Using phenotype microarrays in the assessment of the antibiotic susceptibility profile of bacteria isolated from wastewater in on-site treatment facilities

The scope of the study was to apply Phenotype Biolog MicroArray (PM) technology to test the antibiotic sensitivity of the bacterial strains isolated from on-site wastewater treatment facilities. In the first step of the study, the percentage values of resistant bacteria from total heterotrophic bacteria growing on solid media supplemented with various antibiotics were determined. In the untreated wastewater, the average shares of kanamycin-, streptomycin-, and tetracycline-resistant bacteria were 53, 56, and 42%, respectively. Meanwhile, the shares of kanamycin-, streptomycin-, and tetracycline-resistant bacteria in the treated wastewater were 39, 33, and 29%, respectively. To evaluate the antibiotic susceptibility of the bacteria present in the wastewater, using the phenotype microarrays (PMs), the most common isolates from the treated wastewater were chosen: Serratia marcescens ss marcescens, Pseudomonas fluorescens, Stenotrophomonas maltophilia, Stenotrophomonas rhizophila, Microbacterium flavescens, Alcaligenes faecalis ss faecalis, Flavobacterium hydatis, Variovorax paradoxus, Acinetobacter johnsonii, and Aeromonas bestiarum. The strains were classified as multi-antibiotic-resistant bacteria. Most of them were resistant to more than 30 antibiotics from various chemical classes. Phenotype microarrays could be successfully used as an additional tool for evaluation of the multi-antibiotic resistance of environmental bacteria and in preliminary determination of the range of inhibition concentration.     

Mycobiota of ground red pepper and their aflatoxigenic potential

To investigate contamination of ground red pepper with fungi and mycotoxin, we obtained 30 ground red pepper samples from 15 manufacturers in the main chili-pepper-producing areas in Korea. Fungal contamination was evaluated by spreading diluted samples on potato dextrose agar plates. The total fungi counts ranged from 0 to 7.3 × 10³ CFU/g. In the samples, the genus Aspergillus had the highest incidence, while Paecilomyces was isolated most frequently. The next most frequent genera were Rhizopus, Penicillium, Cladosporium, and Alternaria. Within Aspergillus, A. ruber was predominant, followed by A. niger, A. amstelodami, A. ochraceus, A. terreus, A. versicolor, A. flavus, and A. fumigatus. The samples were analyzed for aflatoxins, ochratoxin A, and citrinin by ultra-perfomance liquid chromatography (UPLC) with a fluorescence detector. Ochratoxin A was detected from three samples at 1.03?2.08 μg/kg, whereas no aflatoxins or citrinin were detected. To test the potential of fungal isolates to produce aflatoxin, we performed a PCR assay that screened for the norB-cypA gene for 64 Aspergillus isolates. As a result, a single 800-bp band was amplified from 10 A. flavus isolates, and one Aspergillus sp. isolate. UPLC analyses confirmed aflatoxin production by nine A. flavus isolates and one Aspergillus sp. isolate, which produced total aflatoxins at 146.88?909.53 μg/kg. This indicates that continuous monitoring of ground red pepper for toxigenic fungi is necessary to minimize mycotoxin contamination.     

Isolation and molecular characterization of the fungal endophytic microbiome from conventionally and organically grown avocado trees in South Florida

Fungal endophytes are the most ubiquitous and highly diverse microorganisms that inhabit the interior of healthy plants. They are important in plant ecology and offer untapped potential to improve plant health and productivity in agroecosystems. The endophytic assemblage of avocado is poorly understood; therefore, surveys of fungal endophytes of Persea americana Mill. (Avocado) in South Florida organic and conventional orchards were conducted. A total of 17 endophytic fungal species were recovered from healthy avocado terminal branches. Endophytic fungal species were identified by rDNA sequencing of the internal transcribed spacer (ITS) region, using UNITE Species Hypotheses to reliably assign a taxon name, and determined as belonging to the genera Alternaria, Cladosporium, Colletotrichum, Corynespora, Diaporthe, Lasiodiplodia, Neofusicoccum, Neopestalotiopsis, Phyllosticta, and Strelitziana. Endophyte community assemblage differed between organic and conventional agroecosystems. This is the first report of Alternaria eichhorniae, Cladosporium tenuissimum, Corynespora cassiicola, Colletotrichum alatae, Diaporthe fraxini-angustifoliae, Lasiodiplodia gonubiensis, Neofusicoccum algeriense, Neofusicoccum andinum, Neopestalotiopsis foedans, Phyllosticta capitalensis, and Strelitziana africana as endophytes of avocado. Evaluation using pathogenicity tests on avocado leaves and terminal branches showed that endophytic fungal isolates did not cause disease symptoms.     

