Comparison of α-acetolactate synthase and α-acetolactate decarboxylase in Lactococcus spp. and Leuconostoc spp.

Summary Cell-free extracts of Leuconostoc and Lactococcus species were tested for their -acetolactate synthase and -acetolactate decarboxylase activities. In Leuconostoc mesenteroides subsp. cremoris, Leuconostoc mesenteroides subsp. mesenteroides and Leuconostoc lactis, the Km of -acetolactate synthase for pyruvate was close to 10 mM whereas it was 30 mM in Lactococcus lactis subsp. lactis biovar. diacetylactis. The Km of -acetolactate decarboxylase for -acetolactic acid was very low (0.3 mM) in Leuconostoc species in comparison to Lactococcus lactis subsp. lactis biovar. diacetylactis (60 mM). In the latter bacterium, -acetolactate decarboxylase showed a sigmoidal dependance upon -acetolactic acid and was activated by the three branchedchain amino acids: leucine, isoleucine and valine.     

Nucleoids and coated vesicles of “Epulopiscium” spp.

We describe here aspects of the anatomy of two “Epulopiscium” morphotypes, unusually large bacteria that are not yet cultured and that reproduce by the internal generation of two or more vegetative daughter cells. Two morphotypes, A and B, which are enteric symbionts of several species of herbivorous surgeonfish (Acanthuridae), were collected around the Great Barrier Reef of Australia, preserved there, and later stained for light microscopy. Some samples were examined by electron microscopy. In both morphotypes, countless discrete nucleoplasms or nucleoids were found to occupy a single shallow layer just beneath the surface all around these organisms. At each end of the morphotype B cells, a membrane-bound compartment containing dense cords of chromatin was observed. When these were found at each end of growing daughter cells, no polar compartments were then found in their mother organism. Electron micrographs of sections of morphotype A symbionts show that their outermost region is composed of tightly packed coated vesicles, each surrounded by a thin, dense, spacious capsule. Near the surface of type A organisms the remains of broken vesicles, broken capsules, and a finely fibrous matrix fuse to form a fabric that serves as the cell wall. Morphotype B organisms, however, were observed to have a distinct, morphologically continuous outer wall. Received: 3 December 1997 / Accepted: 11 June 1998     

Production and localisation of carboxymethylcellulase,xylanase and β-glucosidase fromCellulomonas andMicrococcus spp.

Summary Cellulomonas and Micrococcus spp. grew well at 30°C, pH 7.0, and produced carboxymethylcellulase (CMCase) and xylanase enzymes. Only one species of Micrococcus was able to produce an appreciable amount of -glucosidase. This is the first report where Micrococcus sp., isolated from termite gut, was able to produce all three enzymes (i.e. CMCase, xylanase and -glucosidase) required for degradation of cellulosic and hemicellulosic substrates. Offprint requests to: A. Varma     

Variation analysis of pathogenic Vibrio spp. and Pseudomonas spp. in Changhai mollusc farming waters using real-time PCR assay during 2011–2014

Bacterial disease has caused high mortality of breeding molluscs from 2009 to 2011 in the Changhai area (Dalian, China). Vibrio spp. and Pseudomonas spp. have been detected as major pathogenic agents for aquatic animals in this area. In the present study, four virulence genes including vsm, toxR, aprX and carA were targeted to develop a real-time PCR assay for the quantitative detection of Vibrio splendidus, V. parahaemolyticus, Pseudomonas fluorescens and P. putida, respectively. The sensitivity and specificity of the assays were verified by experimental samples, and the variation tendencies of pathogenic V. splendidus, V. parahaemolyticus, P. fluorescens and P. putida strains from June to September were also detected in mollusc farming waters during 2011–2014. The concentration of V. splendidus increased from June to July, reduced in August, and then increased again in September. The highest count of V. parahaemolyticus appeared in July, and then dramatically decreased from August to September. Conversely, the counts of P. fluorescens and P. putida remained at lower levels from June to August, and then dramatically peaked in September. All four pathogenic bacteria displayed similar fluctuation tendencies of count variation in each year, and their concentrations were found to have a correlation with the average annual temperature. The variation tendency of pathogenic bacteria with temperature suggested that temperature was one of the most important factors to regulate the bacterial growth in a farming area, which could further provide information for the early warning of disease outbreak by using a convenient real-time PCR assay.     

