The influence of EDTA on the composition of alginate synthesized by Azotobacter vinelandii

1. During an investigation of the physiology of Azotobacter vinelandii with particular reference to polysaccharide formation, a suitable medium which was precipitate-free was developed by adding EDTA at a concentration of 50 mg/l to a basal medium containing one of eight different carbohydrates as sole carbon source.
2. Acetylated alginate was always produced by the organism when cultured under defined conditions, regardless of the carbohydrate source incorporated in the basal medium.
3. When EDTA was added to the medium, the bacteria produced acetylated polyuronides with a preponderance of mannuronic acid residues.
4. A comparison of the infrared spectra of the alginate produced by Azotobacter vinelandii and the affect of EDTA upon the mannuronic acid/guluronic acid ratios of the alginate are reported.

     

Predicted effects of increasing salinity on the crustacean zooplankton community of Pyramid Lake,Nevada

Since 1933 the salinity of Pyramid Lake, Nevada, U.S.A., has increased 32% to nearly 5.5‰. We tested the hypothesis that further increases of 1.5 to 2 times (1.5× to 2×) its present salinity would significantly reduce species richness and alter population structures of the existing crustacean zooplankton community. Three strategies were applied: in addition to monitoring zooplankton in semicontrolled indoor microcosms at 1×, 1.5× and 2× and conducting range-finding, acute, and chronic salinity bioassays, the present zooplankton community of Walker Lake (2×) was compared with that existing in Pyramid Lake (1×). Ceriodaphnia quadrangula and Diaphanosoma leuchtenbergianum, both collected from Pyramid Lake, were lacking in Walker Lake. Populations of Cyclops vernalis were significantly lower and those of Diaptomus sicilis and Moina hutchinsoni were significantly higher in Walker Lake than in Pyramid Lake. Densities of Ceriodaphnia and Cyclops were low in microcosms at salinities > 1×. Diaphanosoma could not be maintained in microcosms, regardless of salinity. Numbers of Diaptomus and Moina in microcosms were proportional to salinity level. Short-term LC50 salinities (‰) were as follows: Diaphanosoma, 6.5; Ceriodaphnia, 7.1; Diaptomus, 13.3; Cyclops, 14.8; and Moina, 17.8. Multiple-generation, chronic bioassays were run only on Cyclops and Diaptomus. Three generations of Cyclops were produced at salinities of 4.0 to 8.5‰, but not at 9.8‰ or higher. Diaptomus was unable to complete three generations at salinities ?9.6‰. We speculate that high salinity in Walker Lake may indirectly benefit Diaptomus by negatively affecting predatory Cyclops, and benefit Moina by causing extinction of competing salinity-intolerant Diaphanosoma and Ceriodaphnia. Except for the response of Diaptomus, results from bioassays were in general agreement with results from microcosms and with field data. Untested predator-prey interactions could be responsible for the apparent discrepancy.     

Possible Effects of Glyphosate on Mucorales Abundance in the Rumen of Dairy Cows in Germany

Glyphosate (N-phosphonomethyl glycine) is registered as a herbicide for many food and non-food crops, as well as non-crop areas where total vegetation control is desired. Glyphosate influences the soil mycobiota; however, the possible effect of glyphosate residues in animal feed (soybean, corn, etc.) on animal mycobiota is almost unknown. Accordingly, the present study was initiated to investigate the mycological characteristics of dairy cows in relationship to glyphosate concentrations in urine. A total of 258 dairy cows on 14 dairy farms in Germany were examined. Glyphosate was detected in urine using ELISA. The fungal profile was analyzed in rumen fluid samples using conventional microbiological culture techniques and differentiated by MALDI-TOF mass spectrometry. LPS-binding protein (LBP) and antibodies (IgG1, IgG2, IgA, and IgM) against fungi were determined in blood using ELISA. Different populations of Lichtheimia corymbifera, Lichtheimia ramosa, Mucor, and Rhizopus were detected. L. corymbifera and L. ramosa were significantly more abundant in animals containing high glyphosate (>40 ng/ml) concentrations in urine. There were no significant changes in IgG1 and IgG2 antibodies toward isolated fungi that were related to glyphosate concentration in urine; however, IgA antibodies against L. corymbifera and L. ramosa were significantly lower in the higher glyphosate groups. Moreover, a negative correlation between IgM antibodies against L. corymbifera, L. ramosa, and Rhizopus relative to glyphosate concentration in urine was observed. LBP also was significantly decreased in animals with higher concentrations of glyphosate in their urine. In conclusion, glyphosate appears to modulate the fungal community. The reduction of IgM antibodies and LBP indicates an influence on the innate immune system of animals.     

