Virulence of the entomopathogenic fungus <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> using soybean oil formulation for control of the cotton stainer bug, <Emphasis Type="Italic">Dysdercus peruvianus</Emphasis>

The cotton stainer bug Dysdercus peruvianus (Hemiptera: Pyrrhocoridae) is an insect pest that causes heavy losses in cotton plantations. The need to reduce the use of insecticides for control of this pest has increased steadily, and Metarhizium anisopliae (Ascomycota: Clavicipitaceae) could be an important biopesticide candidate to control this pest. The effect of M. anisopliae on D. peruvianus nymphs and adults using formulations with soybean oil and Agral^® was evaluated. Formulation using 10% soybean oil added to 10⁸ conidia mL^?1 (grown on used and reused rice) was the most effective for nymph and adult, causing 100% mortality 6 and 7 days after exposure, respectively. The SEM analysis of infected insects showed that M. anisopliae conidia were able to adhere anywhere on the exoskeleton, but were more abundant between the joints. Using the same rice for two batches of growth may be an option for improving commercial conidial production of M. anisopliae and may reduce overall costs. Its effect on D. peruvianus adults opens a new possibility for using this fungus as an alternative to chemical pesticides and the use of M. anisopliae in association with integrate pest management.     

Defense Response of Native and Alien Mealybugs (Hemiptera: Pseudococcidae) Against the Solitary Parasitoid Anagyrus sp. nr. pseudococci (Girault) (Hymenoptera: Encyrtidae)

The host behavioral and immune (encapsulation) defenses against the parasitoid Anagyrus sp. nr. pseudococci were compared for five mealybug species with different phylogenetic relationships and geographical origins: i) a Mediterranean native mealybug species, Planococcus ficus, with a long co-evolutionary history with the parasitoid; ii) three alien mealybugs species, Planococcus citri, Pseudococcus calceolariae and Pseudococcus viburni, with a more recent co-evolutionary history; and iii) a fourth alien mealybug species, Phenacoccus peruvianus, with no previous common history with the parasitoid. Three host defense behaviors were registered: abdominal flipping, reflex bleeding and walking away. The native host Pl. ficus and its congener Pl. citri exhibited the lowest probability of defense behavior (0.11?±?0.01 and 0.09?±?0.01 respectively), whereas the highest value was observed in P. viburni (0.31?±?0.02). Intermediate levels of defense behavior were registered for Ps. calceolariae, and Ph. peruvianus. The probability of parasitoid encapsulation was lowest and highest for two alien host species, Ph. peruvianus (0.20?±?0.07) and Ps. viburni (0.86?±?0.05), respectively. The native host Pl. ficus, its congener Pl. citri and Ps. calceolariae showed intermediate values (0.43?±?0.07, 0.52?±?0.06, and 0.45?±?0.09, respectively). The results are relevant with respect to biological control and to understand possible evolutionary processes involved in host range of A. sp. nr. pseudococci.     

Induced responses of <Emphasis Type="Italic">Bougainvillea glabra</Emphasis> Choisy (Nyctaginaceae) against <Emphasis Type="Italic">Phenacoccus peruvianus</Emphasis> Granara de Willink (Hemiptera: Pseudococcidae) attack: preliminary results

