Morphological and molecular description of Blastocrithidia cyrtomeni sp. nov. (Kinetoplastea: Trypanosomatidae) associated with Cyrtomenus bergi Froeschner (Hemiptera: Cydnidae) from Colombia

A new trypanosomatid species, Blastocrithidia cyrtomeni, is herein described using morphological and molecular data. It was found parasitising the alimentary tract of the insect host Cyrtomenus bergi, a polyphagous pest. The morphology of B. cyrtomeni was investigated using light and transmission microscopy and molecular phylogeny was inferred from the sequences of spliced leader RNA (SL rRNA) - 5S rRNA gene repeats and the 18S small subunit (SSU) rRNA gene. Epimastigotes of variable size with straphanger cysts adhering to the middle of the flagellum were observed in the intestinal tract, hemolymph and Malpighian tubules. Kinetoplasts were always observed anterior to the nucleus. The ultrastructure of longitudinal sections of epimastigotes showed the flagellum arising laterally from a relatively shallow flagellar pocket near the kinetoplast. SL RNA and 5S rRNA gene repeats were positive in all cases, producing a 0.8-kb band. The amplicons were 797-803 bp long with > 98.5% identity, indicating that they originated from the same organism. According to the sequence analysis of the SL-5S rRNA gene repeats and the 18S SSU rRNA gene, B. cyrtomeni is different from all other known species or isolates of Trypanosomatidae. Both analyses indicate that among known species, it is most closely related to Blastocrithidia triatomae.

The distribution pattern of Lutzomyia longipalpis (Diptera: Psychodidae) in the peridomiciles of a sector with canine and human visceral leishmaniasis transmission in the municipality of Dracena, São Paulo, Brazil

The specimen distribution pattern of a species can be used to characterise a population of interest and also provides area-specific guidance for pest management and control. In the municipality of Dracena, in the state of São Paulo, we analysed 5,889 Lutzomyia longipalpis specimens collected from the peridomiciles of 14 houses in a sector where American visceral leishmaniasis (AVL) is transmitted to humans and dogs. The goal was to analyse the dispersion and a theoretical fitting of the species occurrence probability. From January-December 2005, samples were collected once per week using CDC light traps that operated for 12-h periods. Each collection was considered a sub-sample and was evaluated monthly. The standardised Morisita index was used as a measure of dispersion. Adherence tests were performed for the log-series distribution. The number of traps was used to adjust the octave plots. The quantity of Lu. longipalpis in the sector was highly aggregated for each month of the year, adhering to a log-series distribution for 11 of the 12 months analysed. A sex-stratified analysis demonstrated a pattern of aggregated dispersion adjusted for each month of the year. The classes and frequencies of the traps in octaves can be employed as indicators for entomological surveillance and AVL control.

Evolution of insecticide resistance in non-target black flies (Diptera: Simuliidae) from Argentina

Black flies, a non-target species of the insecticides used in fruit production, represent a severe medical and veterinary problem. Large increases in the level of resistance to the pyrethroids fenvalerate (more than 355-fold) and deltamethrin (162-fold) and a small increase in resistance to the organophosphate azinphos methyl (2-fold) were observed between 1996-2008 in black fly larvae under insecticide pressure. Eventually, no change or a slight variation in insecticide resistance was followed by a subsequent increase in resistance. The evolution of pesticide resistance in a field population is a complex and stepwise process that is influenced by several factors, the most significant of which is the insecticide selection pressure, such as the dose and frequency of application. The variation in insecticide susceptibility within a black fly population in the productive area may be related to changes in fruit-pest control. The frequency of individuals with esterase activities higher than the maximum value determined in the susceptible population increased consistently over the sampling period. However, the insecticide resistance was not attributed to glutathione S-transferase activity. In conclusion, esterase activity in black flies from the productive area is one mechanism underlying the high levels of resistance to pyrethroids, which have been recently used infrequently. These enzymes may be reselected by currently used pesticides and enhance the resistance to these insecticides.

Large indoor cage study of the suppression of stable Aedes aegypti populations by the release of thiotepa-sterilised males

The sterile insect technique (SIT) is a promising pest control method in terms of efficacy and environmental compatibility. In this study, we determined the efficacy of thiotepa-sterilised males in reducing the target Aedes aegypti populations. Treated male pupae were released weekly into large laboratory cages at a constant ratio of either 5:1 or 2:1 sterile-to-fertile males. A two-to-one release ratio reduced the hatch rate of eggs laid in the cage by approximately a third and reduced the adult catch rate by approximately a quarter, but a 5:1 release drove the population to elimination after 15 weeks of release. These results indicate that thiotepa exposure is an effective means of sterilising Ae. aegypti and males thus treated are able to reduce the reproductive capacity of a stable population under laboratory conditions. Further testing of the method in semi-field enclosures is required to evaluate the mating competitiveness of sterile males when exposed to natural environmental conditions. If proven effective, SIT using thiotepa-sterilised males may be incorporated into an integrated programme of vector control to combat dengue in Cuba.

