Resistance of stored-product insects to phosphine

The objectives of this work were to assess phosphine resistance in insect populations (Tribolium castaneum, Rhyzopertha dominica, Sitophilus zeamais and Oryzaephilus surinamensis) from different regions of Brazil and to verify if the prevailing mechanism of phosphine resistance in these populations involves reduced respiration rates. Sixteen populations of T. castaneum, 15 of R. dominica, 27 of S. zeamais and eight of O. surinamensis were collected from 36 locations over seven Brazilian states. Each population was tested for resistance to phosphine, based on the response of adults to discriminating concentrations, according to FAO standard method. For each insect species, the production of carbon dioxide of the most resistant and of the most susceptible populations was inversely related to their phosphine resistance. The screening tests identified possible phosphine resistant populations. R. dominica and O. surinamensis were less susceptible to phosphine than the other two species. The populations with lower respiration rate showed a lower mortality at discriminating concentration, possibly related to a phosphine resistance mechanism. Phosphine resistance occurs in stored-product insects, in different regions of Brazil, and the resistance mechanism involves reduced respiration rate.

Random Mating Between Two Widely Divergent Mitochondrial Lineages of Cryptolestes ferrugineus (Coleoptera: Laemophloeidae): A Test of Species Limits in a Phosphine-Resistant Stored Product Pest

Effective pest management relies on accurate delimitation of species and, beyond this, on accurate species identification. Mitochondrial COI sequences are useful for providing initial indications in delimiting species but, despite acknowledged limitations in the method, many studies involving COI sequences and species problems remain unresolved. Here we illustrate how such impasses can be resolved with microsatellite and nuclear sequence data, to assess more directly the amount of gene flow between divergent lineages. We use a population genetics approach to test for random mating between two 8 ± 2% divergent COI lineages of the rusty grain beetle, Cryptolestes ferrugineus (Stephens). This species has become strongly resistant to phosphine, a fumigant used worldwide for disinfesting grain. The possibility of cryptic species would have significant consequences for resistance management, especially if resistance was confined to one mitochondrial lineage. We find no evidence of restricted gene flow or nonrandom mating across the two COI lineages of these beetles, rather we hypothesize that historic population structure associated with early Pleistocene climate changes likely contributed to divergent lineages within this species.

Pest risk assessment of insects in sea cargo containers

A survey of the floors of 3001 empty sea cargo containers in storage was undertaken to estimate the quarantine risk of importing exotic insect pests into Australia, with special reference to pests of timber. More than 7400 live and dead insects were collected from 1174 containers. No live infestations of timber-feeding insects were recorded, but feeding damage detected in one floor indicates a low risk of importing colonies of timber pests in containers. The survey collection of dead insects demonstrates that containers are regularly exposed to economically important quarantinable insects, including timber pests (bostrichids, curculionids, cerambycids, siricids and termites), agricultural pests (including Adoretus sinicus, Adoretus sp., Carpophilus obsoletus and Philaenus spumarius), and nuisance pests (vespids and Solenopsis sp.). Stored product pests were found in more than 10% of containers. The assessment of pest risk associated with shipping containers is discussed in terms of the quantity and quality of opportunities for exotic insects to establish via this pathway.

Toxic effects of essential plant oils in adult Sitophilus oryzae (Linnaeus) (Coleoptera, Curculionidae)

Toxic effects of essential plant oils in adult Sitophilus oryzae (Linnaeus) (Coleoptera, Curculionidae). Stored grains are subject to losses in quality nutritional value and in sanitation from the time they are stored to the time they are consumed. Botanical insecticides may offer an alternative solution for pest control. The objective was to test the insecticidal properties of the essential oils of Cymbopogon citratus (leaf), Zingiber officinale (root) and Mentha sp. (leaf). The efficacy of these oils was tested to control the rice weevil, S. oryzae, using hydrodistillation. Chemical analysis of the essential oils was carried out by gas chromatography. Major components of C. citratus were geranial (48%) and neral (31%), of Z. officinale were α-zingibereno (13%), geranial (16%), neral (10%) and α-farneseno (5%) and of Mentha sp. was menthol (92%). Bioassays were carried out by fumigation and topical application. In topical application assays, the essential oil of C. citratus had greater toxicity (LC50 0.027 µL mL-1) and shorter exposure time than the oils of the other two plants. After 24 h and 48 h, 70% and 100% mortality of S. oryzae occurred, respectively. In fumigation assays, essential oil of Z. officinale had a lower LC50 (1.18 µL cm-2) and 70% mortality after 24 h exposure. Therefore, we recommend the use of essential oils of C. citratus and Z. officinale to control the rice weevil S. oryzae.