Production of β-glycosidases (linamarase and amygdalase) and pectolytic enzymes by Penicillium spp.

Fifty-eight strains, representing 31 species of Penicillium, were screened for extracellular -glycosidase (amygdalase/linamarase) and pectolytic (polygalacturonase, pectin lyase) enzymes. One strain each of P. turbatum, P. piceum and P. paxilli showed very high -glycosidase activity and slightly lower activities were found in P. crustosum, P. expansum, P. oxalicum and P. aurantiogriseum. Generally, maximum -glycosidase activity showed reached during the stationary phase of growth. The seven species with highest -glycosidase activity showed different patterns of pectolytic activities, indicating that different species or combinations of species could be selected for different potential applications.L. Brimer is with the Department of Pharmacology and Pathobiology, Royal Veterinary & Agricultural University, 13 Bulowsvej, DK 1870 Frederiksberg C, Denmark; A.R. Cicalini and F. Federici are with the Dipartimento Agrobiologia e Agrochimica, University of Tuscia, Via S.C. de Lellis, I-01100 Viterbo, Italy. M. Petruccioli is with the Dipartimento di Biologia, Difesa e Biotecnologie Agro-Forestali, University of Basilicata, Via N. Sauro, 85, I-85100 Potenza, Italy.     

Fusarium spp. and Fusarium mycotoxins in maize: a problem for Flanders?

Fusarium species cause not only root, stem and ear rot with severe reductions in crop yield, they produce also toxic secondary metabolites (mycotoxins) such as deoxynivalenol (DON) and zearalenone (ZEA). During several growing seasons the presence of Fusarium spp was followed up. DON and ZEA were determined and related to infection levels. The distribution of DON and ZEA in the different plant parts was studied as well as the influence of the ensiling process on the mycotoxin content. More or less important varietal differences in susceptibility for Fusarium spp. could be detected. DON and ZEA were clearly present in most of the analysed samples. No clear relationship could be detected between visual disease symptoms and mycotoxin content. The accumulation of DON and ZEA was different for the analysed aerial plant parts. The ensiling process gave no reduction of the mycotoxin content.     

α-Pyrone and seiricuprolide derivatives from the mangrove-derived fungi Pestalotiopsis spp. PSU-MA92 and PSU-MA119

Investigation of the mangrove-derived fungi Pestalotiopsis spp. PSU-MA92 and PSU-MA119 resulted in the isolation of three new α-pyrones, pestalotiopyrones A–C (1–3), and two new seiricuprolides, pestalotioprolides A (4) and B (5), together with two known compounds. Their structures were identified by analysis of spectroscopic data. Compound 5 was isolated as its diacetate derivative (6). The antibacterial and antifungal activities of 2 were evaluated.     

Recruitment dynamics and fishery characteristics of juvenile goatfishes Mulloidichthys spp. in Hawai‘i

The most common goatfishes in Hawai‘i, Mulloidichthys flavolineatus and M. vanicolensis, comprise a unique resource due to their cultural, ecological and biological significance. These species exhibit pulse-type recruitment to nearshore areas during the summer months. Such pulses of juvenile fishes provide prey for pelagic and nearshore fishes and support a popular directed fishery. However, limited scientific information exists on juvenile stages of these fishes, known locally as oama, despite their contribution to coastal ecology and the extensive nearshore fisheries. Here we resolve growth rates, habitat preferences, hatching dates, size and age structure, as well as fishing catch rates based on new recruits in 2014 and 2015. We sampled 257 M. flavolineatus and 204 M. vanicolensis to compare ecological and fisheries characteristics between species and years. Both show strong habitat segregation, with M. vanicolensis found almost exclusively on hard and M. flavolineatus on soft substrates. Oama recruited in anomalously high numbers in 2014, a trend reflected in a higher catch per unit effort. In contrast, 2015 recruits grew faster, were heavier on average and hatched later than during 2014. Both species have calculated hatch dates in March to July, with M. vanicolensis hatching earlier, recruiting earlier and being consistently larger than M. flavolineatus. This baseline information regarding recruitment and early life-history characteristics can enhance management for other data-limited species that comprise a substantial component of nearshore fisheries in Hawai‘i.     