Avaliação da infestação de insetos-praga associados à batata (Solanum tuberosum L.) sob efeito de nutrientes nitrogenados e potássicos e teores acumulados de aminoácidos livres nas cultivares Achat e Monalisa

Evaluation of insect-pest infestation associated to potato (Solanum tuberosum L.) under effect of nitrogen and potassium fertilizers and the accumulated amount of free aminoacids in Achat and Monalisa cultivars. The objective of this work was to evaluate the occurence of insect-pests on potato plants influenced by dosages of nitrogen and potassium accumulated in plant organs. A total of 169 plants of the Achat and Monalisa cultivars were evaluated to determine the presence-absence of Diabrotica speciosa Germar, 1824 and Agrotis ipsilon Hüfnagel, 1767. The experiment was carried out and executed at the Universidade Federal Fluminense, and the delineation was complete randomized block design, with four replication and nine treatments, using three fertilization level (0; 75 and 150 Kg/ha) with N-urea + KCl. The aminoacid levels were adjusted by the Leucine standard-curve (µg/l), using the Ninhydrin method, at 570 nm. The results showed that the tubercles of Monalisa accumulated high free aminoacid levels with 7,95% in the treatment N1K2 and 7,75% in the N2K1.These treatments, induced the infestation by D. speciosa larvae in 27,03%, when the aminoacid level was 2,01 ± 0,58% (X ± EP), with probability of 0,0196pest infestation increment. The results indicated that the aminoacid level increasing affects the tubercles production, mainly in the Monalisa cultivar, in winter cropping with long dry periods.

Leaf consumption and duration of instars of the cassava defoliator Erinnyis ello (L., 1758) (Lepidoptera, Sphingidae)

The objectives of this research were to evaluate leaf consumption and the developmental time of the larvae of Erynnyis ello (L., 1758) (Lepidoptera, Sphingidae) reared on cassava, in order to obtain information for the integrated management of this pest. The larvae were reared on excised cassava leaves in Petri dishes and later in gerbox, and kept in chambers at 24 ± 2 ºC and 75 ± 10% RH. The total leaf area consumed by the larva to complete its development was 589.67 cm²; each of the five instars consumed, respectively: 1.89 cm²; 5.74 cm²; 17.48 cm²; 76.66 cm²; and 487.90 cm². The consumption by the first three instars was insignificant, and did not reach 5% altogether; the 4th represented 13%; the 5th presented a consumption significantly higher, about 82.7%. The total time for the larval development was 22.61 days, and the duration for each of the five larval instar was, respectively: 4.35; 3.19; 3.32; 4.52; and 4.94 days. The pre-pupal period lasted 2.29 days. Since the highest consumption is by the 5th instar larva, the control should be applied before this age to avoid heavier damages to the cassava crop.

Características biológicas de Nipteria panacea Thierry-Mieg (Lepidoptera, Geometridae), desfolhadora do abacateiro, na região serrana do Espírito Santo, Brasil

Biological characteristics of Nipteria panacea Thierry-Mieg (Lepidoptera, Geometridae), an avocado tree defoliator, in highlands areas of Espírito Santo, Brazil. The objective of this research was to evaluate some biological characteristics of the avocado tree defoliator, Nipteria panacea Thiery-Mieg, in laboratory. This species showed the mean development cycle of 58.1 days, embryonic period of eight days and egg viability 56.3%. Pos-embryonic development lasted 36.5 days with viability of 48.2%, pupal period 11.6 days with viability of 76.0%, and mean longevity of females was 19.5 days with a production of 177 eggs per female. Other parameters were also observed and discussed.

