Contact irritancy and spatial repellency behaviors in Anopheles albimanus Wiedemann (Diptera: Culicidae) collected in Orange Walk,Belize, C.A.

Anopheles albimanus Wiedemann is a vector of malaria in northern Belize. Previous studies have identified behavioral responses in this species when exposed to certain chemicals. We used a high throughput screening system to evaluate the contact irritant and spatial repellency responses exhibited by An. albimanus collected from Orange Walk, Belize, Central America. Anopheles albimanus behavior was significantly altered by a number of key public health insecticides. Significant irritant responses to alphacypermethrin, deltamethrin, permethrin, DDT, and propoxur were recorded, while spatial repellency responses were only elicited by DDT. Malathion resulted in neither a contact irritant or spatial repellency response. These results are in congruence with behavioral patterns previously observed for An. albimanus and for Aedes aegypti in the same system. This study produced baseline data on the behaviors of An. albimanus and confirms that the majority of compounds used as indoor residual sprays may induce premature exiting by An. albimanus in addition to killing the mosquito. This effect would decrease contact with humans and thus disrupt malaria transmission.     

Statistical optimization of media for enhanced azadirachtin production from redifferentiated zygotic embryo cultures of neem (Azadirachta indica A. Juss.)

Callus cultures from zygotic embryos of neem (Azadirachta indica) were initiated and analyzed for azadirachtin production. Medium components were screened and optimized using the statistical techniques of Plackett–Burman and response surface methodology. The Plackett–Burman design, with five medium components (Murashige and Skoog major salts, sucrose, casein hydrolysate, indole-3-acetic acid, and N⁶-benzylaminopurine), was performed to screen the variables that significantly affected azadirachtin production. The three variables—Murashige and Skoog major salts, sucrose, and N⁶-benzylaminopurine—significantly affected azadirachtin production and were significant factors for optimization using response surface methodology. The experimental results were fitted to a second-order polynomial model with a correlation coefficient (R ²) of 0.9582. The optimal concentrations of variables for maximum azadirachtin production were full-strength Murashige and Skoog major salts, 5.68% sucrose, and 10.42 μM N⁶-benzylaminopurine. The maximum azadirachtin production by the predicted model was 5.13 mg/g dry weight, which was in agreement with the actual experimental value of 4.97 mg/g dry weight.     

A methodology based on insecticide impregnated filter paper for monitoring resistance to deltamethrin in Triatoma infestans field populations

The domiciliary presence of Triatoma infestans (Klug) (Hemiptera: Reduviidae) after control interventions was reported in recent years. Toxicological studies showed high levels of resistance to pyrethroids suggesting resistance as one of the main causes of deficient control. The aim of the present study was to develop a protocol to test resistance to deltamethrin in T. infestans collected from the field by discriminate concentration. To evaluate field insects, the effect of age (early vs. later) and nutritional state (starved vs. fed) on the deltamethrin susceptibility of each developmental stage was studied. Topical and insecticide impregnated paper bioassays were used. Using the impregnated paper, the susceptibility to deltamethrin was not affected by the age of the stadium and the nutritional states, and varied with the post‐exposure time and with the different developmental stages. A discriminant concentration of deltamethrin (0.36% w/v) impregnated in filter paper was established for all developmental stages. Finally, the methodology and the discriminant concentration were evaluated in the laboratory showing high sensitivity in the discrimination of resistance. The present study developed a methodology of exposure to insecticide impregnated papers and proposes a protocol to test T. infestans in field populations with the aim to detect early evolution of resistance to deltamethrin.     

Behavioral responses of Aedes aegypti,Aedes albopictus,Culex quinquefasciatus,and Anopheles minimus against various synthetic and natural repellent compounds

The behavioral responses of colony populations of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus to four essential oils (citronella, hairy basil, catnip, and vetiver), two standard repellents (DEET and picaridin), and two synthetic pyrethroids (deltamethrin and permethrin) were conducted in the laboratory using an excito‐repellency test system. Results revealed that Cx. quinquefasciatus and An. minimus exhibited much stronger behavioral responses to all test compounds (65–98% escape for contact, 21.4–94.4% escape for non‐contact) compared to Ae. aegypti (3.7–72.2% escape (contact), 0–31.7% (non‐contact)) and Ae. albopictus (3.5–94.4% escape (contact), 11.2–63.7% (non‐contact)). In brief, essential oil from vetiver elicited the greatest irritant responses in Cx. quinquefasciatus (96.6%) and An. minimus (96.5%) compared to the other compounds tested. The synthetic pyrethroids caused a stronger contact irritant response (65–97.8% escape) than non‐contact repellents (0–50.8% escape for non‐contact) across all four mosquito species. Picaridin had the least effect on all mosquito species. Findings from the current study continue to support the screening of essential oils from various plant sources for protective properties against field mosquitoes.     

