Argentine ant (Hymenoptera: Formicidae) trail pheromone enhances consumption of liquid sucrose solution

We investigated whether the Argentine ant, Linepithema humile (Mayr), trail pheromone, Z9-16:Ald, could enhance recruitment to and consumption of liquid sucrose solutions. All tests were done as paired comparisons with a 10% sucrose solution as food. In the laboratory, mixing 20 microl of a 10-microg/ml solution of the pheromone with 50 microl of the 10% sucrose solution increased the number of ants feeding by >150%. In a field test, we combined the trail pheromone with a 10% sucrose solution in 50-ml vials. These vials were covered with a plastic membrane that has 1.5-mm-diameter holes punched uniformly across its surface. Ants could drink from the holes after the vials were inverted. For half of the vials, 1 microg of the pheromone was put onto the plastic membrane before the vials were filled with a 10% sucrose solution. The remaining vials had no pheromone on the plastic membrane. After 4 h we measured the consumption in each vial. Bait consumption with the pheromone was enhanced by 29%. In a 2nd series of tests, vials were left outside for 24 h. The consumption rate was 33% higher with the pheromone compared with the controls that didn't have pheromone.     

THE EFFECT OF CERTAIN SOIL TREATMENTS ON DIDYMELLA STEM-ROT OF TOMATOES

About 93% of Didymella lycopersici spores were destroyed after 4 weeks incubation in unsterilized soil. A survey of the microflora of glasshouse soil receiving different treatments and inoculated with D. lycopersici showed no clear relation between numbers of any group of organisms and the incidence of stem rot. Sterilized soil was not made toxic to D. lycopersici by the growth of a number of soil microorganisms even after 9 months incubation, but addition of unsterilized soil or of a suspension of unsterilized soil quickly restored toxicity. Direct observations of spores in soil on slides showed that their fate varied with the treatment of the soil before inoculation. With fresh soil or air-dry soil moistened 2 or more days before inoculation, lysis of spores occurred. With air-dry soil moistened and inoculated simultaneously, some spores germinated but growth of germ tubes soon ceased. No direct connexion could be seen between the fate of the spores and soil microorganisms. Addition of glucose to unsterilized soil reduced its toxicity to D. lycopersici. Soils steamed for 1 min. or longer were not toxic to D. lycopersici , but soils steamed for very short periods were as toxic as unsterilized soils although the soil microflora was much reduced.     

Factors associated with Pseudomonas pickettii intrinsic contamination of commercial respiratory therapy solutions marketed as sterile.

Laboratory investigations were conducted to study the growth dynamics of Pseudomonas pickettii in commercial 0.9% sodium chloride solution under various environmental conditions and to determine the retention of these organisms after challenge through a 0.2-micron cartridge filter system. Low numbers of P. pickettii (1 to 10 CFU/ml of test solution) inoculated into commercial vials containing 5 ml of 0.9% sodium chloride solution and 500-ml volumes of 0.9% sodium chloride solution were shown to proliferate over a 168-h incubation period. These organisms demonstrated growth over a wide range of temperatures (15 to 42 degrees C) in this salt solution, and survival studies at 50, 55, and 60 degrees C indicated that this strain was not unusually resistant to heat (with the times required at a given temperature to reduce the surviving microbial population 10-fold [D-values] being 26.0, 1.9, and 0.7 min, respectively). A challenge test demonstrated that P. pickettii organisms were not completely retained by a 0.2-micron cartridge filter. The number of organisms detected increased from 1 CFU/liter of effluent at 1 to 2 min to a maximum of 176 CFU/liter at 4 to 5 min. Our results indicate that P. pickettii can penetrate a 0.2-micron filtration system and that the passage of organisms and subsequent microbial growth in the filter effluent probably are the mechanisms by which these organisms were recovered from "sterile" commercial 0.9% sodium chloride solution.     