Comparação dos sistemas de cultivo nativo e adensado de erva mate, Ilex paraguariensis St. Hil., quanto à ocorrência e flutuação populacional de insetos

This research was carried out in order to compare the occurrence of insects in two maté cultivation systems, native and high tree density. It was performed from August/2000 to September/2001, in a private property in São Mateus do Sul county, in Paraná State, Brazil. Visual inspections of trees and light traps were used to evaluate insect populations in both areas. For Hedypathes betulinus (Klug) (Coleoptera, Cerambycidae), only six adults were observed in the dense area. Based on presence of sawdust at the basis of the trunk, it was obtained that the number of attacked trees did not surpass 11% in either area. For Gyropsylla spegazziniana (Lizer y Trelles) (Hemiptera, Psyllidae), the number of galls per tree was counted and it was observed that the population peak occurred from November to January. For Hylesia spp. (Lepidoptera, Saturniidae) and Thelosia camina Schaus (Lepidoptera, Eupterotidae), the presence of caterpillars on the trees was noticed from September to February, with the population peak in November and December. Adults of Hylesia spp. were more numerous in February and March. Two species that were not previously recorded for Brazil on maté were identified: Hylesia paulex Dognin (83%) and Hylesia remex Dyer (17%), collected with light traps. The maté caterpillar, T. camina was not collected with these traps. Nymphs and adults of Ceroplastes grandis Hempel (Hemiptera, Coccidae) were observed along the year on the branches, with population peak between April and June for the nymphs and from September to November for the adults. It should be considered that despite higher insect incidence in the dense area compared to the native area, the first presents higher yield, and that with a good pest management program the insect problems can be minimized.

Aspectos biológicos de Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera, Bruchidae) em Phaseolus vulgaris L., cv. Carioca (Fabaceae), sob condições de laboratório

Zabrotes subfasciatus is a serious pest of common beans, P. vulgaris L.. In Brazil there are several studies dealing with resistance of bean genotypes to this insect, while other studies have emphasized the utilization of oils and powders from plants to repel their attack. In this paper, fecundity, fertility, pattern of oviposition, life cycle and longevity were evaluated for a Brazilian stock from the Goiás State on P. vulgaris cv. Carioca, at 30ºC and 70% R.H. The mean fecundity was 38 eggs per female and 73% of viability. Egg laying showed an aggregated pattern. Males and females lived an average of 13 and 9 days, respectively. The total life cycle lasted for about 28 days.

Development of a control alternative for the citrus fruit borer, Ecdytolopha aurantiana (Lepidoptera, Tortricidae): from basic research to the grower

All research steps, developed from 1995 to 2000, to synthesize the sex pheromone of Ecdytolopha aurantiana (Lima, 1927) are described, in order to monitoring this pest that causes losses in the order of 50 million dollars per year to citriculture in the State of São Paulo. The basic researches conducted are described, including the development of an artificial diet for the insect, the study of its temperature and humidity requirements, behavioral studies, and synthesis of the male-attracting substance up to the formulation and distribution of the pheromone to the grower, by means of its commercialization. It is a case of success, at a cost of 50 thousand dollars, involving inter- and multidisciplinary researches, which can be adopted to other insect pests in the country.

Analysis of the genetic diversity in Metopolophium dirhodum (Walker) (Hemiptera, Aphididae) by RAPD markers

The emergence of host-races within aphids may constitute an obstacle to pest management by means of plant resistance. There are examples of host-races within cereals aphids, but their occurrence in Rose Grain Aphid, Metopolophium dirhodum (Walker, 1849), has not been reported yet. In this work, RAPD markers were used to assess effects of the hosts and geographic distance on the genetic diversity of M. dirhodum lineages. Twenty-three clones were collected on oats and wheat in twelve localitites of southern Brazil. From twenty-seven primers tested, only four primers showed polymorphisms. Fourteen different genotypes were revealed by cluster analysis. Five genotypes were collected only on wheat; seven only on oats and two were collected in both hosts. Genetic and geographical distances among all clonal lineages were not correlated. Analysis of molecular variance showed that some molecular markers are not randomly distributed among clonal lineages collected on oats and on wheat. These results suggest the existence of host-races within M. dirhodum, which should be further investigated using a combination of ecological and genetic data.

Rearing method of Oryzaephilus surinamensis (L.) (Coleoptera, Silvanidae) on various wheat grain granulometry

Oryzaephilus surinamensis is one of most common insect pest of grains and a variety of stored products, and has been found in high numbers in almost all storage facilities. However, laboratory mass rearing of this insect for bioassays is not a simple task, mainly because of its feeding behavior, small size, and high mobility. Thus, the aim of this work was to develop a simple and efficient laboratory rearing method for O. surinamensis, using wheat kernels milled into different granulometry to obtain large number and standardized population at different life stages for bioassays. The adults were collected from storage grain facilities in the southern region of Brazil and 100 specimens were placed inside glass jars with wheat kernels milled at different grades and kept at 25±0.5ºC and 65±5% relative humidity. The insects were allowed to copulate and lay eggs for 10 days and then removed. The number of eggs, larvae, and pupae was counted at five-day intervals; longevity of the second generation adults was evaluated. The kernels milled at grade 20 were the best medium for offspring production: 89% of eggs by the 5th day; 30.5% larvae by the 10th day; 43% pupae by the 30th day and 63.4% adults at the 46th day. The adults survived up to 450 days. Culturing O. surinamensis under the described conditions, transferring the parental adults by the 10th day after infestation and replacing the media when population builds up will produce enough insects of each stage for various laboratory bioassays.