Pine wilt disease and possible causes of its incidence in Russia

Surveys of forests and stockpiled timber of pine, spruce, larch, and silver fir in 14 administrative subjects of the Russian Federation revealed widespread occurrence of the coniferous wood parasitic nematode Bursaphelenchus mucronatus. Twenty species of bacteria belonging to 13 genera have been detected in 25 B. mucronatus isolates, and their identity has been determined by direct sequencing of the 16S RNA gene. The most frequently occurring were bacteria from the genera Pseudomonas, Stenotrophomonas, Pantoea, Bacillus, Burkholderia, and Serratia. Prevalence of Pseudomonas brenneri and P. fluorescence, which were also found in the nematode dauer larva (L_IV) isolated from the fir sawyer beetle Monochamus urussovi, have also been assessed. Two nematode B. xylophilus isolates from Portugal and one isolate from the United States have been examined, and 10 symbiotic bacteria species have been isolated, including Agrobacterium tumefacience, P. fluorescens, P. brenneri, Rahnella aquatilis, Stenotrophomonas maltophilia, S. rhizophila, and Yersinia mollaretii.     

Identification and Pathogenicity of Fungal Pathogens Causing Black Point in Wheat on the North China Plain

Fungi associated with black point were isolated from three highly susceptible wheat genotypes in the North China Plain. The 21 isolates represented 11 fungal genera. The most prevalent genera were Alternaria (isolation frequency of 56.7%), Bipolaris (16.1%), and Fusarium (6.0%). The other eight genera were Curvularia, Aspergillus, Cladosporium, Exserohilum, Epicoccum, Nigrospora, Penicillium, and Ulocladium; their isolation frequencies ranged from 0.8 to 4.8%. The pathogenicity of the isolates was individually assessed in the greenhouse by inoculating wheat plants with spore suspensions. Ten of the 21 isolates caused significantly higher incidences of black point than that the controls. These isolates belonged to eight fungal species (A. alternata, B. sorokiniana, B. crotonis, B. cynodontis, C. spicifera, F. equiseti, E. rostratum, and E. sorghinum) based on morphological traits and phylogenetic analysis. The average incidences of black point in the eight fungal species were 32.4, 54.3, 43.0, 41.9, 37.2, 38.8, 50.1, and 34.1%, respectively. B. sorokiniana and A. alternata were determined to be the most important pathogens in the North China Plain based on fungal prevalence and symptom severity. This study is the first to identify E. rostratum as a major pathogen causing black point in wheat.     

Itaconic Acid Production by Filamentous Fungi in Starch-Rich Industrial Residues

Several fungi and starch-rich industrial residues were screened for itaconic acid (IA) production. Out of 15 strains, only three fungal strains were found to produce IA, which was confirmed by HPLC and GC–MS analysis. These strains were identified as Aspergillus terreus strains C1 and C2, and Ustilago maydis strain C3 by sequencing of 18S rRNA gene and internal transcribed spacer regions. Cis-aconitate decarboxylase (cad) gene, which encodes a key enzyme in IA production in A. terreus, was characterized from strains C1 and C2. C1 and C2 cad gene sequences showed about 96% similarity to the only available GenBank sequence of A. terreus cad gene. 3-D structure and cis-aconitic acid binding pocket of Cad enzyme were predicted by structural modeling. Rice, corn and potato starch wastes were screened for IA production. These materials were enzymatically hydrolyzed under experimentally optimized conditions resulting in the highest glucose production of 230 mg/mL from 20% potato waste. On comparing the production potential of selected strains with different wastes, the best IA production was achieved with strain C1 (255.7 mg/L) using potato waste. Elemental composition as well as batch-to-batch variation in waste substrates were analyzed. The difference in IA production from two different batches of potato waste was found to inversely correlate with their phosphorus content, which indicated that A. terreus produced IA under phosphate limiting condition. The potato waste hydrolysate was deionized to remove inhibitory ions like phosphate, resulting in improved IA production of 4.1 g/L by C1 strain, which is commercially competitive.     