A taxonomic study of theVaccinium sect.Oxycoccus (Hill) W.D.J. Kock (Ericaceae) in the Czech Republic and adjacent territories

The ploidy level of plants of theVaccinium sect.Oxycoccus (Hill)W.D.J. Koch sampled in the Czech Republic, Germany, Austria and Poland was determined by chromosome counting and/or by flow cytometry. Forty-five characters were measured and scored in the morphometric analysis. Principal component analysis, cluster analysis, canonical discriminant analysis and classificatory discriminant analysis were used in the statistical analyses. Diploid (2n=24), tetraploid (2n=48) and hexaploid (2n=72) populations were confirmed and a new ploidy level — pentaploid hybrid plants (2n=60) — was revealed. Results of the multivariate morphometric analysis support the separation of the two native species. DiploidV. microcarpum (Turcz. exRupr.)Schmalh. differs from the polyploids by smaller size of petals, shorter style and stamens (stamens have long filaments and short anthers), glabrous pedicels, mostly solitary flowers, earlier flowering and by occurrence predominantly inPolytrichum strictum tufts. The low taxonomic significance of some features often used in keys for their separation (shape of fruits, insertion of prophylla, pubescence of filaments) was confirmed.V. oxycoccos L. includes three ploidy levels. The hexaploids represent the most abundant ploidy level in the area studied. They show a slightly bigger size of petals, longer bracts, prophylla, style, sepal tips and wider seeds in comparison with the tetraploids. Pentaploid cranberries are hitherto known only from the Czech Republic. They differ particularly in the low proportion of fully-developed tetrads.     

Intraspecific Variation and Interspecific Differences in the Bacterial and Fungal Assemblages of Blue Tit (Cyanistes caeruleus) and Great Tit (Parus major) Nests

Although interest in the relationship between birds and microorganisms is increasing, few studies have compared nest microbial assemblages in wild passerines to determine variation within and between species. Culturing microorganisms from blue tit (Cyanistes caeruleus) and great tit (Parus major) nests from the same study site demonstrated diverse microbial communities with 32 bacterial and 13 fungal species being isolated. Dominant bacteria were Pseudomonas fluorescens, Pseudomonas putida, and Staphylococcus hyicus. Also common in the nests were the keratinolytic bacteria Pseudomonas stutzeri and Bacillus subtilis. Dominant fungi were Cladosporium herbarum and Epicoccum purpurascens. Aspergillus flavous, Microsporum gallinae, and Candida albicans (causative agents of avian aspergillosis, favus, and candidiasis, respectively) were present in 30%, 25%, and 10% of nests, respectively. Although there were no differences in nest mass or materials, bacterial (but not fungal) loads were significantly higher in blue tit nests. Microbial species also differed interspecifically. As regards potential pathogens, the prevalence of Enterobacter cloacae was higher in blue tit nests, while Pseudomonas aeruginosa—present in 30% of blue tit nests—was absent from great tit nests. The allergenic fungus Cladosporium cladosporioides was both more prevalent and abundant in great tit nests. Using discriminant function analysis (DFA), nests were classified to avian species with 100% accuracy using the complete microbial community. Partial DFA models were created using a reduced number of variables and compared using Akaike’s information criterion on the basis of model fit and parsimony. The best models classified unknown nests with 72.5–95% accuracy using a small subset of microbes (n = 1–8), which always included Pseudomonas agarici. This suggests that despite substantial intraspecific variation in nest microflora, there are significant interspecific differences—both in terms of individual microbes and the overall microbial community—even when host species are closely related, ecologically similar, sympatric, and construct very similar nests.     

pH and thermal stability studies of carboxymethyl cellulase from intergeneric fusants of Trichoderma reesei/Saccharomyces cerevisiae