Basidiochrome – A Novel Siderophore of the Orchidaceous Mycorrhizal Fungi Ceratobasidium and Rhizoctonia spp.

A novel trishydroxamate siderophore, named basidiochrome, was isolated as the principal siderophore from low-iron culture filtrates of Ceratobasidium and Rhizoctonia species which are known as mycorrhizal fungi associated with orchid roots. Ion-exchange chromatography and preparative HPLC yielded a pure compound which contained two components according to GC–MS analysis: l-N⁵-hydroxy-ornithine and 3-methyl-2-cis-pentenedioic acid (3-methyl-cis-glutaconic acid). FTICR-ESI-MS of both the iron-free and ferric form indicated an elemental composition of C₃₃H₄₇N₆O₁₆Fe (MW = 839) for the ferric form of basidiochrome. The connectivity was further elucidated by 2D-NMR techniques (HSQC, HMBC, COSY, NOESY) indicating that basidiochrome is a novel linear tripeptide consisting of three l-N⁵-hydroxy-ornithines each linked to 3-methyl-2-cis-pentenedioic acid residues.     

Are Ureaplasma spp. a Cause of Nongonococcal Urethritis? A Systematic Review and Meta-Analysis

Background

Nongonococcal urethritis (NGU) is the most common male reproductive tract syndrome. Ureaplasmas spp. including U. urealyticum and U. parvum, have been increasingly reported to be implicated in NGU. However, there are still many contradictions about their pathogenic role in NGU.

Aims

The goals of this study were to evaluate the association of Ureaplasmas spp. with NGU, and to compare the prevalence of Ureaplasmas spp. infection in China relative to the world average.

Methods

A systematic review and meta-analysis was conducted following standard guidelines for meta-analysis. The quality of included studies was assessed by Newcastle-Ottawa scale.

Results

A total of seven studies involving 1,507 NGU patients and 1,223 controls were eligible for meta-analysis. There was no significant difference in the Ureaplasma spp. positive rate between the NGU and control groups. However, the U. urealyticum positive rate was significantly higher in NGU patients compared to controls; the U. parvum positive rate was significantly higher in controls compared to NGU patients. Furthermore, within the NGU patient group, the positive rate of U. urealyticum was significantly higher than that of U. parvum, whereas within the control group, the opposite trend was observed. Compared to the world average, a significantly higher positive rate of Ureaplasma spp. was observed in both the NGU and control groups in China.

Conclusions

Our analysis supports that U. urealyticum, but not U. parvum, is an etiological agent in NGU. More detailed studies of these two species in China and the world could contribute to a better understanding of the epidemiology and pathogenesis, and facilitate the development of better strategies for treatment and prevention of NGU.     

Development and validation of qualitative SYBR®Green Real-Time PCR for detection and discrimination of Listeria spp. and Listeria monocytogenes

A combination of four qualitative SYBR®Green qPCR screening assays targeting two levels of discrimination: Listeria genus (except Listeria grayi) and Listeria monocytogenes, is presented. These assays have been developed to be run simultaneously using the same polymerase chain reaction (PCR) programme. The paper also proposes a new validation procedure to specifically validate qPCR assays applied to food microbiology according to two guidelines: the ISO 22118 norm and the “Definition of minimum performance requirements for analytical methods of GMO testing”. The developed assays target the iap, prs and hlyA genes that belong to or neighbour the virulence cluster of Listeria spp. The selected primers were designed to amplify short fragments (60 to 103 bp) in order to obtain optimal PCR efficiency (between 97 and 107 % efficiency). The limit of detection of the SYBR®Green qPCR assays is two to five copies of target genes per qPCR reaction. These assays are highly accurate (98.08 and 100 % accuracy for the Listeria spp. and L. monocytogenes assays, respectively).     