Ocorrência e aspectos biológicos de Anomala testaceipennis Blanchard (Coleoptera, Scarabaeidae)

Os coleópteros da família Scarabaeidae são importantes pragas em áreas de culturas e pastagens, principalmente devido à ação das larvas, as quais danificam o sistema radicular. São escassas as informações sobre esse grupo de pragas, desta forma, o presente trabalho teve por objetivo estudar a ocorrência e o ciclo biológico de Anomala testaceipennis Blanchard, 1856. De novembro de 2005 a novembro de 2006 foram realizadas coletas diárias com uma armadilha luminosa, e em laboratório os adultos foram mantidos em recipientes plásticos, contendo solo e mudas de Brachiaria decumbens, para estudo de biologia. Foram coletados 263 adultos de A. testaceipennis, em quase todos os meses do ano, não sendo registrados apenas em março e junho. Sobre a biologia verificou-se que o período embrionário durou em média 13,2 dias, o 1° instar 26,7 dias, o 2° instar 19,4 dias, o 3° instar 58,2 dias, a fase de pré-pupa 50,2 dias e a fase de pupa 13,6 dias. A longevidade dos adultos foi semelhante para machos e fêmeas, e durou em média 14,1 dias. No laboratório as fêmeas ovipositaram 7,3 ovos em média. O ciclo de ovo a adulto durou 139,4 dias em média.

Feeding and oviposition preferences of Ctenarytaina spatulata Taylor (Hemiptera, Psyllidae) for Eucalyptus spp. and other Myrtaceae in Brazil

Feeding and oviposition preferences of Ctenarytaina spatulata Taylor (Hemiptera, Psyllidae) for Eucalyptus spp. and other Myrtaceae in Brazil. The Australian psyllid, Ctenarytaina spatulata Taylor (Hemiptera, Psyllidae), was first detected in Brazil in 1994, where it was found on drought-affected shoots of Eucalyptus grandis in a plantation located in the northern part of Paraná State. The oviposition and feeding preferences of this psyllid were examined on 19 Eucalyptus species, one Eucalyptus hybrid (Cambiju), three Corymbia species and four native Myrtaceae species (Hexaclames edulis, Marlieria edulis, Plinia trunciflora, and Psydium sp.) under greenhouse conditions. The largest populations of C. spatulata were found on E. robusta and E. pellita, while sizeable infestations were also found on E. urophylla, E. grandis, and the Cambiju hybrid. The plants with the greatest symptoms of damage were E. grandis and E. resinifera. Eucalyptus cinerea, E. benthamii, E. pilularis, and E. dunnii were not infested and E. cloeziana was minimally infested. Among the Corymbia species, the number of eggs of C. spatulata was very low on C. citriodora and C. torelliana. No eggs and nymphs of C. spatulata were found on native Brazilian Myrtaceae. The number of eggs on plants was highly correlated with the subsequent levels of nymphs, suggesting that egg counts can be used as a viable monitoring tool to assist with the integrated management of this pest.

Biological cycle of Tenuipalpus heveae Baker (Acari, Tenuipalpidae) on leaflets of three rubber tree clones

Life cycle of Tenuipalpus heveae Baker (Acari, Tenuipalpidae) on leaflets from three rubber tree clones. The biological cycle of Tenuipalpus heveae Baker, 1945 (Tenuipalpidae), a potential rubber tree pest mite, was studied by the observation of individuals reared on leaflets of the clones GT 1, PB 235 and RRIM 600, in controlled environmental conditions. Three daily observations were done of 60 eggs on leaflets from each clone in order to verify the development of immature stages and the female oviposition. The fertility life table was constructed based in the collected data. Mites reared on PB 235 had faster rate of development, requiring less time in days, to double its population in number (TD), and had the highest values for egg production, female longevity, net reproductive rate (Ro), intrinsic rate of natural increase (r m) and finite rate of increase (λ). Lower reproductive values and the longest time necessary to reach adult stage were recorded for the mites on GT 1. In all studied clones, the deutonymphal phase had the highest viability, while the larval phase had the lowest, highlighted by the survivorship curve that indicated high mortality during this life stage. The clone PB 235 allowed the most suitable conditions for the development of T. heveae, followed by RRIM 600, while GT 1 was the less suitable substratum to rear this mite species.