Quality of beans stored under hermetic conditions

This study aimed to evaluate the quality of beans (Phaseolus vulgaris L.) stored with different moisture contents under hermetic conditions. Beans with 12.3%, 15.7%, and 17.8% moisture content were used in the experiment. They were packed in bag type silos (3 kg), PET bottles (1.5 L), and glass containers (3 L), covered with organza fabric (control), and stored at 25 ºC and 70±5% relative humidity, for 120 days. The evaluated characteristics included moisture content, apparent density, electrical conductivity, germination percentage, cooking time, and bean classification, every 30 days. Except for the electrical conductivity and cooking time, the other characteristics were kept for 120 days in the stored product with moisture contents of 12.3% and 15.7% in hermetic conditions. Beans with 17.8% of the moisture content, electrical conductivity and cooking time increased, and apparent density and germination were reduced. Beans stored in the control, with any of the moisture contents, showed high quality loss during the long storage. It was concluded that it is possible to storage beans with moisture contents of 12.3% and 15.7%, with quality, for until 120 days, in hermetic conditions.

Insect prevention and control in plants processing dry milk /

Cover title.

Caenorhabditis elegans mutants resistant to phosphine toxicity show increased longevity and cross-resistance to the synergistic action of oxygen

Phosphine (hydrogen phosphide, PH3) is the fumigant most widely used to protect stored products from pest infestation. Despite the importance of this chemical, little is known about its mode of action. We have created three phosphine-resistant lines (pre-1, pre-7, pre-33) in the model organism C. elegans, with LC50 values 2, 5, and 9 times greater than the fully susceptible parental strain. Molecular oxygen was shown to be an extremely effective synergist with phosphine as, under hyperoxic conditions, 100% mortality was observed in wild-type nematodes exposed to 0.1 mg/l phosphine, a nonlethal concentration in air. All three mutants were resistant to the synergistic effects of oxygen in proportion to their resistance to phosphine with one mutant, pre-33, showing complete resistance to this synergism. We take the proportionality of cross-resistance between phosphine and the synergistic effect of oxygen to imply that all three mutants circumvent a mechanism of phosphine toxicity that is directly coupled to oxygen metabolism. Compared with the wild-type strain, all three mutants have an extended average life expectancy of from 12.5 to 25.3%. This is consistent with the proposed involvement of oxidative stress in both phosphine toxicity and ageing. Because the wild-type and mutant nematodes develop at the same rate, the longevity is unlikely to be caused by a clk-type reduction in oxidative metabolism, a potential alternative mechanism of phosphine resistance.

Utilização de bioensaios e marcadores moleculares para detecção da resistência de coleópteros de produtos armazenados a inseticidas

The qualitative and quantitative losses caused by stored product insects are of great concern, and since there is only a few active ingredients available for their control it is very important to have a frequent insect resistance monitoring. The objective of this research is to evaluate combination of bioassays and molecular marker techniques to detect insecticide resistance in stored product beetles. The Coleoptera species used for the tests were Sitophilus oryzae (L.) (Curculionidae), Rhyzopertha dominica (F.) (Bostrichidae) and Oryzaephilus surinamensis (L.) (Silvanidae). For the bioassays it was used the impregnated filter paper technique, applying 1 mL of deltamethrin (K-Obiol 25 CE TM) using four concentrations and five replicates, including a control with solvent only. Ten adults of each species were liberated separately on each dish. The mortality was evaluated after 24 h and resistance determined by probit analysis. The samples used for the PCR-RAPD were either in vivo or preserved in 70% ethanol, kept in -18°C freezer. After extraction, quantification and DNA quality analysis, the 25 µL samples had the DNA amplified and tested with six primers. The bioassays showed a crescent mortality proportional to insecticide concentration. The resistance factor for R. dominica, S. zeamais and S. oryzae were: 2,2; 3,2 and 9,2, respectively, compared to the susceptible populations of each species. The PCR-RAPD analysis revealed bands which indicate inter and intraspecific variability in the populations, but it was not possible to correlate them to resistance. The association of bioassay and PCR-RAPD represents a precise and valuable tool for resistance management of stored product insects, but more populations and primers should be tested.

