Epidermal Growth Factor-Mediated Mitogen-Activated Protein Kinase3/1 Pathway Is Conducive to In Vitro Maturation of Sheep Oocytes

Epidermal growth factor (EGF) has been shown to facilitate the in vitro maturation of sheep oocytes, and enhance embryo’s capability for further development. However, such kind of molecular mechanism has not yet been elucidated. In the present study, we investigated the effect of EGF-mediated mitogen-activated protein kinases 3 and 1 (MAPK3/1) pathway on in vitro maturation of sheep oocytes. U0126, a specific inhibitor of MEK (MAPK kinase), was added into the maturation culture medium to block the EGF-mediated MAPK3/1 pathway with different doses. Then, the nuclear maturation of sheep oocytes was examined. Additionally, the effect of EGF-mediated MAPK3/1 on cytoplasmic maturation was examined though in vitro fertilization and embryonic development. The rate of germinal vesicle breakdown (GVBD) after 6 h of culture with 10⁻⁴ mol/l of U0126 (50.4%) was significantly decreased compared with control (67.2%, p < 0.05), and the first polation body (PB1) extrusion rate after 22 h of culture in drug treatment was also significantly inhibited compared with control (28.6% vs. 48.4%, p < 0.05). However, 10⁻⁶ mol/l U0126 had slight effect on oocyte nuclear maturation. The normal distribution rate of α-tubulin in the oocytes after 22 h of in vitro maturation was significantly decreased in the 10⁻⁴ mol/l U0126 group (54%) compared with control (68%, p < 0.05). After in vitro fertilization, the cleavage rate in drug treatments (56.8% in 10⁻⁶ mol/l U0126 group and 42.6% in 10⁻⁴ mol/l U0126 group) was significantly decreased compared with control (72.3%, p < 0.01). The blastocyst rate in 10⁻⁴ mol/l U0126 group (17.6%) was also significantly decreased compared with control (29.9%, p < 0.05). Collectively, these results suggest that EGF-mediated MAPK3/1 pathway is conducive to in vitro maturation of sheep oocytes.     

3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Activity in Ochromonas malhamensis: A System to Study the Relationship between Enzyme Activity and Rate of Steroid Biosynthesis

3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the key regulatory enzyme of the isoprenoid pathway, was found to be predominantly microsomal in Ochromonas malhamensis, a chrysophytic alga. Detection of HMG-CoA reductase requires the presence of 1% bovine serum albumin during cell homogenization, and the activity is stimulated by the presence of Triton X-100. The enzyme has a pH optimum of 8.0 and an absolute requirement for NADPH. When grown in 10 micromolar mevinolin, a competitive inhibitor of HMG-CoA reductase, O. malhamensis shows a 10- to 15-fold increase in HMG-CoA reductase activity (after washing) with little or no effect on cell growth rate. Cultures can be maintained in 10 micromolar mevinolin for months. O. malhamensis produces a large amount (1% dry weight) of poriferasterol, a product of the isoprenoid pathway. The addition of 10 micromolar mevinolin initially blocked poriferasterol biosynthesis by >90%; within 2 days the rate of synthesis returned to normal levels. Immediately after mevinolin was washed from the 2-day culture, there was a transient 2.5-fold increase in the rate of poriferasterol biosynthesis. The rate of poriferasterol biosynthesis and the level of HMG-CoA reductase activity both fell to control levels within hours.     

Immobilization of Pseudomonas cepacia lipase onto electrospun polyacrylonitrile fibers through physical adsorption and application to transesterification in nonaqueous solvent

The lipase of Pseudomonas cepacia was immobilized onto electrospun polyacrylonitrile (PAN) fibers and used for the conversion of (S)-glycidol with vinyl n-butyrate to glycidyl n-butyrate in isooctane. The rate of reaction with the adsorbed lipase was 23-fold higher than the initial material. After 10 recyclings, the initial reaction rate was 80% of the original rate. This system of enzyme immobilization is therefore suitable for carrying out transesterification reactions in nonaqueous solvents.     