Iron plaque enhances phosphorus uptake by rice (Oryza sativa) growing under varying phosphorus and iron concentrations

Both solution culture and pot experiments were performed to investigate (a) the effects of external Fe (II) concentrations and forms on the formation of iron plaque on the roots of rice (Oryza sativa) and subsequent P adsorption on iron plaque and shoot P concentrations and (b) the effects of soil moisture regimes on the formation of iron plaque and P adsorption on root surfaces and P accumulation in shoots. The results showed that iron plaque was significantly increased with increasing Fe²⁺ concentrations in the solution culture. The amounts of P adsorbed on the iron plaque were increased significantly with external Fe²⁺ concentrations. Although shoot P concentration was not significantly affected by Fe²⁺ treatment after incubation for 2 days, it was significantly increased in the Fe‐treated plants compared with Fe‐deprived ones after incubation for 4 days. Soil culture experiment showed that the formation of iron plaque on root surfaces was promoted by exogenous iron, with greater amount of iron plaque being formed by addition of ferric hydroxide than of ferric oxide. Phosphorus adsorption on iron plaque also increased with the addition of iron oxides, and increasing soil P increased the amounts of P associated with the iron plaque and shoot P concentration. The amounts of iron plaque were almost sixfold higher under flooding condition than under field capacity condition. Plants pretreated under flooding condition generally had higher shoot P concentrations when they were transplanted to solutions with varying P levels, and this was most pronounced in the treatment with highest solution P concentration. The results suggest that iron plaque acts as a nutrient reservoir for phosphorus in the rhizosphere and helps enhance P acquisition by rice.     

Changes in water content of two agricultural soils does not alter labile P and C pools

Aims

An incubation study was conducted to investigate how changes in soil water content affect labile phosphorus and carbon pools, mineralisation patterns and microbial community composition.

Methods

Two soils from different climatic histories were subjected to four long-term (15 weeks) soil water regimes (constant field capacity (m); 3 dry-rewet (DRW) cycles evenly spaced (intermittent, int); 3 DRW cycles with a shorter interval after a long dry period (false break, fb); constantly air-dry (d)) (incubation period 1). In the subsequent incubation period 2, a set of cores from each treatment were subjected to one DRW cycle (air-dry for 7 day; field capacity for 14 day) or maintained at field capacity.

Results

Long-term soil water regime altered soil respiration with the largest CO₂ pulse occurring in soil with the longest dry period. However, changing the distribution of the 3 DRW events within incubation period 1 (int/fb) did not alter cumulative CO₂. In addition, DRW during incubation period 2 did not affect cumulative CO₂ in either treatment (m, int, fb, d) (except for Hamilton int). Our results show that carbon and phosphorus availability and the size and community composition of the microbial biomass were largely unaffected by fluctuating soil water content.

Conclusions

Changes in soil water content altered respiration, phosphatase activity and microbial C:P ratio and indicate physiological and/or functional changes in the microbial community. However, it appeared that these would have little impact on plant P availability.     

Coating Soybean Seed with Oxamyl for Control of Heterodera glycines

Oxamyl coated on soybean (Glycine max (L.) Merr. cv. Elgin) seeds in solutions of 20, 40, 80, and 160 mg/ml had no serious deleterious effects on seedling emergence and growth when planted in sterile soil. Seedling emergence on day 3 was less than that of the uncoated control, but by day 7 emergence was equal to, or greater than, the control. Shoot and root growth from seed coated with oxamyl in 40 and 80 mg/ml solutions was greater than that of the control. In soil infested with soybean cyst nematode, Heterodera glycines, shoot weight of soybean plants from seeds coated with oxamyl in 80 mg/ml solution was 11 and 9% greater at weeks 3 and 7, respectively, than from uncoated seeds. Numbers of juveniles (J3 and J4) and adults of H. glycines observed on the roots of plants from oxamyl-coated seeds were 83, 42, and 49% less at weeks 3, 5, and 7, respectively, than numbers on the roots of the untreated control. Numbers of J2 extracted from the roots of plants from oxamyl-coated seeds were 75% less at weeks 5 and 7 than those extracted from roots of uncoated seeds. The numbers of J2 extracted from the soil planted to oxamyl-coated seeds were 51 and 33% less at weeks 5 and 7, respectively, than from soil planted to uncoated seed.     