Legume type and temperature effects on the toxicity of insecticide to the genus Callosobruchus (Coleoptera: Bruchidae)

Three bruchid pest species, Callosobruchus maculatus, Callosobruchus chinensis and Callosobruchus rhodesianus, were studied for their response to insecticide toxicity taking into account the separate and interactive effects of temperature and pre-adult food. The food types used were cowpea (Vigna unguiculata) and mungbean (Vigna radiata). Callosobruchus maculatus was the most tolerant to malathion and the least affected by temperature change while C. rhodesianus was the least tolerant. Over a 4 C range (23, 25, 27 C), there was generally a significant impact of temperature on the tolerance of the three species to the insecticide. The food type on which the insects developed influenced considerably the degree of insecticide tolerance. Callosobruchus maculatus and C. chinensis populations reared onmungbean had higher tolerance to malathion than their counterparts reared on cowpea, but the opposite was observed in C. rhodesianus populations. The food influence in this study suggested an ancestral cause or fitness cost depending on the species. The interaction of food-by-temperature had no significant effect on malathion toxicity to this genus. Correlation analysis showed C. chinensis to be relatively less sensitive to insecticide concentration over the range studied compared with the other two species.

Varietal effects of cowpea, Vigna unguiculata, on tolerance to malathion in Callosobruchus maculatus (Coleoptera: Bruchidae)

The impact of cowpea variety on the response of cowpea bruchid, Callosobruchus maculatus, to malathion was investigated. The interaction of six cowpea varieties (Adamawa Brown, Ife BPC, Ife Brown, Lilongwe, Ntcheu and NCRI-L25) with the geographical strains of C. maculatus (Brazil and Cameroon), temperature (23, 25, 27 C) and insecticide concentration were considered. Cowpea variety (V) had an unpredictable effect on C. maculatus response to malathion. Bruchid populations produced by Ife BPC were the most susceptible to malathion while those yielded by NCRI-L25 were the most tolerant. Regardless of the cowpea variety, the Brazil strain showed higher tolerance than the Cameroon strain. There was significant effect of temperature (T) and insecticide concentration (C) on malathion tolerance in both strains (S). Likewise, there was significant impact of all two-way interactions on cowpea bruchid tolerance except V x C. Significant three-way interactions on C. maculatus tolerance to malathion was only observed in S T V and S T C. The predictability of changing one of the factors on the susceptibility of C. maculatus to insecticide was very low. This study suggests a need to take the insecticide tolerance of insect populations produced by novel varieties into account during plant breeding in addition to factors such as yield and resistance to insect and disease attack.

Efeito de períodos de semeadura e das condições de armazenamento sobre a qualidade de grãos de feijão para o cozimento

A conservação adequada pode manter as características do grão recém-colhido por mais tempo, assegurando o valor comercial do produto estocado. Assim, o objetivo deste trabalho foi avaliar o efeito de períodos de semeadura na qualidade de cozimento de grãos de feijão da cultivar TPS Nobre, quando armazenados em diferentes condições de temperatura, de umidade e tempo de estocagem. O delineamento experimental utilizado foi o inteiramente casualizado, com três repetições. Os tratamentos consistiram da combinação de três períodos de semeadura, com três ambientes de armazenamento e três tempos de armazenamento dos grãos (0, 3 e 6 meses). A interpretação dos resultados permitiu inferir que as porcentagens de grãos normais e de grãos duros revelaram comportamento diferenciado em função do ambiente e do tempo de armazenamento. Constatou-se maior porcentagem de grãos duros após seis meses de armazenamento, realizado sob temperatura média do ar de 12 °C e umidade relativa de 45%-50% e também quando os grãos foram resultantes de semeadura realizada tardiamente (21/10/03). A absorção de água e o tempo de cozimento dos grãos de feijão foram alterados em função do período de semeadura, das condições e do tempo de armazenamento, caracterizando a existência de efeitos da interação genótipos x ambientes na qualidade para o cozimento dos grãos. Sendo assim, maior ou menor tempo para a absorção de água ou para o cozimento podem ser requeridos em função da qualidade dos grãos no momento da colheita ou das condições de temperatura e de umidade de armazenamento.