Autonomic control of heart rate is virtually independent of temperature but seems related to the neuroanatomy of the efferent vagal supply to the heart in the bullfrog, Lithobathes catesbeianus

Bullfrogs, Lithobathes catesbeianus, bearing a femoral artery cannula were held at 3 temperatures (10, 20 and 30 °C) for 24 h. Changes in heart rate were recorded before and after injection of cholinergic and adrenergic antagonists. Normal heart rate doubled for each temperature increment. Adrenergic tone on the heart varied around 20% at all 3 temperatures but cholinergic tone increased from −5% to 10% between 10 and 30 °C. In contrast, cholinergic tone increased from 75% at 5 °C to 329% at 25 °C in Xenopus laevis. Injection of the neural tracer True Blue into the cervical vagus of the bullfrog revealed a single location for vagal preganglionic neurons (VPN) in the dorsal vagal motor nucleus (DVN), while Xenopus had 30% of its VPN in a ventro-lateral group outside the DVN. Broader comparative studies have suggested that differences in the extent of vagal tone may relate to the location of VPN in the brainstem and this may be the case in these amphibians.     

Effects of salinity on the respiration and heart rate of the common mussel, Mytilus edulis L., and the black chiton, Katherina tunicata (Wood)

The rate of oxygen consumption of stepwise acclimated Mytilus edulis L. increased linearly from 30 to 10‰ salinity (S) while that of Katherina tunicata (Wood) was not significantly different between 10 and 30‰ S. Heart rate was 21–22 and 17–18 beats m^?1 in Mytilus edulis and Katherina tunicata, respectively, and no difference was found in the heart rate of either species acclimated stepwise to 10, 20 or 30‰ S. The average oxygen consumption rate of Mytilus edulis exposed to 12 h, 30-10-30 and 10-30-10‰ S cycles of fluctuating salinity was significantly lower than the respective control rate: there was a similar response during the 30-10-30‰ S cycle in Katherina tunicata. The respiration rate of Mytilus edulis and Katherina tunicata declined as salinity deviated from the control salinity and increased as salinity returned to the control salinity. The rate of oxygen consumption by K. tunicata varied directly with the ambient salinity during the 10-30-10‰ S cycle. The average heart rate of Mytilus edulis was significantly lower during cyclic changes in salinity than at the respective control salinities; a similar relationship existed for Katherina tunicata during the 10-30-10‰ S cycle. Heart rate of Mytilus edulis varied in a parallel manner with oxygen consumption during both cycles. Katherina tunicata heart rate was relatively constant and could not be fitted to a regression line during the 10-30-10‰ S cycle. The normalized heart rate increased to 113% of control at 10‰ S of the 30-10-30‰ S cycle and returned to the control rate by 12 h. The oxygen consumption and heart rate of these two species are not directly coupled to regulation of water volume because different responses are observed with respect to salinity although there is poor water volume regulation in both species.     

Microbial hydrolysis of 7-xylosyl-10-deacetyltaxol to 10-deacetyltaxol

Enterobacter sp. CGMCC 2487, a bacterial strain isolated from the soil around a Taxus cuspidata Sieb. et Zucc. plant, was able to remove the xylosyl group from 7-xylosyltaxanes. The xylosidase of this strain was an inducible enzyme. In the bioconversion of 7-xylosyl-10-deacetyltaxol (7-XDT) to 10-deacetyltaxol (10-DT), for the purpose of enhancing the conversion efficiency, the effects of NH₄⁺, oat xylan, temperature, pH value, cell density and substrate concentration on the bioconversion have been systematically investigated. 3.0 mM NH₄⁺, 0.6% oat xylan in the media could enhance the yield of 10-DT; the optimum biocatalytic temperature was 26 °C and optimum pH value was 6.0. The highest conversion rate and yield of 10-DT from 7-XDT reached 92% and 764 mg/L, respectively. In addition, the biocatalytic capacity of the cell cultures remained 66.1% after continuous three batches. These results indicate that converting 7-XDT to 10-DT, a useful intermediate for the semisynthesis of paclitaxel or other taxane-based anticancer drugs by a novel bacterial strain, Enterobacter sp. CGMCC 2487, would be an alternative for the practical application in the future.     

