Survival of several <Emphasis Type="Italic">Rhizobium/Bradyrhizobium</Emphasis> strains on different inoculant formulations and inoculated seeds

The effect of a variety factors on the survival of several rhizobia strains on inoculants and inoculated seeds has been evaluated. Since the rhizobia strains showed different cell-density-evolution patterns on peat-based inoculants and on inoculated seeds, several inoculant formulations with highly effective Rhizobium/Bradyrhizobium strains (for Lupinus, Hedysarum, Phaseolus and Glycine max.) were monitored under the following storage conditions: (a) the inoculants were kept refrigerated (at 4 °C), or (b) at room temperature (25 °C). The effect of water content (30–50%, w/w) in the inoculants as well as that of several seed-coating adhesives were also investigated. Alternative carriers including perlite and vermiculite were tested. For all of the strains, survival on sterile peat-based inoculants was higher than on the corresponding unsterile peat formulation; for the latter, refrigerated storage conditions are recommended to ensure high bacterial densities. The water content of the inoculants had a differential effect on strain survival depending on the sterility of the peat, such that a high water content was more detrimental when unsterilized peat was employed. The best adherent for rhizobia survival was a gum arabic/water solution. Perlite was as effective as peat in maintaining a high population of rhizobia, at least for 6 months of storage. Electronic Publication     

Relationship of in vitro and in planta screening: improving the selection process for biological control agents against Fusarium root rot in row crops

Fusarium root rot in row crops is typically managed by cultural practices and fungicide seed treatments. Biological control using microbial agents is another option but needs further development for improved disease management. Screening to identify biocontrol agents are crucial. However, relationships among the steps and how to improve the screening process are unresolved questions. Strains of Burkholderia (4), Bacillus (5) and Trichoderma (26) were studied in vitro against six Fusarium pathogens. All the bacteria and five selected Trichoderma strains were tested in planta in the greenhouse against diseases of Fusarium graminearum and Fusarium oxysporum. Burkholderia ambifaria C628, Bacillus simplex R180, and all Trichoderma isolates showed high reduction in disease levels in corn, soybean and wheat, ranging from 16 to 63%. Responses of the biocontrol agents during in vitro and in planta screening did not always correlate. In vitro and in planta tests should be considered independently in selecting biocontrol candidates.     

Yersiniabactin and other siderophores produced by clinical isolates of Enterobacter spp. and Citrobacter spp

We analyzed the ability of extraintestinal strains of Enterobacter spp. and Citrobacter spp. to employ different siderophore-mediated strategies of iron acquisition. All strains produced iron-chelating compounds. Cross-feeding assays indicated that most isolates of both Enterobacter spp. and Citrobacter spp. excreted catecholate siderophore enterobactin, less produced aerobactin, and single strains excreted hydroxamates different from aerobactin. Besides, we analyzed if the strains had the ability to produce the siderophore yersiniabactin coded by the Yersinia high-pathogenicity island (HPI). The presence of HPI genes was observed in single isolates of three species: E. cloaceae, E. aerogenes and C. koseri. A detailed polymerase chain reaction analysis revealed differences in the genetic organization of the HPIs; however, in a cross-feeding test we proved that yersiniabactin was produced and the island was functional.     

Classification of black-pigmented anaerobes by pyrolysis mass spectrometry (PMS) and conventional tests (CTs)

Abstract Clinical (66: dental 53; vaginal 4; wound 9) and reference (5^∗) strains of pigmented Gram-negative anaerobic bacilli were examined in pyrolysis mass spectrometry (PMS) and conventional tests (CTs). The strains were identified in CTs as: Prevotella intermedia (48^∗); Pr. melaninogenica (1); Pr. corporis (7); Porphyromonas asaccharolytica (12^∗); P. endodontalis (1^∗) and P. gingivalis (2^∗). Numerical classification based on CTs resolved five clusters comprising strains identified as (I) Pr. corporis , (II) Pr. melaninogenica , (III) Pr. intermedia , (IV) P. gingivalis and (V) P. asaccharolytica and P. endodontalis . Numerical classification based on PMS showed a similar division, with decreasing homogeneity in the order Pr. intermedia, Pr. corporis, P. asaccharolytica , in agreement with the ordering of homogeneity for these species in CTs. PMS clusters corresponding the Porphyromonas spp. were clearly distinct from those of Prevotella spp. PMS and CT classifications disagreed on cluster membership for only six of the strains. PMS identification from blind challenge sets agreed with conventional identification for 64 of 67 strains.     