Polysaccharide production and the possible occurrence of GDP-d-mannose dehydrogenase inAzotobacter vinelandii

During the growth ofAzotobacter vinelandii in batch culture in Burk's 2% glucose medium supplemented with 50mg EDTA per litre, water-insoluble capsular polysaccharide material accumulated in cultures prior to the appearance of water-soluble polysaccharide in the culture medium. On isolation, hydrolysis and chromatography, both these polysaccharides were observed to be composed of carbohydrate monomers having the same chromatographic mobilities as glucose, rhamnose, guluronic acid and mannuronic acid. The activity of GDP-d-mannose dehydrogenase recorded in crude cell-free extracts fromAzotobacter vinelandii, when these polysaccharides were produced, may indicate a close similarity between the biosynthetic pathway of alginate synthesis in marine Phaeophyceae and this soil microorganism.     

Response of the population size and community structure of Paenibacillus spp. to different fertilization regimes in a long-term experiment of red soil

Background and aims

Paenibacillus spp. are widely considered to impact the fertility and health of soil. The aim of this study was to evaluate how different fertilization regimes affect the population size and community structure of Paenibacillus spp. over a long period of time in red soil.

Methods

Soil samples were collected from a long-term experiment and were then analyzed using real-time PCR and PCR-DGGE. The correlation analysis, PCA and RDA were used to explore the relationships among Paenibacillus spp. population, community structure and soil properties in different treatments.

Results

The pH was seriously decreased only by the application of chemical fertilizer. The largest population of Paenibacillus spp. was found in the soil treated with organic fertilizer application, while the richest diversity was observed in the soil treated only with the chemical fertilizer. The Paenibacillus spp., Paenibacillus alkaliterrae, Paenibacillus campinasensis, and Paenibacillus xylanilyticus were found in all treatments. Paenibacillus castaneae was found in the soil treated with NPK, and Paenibacillus pabuli was specifically observed in the lime-amended treatment. Paenibacillus taichungensis and Paenibacillus prosopidis were detected in the soil treated with only chemical fertilizer. Except for the ammonium and pH, all the tested soil fertility parameters (total C, total N, nitrate, available K and available P) could significantly affect both the Paenibacillus spp. population number and diversity. The soil pH was significantly correlated with Paenibacillus spp. diversity only.

Conclusions

Our results indicate that the different long-term fertilization regimes have varied impact on both the Paenibacillus spp. population size and the diversity of the community associated with the soil properties tested. These results can help to enrich the information on the response of beneficial soil microbes to different long-term fertilization regimes.     

On the carrying behaviour of basic South American primates

InCallithrix, Saguinus, Aotus, andCallicebus other group members than the mother participate in infant care. Differences among these species are obvious in respect to the time of being off any caregiver, and in respect to the carrying position (Aotus andCallicebus). Alloparental care, a basic behaviour pattern in these species, has evolved from the parking behaviour of the prosimians. The infants are «parked» at other group members.     

The establishment ofAnagyrus saccharicola Timb. [Hymenoptera: Encyrtidae] in Barbados,West Indies,against the sugarcane mealybug,Saccharicoccus sacchari (Ckll.) (Hemiptera: Coccidae]

During the survey of the natural enemies ofSaccharicoccus sacchari in Barbados in 1968 and 1969, the following six indigenous species of natural enemies were recorded: —Hyperaspis trilineata andNephus sp.(Coccinellidae); Oligota barbadorum (Staphylinidae); an unidentified Cecidomyid(Cecidomyidae); Pseudaphycus mundus (Encyrtidae) andAspergillus sp. (Aspergillacae). Of these,H. trilineata was found in fair numbers in the Westmoreland (St. James) area only;Aspergillus sp. was more abundant during the wet season, while the other species were recorded usually in small numbers from most parishes. As the indigenous natural enemies do not provide effective control of the pest, three exotic predators and one parasite were introduced into Barbados, through the Commonwealth Institute of Biological Control.Cryptolaemus montrouzieri, Hyperaspis sp. andNephus sp. were obtained from India andAnagyrus saccharicola from East Africa. OnlyA. saccharicola became established. It was recovered soon after its release and, augmented by additional releases, spread rapidly. In January 1972, the levels of parasitism in the dry, intermediate and high rainfall areas were: 8.3, 9.0 and 9.7%.     