Screening for potential probiotic from spontaneously fermented non-dairy foods based on in vitro probiotic and safety properties

The aim of this study was to screen potential probiotic lactic acid bacteria from Chinese spontaneously fermented non-dairy foods by evaluating their probiotic and safety properties. All lactic acid bacteria (LAB) strains were identified by 16S rRNA gene sequencing. The in vitro probiotic tests included survival under low pH and bile salts, cell surface hydrophobicity, auto-aggregation, co-aggregation, antibacterial activity, and adherence ability to cells. The safety properties were evaluated based on hemolytic activity and antibiotic resistance profile. The salt tolerance, growth in litmus milk, and acidification ability were examined on selected potential probiotic LAB strains to investigate their potential use in food fermentation. A total of 122 strains were isolated and identified at the species level by 16S rRNA gene sequencing and included 62 Lactobacillus plantarum, 40 Weissella cibaria, 12 Lactobacillus brevis, 6 Weissella confusa, and 2 Lactobacillus sakei strains. One W. cibaria and nine L. plantarum isolates were selected based on their tolerance to low pH and bile salts. The hydrophobicity, auto-aggregation, co-aggregation, and antagonistic activities of these isolates varied greatly. All of the 10 selected strains showed multiple antibiotic resistance phenotypes and no hemolytic activity. The highest adhesion capacity to SW480 cells was observed with L. plantarum SK1. The isolates L. plantarum SK1, CB9, and CB10 were the most similar strains to Lactobacillus rhamnosus GG and selected for their high salt tolerance and acidifying activity. The results revealed strain-specific probiotic properties were and potential probiotics that can be used in the food industry.     

Diversity of endophytic fungal and bacterial communities in <Emphasis Type="Italic">Ilex paraguariensis</Emphasis> grown under field conditions

The composition and diversity of the endophytic community associated with yerba mate (Ilex paraguariensis) was investigated using culture-depending methods. Fungi were identified based on their micromorphological characteristics and internal transcribed spacer rDNA sequence analysis; for bacteria 16S rDNA sequence analysis was used. Fungal and bacterial diversity did not show significant differences between organ age. The highest fungal diversity was registered during fall season and the lowest in winter. Bacterial diversity was higher in stems and increased from summer to winter, in contrast with leaves, which decreased. The most frequently isolated fungus was Fusarium, followed by Colletotrichum; they were both present in all the sampling seasons and organ types assayed. Actinobacteria represented 57.5 % of all bacterial isolates. The most dominant bacterial taxa were Curtobacterium and Microbacterium. Other bacteria frequently found were Methylobacterium, Sphingomonas, Herbiconiux and Bacillus. Nitrogen fixation and phosphate solubilization activity, ACC deaminase production and antagonism against plant fungal pathogens were assayed in endophytic bacterial strains. In the case of fungi, strains of Trichoderma, Penicillium and Aspergillus were assayed for antagonism against pathogenic Fusarium sp. All microbial isolates assayed showed at least one growth promoting activity. Strains of Bacillus, Pantoea, Curtobacterium, Methylobacterium, Brevundimonas and Paenibacillus had at least two growth-promoting activities, and Bacillus, Paenibacillus and the three endophytic fungi showed high antagonistic activity against Fusarium sp. In this work we have made a wide study of the culturable endophytic community within yerba mate plants and found that several microbial isolates could be considered as potential inoculants useful for improving yerba mate production.     