An economical system for liquid scintillation counting

Small glass shell vials (12 × 35 mm minivials), containing 2.0 ml of a dioxane-based scintillation solution plus a ¹⁴C-labeled sample, were placed in a conventional glass, 20-ml count vial and assayed in a scintillation spectrometer. Statistical comparison of counts recorded from ¹⁴C samples prepared both in the minivial system and conventional 20-ml count vials indicated that the two systems were equivalent with sample volumes of 10 and 100 μliters (1600-cpm solution) and 10 μliters (60-cpm solution). Conventional 20-ml glass or plastic count vials were both acceptable as containers for the minivials.There were no significant differences in the counts from samples in minivials placed on-center and off-center in the container vial. Cost per sample was reduced from 24.8 cents (conventional glass vials) to 4.7 cents (minivial system).     

Effect of period of air-dry storage of soils on the subsequent accumulation of mineral nitrogen during incubation

Summary The accumulation of NH₃ and NO₃ during incubation, under standard conditions, of nine mineral soils was determined following one week and 12 weeks air-dry storage.NH₃ accumulation was usually slightly greater whilst NO₃ accumulation was usually less during incubation after 12 weeks than after one week of air-dry storage. However, for the soils as a whole there were only small differences in either constituent accumulating during incubation due to period of air-dry storage.Total mineral nitrogen accumulation (NH₃ ^–+NO₃ ^–N) was very similar for four of the soils, was lower for three of the soils, and was higher for two of the soils during incubation following 12 weeks than following 1 week of air-dry storage. The average values for the soils as a whole were only slightly different between the two periods of air-dry storage.     

Survival of Cowpea Rhizobia in Soil as Affected by Soil Temperature and Moisture

Successful inoculation of peanuts and cowpeas depends on the survival of rhizobia in soils which fluctuate between wide temperature and moisture extremes. Survival of two cowpea rhizobial strains (TAL309 and 3281) and two peanut rhizobial strains (T-1 and 201) was measured in two soils under three moisture conditions (air-dry, moist (−0.33 bar), and saturated soil) and at two temperatures (25 and 35°C) when soil was not sterilized and at 40°C when soil was sterilized. Populations of rhizobia were measured periodically for 45 days. The results in nonsterilized soil indicated that strain 201 survived relatively well under all environmental conditions. The 35°C temperature in conjunction with the air-dry or saturated soil was the most detrimental to survival. At this temperature, the numbers of strains T-1, TAL309, and 3281 decreased about 2 logs in dry soil and 2.5 logs in saturated soil during 45 days of incubation. In sterilized soil, the populations of all strains in moist soil increased during the first 2 weeks, but decreased rapidly when incubated under dry conditions. The populations did not decline under saturated soil conditions. From these results it appears that rhizobial strains to be used for inoculant production should be screened under simulated field conditions for enhanced survival before their selection for commercial inoculant production.     

Release of fossil methane from mineral soil particles, and its implication for estimation of methane oxidation in a mineral subsoil

In a preliminary experiment we found that methane evolved from a sandy subsoil during aerobic incubation of shaken soil slurries. In the study presented here the methane was found to be released from the sand particles by mechanical weathering, caused by the grinding effect of the shaking. Large amounts of gas (about 0.5 ml gas g^–1 soil) were extracted by intense grinding of the soil in gas tight serum vials. Methane was the main hydrocarbon in the emitted gas, but also a considerable amount of ethane was present, as well as minor amounts of heavier hydrocarbons (up to C₆). The ¹³C-values of the emitted methane and ethane were –33 and –29 , respectively. Together these results demonstrate a thermogenic origin of the gas. This paper also reports the results of an incubation experiment where possible methane oxidation was looked for. If a possible release of methane is not accounted for, methane oxidation may be overlooked, as illustrated in this paper. Methane consumption was detected only in soil from 40 cm, in contrast to soil sampled at 100 cm and deeper where a slight production was measured. When methane oxidation was inhibited by dimethyl-ether, a significant release of methane was seen. The release was probably caused by chemical weathering. When this methane release was taken into account, methane oxidation was found to be present at all measured depths (40 to 200 cm). Fertilization with urea inhibited the methane oxidation at 40 cm but not at deeper layers. It is hypothesized that ammonia oxidizing bacteria were the main methane oxidizers in this mineral subsoil (deeper than 1 m), and that oxidation of methane might be a survival mechanism for ammonia oxidizers in ammonia limited environments.     