The Production of β-Fructofuranosidase from Bifidobacterium spp.

The productions of β-fructofuranosidase from Bifidohacterium longum A1, B. adolescentis G1, and four other strains of Bifidobacteria were investigated. All strains used in this study were grown in modified BL broth containing a mixture of fructooligosaccharides [1^F (1-β-D-fructofuranosyl)_n-1sucrose, GF_n (n = 2 – 5)] as the only carbon source. Hydrolyses of 1-kestose, sucrose, and inulin were detected in the extract of the cell. The highest activity on 1-kestose was detected in the extract of B. longum A1 followed by B. adolescentis G1. The other extracts weakly attacked 1-kestose. The relative activities of the extract of B. adolescentis G1 for 1-kestose, nystose, 1^F-fructosylnystose, sucrose, and inulin were 100, 82.5, 50.8, 28.3, and 15.0, respectively. The relative activities for various substrates differed from invertases (yeast β-fructofuranosidases) and exo-inulinase from Penicillium trzehinskii.     

食蚊鱼（Gambusia spp.）入侵生态学研究进展

食蚊鱼(Gambusia spp.)是原产于北美洲的著名入侵物种,它对蚊类幼虫具有很强的捕食压力,被作为疟疾防治生物工具而在全世界温带和热带地区扩散.近20多年来的研究认为,食蚊鱼通过食物竞争和捕食等机制威胁引入地的无脊椎动物、鱼类、两栖类的生存,显著影响引入地的生物多样性.它们具有广泛的生境适应性、生长迅速、卵胎生、高生殖率、对生殖生态条件无特殊要求等特点.另外,它们的个体野外寿命不超过2a,种群更新速度快,种群内形成春季和夏季两个繁殖群体,并具有不同的繁殖生物学特点,因而形成复杂的世代结构.遗传上,雌鱼具有混交及能长期贮存精子的特点,能快速建立种群并克服奠基效应.这些种群通过快速适应性进化而形成一定规模的地理群体.当前,由于食蚊鱼在预防疟疾的工作中仍具有不可替代的作用,它会借助于人力的作用而继续扩散.为减轻它对非目标地区和非目标生物的影响,应进一步深入开展其入侵生态学研究.     

Mortality of Herring Eggs on Different Algal Substrates (Furcellaria spp. and Cladophora spp.) in the Baltic Sea – An Experimental Study

In the Baltic Sea, herring (Clupea harengus membras) spawns in the littoral zone, where its eggs are attached to algae or vascular plants. Field studies indicate that egg mortality can be very high (up to 100%) in eggs that are attached to red algae (Rajasilta et al., 1989, 1993). Because high mortality can be due to allelochemical effects of the algae, we studied the mortality of herring eggs on different algal substrates experimentally. Four types of substrates were tested: fresh Cladophora and Furcellaria, and Furcellaria that had decomposed six days or 23 days. The incubation time in the experiments was 3 days and incubation temperature 12–13 ^oC (ca. 700–800 h-degrees). The results were in accordance with observations made in field studies and indicated significant differences among the substrate types. In eggs attached to fresh Cladophora, mortality was significantly lower (mean=2.8%; n=20) than in those attached to Furcellaria, independently of the treatment of the algae. The highest values of mortality (mean=14.4%; n=20) were found in eggs attached to Furcellaria that had decomposed over a six days’ period. This suggested that Furcellaria contain some chemical substances, which can cause mortality in herring embryos and the effect seems to be dependent on the state of decomposition of the algae.     