Strain differentiation of airborne opportunistic microorganisms within a municipal landfill area as assessed by PCR MP method

A municipal landfill is the site where occurs differentiation of microorganisms inclusive of several hazardous to human health. The aim of this study was to evaluate by a PCR melting profile (PCR MP) technique the level of genetic intraspecies relatedness of strains, representing several opportunistic bacteria and fungi commonly found in bioaerosol in the landfill site. In total, 27 strains representing four bacterial species (i.e. Escherichia coli, Proteus mirabilis, Staphylococcus sciurii, S. xylosus) and 36 fungal strains belonging to Aspergillus fumigatus and A. flavus were isolated from air samples collected by an Anderson impactor within the landfill area. The PCR melting profile approach clearly indicated that except E. coli, represented by one genotype, other microbial species underwent significant genetic variability in the active sector and surrounding the landfill forest and field areas. Although the genetic relatedness of some strains could testify to distribution of microbes from the active sector to the surroundings in the past, the bacterial and fungal isolates indicated site-specific genetic fingerprints. This is the first report on the distribution of airborne opportunistic microbial species within the landfill area, performing a comparison of their genotypes and evaluation of genetic relatedness between the isolates using the PCR MP method.     

Antimicrobial properties of endophytic actinomycetes isolated from <Emphasis Type="Italic">Combretum latifolium</Emphasis> Blume,a medicinal shrub from Western Ghats of India

Endophytic actinomycetes were isolated from Combretum latifolium Blume (Combretaceae),Western Ghats of Southern India and identified by its characteristic culture morphology and molecular analysis of 16S rRNA gene sequences. In this survey of endophytic actinomycetes, a total of 117 isolates representing 9 different genera of endophytic actinomycetes were obtained using four different isolation media and several of them seemed to be novel taxa. Streptomyces genera (35%) was the most frequently isolated strains, followed by Nocordiopsis (17%) and Micromonospora (13%). ISP-4 medium recovered more isolates (47%) when compared to rest of the media used. Preliminary antibacterial activity of the isolates was carried out by confrontation test. Ethyl acetate fraction of selected isolates in disc diffusion assay exhibited broad spectrum antimicrobial activity against test human pathogens. All Streptomyces spp. strains displayed significant antimicrobial activity against test pathogens. Strain CLA-66 and CLA-68 which are Nocordipsis spp. inhibited both bacterial and fungal pathogens where as other isolates inhibited atleast three test human pathogens in disc diffusion assay. Antimicrobial screening of endophytic actinomycetes from this host may represent a unique potential niche for antimicrobial compounds of industrial and pharmaceutical applications. This work is the first comprehensive report on incidence of potential endophytic actinomycetes inhabiting C. latifolium Blume.     

Monitoring of ochratoxin A and ochratoxin-producing fungi in traditional salami manufactured in Northern Italy

Fungi have a crucial role in the correct maturation of salami, but special attention should be addressed to the production of the nephrotoxic, immunotoxic, and carcinogenic mycotoxin ochratoxin A (OTA). In a monitoring study conducted in Northern Italy, OTA was detected by liquid chromatography coupled with mass spectrometry in 13 out 133 samples of traditional salami (9.8% of the total count). Mycological analysis of these samples yielded 247 fungal isolates which were identified to species level. The most frequent species were Penicillium nalgiovense, P. solitum, and P. chrysogenum. P. nordicum, an OTA-producing species commonly found in proteinaceous food, was not found in these samples. Three isolates were found to be Aspergillus westerdijkiae, an OTA-producing species. In order to check the results of the microbiological identification, 19 different strains of Aspergillus and 94 of Penicillium were tested for the presence of a sequence common to OTA-producing fungi by real-time PCR. None of the studied isolates, including the three A. westerdijkiae, possessed the otanpsPN target which is common to OTA-producing strains. Two out of three isolates of the A. westerdijkiae were also PCR-negative for the otanpsPN gene and did not produce OTA in culture. Conversely, this target sequence was amplified from the DNA purified from 14 salami casings including three casings harboring A. westerdijkiae. The amplification of sequences specific for OTA-producing strains performed on total genomic DNA extracted directly from salami casings provided a more suitable approach than PCR analysis of isolates from salami for the OTA-related otanpsPN gene to evaluate the risk of OTA contamination.     