Occurrence of Phytophthora plurivora and other Phytophthora species in oak forests of southern Poland and their association with site conditions and the health status of trees

Phytophthora plurivora and other Phytophthora species are known to be serious pathogens of forest trees. Little is known, however, about the presence of P. plurivora in Polish oak forests and their role in oak decline. The aims of this study were to identify P. plurivora in healthy and declining Quercus robur stands in southern Poland and to demonstrate the relationship between different site factors and the occurrence of P. plurivora. In addition, the virulence of P. plurivora and other Phytophthora species was evaluated through inoculations using 2-year-old oak seedlings. Rhizosphere soil was investigated from 39 oak stands representing different healthy tree statuses. The morphology and DNA sequences of the internal transcribed spacer regions (ITS) of the ribosomal DNA and the mitochondrial cox1 gene were used for identifications. P. plurivora, an oak fine root pathogen, was isolated from rhizosphere soil samples in 6 out of 39 stands. Additionally, Phytophthora cambivora, Phytophthora polonica and Phytophthora rosacearum-like were also obtained from several stands. The results showed a significant association between the presence of P. plurivora and the health status of oak trees. Similar relationships were also observed for all identified Phytophthora species. In addition, there was evidence for a connection between the presence of all identified Phytophthora species and some site conditions. Phytophthora spp. occurred more frequently in declining stands and in silt loam and sandy loam soils with pH?≥?3.66. P. plurivora and P. cambivora were the only species capable of killing whole plants, producing extensive necrosis on seedling stems.     

The influence of repetitive thermal stresses on the dominance of reef-building Acropora spp. (Scleractinia) on coral reefs of the Maldive Islands

The distribution and size-age structure of Acropora corals were studied in two Maldivian atolls that differ in their geographic position and sea surface temperature regimes. The frequency and strength of thermal anomalies for the last 2 decades had a significant influence on the abundance, mortality rates, and age structures of acroporid communities. The long-term temperature amplitude was higher and the maxima were more pronounced in the northernmost Ihavandippolu Atoll than those in the equatorial South Huvadhoo Atoll. These differences resulted in a 10.4% mean cover of Acropora at Ihavandippolu Atoll, whereas the Acropora cover in the South Huvadhoo Atoll reached 59.5%. In the northern atoll, the coral mortality rate after the 2010 thermal anomaly was 3 times higher than that in the southern atoll. Younger acroporid colonies (up to 2 years old) dominated the northern atoll reefs, while the southern atoll showed a high proportion of older mature colonies. In both atolls, healthy table colonies of Acropora cytherea with a disk diameter greater than 2 m were observed that apparently survived three thermal anomalies since 1998. The mechanisms of acclimatization of Acropora and the prospects for its dominance in the Maldives under changing environmental conditions are discussed.     

Influence of bovine lactoferrin on the growth of selected probiotic bacteria under aerobic conditions

Bovine lactoferrin (bLf) is a natural glycoprotein, and it shows broad-spectrum antimicrobial activity. However, reports on the influences of bLf on probiotic bacteria have been mixed. We examined the effects of apo-bLf (between 0.25 and 128 mg/mL) on both aerobic and anaerobic cultures of probiotics. We found that bLf had similar effects on the growth of probiotics under aerobic or anaerobic conditions, and that it actively and significantly (at concentrations of >0.25 mg/mL) retarded the growth rate of Bifidobacterium bifidum (ATCC 29521), B. longum (ATCC 15707), B. lactis (BCRC 17394), B. infantis (ATCC 15697), Lactobacillus reuteri (ATCC 23272), L. rhamnosus (ATCC 53103), and L. coryniformis (ATCC 25602) in a dose-dependent manner. Otherwise, minimal inhibitory concentrations (MICs) were 128 or >128 mg/mL against B. bifidum, B. longum, B. lactis, L. reuteri, and L. rhamnosus (ATCC 53103). With regard to MICs, bLf showed at least four-fold lower inhibitory effect on probiotics than on pathogens. Intriguingly, bLf (>0.25 mg/mL) significantly enhanced the growth of Rhamnosus (ATCC 7469) and L. acidophilus (BCRC 14065) by approximately 40–200 %, during their late periods of growth. Supernatants produced from aerobic but not anaerobic cultures of L. acidophilus reduced the growth of Escherichia coli by about 20 %. Thus, bLf displayed a dose-dependent inhibitory effect on the growth of most probiotic strains under either aerobic or anaerobic conditions. An antibacterial supernatant prepared from the aerobic cultures may have significant practical use.     