Enumeration, isolation and identification of diazotrophs from Korean wetland rice varieties grown with long-term application of N and compost and their short-term inoculation effect on rice plants

AIM: This study has been aimed (i) to isolate and identify diazotrophs from Korean rice varieties; (ii) to examine the long-term effect of N and compost on the population dynamics of diazotrophs and (iii) to realize the shot-term inoculation effect of these diazotrophs on rice seedlings. METHODS AND RESULTS: Diazotrophic and heterotrophic bacterial numbers were enumerated by most probable number method and the isolates were identified based on morphological, physiological, biochemical and 16s rDNA sequence analysis. Long-term application of fertilizer N with compost enhanced both these numbers in rice plants and its environment. Bacteria were high in numbers when malate and azelaic acids were used as carbon source, but less when sucrose was used as a carbon substrate. The combined application promoted the association of diazotrophic bacteria like Azospirillum spp., Herbaspirillum spp., Burkholderia spp., Gluconacetobacter diazotrophicus and Pseudomonas spp. in wetland rice plants. Detection of nifD genes from different diazotrophic isolates indicated their nitrogen fixing ability. Inoculation of a representative isolate from each group onto rice seedlings of the variety IR 36 grown in test tubes indicated the positive effect of these diazotrophs on the growth of rice seedlings though the percentage of N present in the plants did not differ much. CONCLUSIONS: Application of compost with fertilizer N promoted the diazotrophic and heterotrophic bacterial numbers and their association with wetland rice and its environment. Compost application in high N fertilized fields would avert the reduction of N(2)-fixing bacterial numbers and their association was beneficial to the growth of rice plants. SIGNIFICANCE AND IMPACT OF THE STUDY: The inhibitory effect of high N fertilization on diazotrophic bacterial numbers could be reduced by the application of compost and this observation would encourage more usage of organic manure. This study has also thrown light on the wider geographic distribution of G. diazotrophicus with wetland rice in temperate region where sugarcane (from which this bacterium was first reported to be associating and thereon from other plant species) is not cultivated.     

The use of probiotics in shrimp aquaculture

Shrimp aquaculture, as well as other industries, constantly requires new techniques in order to increase production yield. Modern technologies and other sciences such as biotechnology and microbiology are important tools that could lead to a higher quality and greater quantity of products. Feeding and new practices in farming usually play an important role in aquaculture, and the addition of various additives to a balanced feed formula to achieve better growth is a common practice of many fish and shrimp feed manufacturers and farmers. Probiotics, as 'bio-friendly agents' such as lactic acid bacteria and Bacillus spp., can be introduced into the culture environment to control and compete with pathogenic bacteria as well as to promote the growth of the cultured organisms. In addition, probiotics are nonpathogenic and nontoxic microorganisms without undesirable side-effects when administered to aquatic organisms. These strains of bacteria have many other positive effects, which are described in this article.     

中国特产雉类—褐马鸡,藏马鸡和蓝马鸡卵壳的电镜观察

迄今为止,在国内有关野生鸟类卵壳的超微结构报道尚少。我们对中国特产鸡类——三种马鸡卵壳的表皮、气孔、木栅层、锥体层、乳头结、壳膜和基帽进行了扫描巨镜观察,并对结果进行了比较、讨论。认为三种马鸡不同结构的形态,对研究它们的亲缘关系提供了新的价值。     

Sources of resistance to stem nematode, Ditylenchus dipsaci (Kühn) Filipjev, in species of Medicago and Trifolium

The putatively resistant lucerne cv. Euver was as susceptible to 11 English and 11 French populations of ‘lucerne race’ stem nematode, Ditylenchus dipsaci, as the susceptible cv. Europe. The ‘resistant’ cv. Vertus showed some resistance to five of the English and four of the French populations but was just as susceptible as Europe to three English and four French populations. Twelve annual species of Medicago were very resistant to a mixture of two English and two French populations of the nematode. None of 13 perennial spp. of Medicago was very resistant to the same mixture of nematodes but resistant plants were identified in M. borealis, M. dzhawakhetica, M. glutinosa and M. romanica. Twenty six spp. of Trifolium were resistant to a mixture of two English and two French populations of ‘red clover race’ stem nematode.     