Degradation capacities of bacteria and yeasts isolated from the gut of <Emphasis Type="Italic">Dendroctonus rhizophagus</Emphasis> (Curculionidae: Scolytinae)

Bark beetles (Curculionidae: Scolytinae) feed on the xylem and phloem of their host, which are composed of structural carbohydrates and organic compounds that are not easily degraded by the insects. Some of these compounds might be hydrolyzed by digestive enzymes produced by microbes present in the gut of these insects. In this study, we evaluated the enzymatic capacity of bacteria (Acinetobacter lwoffii, Arthrobacter sp., Pseudomonas putida, Pseudomonas azotoformans, and Rahnella sp.) and yeasts (Candida piceae, Candida oregonensis, Cyberlindnera americana, Zygoascus sp., and Rhodotorula mucilaginosa) isolated from the Dendroctonus rhizophagus gut to hydrolyze cellulose, xylan, pectin, starch, lipids, and esters. All isolates, with the exception of C. piceae, showed lipolytic activity. Furthermore, P. putida, P. azotoformans, C. americana, C. piceae, and R. mucilaginosa presented amylolytic activity. Esterase activity was shown by A. lwoffii, P. azotoformans, and Rahnella sp. Cellulolytic and xylanolytic activities were present only in Arthrobacter sp. and P. azotoformans. The pectinolytic activity was not recorded in any isolate. This is the first study to provide evidence on the capacity of microbes associated with the D. rhizophagus gut to hydrolyze specific substrates, which might cover part of the nutritional requirements for the development, fitness, and survival of these insects.     

Diversity of <Emphasis Type="Italic">Colletotrichum</Emphasis> spp. isolated from chili pepper fruit exhibiting symptoms of anthracnose in Thailand

Colletotrichum spp. are causal agents of anthracnose disease in chili fruits and other tropical crops. The disease is increasing in chili fruits in Thailand and significantly reduces fruit quality and fruit production. Forty-eight isolates of Colletotrichum spp. associated with chili anthracnose were collected from different areas of Thailand during 2010–2015. Based on morphological characteristic identification, 10 isolates were shown to belong to the C. gloeosporioides species complex, 24 isolates belong to the C. acutatum species complex and 14 isolates to C. capsici. For molecular identification, two primer sets, ITS1/ITS4 and ACT528/ACT738, were used for amplification of the internal transcribed spacer of rRNA gene (ITS1–5.8S–ITS2) and partial region actin gene (ACT), respectively. The phylogenetic analysis of individual and combined ITS region and actin nucleotide sequences identified the collected isolates into 4 species: C. gloeosporioides, C. siamense, C. acutatum and C. capsici. The pathogenicity test demonstrated that all four species were pathogenic on intact unwounded and healthy fruits. These results indicated that C. capsici, C. acutatum, C. gloeosporioides and C. siamense were the causal agents of chili anthracnose disease.     

The key role of lignin decomposing fungi in the decay of roofs thatched with water reed

We investigated the involvement of microorganisms in the rapid reed decay of roofs thatched with water reed. Numerous bacteria and fungi were isolated by enrichment cultures from reed samples and from fungal fruit bodies on roofs. All strains were characterised in respect to their abilities to degrade cellulose, hemicelluloses and the lignin model substance Poly-R-478. Only 15 of the 92 isolated bacterial strains were capable of degrading cellulose and hemicelluloses. However, nearly all 61 of the identified fungal isolates had these abilities. Nevertheless, only 14 of the isolated fungal strains as well as a reference isolate of Trametes versicolor were capable of degrading Poly-R-478. The ability of the microorganisms to attack complete reed was assessed using a newly developed test system which measures the loss of dry weight during the incubation. A significant loss of dry weight was apparent only in tests using the ligninolytic fungi Pycnoporus cinnabarinus, Trametes versicolor, Phlebia tremellosa and some Mycena species, but not in the case of the majority of cellulolytic bacteria and fungi. From these results, we conclude that ligninolytic fungi are capable of destroying complete reed structure and that they play the key role in the process of the rapid decay of roofs thatched with reed. Directly after the initial lignin attack, cellulose and hemicellulose were degraded to a great extent, evidenced by the large loss of dry weight (up to 72 %), which significantly exceeds the lignin content of reed (ca. 15 %). However, after the initial attack by ligninolytic fungi, bacteria or other fungi capable of degrading cellulose and hemicelluloses may contribute to the progressive decay of reed under natural conditions. Furthermore, we show that the rate of decay depends on the source of the reed and on the reed quality.     