Improving Heat Transfer at the Bottom of Vials for Consistent Freeze Drying with Unidirectional Structured Ice

The quality of lyophilized products is dependent of the ice structure formed during the freezing step. Herein, we evaluate the importance of the air gap at the bottom of lyophilization vials for consistent nucleation, ice structure, and cake appearance. The bottom of lyophilization vials was modified by attaching a rectified aluminum disc with an adhesive material. Freezing was studied for normal and converted vials, with different volumes of solution, varying initial solution temperature (from 5°C to 20°C) and shelf temperature (from ?20°C to ?40°C). The impact of the air gap on the overall heat transfer was interpreted with the assistance of a computational fluid dynamics model. Converted vials caused nucleation at the bottom and decreased the nucleation time up to one order of magnitude. The formation of ice crystals unidirectionally structured from bottom to top lead to a honeycomb-structured cake after lyophilization of a solution with 4% mannitol. The primary drying time was reduced by approximately 35%. Converted vials that were frozen radially instead of bottom-up showed similar improvements compared with normal vials but very poor cake quality. Overall, the curvature of the bottom of glass vials presents a considerable threat to consistency by delaying nucleation and causing radial ice growth. Rectifying the vials bottom with an adhesive material revealed to be a relatively simple alternative to overcome this inconsistency.     

Radioimmunoassay of prostaglandin F2α using sephadex G-75 gel chromatography

The development of a “bound—free” separation technique and its application to the radioimmunoassay of prostaglandin F_2α is described. The method is simple, rapid, free of non-specific binding and could be performed either at 4°C or at room temperature. A total of 100 tubes could be subjected to “bound—free” separation in 30 min at 4°C. The bound fraction is collected directly into scintillation vials. The total column length was 9.5 cm, of which the bed volume was 2.5 ml. The PGF_2α radioimmunoassay incubation volume of 0.3 ml when bedded in required 1.4 ml of elution buffer to elute the antibody-bound fraction. The free fraction was washed out with 4.0 ml of buffer and the columns were ready for further use. A standard curve of high sensitivity (5 pg) and good reproducibility (CV %: intra-assay = 6.54; inter-assay = 9.68) was obtained.     

Acetylene reduction assay for nitrogen fixation under field conditions in remote areas

Summary A method for intensively sampling soil for nitrogen fixation potential using acetylene reduction assay is discussed. Acetylene was generated from calcium carbide. Soil cores were incubated in Mason jars with specially adapted lids. Air samples from the jars were stored and transported over dry KOH in 10 ml serum vials. The method overcomes many problems associated with other sampling procedures, and produces statistically reproducible data.Contribution #4 — Devon Island IBP Project and CCIBP contribution #173.Contribution #4 — Devon Island IBP Project and CCIBP contribution #173.     

Statistical procedure for evaluating the sensitivity of Limulus amoebocyte lysate by using a reference lysate.

A study was designed to estimate variability of the Limulus amoebocyte lysate test by comparing a reference lysate against itself. Three technicians performed parallel tests, i.e., titrated side by side, the contents of two vials of reference lysate on 4 different days using 24 vials of the United States reference lysate and 12 vials of the United States reference endotoxin. Each parallel test was replicated three times. From the sensitivity endpoints, ratios were calculated for each parallel test. These ratios were converted to the logarithm for estimating variability among technicians and among vials of endotoxin. By using the overall variability of log ratios, a statistical procedure was developed to evaluate the sensitivity of each lot of licensed lysate submitted to the Bureau of Biologics for release.     

Indirect effect of abscisic acid on the induction of secondary dormancy in lettuce seeds