Composition of the cell wall and other fractions of the autolyzed yeast form of Histoplasma capsulatum

Comparison of two strains ofHistoplasma capsulatum yielded data differing only in quantification, and the constituents observed and identified were galactose, glucose, mannose, glucosamine and amino acids. A comparison of hydrochloric acid and formic acid hydrolyses ofH. capsulatum fractions indicated hydrochloric acid to be of more value than 88 per cent formic acid hydrolysis for composition analyses. The removal of formyl esters from formic acid hydrolysates was found necessary and was accomplished byN HCl hydrolysis for 30 min. Two derivative artifacts were observed with formic acid hydrolysis; D-1, which was refractory to subsequent HCl hydrolysis, and D-2, which disappeared after HCl hydrolysis. Another artifact, D-3, was observed with 6N HCl hydrolysis of histoplasma cell wall fractions. The following conditions of hydrolysis were found to be useful: (1) glucose release was measured after hydrolysis inN HCl for 4 hr; (2) glucosamine release was measured after hydrolysis in 6N HCl for 9 hr; (3) amino acid release was accomplished by 6N HCl hydrolysis for 18 hr; and (4), hexoses released were determined by gas liquid chromatography (GLC) after hydrolysis in bothN HCl and in 88 per cent formic acid for 24 hr, followed byN HCl for 30 min. Several different types of carbohydrate polymers have been reported in the parasitic yeast form ofH. capsulatum. There is general agreement on the occurrence of amino acids as protein (8, 12, 13), chitin (7, 19) and several hexoses, including glucose and glucosamine, which are found in cell wall polymers (7, 8, 11–16, 19, 20, 24). The presence of uronic acid was also reported (14, 15), but not confirmed, by Domer, Hamilton & Harkin (8), and mannose was not found by all investigators (12). We undertook a study of graded acid hydrolyses and of composition analysis of the autolysis products of the yeast form by various procedures in order to add further to the above information.     

Prey range of the predatory ladybird Cryptolaemus montrouzieri

The prey range of Cryptolaemus montrouzieri was studied in the laboratory to investigate whether the mealybug destroyer can contribute to the suppression of other pest insects besides mealybugs and to assess its potential impact on non-mealybug populations as part of an environmental risk assessment for its use in biological control. Prey tested in these experiments were: tobacco aphid Myzus persicae nicotianae (Sulzer)(Hemiptera: Aphididae), pea aphid Acyrthosiphon pisum (Harris)(Hemiptera: Aphididae), tobacco whitefly Bemisia tabaci (Gennadius)(Hemiptera: Aleyrodidae), southern green stinkbug Nezara viridula (L.)(Hemiptera: Pentatomidae) eggs, western flower thrips Frankliniella occidentalis (Pergande)(Thysanoptera: Thripidae), two-spotted ladybird Adalia bipunctata (L.)(Coleoptera: Coccinellidae) eggs and eggs of the greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae). Larval survival was high to moderate when C. montrouzieri was provided with hemipteran prey and poor to zero when the ladybirds were provided with non-hemipteran prey. Females reared on M. persicae and A. pisum produced similar numbers of eggs as their counterparts fed the citrus mealybug Planococcus citri (Risso)(Hemiptera: Pseudococcidae), but fecundity was significantly lower when the ladybirds were reared on B. tabaci nymphs or on A. bipunctata eggs. Prey species that were found to be less suitable for immature development of C. montrouzieri could still be an adequate food source for reproduction and survival of adult ladybirds. For example, only 8 % of the predator larvae reached the adult stage when provided with A. bipunctata eggs, but females that had developed on eggs of the Mediterranean flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) and that were supplied with A. bipunctata eggs from adult emergence on, were only 35 % less fecund than females provided with mealybugs in their adult life. The results are discussed in relation to the development of a suitable methodology for prey/host range testing in the framework of an environmental risk assessment for arthropod natural enemies.     