Molecular profiling of fungal assemblages in the healthy and infected roots of <Emphasis Type="Italic">Decalepis arayalpathra</Emphasis> (J. Joseph &amp; V. Chandras) Venter,an endemic and endangered ethnomedicinal plant from Western Ghats,India

Decalepis arayalpathra, an endangered, endemic ethnomedicinal plant from southern Western Ghats, India, is targeted for its aromatic and medicinal properties. This study aimed at to identify fungal endophyte populations associated with healthy and diseased roots of this perennial shrub. Healthy and rotted root samples of D. arayalpathra were collected, fungal endophytes assemblages were identified both by culture-dependent and culture-independent approaches, further sequenced and the retrieved sequences were analysed with the reference sequences in GenBank to know their phylogenetic relationships. Analysis of the ITS rDNA region generated 24 different Ascomycota and three Basidiomycota taxa. Trichoderma sp. was most abundant in healthy and diseased root samples, while Penicillium and Aspergillus were confined to healthy roots. Furthermore, Fusarium solani, Fusarium oxysporum and Mucor velutinosus were found to be the most frequent fungi identified from the rotted root samples, thus substantiated to be the cause for D. arayalpathra decline in the wild. Interestingly, the strains assigned to Fusarium sp. were isolated from diseased roots showing typical clearly visible symptoms, such as a severe brown discolouration on the taproot. Molecular profiling of all the pure fungal isolates, viz., Trichoderma, Penicillium, Aspergillus, Fusarium and Mucor, revealed high sequence similarities (≥ 98 %) to corresponding reference sequences. Sequencing of Trichoderma pure cultures isolated from healthy and diseased roots revealed sequence similarities to Trichoderma harzianum, T. hamatum, T. koningiopsis, T. asperellum, T. pubescens and Hypocrea sp. This confirms the morphological examinations, as Hypocrea is the teleomorph stage of Trichoderma sp. This study signifies the first work pertaining to the taxonomy of the fungal endophytic community of D. arayalpathra, and the results reported in this work may help to ascertain the cause of root rot disease often perceived in D. arayalpathra. Also, it could be useful to identify the promising endophytic communities against the root rot diseases occurring in D. arayalpathra.     

Screening and molecular identification of lactic acid bacteria from <Emphasis Type="Italic">gari</Emphasis> and <Emphasis Type="Italic">fufu</Emphasis> and <Emphasis Type="Italic">gari</Emphasis> effluents

Bacterial strains were isolated from cassava-derived food products and, for the first time, from cassava by-products, with a focus on gari, a flour-like product, and the effluents from the production processes for gari and fufu (a dough also made from cassava flour). A total of 47 strains were isolated, all of which were tested to determine their resistance to acidic pH and to bile salt environments. Four of the 47 isolates tested positive in both environments, and these four isolates also showed antibacterial behaviour towards both Gram-positive and Gram-negative microbial pathogens (i.e. Methicillin-resistance Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Salmonella enteritidis, Escherichia coli, Escherichia coli (O157), Yersinia enterocolitica). In most cases, the antibacterial activity was related to bacteriocin production. Molecular identification analysis (16S rDNA and randomly amplified polymorphic DNA-PCR) revealed that the four isolates were different strains of the same species, Lactobacillus fermentum. These results demonstrate that bacteria isolated from cassava-derived food items and cassava by-products have interesting properties and could potentially be used as probiotics.     