Are Helicoverpa armigera and Tetranychus spp. populations still susceptible to pesticides? A Zimbabwean study

The African bollworm, Helicoverpa armigera (Hübner), and the cotton red spider mite, Tetranychus spp., are important pests of cotton in Zimbabwe. A study to assess H. armigera resistance to fenvalerate 20 EC (Cyano (3-phenoxyphenyl) methyl 4-chloro-α-(1-methylethyl)benzene acetate) and Tetranychus spp. resistance to amitraz 20 EC (n’-(2,4-dimethylphenyl)-n-[[(2,4-dimethylphenyl)imino]methyl]-n-) was conducted at the Cotton Research Institute (CRI) during the 2005/06 cotton-growing season. Field populations of H. armigera and Tetranychus spp. were collected from some of Zimbabwe’s major cotton-growing areas of Sanyati, Umguza, Chisumbanje, Chinhoyi and the CRI in Kadoma and exposed to bioassays. The African bollworm leaf disc technique and the red spider mite attached leaf-dipping technique were used to assess responses of the African bollworm and red spider mite to the pesticides. Susceptible laboratory populations served as the standard populations and their responses were compared with those of the field populations. The graphical method and MSTAT-C probit analysis computer program were used to calculate LC₅₀ values. Although the CRI field population, used as a reference population for the registration of H. armigera insecticides, had the highest LC₅₀ value (graphical?=?0.000100000; MSTAT-C?=?0.000088195) compared with all the other field populations, overall log-dose probit bioassays on all field-collected strains of the bollworm showed no resistance to the pyrethroid (RFs?=?0.04–0.54-fold). Tetranychus spp. showed very low levels of resistance (RFs?=?1.26–2.00-fold). Continuous monitoring of major cotton pests, especially H. armigera and Tetranychus spp., from all cotton districts of Zimbabwe is vital for early detection of resistance development.     

Ants and extrafloral nectaries: no evidence for plant protection in Helichrysum spp. — ant interactions

Summary Characterstics of Australian endemic Helichrysum bracteratum and H. viscosum suggest that foraging ants act as guards of developing flowerheads, protecting capitula from seed predators: (1) extrafloral nectar is secreted from leaves subtending the capitula and from bracts encircling the floral disc during pre- to post-flowering periods; (2) capitula are attended by ants; and, (3) encounters between ants and other capitula visitors, including predispersal seed predators such as Tephritis sp. (Diptera), can be frequent. In experiments to test the ant-guard hypothesis, exclusion of ants from plants increased abundance of other insects on the developing capitula. The difference between ant-access and ant-exclusion treatments was related to ant abundance on the access plants. These effects were statistically significant in spite of the large variation in insect activity between sites and through the season.The increased abundance of insects on capitula following ant-exclusion did not, however, result in significant increases in the number of adult seed predators observed on capitula, the number of immature seed predators in capitula, or capitula damage as estimated between ant-access and exclusion treatments of either H. bracteatum or H. viscosum. Further, the ant-exclusion treatment on H. bracteatum had no significant influence on pollination as measured by seed set or on the degree of parasitism of Tephritis sp. by Megastigmus sp. Site and season most strongly affected numbers of immature seed predators and damage to capitula.We discuss these findings in relation to the ant-guard hypothesis and suggest that generalization of the protection hypothesis to all plants with extrafloral nectaries is premature.     

Production of cyclomaltononaose (δ-cyclodextrin) by cyclodextrin glycosyltransferases from Bacillus spp. and bacterial isolates

The conversion of soluble starch to cyclomaltohexaose (α-CD), cyclomaltoheptaose (β-CD), cyclomaltooctaose (γ-CD) and cyclomaltononaose (δ-CD) by cyclodextrin glycosyltransferases (E.C. 2.4.1.19) from Bacillus spp. and bacterial isolates was studied. The results show that δ-CD was formed by all the enzymes investigated in the range of 5%–11.5% of the total amount of α-, β-, γ-, and δ-CD produced. Received: 17 February 1998 / Received revision: 18 May 1998 / Accepted: 21 May 1998