Electrochemical checking of aerobic isolates from electrochemically active biofilms formed in compost

Aims:  To design a cyclic voltammetry (CV) procedure to check the electrochemical activity of bacterial isolates that may explain the electrochemical properties of biofilms formed in compost.
Methods and Results:  Bacteria catalysing acetate oxidation in garden compost were able to form electrochemically active biofilms by transferring electrons to an electrode under chronoamperometry. They were recovered from the electrode surface and identification of the isolates using 16S rRNA sequencing showed that most of them were Gammaproteobacteria, mainly related to Enterobacter and Pseudomonas spp. A CV procedure was designed to check the electrochemical activity of both groups of isolates. Preliminary CVs suggested that the bacteria were not responsible for the catalysis of acetate oxidation. In contrast, both groups of isolates were found to catalyse the electrochemical reduction of oxygen under experimental conditions that favoured adsorption of the microbial cells on the electrode surface.
Conclusions:  Members of the genera Enterobacter and Pseudomonas were found to be able to catalyse the electrochemical reduction of oxygen.
Significance and Impact of the Study:  This study has shown the unexpected efficiency of Enterobacter and Pseudomonas spp. in catalysing the reduction of oxygen, suggesting a possible involvement of these species in biocorrosion, or possible application of these strains in designing bio-cathode for microbial fuel cells.     

Evaluation of persistence and effectiveness of bacterial cell suspensions and culture filtrates against M. incognita on brinjal

Keeping in view the staid health and ecological apprehensions coupled with the use of pesticides, entomopathogenic nematodes have the potential to supersede pesticides for the management of various pests. Brinjal plants are the most seriously affected by Meloidogyne incognita. The main objective of this study was to evaluate the persistence effectiveness of bacterial cell suspensions (Xenorhabdus and Photorhabdus spp.) and their culture filtrates in soil up to 7, 14 and 21?days and their response against M. incognita as a source of biological control for nematode management. In a life cycle study, Xenorhabdus and Photorhabdus spp., isolated from Steinernema asiaticum and Heterorhabditis bacteriophora, were proved more effective in influencing the life cycle of RKNs. It was found that all the treatments of bacterial cell suspensions and their culture filtrates at all persistent times proved effective in reducing the number of females and egg masses as compared to control. It delayed penetration of nematode juveniles (J2) into host roots. It was concluded that persistence effectiveness of bacteria and their metabolites decreased in soil with time.     

Prevalence of gastrointestinal parasites in captive non‐human primates of twenty‐four zoological gardens in China

Captive primates are susceptible to gastrointestinal (GIT) parasitic infections, which are often zoonotic and can contribute to morbidity and mortality. Fecal samples were examined by the means of direct smear, fecal flotation, fecal sedimentation, and fecal cultures. Of 26.51% (317/1196) of the captive primates were diagnosed gastrointestinal parasitic infections. Trichuris spp. were the most predominant in the primates, while Entamoeba spp. were the most prevalent in Old World monkeys (P < 0.05). These preliminary data will improve the management of captive primates and the safety of animal keepers and visitors.     

Ecological strategies of Dalbulus leafhoppers

Abstract. 1. Developmental times from egg to adult eclosion were established for eight Dalbulus leafhopper species on maize (Zea mays) and its perennial, gamagrass relative, Tripsacum dactyloides. Experiments were conducted with a 16 h day (25–27°C) and an 8 h night (21–22°C).
2. Developmental times were shortest for the maize specialists, c. 26 days for D. maidis and c. 27 days for D. elimatus, and longest for the Tripsacum specialists, c. 32 days for D. tripsacoides, c. 34 days for D. quinquenotatus and c. 40 days for D. guzmani. Developmental times were intermediate for D. gelbus, D. longulus and D. guevarai, species that utilize both maize and Tripsacum as field hosts.
3. For all species, development was significantly faster on maize (species mean 29 days) than on T. dactyloides (species mean 34 days).
4. D. maidis and D. elimatus developed significantly faster (c. 3 days) on maize than on its close perennial relative, Z. diploperennis.
5. As measured by a 48 h oviposition period, D. maidis, D. elimatus and D. gelbus produced significantly more progeny on maize than on gamagrass. D. quinquenotatus produced more progeny on T. dactyloides than on maize. D. maidis and D. elimatus produced more progeny on maize than on Z. diploperennis.
6. These studies provide support for the hypothesis which predicts that herbivorous insects that specialize on annual plants will have a shorter generation time and be more fecund than perennial specialists. The genus Dalbulus appears to be composed of species that fit an r-K continuum, with maize specialists and Tripsacum specialists at opposite ends of the continuum.     