Plants respond to herbivorous attack through a defence system that includes structural barriers, release of toxic chemicals, and attraction of natural enemies of the target pests, etc. In this study, plants of Bougainvillea glabra Choisy (Nyctaginaceae) were artificially infested with the obligate phloem-feeding insect pest Phenacoccus peruvianus Granara de Willink (Hemiptera: Pseudococcidae) and compared with control uninfested plants. Leaf samples were collected at 2, 6, 12, 24, 48, 96 and 144 h after infestation to monitor the plant response. Proline (Pro), malondialdehyde (MDA), chlorophyll (Chl_a, Chl_b, and Chl_a+b), and total carotenoid (Car) contents, as well as the activities of catalase (CAT) and phenylalanine ammonia-lyase (PAL) antioxidant enzymes, were measured at each sampling time to better understand the mechanism of plant defences. Proline content was the highest at 2 h after infestation and remained high throughout the experiment, while MDA content differed significantly only at 12 h after infestation. The value of chlorophyll was higher in the control plants, indicating that insect attack seriously compromised the photosynthetic activity of infected plants. Enzymatic activities showed significant increases, CAT of about 50% after 24 h and PAL of about 43% after 6 h. This study will be useful for understanding Bougainvillea plant defence against mealybugs and for showing that this ornamental species is able to activate enzymatic and molecular mechanisms in response to insect attack.     

Suitability of five mealybug species (Hemiptera,Pseudococcidae) as hosts for the solitary parasitoid Anagyrus sp. nr. pseudococci (Girault) (Hymenoptera: Encyrtidae)

Anagyrus sp. nr. pseudococci is an endoparasitoid which has been used as a biological control agent of mealybug pests. In this study, we compared the suitability of five mealybugs species with different phylogenetic relationships and geographical origins as hosts of this parasitoid. The selected mealybugs were: (1) a Mediterranean-native species, Planococcus ficus, sharing a long co-evolutionary history with the parasitoid; (2) three exotic species, the Afrotropical Planococcus citri, the Australasian Pseudococcus calceolariae and the Neotropical Pseudococcus viburni, with a recent history; and (3) the Neotropical Phenacoccus peruvianus, with no previous common history with the parasitoid. Host suitability was assessed based on different fitness parameters, such as body size, developmental time, emergence rate and sex ratio. The parasitoid was able to complete development in all mealybug species. Nevertheless, its emergence rate significantly varied among mealybug species, with the highest values observed in Pl. ficus and Pl. citri, intermediate values in Ps. calceolariae and the lowest ones in Ps. viburni and Ph. peruvianus. The body size of adult wasp females varied with host suitability and was positively correlated with other measures of parasitoid fitness, including the emergence rate and the sex ratio. The parasitoid developmental time differed among mealybug species but did not correlate with any other measure of fitness. A female biased sex ratio was found in the parasitoid progeny emerged from all mealybug species, except in Ps. viburni and Ph. peruvianus. There was a direct relationship between the proportion of females in the parasitoid progeny and the emergence rate.     

Host selection behaviour and specificity of the solitary parasitoid of mealybugs Anagyrus sp. nr. pseudococci (Girault) (Hymenoptera,Encyrtidae)

The host selection behaviour of Anagyrus sp. nr. pseudococci was compared in no-choice tests among five mealybug species of different geographical and phylogenetic origin, including the Mediterranean native host, Planococcus ficus and four exotic mealybug species, one of the same genus, Pl. citri, two Pseudococcus species, Ps. calceolariae and Ps. viburni and a more distant one, Phenacoccus peruvianus. All five studied mealybug species were recognised by the parasitoid as potential hosts and parasitised, but the behavioural pattern of host recognition, host handling and the level of host acceptance of Anagyrus sp. nr. pseudococci significantly varied among the five studied species, indicating a clear preference for the two Planococcus species, Pl. ficus in particular. The results suggest that A. sp. nr. pseudococci has a broader host range and a more generalist behaviour in comparison with other Anagyrus species. Practical implications of the findings are discussed.     