Parasitoids of the corn borerOstrinia furnacalis [Lep.: Pyralidae] in the Mariana Islands

A survey of the parasitoids of the Asian corn borer,Ostrinia furnacalis (Guenee), in the Mariana Islands showed that few of the borers were parasitized. Parasitoids were reared from only 30 pupae out of more than 4 500 sampled. Three species of pupal parasitoids were found:Xanthopimpla punctata (F.),Brachymeria albotibialis (Ashmead) andTetrastichus? inferens Yoshimoto. No parasitoids were reared from mature larvae. The only egg parasitoid observed wasTrichogramma chilonis Ishii. During the month prior to the sweet corn harvest, which was the peak period of moth oviposition, about 40% of the egg masses had at least one egg parasitized. An average of about 35% of the eggs within a mass were parasitized.     

Location of the genes coding for 18S and 28S ribosomal RNA in the human genome

³H-rRNA obtained from Xenopus laevis tissue cultured cells, or a ³H-cRNA made from Xenopus ribosomal DNA, was used for heterologous in situ hybridisation with human lymphocyte metaphase chromosomes. Prior to hybridisation, chromosome spreads were stained with Quinacrine and selected cells showing good Q-banding photographed; the same cells were then rephotographed after autoradiography and pairs of photographs for each cell were used to make dual karyotypes. The chromosomes within each karyotype were divided into equal sized segments (approx. 0.7 μ), with a fixed number of segments for each chromosome type. The distribution of silver grains between segments showed that the ³H-RNAs hybridised specifically to the nucleolar organising regions of the D and G group chromosomes with no other sites of localised labelling in the complement. Control experiments showed no localisation, with insignificant labelling, when metaphase spreads were incubated in a mixture containing Xenopus ³H-rRNA and competing cold human (HeLa) rRNA. Filter hybridisation experiments on isolated human DNA showed that the Xenopus derived ³H-RNAs hybridised to a fraction of human DNA which was on the heavy side of the main DNA peak and that these RNAs were competed out in the presence of excess cold human rRNA, confirming the specificity of the heterologous hybridisation. In situ hybridisation experiments were also carried out on cells from individuals with one chromosome pair showing heteromorphism for either a very long stalk (nucleolar constriction) subtending a satellite, or a large satellite. It was shown that the chromosome with the large stalk hybridised four times as much ³H-rRNA as its homologue, whereas differences in the sizes of the subtended satellites did not materially affect hybridisation levels indicating that rDNA is located in the stalks and not the satellites. The amount of ³H-rRNA hybridised differs between chromosomes and individuals; these differences are heritable and rDNA can be detected by in situ hybridisation in all three chromosomes number 21 in cells from Down's patients and in translocated chromosomes conta.ining a nucleolar constriction. Different D and G group chromosomes which hybridised equal amounts of ³H-rRNA participated in rosette associations at metaphase in a random fashion in some individuals and in a non-random fashion in others. In all individuals studied chromosomes with large amounts of rDNA were not found to be preferentially involved in association. It was therefore concluded that the probability of a chromosome being involved in the formation of a common nucleolus is not a simple function of its rDNA content and other possible factors are considered.     

Production of the mosquito-parasitic fungus,Coelomomyces dodgei,through synchronized infection and growth of the intermediate copepod host,Cyclops vernalis

High yields of the copepodCyclops vernalis infected with the mosquito-parasitic fungusCoelomomyces dodgei Couch & Dodge were obtained by infecting nauplii in large synchronously developing populations. Exposure of 2000 48 or 72 h old nauplii to 6×10³ sporangia at the time of meiospore release yielded ca. 1500 infected copepods. Based on yields of infected copepods, susceptibility ofC. vernalis toC. dodgei decreased as copepods developed. Infection rates were 75% for copepods exposed as 48 or 72 h old nauplii but declined to 32 and 9.6%, respectively, for those exposed as copepodids or adults. The relevance of these results for domestication of other species ofCoelomomyces and studies on non-target organisms is discussed, and improved procedures for routine production ofC. dodgei are described.