Insecticidal and neuroblocking potencies of variants of the imidazolidine moiety of imidacloprid-related neonicotinoids and the relationship to partition coefficient and charge density on the pharmacophore

The pharmacophore of the neonicotinoid insecticide imidacloprid, nitroiminoimidazolidine, was modified to heterocycles such as thiazolidine, pyrrolidine, dihydroimidazole, dihydrothiazole, and pyridone conjugated to nitroimine (=NNO2) or nitromethylene (=CHNO2). Their 6-chloro-3-pyridylmethyl or 5-chloro-3-thiazolylmethyl derivatives were examined for insecticidal activity against the American cockroach by injection and for neuroblocking activity using the cockroach ganglion. Most of the compounds having the neonicotinoidal pharmacophore exhibited insecticidal activity at the nanomolar level, which was enhanced in the presence of synergists, and high neuroblocking activity at the micromolar level. Quantitative analysis for the compounds showed that the neuroblocking potency is proportional both to the Mulliken charge on the nitro oxygen atom and to the partition coefficient log P value. The equation for the insecticidal versus neuroblocking potencies indicated that both potencies are related proportionally with each other when the other factors are the same.     

Determination of theaflavins including methylated theaflavins in black tea leaves by solid-phase extraction and HPLC analysis

A quantitative method for four theaflavins and two methylated theaflavin derivatives in black tea leaves was developed by solid-phase extraction and a high-performance liquid chromatographic method with photodiode array detection. The theaflavins in black tea leaves were extracted three times with 40 vol 50% aqueous ethanol (mg dry tea powder/mL) containing 2% ascorbic acid. The ethanol extracts were diluted 4-fold with distilled water. All diluted extracts were directly applied to the solid-phase C18 cartridge column without concentration. The fraction of theaflavins was obtained by 40% ethanol extraction after rinsing with water followed with 15% ethanol extraction. An aliquot of theaflavins after concentration was injected onto an ODS C18 reversed-phase column, and four theaflavins and two methylated theaflavins were sufficiently separated by a linear gradient system using distilled water and acetonitrile with 0.5% acetic acid. This analytical method is sensitive for the determination of a small amount of methylated theaflavins, since various interfering substances produced during the fermentation process were eliminated in advance by solid-phase extraction. Using this analytical method, we also demonstrated that methylated theaflavins were easily produced during the manufacture of black tea.     

Molecular Characterization and Diagnosis of QoI Resistance in Cucumber and Eggplant Fungal Pathogens

ABSTRACT The molecular mechanism of QoI fungicide resistance was studied using isolates of cucumber Corynespora leaf spot fungus (Corynespora cassiicola) and the eggplant leaf mold (Mycovellosiella nattrassii). In both pathogens, a mutation at position 143 from glycine to alanine (G143A) was detected in the cytochrome b gene that encodes for the fungicide-targeted protein. Moreover, the nucleotide sequence at amino acid position 143 was converted from GGT or GGA in sensitive (wild-type) to GCT or GCA in resistant (mutant-type) isolates. The methods of polymerase chain reaction restriction fragment length polymorphism commonly used for QoI resistance monitoring were employed successfully, leading to the amplified gene fragment from resistant isolates being cut with the restriction enzyme ItaI. However, heteroplasmy (the coexistence of wild-type and mutated alleles) was found when the resistant isolates of C. cassiicola, M. nattrassii, and Colletotrichum gloeosporioides (strawberry anthracnose fungus) were subcultured in the presence or absence of QoI fungicides. QoI resistance of cucumber powdery and downy mildew isolates persisted for a few years following the removal of the selection pressure imposed by the fungicide under both laboratory and commercial greenhouse conditions. The proportion of mutated sequences in cytochrome b gene decreased over time in the pathogen population. The protective efficacy of the full dose of azoxystrobin decreased when the populations of powdery and downy mildews contained resistant isolates at 10%. Using FMBIO, a fluorescence bio-imaging analyzer, the mutant allele from the QoI-resistant isolates could be detected at the level of 1%, whereas the detection sensitivity of ethidium-bromide-stained gels was approximately 10 times lower.     

Settling and raising postlarval abalone Haliotis diversicolor supertexta (Lischke) on microparticulate diets embedded in a layer of alginate

Two 21 day experiments were conducted to determine whether microparticulate diets could be used to substitute for diatoms for raising (Experiment 1) as well as settling and on‐growing (Experiment 2) Haliotis diversicolor supertexta postlarvae. The experiments were conducted in 10 L flow‐through tanks and each diet had three replicate tanks. In experiment 1, three experimental microparticulate diets supplied by a commercial feed company, labelled SF‐15, SF‐15 (T) and JFY‐13–1, were used, and in experiment 2, SF‐15 and SF‐15 (T). Artificial diets were compared to diatoms in terms of supporting growth and survival. The survival rate for experiment 1 was lowest for JFY‐13–1 (0%) and highest for diatoms (12.9±0.8%). The final size of postlarvae fed SF‐15 and SF‐15 (T) was significantly greater than that of postlarvae fed diatoms, 973±193 μm and 996±220 μm compared to 786±141 μm. In experiment 2, a total number of 501 larvae settled onto plates covered with SF‐15, 254 on SF‐15 (T) and 1085 on diatoms. The highest survival rate was 33.1±6.9% for SF‐15 and the lowest was 17.3±5.0% for diatoms. However, the final length of postlarvae fed diatoms was significantly greater than SF‐15 and SF‐15 (T); 1018±379 μm compared to 728±89 μm and 717±90 μm. The artificial micro diets used in this study were comparable to diatoms in terms of grow and survival of postlarval H. diversicolor supertexta.     

Isolation of chicken taste buds for real‐time Ca2+ imaging

We isolated chicken taste buds and used a real‐time Ca²⁺ imaging technique to investigate the functions of the taste cells. With RT‐PCR, we found that isolated chicken taste bud‐like cell subsets express chicken gustducin messenger RNA. Immunocytochemical techniques revealed that the cell subsets were also immunopositive for chicken gustducin. These results provided strong evidence that the isolated cell subsets contain chicken taste buds. The isolated cell subsets were spindle‐shaped and approximately 61–75 μm wide and 88–98 μm long, and these characteristics are similar to those of sectional chicken taste buds. Using Ca²⁺ imaging, we observed the buds' response to 2 mmol/L quinine hydrochloride (a bitter substance) and their response to a mixture of 25 mmol/L L‐glutamic acid monopotassium salt monohydrate and 1 mmol/L inosine 5′‐monophosphate disodium salt, umami substances. The present study is the first morphological demonstration of isolated chicken taste buds, and our results indicate that the isolated taste buds were intact and functional approaches for examining the taste senses of the chicken using Ca²⁺ imaging can be informative.     

Incentive for shifts in water management systems by shrimp culturists in southern Thailand

ABSTRACT:   Water management systems and other business situations of shrimp culturists were surveyed in two districts in southern Thailand. There were three types of water management systems in southern Thailand, namely closed, semiclosed and open systems, categorized by the frequency of exchange of pond water. Shrimp culturists in those districts tended to shift their system from the open system to the closed system of their own accord, and the average net income ratio decreased due to the change of water management system in a district from more than 2 to less than 1. The reason for this shift, in spite of the decrease in the profit rate, was prevention of infectious disease caused by water exchange. In conclusion, it was proven that independent shrimp culturists, such as medium and small scale shrimp pond owners in southern Thailand, would accept short-term decreases in profit rate in order to stabilize production.