Fitness and life history parameters of <Emphasis Type="Italic">Leptomastix epona</Emphasis> and <Emphasis Type="Italic">Pseudaphycus flavidulus</Emphasis>, two parasitoids of the obscure mealybug <Emphasis Type="Italic">Pseudococcus viburni</Emphasis>

Fitness and life table parameters of two endoparasitoids of the obscure mealybug Pseudococcus viburni (Signoret), the solitary Leptomastix epona (Walker) and the gregarious Pseudaphycus flavidulus (Brèthes), were examined in relation to temperature and host size with a view to determine the efficacy of the parasitoids as biocontrol agents of the pest. Three temperature levels (21°C, 26°C and 31°C) and two host sizes classes (small, which mostly comprised third instar nymphs and large, which consisted of female adults) were studied. The lower developmental threshold and thermal constant of the host and the parasitoids were found similar so the coincidence of pest and parasitoids is likely. The rate of development of the parasitoids increased with a linear trend as the temperature increased from 21°C to 31°C. Temperature had a significant effect on mummification in both parasitoid species and on successful parasitism by P. flavidulus. Host size had a significant effect on the mummification caused by L. epona and on the proportion of the male offspring which emerged as well as on the successful parasitism by P. flavidulus. Life table parameters of the parasitoids were estimated in small and large hosts at 26°C in the laboratory. Both parasitoids achieved a greater intrinsic rate of natural increase and gross reproductive rate in addition to a shorter generation and doubling time in large mealybugs compared with small ones. Consequently, large hosts are expected to have a higher impact on the rise of the parasitoids population and the potential of the parasitoids to control the mealybug population improves with the increase of host size. Handling Editor: Torsen Meiners.     

First record of the insect pathogenic alga Helicosporidium sp. (Chlorophyta: Trebouxiophyceae) infection in larvae and pupae of Rhizophagusgrandis Gyll. (Coleoptera, Rhizophaginae) from Turkey

The predator beetle Rhizophagus grandis Gyll. (Coleoptera, Rhizophaginae) is one of the most important biological control agents, mass-bred and used to suppress populations of an important pest: the great spruce bark beetle, Dendroctonus micans. The achlorophyllous alga Helicosporidium sp. was first discovered in the pest. Later it was also found in the predator, but only in the adults. In this study, the pathogenic alga Helicosporidium sp. was discovered in larvae and early pupae of R. grandis for the first time. The morphological characteristics of the pathogenic alga were revealed by light and electron microscopy. Infection rates of Helicosporidium sp. in the larvae and pupae of R. grandis were 23.5% and 6.25%, respectively.     

Karyomorphological effects of two new oil formulations on Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

The cotton bollworm Helicoverpa armigera is a serious pest of many economically important crops. Since this pest has become resistant to the conventional synthetic insecticides, newer compounds and formulations are being developed against this insect pest. Many natural compounds isolated from the plants were tested against this pest. Among them Hyptis suaveolens and Melochia corchorifolia showed insecticidal properties against H. arnigera. Based on bioefficacy studies, caryophyllene and β-sitosterol were isolated from H. suaveolens and M. corchorifolia respectively. The isolated natural compounds were further developed as formulations in various combinations with neem (Azadirachta indica) and karanj (Pongamia pinnata) oils. The present study indicated that the formulations influenced the karyomorphology of H. armigera.     

Molecular and Morphological Identification of Mealybug Species (Hemiptera: Pseudococcidae) in Brazilian Vineyards

Mealybugs (Hemiptera: Pseudococcidae) are pests constraining the international trade of Brazilian table grapes. They damage grapes by transmitting viruses and toxins, causing defoliation, chlorosis, and vigor losses and favoring the development of sooty mold. Difficulties in mealybug identification remain an obstacle to the adequate management of these pests. In this study, our primary aim was to identify the principal mealybug species infesting the major table grape-producing regions in Brazil, by morphological and molecular characterization. Our secondary aim was to develop a rapid identification kit based on species-specific Polymerase Chain Reactions, to facilitate the routine identification of the most common pest species. We surveyed 40 sites infested with mealybugs and identified 17 species: Dysmicoccus brevipes (Cockerell), Dysmicoccus sylvarum Williams and Granara de Willink, Dysmicoccus texensis (Tinsley), Ferrisia cristinae Kaydan and Gullan, Ferrisia meridionalis Williams, Ferrisia terani Williams and Granara de Willink, Phenacoccus baccharidis Williams, Phenacoccus parvus Morrison, Phenacoccus solenopsis Tinsley, Planococcus citri (Risso), Pseudococcus viburni (Signoret), Pseudococcus cryptus Hempel, four taxa closely related each of to Pseudococcus viburni, Pseudococcus sociabilis Hambleton, Pseudococcus maritimus (Ehrhorn) and Pseudococcus meridionalis Prado, and one specimen from the genus Pseudococcus Westwood. The PCR method developed effectively identified five mealybug species of economic interest on grape in Brazil: D. brevipes, Pl. citri, Ps. viburni, Ph. solenopsis and Planococcus ficus (Signoret). Nevertheless, it is not possible to assure that this procedure is reliable for taxa that have not been sampled already and might be very closely related to the target species.     