The combined effect of pH and temperature on carboxymethyl cellulase from two intergeneric fusants (M 14 and M 62) of Trichoderma reesei QM 9414/Saccharomyces cerevisiae NCIM 3288 was studied using response surface methodology. A central composite design for two variables was employed for the optimization studies. This study was compared with similar studies carried out with Trichoderma reesei QM 9414. The optimal pH and temperature for the enzymes derived from these organisms were: for the fusant M 14—pH 5.7 and 41.7°C, for the fusant M 62—pH 5.3 and 43°C, and for Trichoderma reesei QM 9414—pH 4.31 and 38.3°C. Received 5 May 1997/ Accepted in revised form 17 July 1998     

Metabolic fingerprinting to discriminate diseases of stored carrots

Volatile metabolites from headspace gas of carrot cv. Vita‐treat inoculated with water or four different pathogens Botrytis cinerea, Erwinia carotovora subsp. carotovora, Aspergillus niger and Fusarium avenaceum were profiled using gas chromatography and mass spectrometry to develop a technology to discriminate diseases. The inoculation of carrot roots with water or different pathogens released a total of 137 different volatile metabolites. Among them, 39 compounds were relatively consistent and 11 were specific to one or more diseases/inoculations. E. carotovora subsp. carotovora produced seven disease‐specific metabolites: 1‐butanol, 3‐methyl; 1‐pentanol; 1‐propanol, 2‐methyl; 2,3‐butanedione; boronic acid, ethyl; butane, 1‐methoxy‐3‐methyl; and ethane, ethoxy. Some metabolites were disease/inoculation discriminatory and were not detected in all treatments: 1,2‐dimethoxy‐ethene was common in carrots inoculated with E. carotovora subsp. carotovora and B. cinerea, while 2‐butanone, 3‐chloro‐4‐hydroxy‐1,4‐diphenyl was common in carrots inoculated with E. carotovora subsp. carotovora, F. avenaceum and water‐inoculated control. The significant mass ions, based on univariate analysis, from a total of 150 (46–195 m/z) and compounds from a total of 32 were further subjected to stepwise discriminant analysis and discriminant analysis. The models for 3 days after inoculation (DAI) were better than those for 6 DAI and 3 + 6 DAI, where up to 90% of the observations were correctly classified into respective inoculations. The disease‐discriminatory compounds from different diseases/inoculations and discriminant analysis models developed here have the potential for the early detection and discrimination of postharvest diseases of carrot cv. Vita‐treat, after validation under commercial conditions.     

The white-spotted longicorn beetle, <Emphasis Type="Italic">Anoplophora malasiaca</Emphasis> (Coleoptera: Cerambycidae), with a blueberry as host plant,utilizes host chemicals for male orientation

Volatile chemicals from Citrus and Salix host plants evoke orientation behavior in males of the species Anoplophora malasiaca (Thomson) (Coleoptera: Cerambycidae). These chemicals are emitted from wounded branches. We hypothesized that when released, these chemicals may indicate the presence of an individual to other conspecifics. Insects that originate from different host plants may use different plant chemicals from their own host to communicate with conspecifics. To further explore this theory, we investigated this communication system in a population of A. malasiaca from a third host plant, blueberry (Vaccinium spp.). Males from a blueberry host (Vaccinium population) were attracted to the odor of wounded Vaccinium branches when released near a female model in the laboratory, as has been observed in males found on Citrus and Salix host plants. The Vaccinium branch extract that was attractive to the males was separated into six fractions, of which two were active. Three active compounds were subsequently identified: β-caryophyllene and sulfur from the hexane fraction, and (E)-phytol in the weakly polar fraction. The latter two active compounds of Vaccinium branches were different from those found in Citrus and Salix.     

Marker-assisted selection and evaluation of the QTL for stigma exsertion under japonica rice genetic background

Stigma exsertion is one of the important traits which contribute to the efficient improvement of commercial seed production in hybrid rice. In order to understand the genetic factors involved in the stigma exsertion of an indica variety—IR24—a QTL analysis was conducted using the F₂ population between a japonica variety—Koshihikari—and a breeding line showing exserted stigma selected from the backcross population between IR24 as a donor and japonica varieties. As a result, a highly significant QTL (qES3), which had been predicted in the recombinant inbred population of IR24, was confirmed at the centromeric region on chromosome 3. qES3 increases about 20% of the frequency of the exserted stigmas at the IR24 allele and explains about 32% of the total phenotypic variance. A QTL near-isogenic line for qES3 increased the frequency of the exserted stigma by 36% compared to that of Koshihikari in a field evaluation, which suggests that qES3 is a promising QTL for the development of a maternal line for hybrid rice. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users. Maiko Miyata and Toshio Yamamoto contributed equally to this study.     