Using combined measurements of gas exchange and chlorophyll fluorescence to investigate the photosynthetic light responses of plant species adapted to different light regimes

One broad-leaved pioneer tree, Alnus formosana, two broad-leaved understory shrubs, Ardisia crenata and Ardisia cornudentata, and four ferns with different light adaptation capabilities (ranked from high to low, Pyrrosia lingus, Asplenium antiquum, Diplazium donianum, Archangiopteris somai) were used to elucidate the light responses of photosynthetic rate and electron transport rate (ETR). Pot-grown materials received up to 3 levels of light intensity, i.e., 100%, 50% and 10% sunlight. Both gas exchange and chlorophyll (Chl) fluorescence were measured simultaneously by an equipment under constant temperature and 7 levels (0?C2,000 ??mol m^?2 s^?1) of photosynthetic photon flux density (PPFD). Plants adapted to-or acclimated to high light always had higher light-saturation point and maximal photosynthetic rate. Even materials had a broad range of photosynthetic capacity [maximal photosynthetic rate ranging from 2 to 23 ??mol(CO₂) m^?2 s^?1], the ratio of ETR to gross photosynthetic rate (P _G) was close for A. formosana and the 4 fern species when measured under constant temperature, but the PPFD varied. In addition, P. lingus and A. formosana grown under 100% sunlight and measured at different seasonal temperatures (15, 20, 25, and 30°C) showed increased ETR/P _G ratio with increasing temperature and could be fitted by first- and second-order equations, respectively. With this equation, estimated and measured P _G were closely correlated (r ² = 0.916 and r ² = 0.964 for P. lingus and A. formosana, respectively, p<0.001). These equations contain only the 2 easily obtained dynamic indicators, ETR and leaf temperature. Therefore, for some species with near ETR/P _G ratio in differential levels of PPFD, these equations could be used to simulate dynamic variation of leaf scale photosynthetic rate under different temperature and PPFD conditions.     

Influence of replacement rate on the welfare of sows introduced to a large dynamic group

This experiment investigated the effects of replacing 10, 20, 30 or 40% of a dynamic group of forty sows on the welfare of newly-introduced animals. The experiment was replicated five times, using a total of 200 multiparous sows. Replacements took place at 3 week intervals, and 3 days prior to sows being added to the group the same number of animals were removed from the group. Sows were added to the dynamic group as pre-formed groups of four animals which had resided together for a period of 5 weeks beginning directly after their piglets were weaned. Replacement rate did not appear to influence overall levels of aggression to which newly-introduced animals were exposed on the day of mixing, however aggression among newly-introduced animals increased significantly as replacement rate increased between 20 and 40% (P<0.05). Injury levels measured 1 week post mixing did not differ significantly between treatments. Newly-introduced sows in the 10% replacement rate treatment spent more time in slatted areas and less time in kennel areas during the first week post mixing than newly-introduced sows in other treatments (P<0.05). In addition, during the first week post mixing when sows in the 10% replacement rate treatment entered kennel areas they spent less time lying down (P<0.05), and when they did lie down they spent less time lying in contact with other animals and more time lying close to the wall (P<0.05) compared to newly-introduced sows in other treatments. It is suggested that this type of behaviour represents intimidation from resident sows. Newly-introduced sows in the 20, 30 and 40% replacement rate treatments showed similar lying behaviour. In all treatments the newly-introduced sows showed a significant increase in the time spent in kennel areas, and the time spent lying in contact with ‘resident’ sows during the first week post mixing, which suggests that the process of social integration with the resident group had begun. The results of the present study suggest that the welfare of newly-introduced sows in dynamic groups may be compromised when replacement rates of 10% are used. There appears to be no additional welfare benefit associated with increasing replacement rate above 20%.     