Life tables for the guava psyllid Triozoida limbata in southeastern Brazil

The aim of this study was to identify, quantify, and determine the importance of biological control agents of the psyllid Triozoida limbata (Enderlein) (Hemiptera: Sternorrhyncha: Triozidae). Ecological life tables were developed during four periods to determine the critical stage and key mortality factors of this pest. Predation and parasitism were found to be the two major factors driving the population dynamics of this pest. The major predators of eggs and nymphs of first and second instars were coccinellids, predatory thrips, lacewings, ants, and spiders, while the major predators of third, fourth, and fifth instars were several species of predatory wasps and syrphids. The endoparasitoid Psyllaephagus sp. (Hymenoptera: Encyrtidae) parasitized fifth instar psyllids. The fifth instar was found to be the critical mortality stage of T. limbata, and key mortality factors include the parasitoid Psyllaephagus sp. and predatory wasps.     

Adaptation of five columnar cactus species to various conditions in the negev desert of Israel

Five species of columnar cacti were examined with the aim of introducing new orchard crops to the Negev desert of Israel. The species comprisedStenocereus gummosus, S. griseus, S. thurberi andPachycereus pringlei, all found in semi-arid regions of Mexico, andCereus peruvianus, which is grown in sub-tropical regions. Young seedlings were planted at four sites in the Negev. The sites differ in climatic conditions and water quality. Growth data, expressed as total stem length and stem biomass, was obtained after five to six years in the orchards. They showed thatC. peruvianus grew best in the site with moderate temperatures and good-quality water, whereas the other species also did well under more extreme conditions of high temperatures and brackish water. Growth of all species was significantly retarded at the site having water with the highest NaCl content. In the sixth year a severe drop in temperature at one of the sites caused extensive injury toC. peruvianus andS. griseus plants, raising doubts as to the suitability of that site for their cultivation. At this stageC. peruvianus is the most promising candidate for domestication. Its growth rate was much higher than that of the other species, yield was precocious and fruit of good quality. Stem tissue ion concentrations (dry weight basis) showed that accumulation of K⁺was similar at all four sites and that Na⁺and Ct were significantly higher (probably in toxic concentrations) at the site with the highest concentrations ofNa⁺and Cl~ ions in the water. At all sites Cl~ concentration inC. peruvianus was much lower than that in the other species indicating that the exclusion of Cl^- from the stems was not correlated to salt tolerance.     

Insecticidal control of the mealybug Phenacoccus gossypiphilous (Hemiptera: Pseudococcidae), a new pest of cotton in Pakistan

The mealybug Phenacoccus gossypiphilous (Stanley) played havoc with the cotton crop in Pakistan during 2005. To control this pest, insecticides of different groups were evaluated in both the laboratory and in field conditions. In the laboratory, bifenthrin, profenofos and chlorpyrifos proved to be the best insecticides for mealybug control, based on their susceptibility with the leaf dip method for their LC₅₀. In field conditions, the recommended application rates of methomyl, profenofos and chlorpyrifos provided the best control: the lethal time studies proved their efficiency for better and timely control of this sporadic pest. The present study has shown that the insecticides tested, in particular profenofos, chlorpyrifos, methomyl and bifenthrin, provide satisfactory control of the cotton mealybug. The control of the insect pest complex throughout the cotton crop predominantly depends on wise and justified use of these chemicals, and necessitates development of an integrated pest management strategy.     