Management of <Emphasis Type="Italic">Sclerotium rolfsii</Emphasis> with integration of non-conventional chemicals,vermicompost and <Emphasis Type="Italic">Pseudomonas syringae</Emphasis>

Two non-conventional chemicals, ZnSO₄ (10⁻⁴ mM) and oxalic acid (4 mM) were tested (alone as well as in combination with seeds bacterized with Pseudomonas syringae strain PUR46 and vermicompost substitution in the potting soil), for their ability to suppress collar rot of chickpea (Cicer arietinum) caused by Sclerotium rolfsii under greenhouse conditions. ZnSO₄ and oxalic acid were applied as pre-inoculation foliar spray on chickpea and subsequently challenged with S. rolfsii. Both the chemicals provided significant protection to chickpea compared to control (100% plant mortality) when used alone as well as in combination with PUR46 and vermicompost. However, ZnSO₄ was more effective than oxalic acid against S. rolfsii. Amongst the treatments tried, plant mortality was least when ZnSO₄ was used in combination with seed bacterization with PUR46 and 25% vermicompost substitution. The findings indicate the utility of integration of the above factors in managing collar rot efficiently.     

Ecological differentiation of Central EuropeanBolboschoenus taxa and their relationship to plant communities

The taxa of theBolboschoenus maritimus group occurring in Central Europe were studied with respect to morphological differentiation, plant communities and soil conditions. Four morphological types ofBolboschoenus were distinguished, differing in habitat conditions, dispersal and distribution: type 1 —Bolboschoenus maritimus subsp.maritimus with narrow triangular fruits; type 2 —B. maritimus subsp.maritimus with wider triangular fruits, type 3 —B. maritimus subsp.compactus with round convex fruits; and type 4 —B. planiculmis auct. with concave fruits. Strong ecological differentiation of these types was found compared with relatively weak morphological diagnostic features. High discriminant efficiency appeared in the relationship to soil chemistry. Differences were also found in habitat-type spectra of individualBolboschoenus morphological types, which was reflected in their affinities to plant communities. Nomenclature, taxonomic classification, and distribution of these taxa in Europe need thorough revision.     

Isolation and Characterization of Antifungal Substances from Burkholderia sp. Culture Broth

A new antagonistic Burkholderia strain, designated MP-1 and producing antifungal activities against various filamentous plant pathogenic fungi, was isolated from the rhizoshere in the Naju area. Cultural characteristic studies strongly suggested that this strain belongs to the genus Burkholderia. The nucleotide sequence of the 16S rRNA gene (1491 pb) of strain MP-1 exhibited close similarity (99% to 100%) with other Burkholderia 16S rRNA genes. Extraction of fermentation broth of Burkholderia sp. MP-1 and various separations and purification steps led to isolation of four pure active molecules. The chemical structure of these four compounds—named phenylacetic acid, hydrocinnamic acid, 4-hydroxyphenylacetic acid, and 4-hydroxyphenylacetate methyl ester—was established on the basis on their gas chromatography–electron impact–mass spectrometry (GC-EI-MS) and trimethylsilation GC-EI-MS data. The four isolated compounds inhibited filamentous fungal growth on potato dextrose agar medium supplemented with 100 mg/L of phenylacetic acid, hydrocinnamic acid, 4-hydroxyphenylacetic acid and 4-hydroxyphenylacetate methyl ester individually.     

Synthesis and pharmacological activity of amides and the ozonolysis product of maleopimaric acid

The synthesis of a new group of maleopimaric acid amides containing fragments of methyl ethers of amino acids, aliphatic amines, imidazole, and N-methylpiperazine was carried out. The ozonolysis of methylmaleopimarate occurs via the cleavage of the double bond C18(19) and the opening of an anhydrous ring with the formation of secotriacid. As a result of the screening of the anti-inflammatory and antiulcer activity of maleopimaric acid derivatives, new effective compounds such as maleopimaric acid and its methyl ether, a product of ozonolysis—diterpenic secotriacid—and maleopimaric acid amide with L-leucine were found. An important advantage of the studied compounds is the low toxicity and the presence of bidirectional activity in the absence of adverse effects on the animal.