Antagonistic yeasts and thermotherapy as seed treatments to control Fusarium fujikuroi on rice

Bakanae disease, caused by Fusarium fujikuroi, is the most important seedborne disease of rice. Biological control and physical treatments can be effective tools to control seedborne diseases. Sixty-two isolates of yeasts and yeast-like fungi were obtained from different rice seeds. Four yeast isolates were selected in dual culture assays for mycelial growth inhibition, and in seed tests for reduction of infection rate. The isolates R23 and R26 were identified as Metschnikowia pulcherrima, the isolate R9 as Pichia guilliermondii, and the isolate SB1 as Sporidiobolus pararoseus. Rice seeds treated with P. guilliermondii R9, M. pulcherrima R23 and R26 significantly reduced the infection rate of F. fujikuroi, compared to some commercial biofungicides. The four selected yeasts reduced the bakanae disease severity in rice plants grown in greenhouse trials. Antagonist seed dressing resulted in reduction of the disease index from 93.0% in the untreated control to 20.0% in P. guilliermondii R9 treated seeds, and to 28.5% in M. pulcherrima R23 treated seeds. Selected antagonists were also used in combination with thermotherapy, which contributed to increase their efficacy. Thus, P. guilliermondii R9 and M. pulcherrima R23 combined with thermotherapy at 60 °C for 10 min decreased the bakanae disease index below 5%, and improved the seed germination rate compared to the single treatments, showing a seed priming effect. This is the first report about the use of antagonistic yeasts for seed dressing of rice to control F. fujikuroi on rice seeds, and biological treatment may be improved through combination with thermotherapy.     

Effects of naloxone on renal responses to noxious stimuli in the cynomolgus monkey

Behavioral influences on excretion of Na⁺ and H₂O were studied during saline diuresis in three unanesthetized adult female cynomolgus monkeys. During control infusions of isotonic saline, the average rates of urine and Na⁺ excretion were 210μl/kg/min and 36.1μl/kg/min, respectively, and the average rate of inulin clearance was 4.6 ml/kg/min. Intermittent exposure to an electrical stimulus applied to the monkey's tail for a 30-min period modestly reduced rates of excretion of Na⁺ and H₂O; these reductions were 58% and 56% of baseline values respectively during the first 10 min, but excretion rates returned to baseline values or exceeded them by the end of the 30-min period. The effects of naloxone hydrochloride (10 mg/kg), an opiate antagonist, were studied by administering the drug immediately before the period of electrical-stimulus delivery. After naloxone, the electrical-stimulus markedly reduced the rates of Na⁺ and H₂O excretion to 29% and 31% of baseline values during the first 10 min, and delayed the return to baseline values. Inulin clearance was not altered significantly by the electrical stimulus in the absence of naloxone, but was decreased to 32% of the baseline rate during the first 10 min of exposure to the electric stimulus in the presence of naloxone. Naloxone had similar effects on rates of Na⁺ and urine excretion in response to 30 min of 108 dBA noise. These results show that renal responses to noxious environmental stimuli (electrical stimulus or noise) can be altered by naloxone.     

Sensitivity of Plasmodium falciparum to Antimalarial Drugs in Hainan Island,China

Pyronaridine and artesunate have been shown to be effective in falciparum malaria treatment. However, pyronaridine is rarely used in Hainan Island clinically, and artesunate is not widely used as a therapeutic agent. Instead, conventional antimalarial drugs, chloroquine and piperaquine, are used, explaining the emergence of chloroquine-resistant Plasmodium falciparum. In this article, we investigated the sensitivity of P. falciparum to antimalarial drugs used in Hainan Island for rational drug therapy. We performed in vivo (28 days) and in vitro tests to determine the sensitivity of P. falciparum to antimalarial drugs. Total 46 patients with falciparum malaria were treated with dihydroartemisinin/piperaquine phosphate (DUO-COTECXIN) and followed up for 28 day. The cure rate was 97.8%. The mean fever clearance time (22.5±10.6 hr) and the mean parasite clearance time (27.3±12.2 hr) showed no statistical significance with different genders, ages, temperatures, or parasite density (P>0.05). The resistance rates of chloroquine, piperaquine, pyronarididine, and artesunate detected in vitro were 71.9%, 40.6%, 12.5%, and 0%, respectively (P<0.0001). The resistance intensities decreased as follows: chloroquine>piperaquine>pyronarididine>artesunate. The inhibitory dose 50 (IC₅₀) was 3.77×10^-6 mol/L, 2.09×10^-6 mol/L, 0.09×10^-6 mol/L, and 0.05×10^-6 mol/L, and the mean concentrations for complete inhibition (CIMC) of schizont formation were 5.60×10^-6 mol/L, 9.26×10^-6 mol/L, 0.55×10^-6 mol/L, and 0.07×10^-6 mol/L, respectively. Dihydroartemisinin showed a strong therapeutic effect against falciparum malaria with a low toxicity.     