Organophosphate resistance in the maize weevil Sitophilus zeamais: Magnitude and behavior

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of temperature on development and population growth of Acarophenax lacunatus (Cross and Krantz) (Prostigmata: Acarophenacidae) on Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

The parasitic mite Acarophenax lacunatus kills the eggs upon which it feeds and seems to have potential as a biological control agent of stored grain pests. The lack of biological studies on this mite species led to the present study carried out in laboratory conditions at eight different temperatures (ranging from 20 to 41°C) and 60% relative humidity using Rhyzopertha dominica as host. The higher the temperature, the faster: (1) the attachment of female mites to the host egg (varying from 1 to 5 h); (2) the increase in body size of physogastric females (about twice faster at 40°C than at 20°C); and (3) the generation time (ranging from 40 to 220 h). In addition, the higher the temperature, the shorter the maximum female longevity (ranging from about 75 to 300 h). The two estimated temperature thresholds for development of A. lacunatus on R. dominica were 18 and 40°C. The average number of female and male offspring per gravid mite were 12.8 and 1.0, respectively, with sex ratios (females/total) ranging from 0.91 to 0.94 (maximum at 30°C). The net reproductive rate and intrinsic rate of increase also presented maximum values at 30°C (12.1 and 0.04, respectively).

Actividad insecticida de Ricinus Communis L. sobre Plodia Interpunctella Hbn. (Lepidoptera: Phycitinae).

La aplicación de insecticidas sintéticos como principal sistema de control de plagas de granos y productos almacenados ha originado el desarrollo de poblaciones de insectos resistentes a dichos químicos, la contaminación del medio ambiente y la acumulación de sustancias tóxicas en los alimentos. En este trabajo se evaluaron los efectos de la aplicación de molido de hojas de ricino sobre larvas de la «polilla de las harinas» (Lepidoptera: Phycitinae). Los molidos vegetales se obtuvieron a partir de hojas de Ricinus communis L. secadas en estufa a 40 ± 2 °C durante 48 horas y posteriormente molidas hasta lograr un polvo de textura similar a la harina de maíz, material con el que se mezcló a fin de lograr concentraciones de 5, 10 y 15 % y un testigo sin ricino. Las unidades experimentales consistieron en cajas de Petri con seis larvas de primer y segundo estadio y se efectuaron cinco repeticiones por tratamiento (n=120). Se registró el número de larvas, pupas y adultos vivos, cada cuatro días, hasta que las larvas sobrevivientes llegaron al estado adulto. Se calcularon los porcentajes de eficacia mediante la fórmula de Abbott. Los resultados se evaluaron por ANVA y test de Tukey. Se determinaron el tiempo efectivo medio (TE50) y concentración efectiva media (CE50) por el método Probit. Los resultados de mortalidad indicaron que la concentración al 15 % superó significativamente al resto de los tratamientos y al testigo.

Determining phosphine resistance in rust red flour beetle, Tribolium castaneum (Herbst.) (Coleoptera : Tenebrionidae) populations from Turkey

Fumigation with phosphine gas is the primary method of controlling stored grain pests. In Turkey, phosphine has been used extensively since the 1950's. Even though high levels of phosphine resistance have been detected in several key stored products pests across the world, it has never been studied in Turkey despite this long history of phosphine use. High-level phosphine resistance has been detected and genetically characterised previously in the rust red flour beetle, Tribolium castaneum in other countries. Since this pest is also a common problem in stored grain environment in Turkey, the current study was undertaken for the first time, to investigate the distribution and strength of phosphine resistance in T. castaneum. Four strains of T. castaneum were tested through bioassays for determining the weak and strong phosphine resistance phenotypes on the basis of the response of adults to discriminating phosphine concentrations of 0.03 mg/L and 0.25 mg/L, for 20 hour exposures respectively. Phenotype testing showed all strains exhibited some level of phosphine resistance with a maximum level of 196 fold. Sequencing and genetic testing of seven field-collected strains showed that all of them carried a strong resistance allele in at the rph2 locus similar to the one previously reported. Our results show that strong resistance to phosphine is common in Turkish strains of T. castaneum.