Diversity of Oil-Degrading Microorganisms in the Gulf of Finland (Baltic Sea) in Spring and in Summer

Diversity of the oil-degrading microbial strains isolated from the water and sediments of the Gulf of Finland (Baltic Sea) in winter and in summer was studied. Substrate specificity of the isolates for aliphatic and aromatic hydrocarbons was studied. The isolates belonged to 32 genera of the types Proteobacteria (alpha-, beta-, and gammaproteobacteria), Actinobacteria,Firmicutes, and Bacteroidetes. Seasonal variations of the oil-degrading microbial communities was revealed. The presence of the known genes responsible for the degradation of oil aliphatic and aromatic hydrocarbons was determined. The alkB sequence of the alkane hydroxylase gene was found in ~16% of the studied strains. The sequence of the phnAc phenanthrene 3,4- dioxygenase was found in Sphingobacterium sp. and Arthrobacter sp. isolates retrieved in winter and summer. In five Pseudomonas sp. strains from winter samples, the classical operons of naphthalene degradation (nah) were localized in catabolic plasmids, of which three belonged to IncР-9, one, to IncР-7, and two to an unidentified incompatibility group. Burkholderia and Delftia strains contained the operons for naphthalene degradation via salicylate and gentisate (nag). The presence of nag genes has not been previously reported for Delftia spp. strains. The sequences of the nagG salicylate 5-hydroxylase gene were also found in Achromobacter, Sphingobacterium, and Stenotrophomonas strains.     

Genetic and ecological differences between <Emphasis Type="Italic">Asphondylia yushimai</Emphasis> and the ivy gall midge, <Emphasis Type="Italic">Asphondylia</Emphasis> sp. (Diptera: Cecidomyiidae), with a new distribution record of the former from Hokkaido and South Korea

The soybean pod gall midge, Asphondylia yushimai, is known to utilize Laurocerasus zippeliana (Rosaceae) and Osmanthus heterophyllus (Oleaceae) as autumn–spring hosts. In addition, ivy, Hedera rhombea (Araliaceae), was thought to be a candidate for an additional autumn–spring host. However, our genetic analysis indicated that no haplotypes of the ivy fruit gall midge, Asphondylia sp., were identical to any of the haplotypes of A. yushimai. Furthermore, the life-history traits of the ivy fruit gall midge, such as voltinism, host-plant range, lower development threshold temperature (LDT), and developmental speed, were clearly different from those of A. yushimai. Thus, the results from genetic analysis and life-history traits revealed that the ivy fruit gall midge was not identical to A. yushimai and that H. rhombea is not an additional autumn–spring host plant for A. yushimai. We also discovered through morphological observation and genetic analysis that A. yushimai is distributed in Hokkaido and South Korea, and that the ivy fruit gall midge exhibits host plant alternation, utilizing both the fruit of Phytolacca americana (Phytolaccaceae) and the flower buds of Paederia foetida (Rubiaceae) as spring–autumn hosts.     

Isolation and characterization of bacteriocin-like inhibitory substances from lactic acid bacteria isolated from Azerbaijan cheeses

The search for lactic acid bacteria (LAB) producing bacteriocin-like inhibitory substances (BLISs) was performed in traditional Azerbaijan cheeses. Eight strains have been isolated and identified, four of which (S1a, S2, Q2b, and M1b) were identified as Lactobacillus buchneri. The remaining four strains (M3a, A4b, Q4a, and A3) were identified as L. pentosus, L. brevis, L. fermentum, and L. collinoides, respectively. Antimicrobial agents of all isolated LAB strains suppressed the growth of Lactobacillus bulgaricus 340, Escherichia coli, Enterococcus durans, and Listeria innocua. Saccharomyces cerevisiae and Candida pseudotropicalis were resistant to these BLISs. Proteinase K and trypsin completely inactivated BLIS whereas lipase and amylase partially inactivated BLIS.     

Antagonistic Activity of Lactic Acid Bacteria <Emphasis Type="Italic">Lactobacillus</Emphasis> spp. against Clinical Isolates of <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis>

The screening of three strains of lactic acid bacteria identified as Lactobacillus rhamnosus, Lactobacillus reuteri, and Lactobacillus helveticus showed significant antagonistic activity against Klebsiella pneumoniae strains characterized by multiple antibiotic resistance. Lactobacilli cocultivated with the Klebsiella strains inhibited their growth 20 to 86% on the first and second days, respectively. Exoproteome analysis of L. rhamnosus cocultivated with K. pneumoniae revealed the induction of peptidoglycan hydrolases, including extracellular lytic transglycosylases, family II (MltA), and endopeptidases capable of disrupting the peptidoglycan bacterial cell wall.