EGFR Mutations in Indian Lung Cancer Patients: Clinical Correlation and Outcome to EGFR Targeted Therapy

Screening for EGFR mutation is a key molecular test for management of lung cancer patients. Outcome of patients with mutation receiving EGFR tyrosine kinase inhibitor is known to be better across different ethnic populations. However, frequency of EGFR mutations and the clinical response in most other ethnic populations, including India, remains to be explored. We conducted a retrospective analysis of Indian lung cancer patients who were managed with oral tyrosine kinase inhibitors. Majority of the patients in the study had adenocarcinoma and were non-smokers. 39/111 patients tested positive for EGFR kinase domain mutations determined by Taqman based real time PCR. The overall response to oral TKI therapy was 30%. Patients with an activating mutation of EGFR had a response rate of 74%, while the response rate in patients with wild type EGFR was 5%, which was a statistically significant difference. Progression free survival of patients with EGFR mutations was 10 months compared to 2 months for EGFR mutation negative patients. Overall survival was 19 months for EGFR mutation patients and 13 months for mutation negative patients. This study emphasizes EGFR mutation as an important predictive marker for response to oral tyrosine kinase inhibitors in the Indian population.     

QTLs for resistance to Phomopsis seed decay are associated with days to maturity in soybean (Glycine max)

Phomopsis seed decay (PSD), primarily caused by Phomopsis longicolla, is a major contributor to poor soybean seed quality and significant yield loss, particularly in early maturing soybean genotypes. However, it is not yet known whether PSD resistance is associated with early maturity. This study was conducted to identify quantitative trait loci (QTLs) for resistance to PSD and days to maturity using a recombinant inbred line (RIL) population derived from a cross between the PSD-resistant Taekwangkong and the PSD-susceptible SS2-2. Based on a genetic linkage map incorporating 117 simple sequence repeat markers, QTL analysis revealed two and three QTLs conferring PSD resistance and days to maturity, respectively, in the RIL population. Two QTLs (PSD-6-1 and PSD-10-2) for PSD resistance were identified in the intervals of Satt100–Satt460 and Sat_038–Satt243 on chromosomes 6 and 10, respectively. Two QTLs explained phenotypic variances in PSD resistance of 46.3 and 14.1 %, respectively. At the PSD-6-1 QTL, the PSD-resistant cultivar Taekwangkong contributed the allele with negative effect decreasing the infection rate of PSD and this QTL does not overlap with any previously reported loci for PSD resistance in other soybean genotypes. Among the three QTLs for days to maturity, two (Mat-6-2 and Mat-10-3) were located at positions similar to the PSD-resistance QTLs. The identification of the QTLs linked to both PSD resistance and days to maturity indicates a biological correlation between these two traits. The newly identified QTL for resistance to PSD associated with days to maturity in Taekwangkong will help improve soybean resistance to P. longicolla.     

