coli was significantly higher in offal than that in meat in domestic beef and domestic pork (p<0.05). In the isolates, 1.0% (6/631 strains) of C. jejuni and 36.2% (55/152 strains) of C. coli were EM resistant, with 41.5% (262/631 strains) of C. jejuni and 65.1% (99/152 strains) of C. coli being ciprofloxacin resistant. A2075G mutation of the 23S rRNA gene was confirmed in all EM-resistant strains.

[Prevalence of Cereulide-Producing Bacillus cereus in Pasteurized Milk].

To recognize the risk of Bacillus cereus in pasteurized milk, we investigated the prevalence of B. cereus and the rate of the production of cereulide from B. cereus isolates. B. cereus was found in 66 out of 101 (65.3%) domestically pasteurized milk samples in Japan. The ces gene was identified in 3 out of 90 B. cereus isolates that were isolated from three samples (one product) among the 101 samples. The ces gene positive isolate, the reference strain F4810/72 and a B. cereus isolate collected in a food poisoning incident were shown the productivity of cereulide using an LC-MS/MS analysis. The LC-MS/MS analysis was confirmed the ability of identification and quantification of cereulide produced in the milk samples. In this study, it was shown that B. cereus strains are prevalent in pasteurized milk, some of these strains produce cereulide, and confirmed usefulness of LC-MS/MS analysis to detect cereulide in milk.

Diagnosis of Leiomyosarcoma after Uterine Artery Embolization for Multiple Leiomyomas.

Uterine sarcoma is significantly rarer than leiomyoma and has poor prognosis. Moreover, the diagnosis of leiomyosarcoma is difficult because its symptoms, including pelvic pain, uterine mass, and/or uterine bleeding, are very similar to those of leiomyoma. There are a few cases of leiomyosarcoma wherein leiomyoma was treated with uterine artery embolization (UAE); these reports revealed that the symptoms of hypermenorrhea or/and pelvic pain persisted even after UAE. Symptoms persisting even after UAE treatment for leiomyomas, especially multiple leiomyomas, should be investigated to rule out leiomyosarcoma. Therefore, long-term follow-up is needed. Here, we describe a case of a 39-year-old woman diagnosed with leiomyosarcoma 3 years after undergoing UAE for multiple leiomyomas.

[Salmonella Serovars Isolated from Retail Meats in Tokyo, Japan and Their Antimicrobial Susceptibility].

To identify the risk of Salmonella in meat, we investigated the prevalence of Salmonella, serovars and their antimicrobial susceptibility patterns. Salmonella was found in 353 out of 849 (41.6%) chicken, 9 out of 657 (1.4%) pork, 1 out of 517 (0.2%) Beef, 6 out of 8 (75.0%) chicken offal, 43 out of 142 (30.3%) pork offal and 4 out of 198 (2.0) beef offal samples collected from retail meats in Tokyo, Japan between 2009 and 2017. Salmonella Infantis was the most common serovar, followed by S. Schwarzengrund in the isolates from domestic chicken meats. The prevalence rate of S. Infantis decreased while that of S. Schwarzengrund increased by the year. Apart from this, the most prevalent serovars were S. Heidelberg in the imported chicken meat isolates, S. Typhimurium and Salmonella O4:i:- in pork, and S. Derby in beef isolates. Antimicrobial testing revealed high resistance to tetracycline (TC) in all meat sample isolates; however, all the isolates were sensitive to carbapenem and fluoroquinolone. Fourteen cefotaxime (CTX) resistant strains, seven extended spectrum ß-lactamase (ESBL) producing strains and twenty-three AmpC producing strains were isolated from chicken meat samples. These findings indicate that the serovar and antimicrobial susceptibility varied among meat samples.

[Prevalence of Antimicrobial Resistance in Escherichia coli Isolated from Retail Meat in Tokyo, Japan].

This study aimed to survey the trend of antimicrobial resistance in Escherichia coli obtained from retail meat. We examined the susceptibilities of 1,115 E. coli isolates obtained from chicken, beef, pork, venison, and wild boar meat from 2011 to 2017 in Tokyo to 14 antimicrobials (ampicillin, cefotaxime (CTX), streptomycin, gentamicin, kanamycin, tetracycline (TC), chloramphenicol, nalidixic acid, ciprofloxacin, sulfamethoxazole-trimethoprim, fosfomycin, amikacin, imipenem, and meropenem). Of all the tested isolates, 18.7% (135/721) isolates from chicken, 77.0% (117/152) from beef, 46.6% (89/187) from pork, 100% (28/28) from venison, and 92.6% (25/27) from wild boar meat were susceptible to all tested antimicrobials. Furthermore, TC resistance was the most common, with rates as high as 56.7% (409/721) and 40.6% (76/187) in the isolates from chicken and pork, respectively. CTX resistance was detected in 4.9% (25/506) of the isolates from domestic chicken and 23.7% (51/215) of the isolates from imported chicken. Moreover, CTX resistance rate in isolates from domestic chicken was significantly lower in 2016 (0.9%, 1/111) and in 2017 (0.8%, 1/121) than in 2012 (10.6%, 17/161). In conclusion, E. coli isolates from retail meat were most commonly resistant to TC, and CTX resistance was higher in E. coli isolates from imported chicken than in E. coli isolates from domestic chicken.