Culturable bacterial microbiota of Plagiodera versicolora (L.) (Coleoptera: Chrysomelidae) and virulence of the isolated strains

Plagiodera versicolora (Laicharting, 1781) (Coleoptera: Chrysomelidae) is an important forest pest which damages many trees such as willow, poplar, and hazelnut. In order to find new microbes that can be utilized as a possible microbial control agent against this pest, we investigated the culturable bacterial flora of it and tested the isolated bacteria against P. versicolora larvae and adults. We were able to isolate nine bacteria from larvae and adults. The isolates were characterized using a combination of morphological, biochemical, and physiological methods. Additionally, we sequenced the partial sequence of the 16S rRNA gene to verify conventional identification results. Based on characterization studies, the isolates were identified as Staphylococcus sp. Pv1, Rahnella sp. Pv2, Rahnella sp. Pv3, Rahnella sp. Pv4, Rahnella sp. Pv5, Pantoea agglomerans Pv6, Staphylococcus sp. Pv7, Micrococcus luteus Pv8, and Rahnella sp. Pv9. The highest insecticidal activity against larvae and adults was obtained from M. luteus Pv8 with 50 and 40 % mortalities within 10 days after treatment, respectively. Extracellular enzyme activity of the bacterial isolates such as amylase, proteinase, lipase, cellulose, and chitinase was also determined. Consequently, our results show that M. luteus Pv8 might be a good candidate as a possible microbial control agent against P. versicolora and were discussed with respect to biocontrol potential of the bacterial isolates.     

The formicid ant,Plagiolepis sp., as a predator of the coffee twig borer,Xylosandrus compactus

The coffee twig borer (Xylosandrus compactus Eichhoff) is an economically important pest of Robusta coffee in Uganda. In this study, a formicid ant, Plagiolepis sp., found in X. compactus galleries at the National Coffee Research Institute in 2014, was evaluated for potential to provide biological control of the twig borer. In a Petri dish feeding bioassay, Plagiolepis sp. preyed on all stages of X. compactus except adults within 24 h. In field bioassays where Plagiolepis sp. was caged over X. compactus-infested twigs for one month in muslin sleeves, the predator colonized pest galleries and eliminated all life stages of X. compactus as opposed to the untreated control. In a survey of Plagiolepis sp. in 11 districts of eastern, central and western Uganda, the ant was present in nine of the districts with highest levels of colonization (over 18%) of X. compactus galleries in Luwero district in the central Lake Victoria crescent agroecological zone. These results appear to confirm that Plagiolepis sp. is an indigenous predator of X. compactus which invades pest galleries and feeds on the pest in the field. For prospective utilization of Plagiolepis sp. as a biological control agent of X. compactus, studies on the biology of Plagiolepis sp., its mass rearing protocols and factors favoring its proliferation in the field are highly recommended.     

Possibilities for biological control ofAntigastra catalaunalis [Lep.: Pyralidae], a new pest of sesame in the western hemisphere

The sesame leafroller,Antigastra catalaunalis (Duponchel), is a recently introduced pest of sesame,Sesamum indicum L. (Pedaliaceae), in the New World. It is presently the chief pest of this crop in Colombia. Four parasites have been found attackingA. catalaunalis in Colombia:Euplectrus sp.[Hym.: Eulophidae], Bracon sp.(Hym.: Braconidae], Spilochalcis sp., andBrachymeria sp.[Hym.: Chalcididae], but they do not appear to exercise significant control of the insect. A literature search resulted in a list of 19 insect and 1 nematode parasites. Some of these parasites are widespread and well known, and an attempt should be made to have them sent to Colombia. Some also parasitize other important pests in that country.     