蚯蚓对湿地植物光合特性及净化污水能力的影响

以香蒲、芦苇和美人蕉为研究对象,并以土壤+沙子+有机质混合物为供试基质模拟人工湿地处理污水,采用向基质中加入蚯蚓与未加入蚯蚓2种处理。研究加入蚯蚓后,香蒲、芦苇和美人蕉光合速率、蒸腾速率、SPAD值和水分蒸发、蒸腾量的变化及其对净化污水能力的影响。结果表明:与未加入蚯蚓相比,加入蚯蚓后,香蒲、芦苇和美人蕉的净光合速率、蒸腾速率、SPAD值和水分蒸发、蒸腾量均增加,其中芦苇的净光合速率、蒸腾速率和水分蒸发、蒸腾量增加达到显著水平(P <0.05),而香蒲的水分蒸发、蒸腾量增加也达到显著水平(P <0.05);加入蚯蚓后,香蒲、芦苇和美人蕉对COD_Mn、NH₄⁺-N、NO₃^--N、TN和TP的去除率均增加,且香蒲和芦苇对COD_Mn的去除率显著增加 (P <0.05)。加入蚯蚓后,香蒲、芦苇和美人蕉的SPAD值均增加,说明蚯蚓能提高湿地植物对氮的吸收,增加植株中的氮含量,促进湿地植物的光合速率和蒸腾速率从而提高对污水的净化能力。     

Enkephalins modulate the responsiveness of rat atria in vitro to norepinephrine

Potential interactions between opiate peptides and catecholamines in mammalian heart were examined using isolated spontaneously beating rat atria as a test system. Methionine-enkephalin (ME), leucine-enkephalin (LE), phe-met-arg-phe amide (FMRFamide), D-ala², N-methyl-phe⁴, met (O)⁵-ol-enkephalin (FK 33-834), methionine-enkephalin arg⁶ arg⁷ (ME arg⁶ arg⁷) and β-endorphin had no effect on basal beating rate of isolated atria at all concentrations up to 10^?5 M. The positive chronotropic effect of norepinephrine (NE) on atrial rate is, however, significantly attenuated by enkephalin peptides. Thus, the maximal chronotropic effect of NE (an increase from 317±7.0 to 473±7.3 beats per minute (bpm) in 250 gm rats at a dose of 10^?5 M NE) is decreased by 42% in the presence of 10^?7 m ME. The action of ME is completely blocked by addition of 10^?7 M naloxone, which by itself has no effect on NE-induced positive chronotropy or basal beating rate. The dose-effect curve for ME attenuation of NE-induced positive chronotropy is bell-shaped, i.e., both 10^?8 M and 10^?5 M ME have no significant effect on NE positive chronotropy. Other enkephalin peptides acted in a similar manner to ME; LE (10^?7 M) and FK 33-834 (10^?8 M) decreased maximal NE-induced positive chronotropy 42 and 27%, respectively. The molluscan cardioexcitatory peptide FMRFamide (10^?7 M) also decreased maximal NE positive chronotropy, about 30%. In contrast, β-endorphin did not significantly affect NE stimulation of atrial rate. We conclude that enkephalins can modulate the noradrenergic responsiveness of rat atria in vitro. The possible physiological relevance of this interaction is discussed.     

A covalent complex between horse heart cytochrome c and yeast cytochrome c peroxidase: kinetic properties

The kinetic properties of a 1:1 covalent complex between horse-heart cytochrome c and yeast cytochrome c peroxidase (ferrocytochrome-c:hydrogen-peroxide oxidoreductase, EC 1.11.1.5) have been investigated by transient-state and steady-state kinetic techniques. Evidence for heterogeneity in the complex is presented. About 50% of the complex reacts with hydrogen peroxide with a rate 20–40% faster than that of native enzyme; 20% of the complex exists in a conformation which does not react with hydrogen peroxide but converts to the reactive form at a rate of 20 ± 5 s⁻¹; 30% of the complex does not react with hydrogen peroxide to form the oxidized enzyme intermediate, cytochrome c peroxidase Compound I. Intramolecular electron transfer between covalently bound ferrocytochrome c and an oxidized site in cytochrome c peroxidase Compound I is too fast to measure, but a lower limit of 600 s⁻¹ can be estimated at 5°C in a 10 mM potassium phosphate buffer at pH 7.5. Free ferrocytochrome c reduces cytochrome c peroxidase Compound I covalently bound to ferricytochrome c at a rate 10⁻⁴ to 10⁻⁵-times slower than for free Compound I. The transient-state ferrocytochrome c reduction rates of Compound I covalently linked to ferricytochrome c are about 70-times too slow to account for the steady-state catalytic properties of the 1:! covalent complex. This indicates that hydrogen peroxide can interact with the 1:1 complex at sites other than the heme of cytochrome c peroxidase, generating additional species capable of oxidizing free ferrocytochrome c.     