During temporary incubation at 25°C in buffered solutions (pH 4.0) of abscisic acid (ABA) seeds of lettuce ( Lactuca sativa L. cv. Olof) lost the red-light initiated ability to germinate in buffer. The development of secondary dormancy required an inhibitory ABA content in the seeds during a number of days. A temporary incubation in ABA during 24 h met these requirements only if the solution was about 100-fold more concentrated than during continuous incubation. Studies with 2-¹⁴C-ABA showed that the amount of ABA which had penetrated in 24 h was reduced by a factor 100 within 3 to 4 days during subsequent incubation in buffer. Both leaching and metabolic changes were involved in the reduction process. The nature of the metabolic products remained obscure. A shift to 2°C after incubation in ABA prevented the induction of secondary dormancy, but inhibited ABA metabolism. ABA did not interfere with the induction rate of secondary dormancy, and it was not required to maintain the state of dormancy. The sole function of ABA was the non-specific inhibition of germination, which indirectly facilitated the development of an ABA independent secondary dormancy. – The level of endogenous ABA was compared to the amount of ABA found in the embryo during and after incubation in ABA solutions marked with 2-¹⁴C-ABA. The level of endogenous ABA in air-dry seeds (0.11 ng/mg dry weight) corresponded to the minimal level at which penetrated ABA inhibited germination. This level had to be present at least during 4 to 5 days to inhibit the effect of red light. Since endogenous ABA was quickly reduced upon imbibition, a regulatory function of endogenous ABA in the inhibition of red light induced germination can be ruled out. A function in the temporary inhibition of dark germination and, consequently, in the development of secondary light irresponsiveness cannot be excluded, however.     

Temporary flooding increases iron phytoavailability in calcareous Vertisols and Inceptisols

Temporary soil flooding before cultivation alleviates iron chlorosis in crops grown on some calcareous Mexican Vertisols. In order to investigate the effectiveness of such practice we carried out experiments with ten calcareous Vertisols from Mexico and eight calcareous Inceptisols from Spain. In an incubation experiment, we studied the release of Fe²⁺ into the solution of soil suspensions in sealed vials with 5 m M CaCl₂. In a pot experiment, we measured the leaf SPAD value (i.e. an estimate of leaf chlorophyll concentration) of lupin and strawberry sequentially grown on a soil-sand mixture previously flooded for 30 days (SPAD_f value) and on a non-flooded (control) mixture (SPAD_c value). The amount of Fe²⁺ released by the soil at day 58 and the increase in oxalate-extractable Fe (Fe_o) upon incubation in vials were larger on average for the Inceptisols than for the Vertisols. The SPAD_c values for lupin and strawberry were (i) larger for the Vertisols than for the Inceptisols (probably because the Vertisols contain little carbonate and induce less Fe chlorosis than the Inceptisols) and (ii) correlated with Fe_o, and with citrate/ascorbate- and DTPA-extractable Fe (Fe_ca, Fe_DTPA). The SPAD_f-SPAD_c differencewas (i) much larger for the Inceptisols than for the Vertisols and (ii) correlated with the increases in Fe_o and Fe_ca caused by flooding and with the amount of Fe²⁺ released in the incubation experiment. We hypothesize that the weak response of the Vertisols to flooding was partly a result of their history including flooding episodes in the field, so a steady state had been reached in which the pool of Fe compounds undergoing reductive dissolution and reprecipitating upon oxidation as poorly crystalline Fe oxides (the main source of phytoavailable Fe) remained relatively constant and thus changed little after pot flooding. The Inceptisols, which had never been flooded in the field, were capable of releasing Fe from sources other than poorly crystalline Fe oxides upon flooding, thus making this treatment effective against Fe chlorosis. Our results point to the need to further study those soil chemical and mineralogical properties that are related to increases in Fe phytoavailability upon temporary soil flooding.     

Statistical procedure for evaluating the sensitivity of Limulus amoebocyte lysate by using a reference lysate.

A study was designed to estimate variability of the Limulus amoebocyte lysate test by comparing a reference lysate against itself. Three technicians performed parallel tests, i.e., titrated side by side, the contents of two vials of reference lysate on 4 different days using 24 vials of the United States reference lysate and 12 vials of the United States reference endotoxin. Each parallel test was replicated three times. From the sensitivity endpoints, ratios were calculated for each parallel test. These ratios were converted to the logarithm for estimating variability among technicians and among vials of endotoxin. By using the overall variability of log ratios, a statistical procedure was developed to evaluate the sensitivity of each lot of licensed lysate submitted to the Bureau of Biologics for release.     

