Endophytic bacterium Pseudomonas protegens suppresses mycelial growth of Botryosphaeria dothidea and decreases its pathogenicity to postharvest fruits.

Apple (Malus domestica Borkh.), one of the most economically important fruits widely consumed worldwide, has been suffering from apple ring rot caused by Botryosphaeria dothidea, which dramatically affects its quality and yield. In the present study, we demonstrated that Pseudomonas protegens, isolated from Chinese leek (Allium tuberosum), significantly suppressed the mycelial growth and propagation of B. dothidea, respectively, further displayed a considerably inhibitory effect on the apple ring rot of postharvest fruits. In addition, P. protegens significantly improved the total soluble solid/titrable acidity (TSS/TA) ratio and soluble sugar/titrable acidity (SS/TA) ratio and drastically maintained the fruit firmness. Further analysis manifested that P. protegens substantially induced the defense-related genes such as MdGLU, MdPAL, MdPOD, MdCAL, and transcription factors related to the resistance to B. dothidea, including MdWRKY15, MdPUB29, MdMyb73, and MdERF11 in apple fruits. Meanwhile, P. protegens considerably restrained the expressions of the pathogenicity-related genes in B. dothidea, including the BdCYP450, BdADH, BdGHY, BdATS, Bdα/ß-HY, and BdSTR. By inference, P. protegens inhibited the apple ring rot on postharvest fruits by activating the defense system of apple fruit and repressing the pathogenic factor of B. dothidea. The study provided a theoretical basis and a potential alternative to manage the apple ring rot on postharvest fruits.

Contamination of Cronobacter spp. in Chinese Retail Spices.

Crononbacter spp. is an opportunistic foodborne pathogen that causes infections in neonates, infants, and immunocompromised adults. Although the contamination of spices with Cronobacter has been previously reported in some countries, there have been no studies on Cronobacter contamination in China. Therefore, this study aimed to investigate the prevalence of Cronobacter spp. in Chinese retail spices. Fifty-six packaged Chinese spices were collected from different markets, and 32 of these were found to be contaminated with Cronobacter. Five species were identified from the 54 isolates of the 32 positive samples: Cronobacter sakazakii (n = 35), Cronobacter muytjensii (n = 8), Cronobacter malonaticus (n = 6), Cronobacter turicensis (n = 3), and Cronobacter dublinensis (n = 2). Pulsed-field gel electrophoresis demonstrated high genetic diversity, as 53 PFGE profiles were revealed for the 54 isolates. Multilocus sequence typing analysis revealed 46 sequence types, and of these, 26 were newly identified. Most of the isolates were sensitive to antibiotics (n = 15), with the exception of cefazolin. This study revealed that the contamination of Chinese retail spices by Cronobacter spp. poses a potential risk to the consumer.

Application of multiple-locus variable number tandem repeat analysis to identify outbreak-associated Neisseria meningitides serogroup C sequence type 4821 in China.

Neisseria meningitidis (N. meningitidis) serogroup C sequence type (ST)-4821 caused an outbreak in 2010 in Shandong province of China. Twenty-one non-outbreak-associated strains were isolated, along with twenty-eight N. meningitides serogroup C ST-4821 isolates. Therefore, it's essential to identify and clarify characterization of the real outbreak-associated strains with a rapid method during an outbreak investigation. In this study, multiple-locus variable number tandem repeat analysis (MLVA) was applied to analyze 84 N. meningitidis strains, among which 58 were recovered from two outbreaks and 26 were sporadic isolates. Three MLVA schemes with different combination of VNTR loci were tested, and two of them were suitable for isolates from China: scheme 2 with six loci was found to separate ST into finer resolution, and scheme 3 with five loci can be used to identify outbreak-associated isolates from the same outbreak that caused by N. meningitidis serogroup C ST-4821.

Clonal characteristics of invasive Neisseria meningitidis following initiation of an A + C vaccination program in China, 2005-2012.

OBJECTIVES: To reveal the population structure of invasive Neisseria meningitidis isolates after the initiation of an A + C vaccination program in China. METHODS: Multilocus sequence typing (MLST) and PorA typing were used to characterize 238 invasive N. meningitidis isolates collected in China between 2005 and 2012. RESULTS: During this period, sequence type (ST)-5, ST-4821 and ST-11 complexes were dominant among serogroups A, C and W, accounting for 100%, 98.5% and 100% of each serogroup, respectively. P1.20,9, P1.7-2,14 and P1.5,2 were the dominant PorA types of serogroups A, C and W, respectively. Serogroup B showed high genetic diversity with two dominant lineages: ST-4821 complex and ST-5662 subgroup. CONCLUSIONS: The population of 238 invasive N. meningitidis isolates was primarily composed of a select group of recognized hypervirulent lineages. Among these clonal complexes, ST-7 serogroup A and ST-11 serogroup W are distributed globally, and other three clones exist only in China.

Genetic diversity of Neisseria meningitidis serogroup C ST-4821 in China based on multiple-locus variable number tandem repeat analysis.

Neisseria meningitidis sequence type (ST)-4821 was first reported in China in 2003, and a new hyper-virulent lineage has been designated as the ST-4821 complex. A large number of N. meningitidis ST-4821 strains have been identified in China since 2003; however, the microevolution characteristics of this complex are unclear. Different combinations of variable number of tandem repeats (VNTR) loci were used in multiple-locus VNTR analysis (MLVA) to analyze 118 N. meningitidis serogroup C ST-4821 strains isolated from seventeen provinces between 2003 and 2012. Additionally, MLVA with five VNTR loci was performed due to its high discriminatory power. One hundred and eighteen isolates were found to comprise 112 subtypes based on MLVA, and 16 outbreak-associated strains were clustered into one group. These data indicate a high level of diversity for N. meningitidis ST-4821 due to microevolution in the last decade. In addition, the results revealed high similarity between isolates from the same geographic origins, which is helpful when monitoring the spread of N. meningitidis serogroup C ST-4821 and will provide valuable information for the control and prevention of bacterial meningitis in China.

Sacrificial adhesion promotion layers for copper metallization of device structures.

The adhesion of copper films to adjacent device layers including TiN, Ta, and TaN diffusion barriers is a crucial reliability issue for integrated circuits. We report that ultrathin layers of poly(acrylic acid) (PAA) prepared on barrier surfaces or on the native oxide of Si wafers dramatically increase the interfacial adhesion of Cu films deposited by the H2 assisted reduction of bis(2,2,7-trimethyloctane-3,5-dionato)copper in supercritical carbon dioxide. Similar improvements were achieved on Si wafers using a simple vapor phase exposure of the substrate to acrylic acid prior to metallization. The deposited films and the substrate/Cu interfaces were analyzed by X-ray photoelectron spectroscopy (XPS), electron microscopy, atomic force microscopy, and variable-angle spectroscopic ellipsometry. No trace of the adhesion layer was detected at the interface, indicating it was sacrificial at the deposition conditions used. Moreover, the presence and subsequent decomposition of the PAA layer during deposition substantially reduced or eliminated metal oxides at the substrate interface. For depositions on PAA-treated Si wafers, copper was present primarily as Cu0 at the interface and Si was present only as Si0. On PAA-treated Ta substrates, XPS analysis indicated Ta was present primarily as Ta0 at the metallized interface whereas Ta2O5 dominated the interface of samples prepared without the adhesion layers. The technique can be extended to patterned substrates using adsorption of acrylic acid or thermal/UV polymerization of acrylic acid.