Combined effect of microbial and chemical control agents on subterranean termites

Termite mortality was measured when fungi were combined with bacteria or a chemical termiticide to determine whether a synergistic effect occurred. The fungus Beauveria bassiana was combined with the non-repellant chemical termiticide imidacloprid. Of the three B. bassiana strains tested one, B. bassiana ATCC 90519, was sufficiently pathogenic on its own that the advantage of a supplementary chemical treatment was marginal. The mortality caused by another fungal strain, B. bassiana ATCC 26037, was improved in combination with imidacloprid at both of the tested chemical concentrations over the first 14 days. The remaining fungal strain, B. bassiana ATCC 90518, demonstrated an overall mortality rate in combination with imidacloprid of 82.5%, versus a rate of 65.0% for the fungus alone. The fungus Isaria fumosorosea (Ifr) was combined with the bacterium Bacillus thuringiensis (Bt). On day 5, Ifr, Bt, and the combined treatment at a 10⁶ spores or cells/ml dosage caused 8.8%, 22.5%, and 15.0% mortality, respectively. The Bt and combined mortality rates are not significantly different. Control mortality on day 5 was 5.0%. On day 13 the combined 10⁶ treatment mortality rate was 91.3%, which was significantly higher than all other treatments: control at 17.5%, Ifr at 36.3% and Bt at 35.0%. When Ifr and Bt were applied at a 10⁹ spores or cells/ml dosage, Ifr alone caused a mortality rate of 97.5% as early as day 5. The combination with Bt could not significantly increase the effectiveness of this dosage. These data demonstrate the potential for synergistic effects of fungal and chemical treatment methods, thereby broadening the use of microbial control agents and reducing the quantity of chemical agents necessary to effect control.     

The rate of nucleic acid annealing to cytological preparations is increased in the presence of dextran sulfate

A hybridization buffer containing 10% dextran sulfate has been used to increase the rate of in situ nucleic acid reannealing to cytological preparations.     

Attractive Effects of American Serpentine Leafminer, Liriomyza trifolii (Burgess), to Light-Emitting Diodes

Phototactic responses of Liriomyza trifolii adults to six different light-emitting diodes (LEDs) were investigated, and their responses were compared to that using a luring lamp (BLB) under laboratory conditions. Based on the attraction rate under optimal light conditions, the green LED (520?±?5 nm) showed the highest attraction rate (99.7 %), followed by the yellow LED (590?±?5 nm, 96.1 %), the red LED (625?±?10 nm, 91.4 %), the blue LED (470?±?10 nm, 91.2 %), the UV LED (365 nm, 71.0 %), and the IR LED (730 nm, 5.6 %). Moreover, the green LED was approximately 1.4 times more attractive than BLB (71.1 %) to L. trifolii adults. These results suggest that the green LED was the most useful for monitoring of L. trifolii adults under optimal conditions.     

Persistence of spores of Pleistophora schubergi (Cnidospora: Microsporida) in the field,and their application in microbial control

Spores of Pleistophora schubergi, when applied to oak trees in the field at 2 × 10⁸ spores/ml with a uv protectant, “Shade,” infected 88% of Anisota senatoria larvae at 4 days after spray application. Spores without the uv protectant infected only 10% of the larvae at 4 days after application. When the spores were applied at the rate of 2 × 10⁸ and 2 × 10⁷ spores/ml in the field, 96 and 72% of the A. senatoria larvae and 100 and 100% of the Symmerista canicosta larvae were infected 14 days after spray application.