Patterns of interspecific associations of stem gallers on willows

Abstract.  The pattern of interspecific associations of three stem-galling sawfly species ( Euura atra , E. elaeagnos , E. purpureae ) and three stem-galling gallmidge species ( Rabdophaga sp. 3–5) was investigated on five willow taxa ( Salix alba , S. fragilis , S.  ×  rubens , S. elaeagnos , S. purpurea ) at five natural sites in Central Europe. The willow species harboured specific species associations of two stem gallers, each pair consisting of one Euura and one Rabdophaga species. The stem gallers were patchily distributed and their densities varied significantly among willow host plant species, host plant individuals, and host plant sexes. Four of the six species showed a significant increase in galling rate with shoot length. The other two species were the sawfly and cecidomyiid pair that induce galls on S. purpurea . The preference of stem gallers to longer shoots was generally not related to higher larval performance in terms of survival. Only one species, Rabdophaga sp. 5, was found to be more abundant on male plants. The correlation of densities of the species pairs of stem gallers was independent of willow sexes. Species pairs of stem gallers co-occurring on the same willow species tended to attack different shoots within the same host plant individual. When species pairs co-occurred on shoots they were usually found in similar densities as when occurring alone on shoots. The stem-galling sawflies usually formed galls at the basal part of a shoot, whereas the gallmidge R . sp. 5 ( R . sp. 3 and R . sp. 4 showed no clear tendency) preferred the middle or distal part of a shoot. This is interpreted with differences of their phenology and oviposition period.     

Multilocus genetic relationships between clinical and environmental Aeromonas strains

Allelic variations in the chromosomal genome of 120 isolates of motile Aeromonas (A. hydrophila, A. sobria and A. caviae) and eight reference strains with one Plesiomonas shigelloides were assessed by analysis of electrophoretically demonstrable polymorphism in 16 genes encoding metabolic enzymes. The strains were collected from humans (n = 59) and from the aquatic environment (n = 61) in canton Tessin, Switzerland. Clustering of the electrophoretic types (ET) from a matrix of pairwise genetic distances, based on the 16 enzyme loci, confirmed the genetic distinctness of the three species. Furthermore, A. hydrophila and A. sobria were divided in three and two groups respectively. For each species clinical strains were well differentiated from those collected in the environment.     

Malachite green agar,a new selective medium for Fusarium spp.

Malachite Green Agar 2.5 ppm (MGA 2.5) is a potent selective medium for isolation and enumeration of Fusarium spp. It has been tested with pure and mixed cultures as well as in naturally contaminated samples. The recoveries of Fusarium species in MGA 2.5 were the same as the recoveries obtained in Nash and Snyder medium. However, this medium is a more selective culture medium for Fusarium spp. than Nash and Snyder medium, because it does not allow the development of colonies belonging to other fungal genera. MGA 2.5 is simple to prepare and less hazardous than other Fusarium selective media containing pentachloronitrobenzene (PCNB). This revised version was published online in June 2006 with corrections to the Cover Date.     

Relatedness of penicillin-binding proteins from various Listeria species

The heterogeneity of penicillin-binding proteins (PBPs) of five Listeria species was investigated. Similarities in the overall PBP pattern were found between those of L. welshimeri and L. innocua, and between L. ivanovii and L. seeligeri, and all were distinct from the PBPs of L. monocytogenes. In all species, however, the primary target for beta-lactam antibiotics, as identified in L. monocytogenes recently, appeared highly conserved. In addition, the low-Mr PBP 5 was biochemically very similar in all strains and contained identical binding properties to beta-lactam compounds, suggesting that this protein may play an important role. All other PBPs varied considerably in their penicilloyl-peptide pattern, indicating differences in their amino acid sequences.