Parasitoid associated with Liriomyza huidobrensis (Diptera: Agromyzidae) outbreaks in tomato fields in Brazil

1. The knowledge of natural factors that affect pest populations is essential in predicting the occurrence of pest outbreaks and in developing integrated pest management programmes. Natural enemies, climatic elements and host plants are among the most important factors affecting pest dynamics.
2. Tomato (Solanum lycopersicum) is the second most consumed vegetable worldwide. The pea leaf miner Liriomyza huidobrensis (Diptera: Agromyzidae) is a major pest to the tomato in Brazil.
3. This study aimed to determine the main natural factors that regulate L. huidobrensis populations in tomato fields in Brazil.
4. Liriomyza huidobrensis densities were evaluated by directly counting the number of active mines on the basal leaf of the middle section of the plant canopy, and predators and parasitoids were assessed using the leaf‐beating‐against‐a‐tray technique. Eight commercial tomato fields were assessed over two years. The phenological growth stages of the tomato plants (vegetative and reproductive) were noted during the assessments.
5. Liriomyza huidobrensis populations peaked between the middle and end of the planting season.
6. Opius sp. (Hymenoptera: Braconidae) was the main natural enemy of L. huidobrensis.
7. Our results suggest that phenological growth stage and Opius sp. are associated with population dynamics of L. huidobrensis in tomato fields.
8. Therefore, integrated pest management programmes should aim to preserve populations of the parasitoid Opius sp.

     

Mantle defensive response of marine pulmonate Trimusculusperuvianus

The marine pulmonate Trimusculusperuvianus (Sowerby, 1835) which is found in caves or narrow crevices along the Chilean coast was studied to determine a possible chemical defense against the intertidal key predator Heliasterhelianthus. T.peruvianus releases a white secretion through the extended mantle animal covers, to contact with the tube feet of starfish. This behaviour significantly decreases predation of gastropod and it has not been described previously for members of the Trimusculidae family.     

Fungi associated with a natural epizootic in Fiorinia externa Ferris (Hemiptera: Diaspididae) populations

Stands of eastern hemlock [(Tsuga canadensis (L.) Carrière] in the northeastern United States are in decline, in part from the attack of elongate hemlock scale, Fiorinia externa Ferris (Hemiptera: Diaspididae). From 2001 to the present, a natural epizootic has been observed in populations of F. externa. Initially discovered at the Mianus River Gorge Preserve in Bedford, New York, the epizootic has also been detected in Pennsylvania, New Jersey and Connecticut. Understanding and assessing the identity of the pathogenic micro‐organisms responsible for this natural mortality is crucial for developing biological controls for this pest. We have isolated and taxonomically and genetically identified entomopathogens, phytopathogens and endophytic fungi associated with F. externa. Isolates of the following were obtained: Colletotrichum sp., Lecanicillium lecanii, Beauveria bassiana, Metarhiziopsis microspora, Myriangium sp., Mycosphaerella sp. anamorph, Nectria sp., Botrytis sp., Phialophora sp. and Fusarium sp.     

Comparison of Phenacoccus solenopsis specimens from different regions of Pakistan using COI molecular barcoding (Hemiptera: Pseudococcidae)

Because correct identification of insects is crucial for pest management involving chemical or biological control agents, we have used a molecular approach to identify and characterize specimens of the cotton pest Phenacoccus solenopsis Tinsley (Sternorrhyncha: Pseudococcidae) present in different regions of Pakistan. The specimens were analyzed through DNA sequence analysis of their mitochondrial COI (mtCOI) gene using an improved procedure that could distinguish between the pest and its associated parasitoid. Our analysis showed no variation among the mealybug specimens from different geographical locations of Pakistan and confirmed that this is the same species and haplotype that is infesting cotton plants in other parts of Asia. This information will assist in the development of biological control programs against P. solenopsis in Pakistan and other Asian countries.