Detection of the mcr-1 gene in colistin-resistant Escherichia coli from retail meat in Japan.

In this study, the presence of the mcr-1 gene in Escherichia coli from retail meat in Japan was investigated. Nine E. coli isolates (eight from chickens and one from pork) carried the mcr-1 gene on the plasmid. In six isolates from domestic chickens, mcr-1 was located on the IncI2 plasmid, which is approximately 60 kb in size. In the remaining three isolates from imported chicken and pork, mcr-1 was located on the IncX4 plasmid (30 kb).

Natural killer cell activity in women with recurrent miscarriage: Etiology and pregnancy outcome.

This study aimed to evaluate whether natural killer (NK) cell activity was associated with the etiology of recurrent miscarriage (RM), and to evaluate the predictive value of NK cell activity for outcomes of following pregnancies in women with RM. Peripheral NK cell activity was measured in 160 non-pregnant women with a history of two or more miscarriages. This activity was compared according to the etiology of RM and to pregnancy outcomes in women who became pregnant. NK cell activity in women with unexplained RM was significantly higher than that in those with known etiologies of RM. NK cell activity in women whose next pregnancies ended in miscarriage of fetuses with a normal chromosome karyotype (MN) was higher than that in those with live births (p<0.05). Women with NK cell activity ≥33% had a higher risk for MN (relative risk 3.4, 95% confidence interval 1.3-8.7). An increase in peripheral NK cell activity was associated with MN. This increase might be involved in the pathophysiology underlying RM.

Prevalence and contamination levels of listeria monocytogenes in ready-to-eat foods in Tokyo, Japan.

We surveyed prevalence and contamination levels of Listeria monocytogenes in ready-to-eat foods between 2000 and 2012 in Tokyo. L. monocytogenes was isolated from 52 (1.7%) out of 2,980 samples. Comparing the prevalence in the study period, 2.2% were positive in the former period (2000-2005) and 1.2% in the latter (2006-2012). Using the most probable number (MPN) technique, 32 samples were contaminated with fewer than 0.3 L. monocytogenes/g, 10 samples with 0.3-1.0/g and 4 samples with more than 1.0/g (the maximum was 2.3/g). The most common serovar was 1/2a, followed by 1/2b, 4b and 1/2c. We revealed that ready-to-eat foods in Tokyo were contaminated with L. monocytogenes, although the contamination levels were low.

Multiplex PCR serogrouping of Listeria monocytogenes isolated in Japan.

PCR serogrouping methods were used to examine strains of L. monocytogenes isolated in Japan. Among 187 strains, 99.5% were classified into 4 PCR serogroups corresponding to conventional serotypes. Only one isolate had a new PCR profile, which may be a variant of serogroup IVb.

Medium-dose intravenous immunoglobulin therapy for women with six or more recurrent miscarriages.

This study aimed to evaluate changes in natural killer (NK) cell activity and the percentage of monocytes in women with recurrent miscarriage who received medium-dose intravenous immunoglobulin (IVIg) therapy. Fourteen women with a history of six or more recurrent miscarriages of unexplained etiology received 60-g IVIg therapy (20 g daily, for three days) during early gestation. NK cell activity in the peripheral blood decreased to 12% one week after therapy compared with before therapy (median, 22%, P < 0.001) and the percentage of monocytes increased from 5.2% to 7.5% (P < 0.005). Four pregnancies ended in live births of healthy neonates, whereas the other ten pregnancies ended in miscarriages. Excluding one miscarriage with a chromosomal abnormality, the live birth rate was 30.8% (4/13). The rate of reduction of NK cell activity in the success group (-58.8%) tended to be greater than that in the failure group (-14.8%, P = 0.057).

Microbial conversion of glucose to a novel chemical building block, 2-pyrone-4,6-dicarboxylic acid.

2-Pyrone-4,6-dicarboxylic acid (PDC) is a catabolic intermediate in Sphingobium sp. SYK-6 (previously characterized as Sphingomonas paucimobilis SYK-6), which is a degrader of lignin-derived aromatic compounds. Recently, PDC has been also characterized as a novel starting material for several potentially useful synthetic polymers. In a previous study, we constructed a biosynthetic system in which PDC was generated efficiently from a chemically synthesized compound, protocatechuate. In order to develop an alternative system for production of PDC, we tried to generate it from glucose, which is a low-cost sugar that can be obtained from abundant cellulosic wastes and biomass crops. We designed a metabolic bypass to PDC from the shikimate pathway in recombinant Escherichia coli cells. PDC accumulated in the medium of recombinant E. coli cells that had been transformed with genes isolated from Emericella niger, E. coli, Pseudomonas putida, and Sphingobium sp. SYK-6. The yield of PDC depended on the combination of genes that we introduced into the cells and on the specific of host strain. Under optimal conditions, the yield and titer of PDC were, respectively, 17.3% and 0.35 mg/l when the concentration of glucose was 2 g/l and the culture volume was 50 ml. Our results open up the possibility of novel utilization of biomass as the source of a useful chemical building block.