Sporulation of Clostridium perfringens (welchii) in Four Laboratory Media

S ummary . Sporulation of 7 strains of Clostridium perfringens ( welchii ) was investigated in 4 laboratory media. A method to induce rapid and simultaneous sporulation was attempted which involved obtaining a purely vegetative culture to inoculate the test media. Heat resistance of spores produced in the individual media by each of 4 selected strains was investigated. The clean spores for the heating tests were obtained by a special procedure which included chilling to 6° for a minimum of 1 week immediately following the usual incubation period, then centrifuging, resuspending to volume in 0.85% NaCl solution and pasteurizing at 75° for 20 min before subjecting to the heating tests. Morphology of each strain was studied using stained microscopic preparations from the 24 h sporulating cultures.
In the Ellner medium spore counts approaching 10⁷/ml were recorded and this medium appeared to be the most efficient when judged in terms of numbers of spores produced. In other media the counts were in the range 10⁴-10⁵ spores/ml. Cooked meat medium yielded slightly higher spore counts than did either SEC broth or modified Wagenaar & Dack medium, the latter contained in a dialysis sac apparatus. A period of chilling to 6° for a minimum of 1 week following incubation enhanced maturation in all cultures except those grown in SEC broth for 24 h or 15 days and those grown 15 days in the modified Wagenaar & Dack medium.
Considerable heat resistance, expressed as percentage spore survival, was recorded for spores of 4 strains when heated at 80°, and heat resistance generally increased with lengthening of incubation time for the culture. Survival of spores heated at 100° for 10 min was usually less than 0.01% but spores in SEC broth after 15 days showed a somewhat greater heat resistance than the others. In no instance did total destruction of spores occur at 100°.     

Reactivity and fate of benzene and formaldehyde in culture medium with and without fetal calf serum; relevance to in vitro mutagenicity testing

Gas chromatographic-mass spectrometric analyses were performed to determine the reactivity and fate of benzene (BEN) and formaldehyde (FA) in culture medium. BEN (solubility in water: 500 ppm) does not react with culture medium, either with or without fetal calf serum, but its volatility, even in closed vials, is so great that 90% of a 250-ppm solution is lost to the head space after 1 h at 24°C. FA, as a 37% aqueous solution, is a complex mixture that changes composition after 15-min incubation at 38°C. FA is extremely reactive in culture medium containing fetal calf serum, and is much less reactive with medium components in the absence of serum. There is a dramatic increase in the number of daughter products in FA-treated medium over time, such that those seen immediately after FA is added to medium have been replaced after 60-min incubation (38°C in closed vials) by many other interaction products. Methods ensuring maximum solubilization and minimal volatilization of BEN during exposure are essential for obtaining reproducible data on the mutagenic potential of BEN. The volatilization of FA from stock formalin solutions, and, more importantly, the interaction product(s) formed by this highly reactive compound with medium components, especially those in serum, are probably the critical aspects of an effective testing protocol for FA.     

Leukocytes Cultured from Small Inocula of Whole Blood and the Preparation of Metaphase Chromosomes by Treatment with Hypotonic KCL

Leukocytes were cultured from 0.2 ml of whole blood inoculated into 5 ml portions of a medium consisting of Eagle's basal amino acids and vitamins at double strength in Earle's balanced salt solution brought to pH 7.0 with 7.5% NaHCO₃, and containing additives: glutamine, 2 mM; penicillin, 100 units/ml; streptomycin, 100 μg/ml; phenol red, 7 μg/ml; fetal or newborn agammaglobulin bovine serum, 15%; phytohemagglutinin M, 2%; and U.S.P. heparin sodium, 20,000 units/liter. Cultures were incubated in closed 60 × 28 mm screw-cap vials, in a gas phase initially of room air, for 3 days at 37 C, with colchicine to make 0.2 μg/ml added for the final 3-5 hr. After incubation, the cells were separated from the medium by centrifugation, the medium replaced by 0.075 M KCI plus 16 U.S.P. units/ml of heparin sodium at 37 C, cells resuspended and allowed to incubate 10 min. Removal of the hypotonic KCI was followed by fixation in methanol-acetic acid, 3:1 (changed twice), spreading cells on slides by the air-drying method, and staining with 1% natural orcein (G. T. Gurr) in 60% acetic acid. Dehydration and covering completed the preparation. KCI, 0.075 M, has been used advantageously in the above way and for cells cultured by other means from skin and other organs of man and other mammals. Combined advantages of the method are: culture of leukocytes from small volumes of whole blood, with very few failures to obtain mitotic cells; a medium which can be stored frozen in culture vials, and which in a simpler form is usable for long term culture of other cell types; and the use of KCI for hypotonic treatment.