Feeding and defecation of Rhodnius (hemiptera: Reduviidae) fed human blood

Feeding and defecation behavior of Rhodnius prolixus Stal, 1859, R. robustus Larrousse, 1927, R. neivai Lent, 1953 and R. pictipes Stal, 1872, artificially fed on human blood, were studied under laboratory conditions. In all species, first instar nymphs did not defecate in the first 30 minutes after feeding. R. pictipes did not accept artificial feeding but fed directly on humans. Nymph and adult R. prolixus had a higher defecation index (DI) than other species; third instar nymphs had the highest DI = 1.62. In all instars, most individuals accepted the food in 3 Pounds minutes and finished feeding in less than 15 minutes.     

Changes associated with laboratory rearing in antennal sensilla patterns of Triatoma infestans, Rhodnius prolixus, and Rhodnius pallescens (Hemiptera, Reduviidae, Triatominae)

We examined changes in the array of antennal sensilla of three species of Triatominae (Triatoma infestans, Rhodnius prolixus, and R. pallescens) following their establishment for different periods in laboratory culture. In each case, the laboratory colonies were compared with conspecific samples taken directly from the field, by quantitative analysis of the sensilla arrays on the three distal segments of the antenna in terms of the densities of three types of chemoreceptors (basiconics and thick and thin walled trichoids) and one type of mechanoreceptor (bristles). Sensilla densities were compared by ANOVA or non-parametric tests, and by multivariate discriminant analysis. Strains of the same species reared in different laboratories showed significant differences in their sensilla arrays, especially when compared to field-collected material from the same geographic origin. A Bolivian strain of T. infestans reared in the laboratory for 15 years and fed at monthly intervals, showed greatest differences from its conspecific wild forms, especially in terms of reductions in the number of chemoreceptors. By contrast, an Argentine strain of T. infestans reared for 25 years in the laboratory and fed weekly, showed a relative increase in the density of mechanoreceptors. A Colombian strain of R. prolixus reared for 20 years and fed weekly or fortnightly, showed only modest differences in the sensilla array when compared to its wild populations from the same area. However, a Colombian strain of R. pallescens reared for 12 years and fed fortnightly, did show highly significant reductions in one form of chemoreceptor compared to its conspecific wild populations. For all populations, multivariate analysis clearly discriminated between laboratory and field collected specimens, suggesting that artificial rearing can lead to modifications in the sensory array. This not only supports the idea of morphological plasticity in these species, but also suggests caution in the use of long-established laboratory material for experimental studies designed to extrapolate the natural behaviour and physiology of these species.     

Defecation index and reproductive success of Triatoma maculata (Hemiptera: Reduviidae) under laboratory conditions

The reproductive and defecating behavior of Triatoma maculata (Erichson 1848) was studied on animals from an university culture in Venezuela. This species does not reach the importance of Rhodnius prolixus Stal 1859 as Chagas disease vector in Venezuela. This study addressed the role of defecating frequency, an index of how dangerous the animals are for the human population, and its relationship with why T. maculata is a less important vector than R. prolixus. Human blood was fed to the insects through an artificial feeding device. The 2nd and 3rd instar nymphs defecated more frequently (Id= 0.6, n=40), and our Vth instar nymphs did not complete sexual differentiation. Fertility was 55% (n=865) and fecundity 8 eggs/female/week (n=26). Egg incubation lasted 22 days (n=477). Female longevity was 51 days (n=26). Intermould time grew progressively from 35 days for 1st to 40 days for 4th instar nymphs (n=40). Mould percentage varied from 0% for Vth to 63% for 3rd instar nymphs. Mortality varied from 8% for 3rd to 100% for Vth instar nymphs. These results support evidences explaining the lesser vectorial capacity and low density of T. maculata in human domiciles, including reduced reproduction and defecation when the animal feeds on human blood.     

Intravital microscopy and image analysis of Rhodnius prolixus (Hemiptera: Reduviidae) hematophagy: The challenge of blood intake from mouse skin

Hematophagous insects transmit many of the most dangerous parasitic diseases. The transmission usually occurs during hematophagy or just after as this is when the vector and the host are in contact. The contact time is determined by the feeding performance of the insect in each host. In triatomines, feeding performance interferes with both their life cycle and the vectorial competence to transmit the hemoflagellate Trypanosoma cruzi. Triatomine bugs are vessel feeders, obtaining their blood meals directly from the vessels (venules or arterioles) of their vertebrate hosts. The host blood intake rate is not constant during the feeding, and the sucking frequency of triatomines tends to be higher and to contain fewer interruptions in pigeons than in mice. To identify the difficulties encountered by triatomine bugs in obtaining blood meals from mouse skin, we used intravital microscopy techniques associated with electromyograms of the cibarial pump. To monitor the vibration of the cannulated vessels and the blood flow through the head of the insect during the engorgement phase, we introduced a novel method for image analysis. The mean number of vessels used during a Rhodnius prolixus blood meal was 3.4 ± 1.2, and the insects fed more in venules (63%) than in arterioles (37%). An important increase in vascular permeability was observed throughout the feeding. Platelet aggregation, rolling and leukocyte adherence were analyzed on the venular endothelium, showing remarkable increases for some time following the R. prolixus feeding. The reduction in sucking frequency that was observed during insect feeding was likely due to the increased cibarial pump filling time. The monitoring of the vessel wall pulsation also permitted the registration of regurgitation-like movements during blood pumping, with these movements being recorded mostly during the second half of the feeding. The evaluation of blood flow through the head of the insect suggested that the regurgitation-like movements were not true regurgitations and were caused by abrupt difficulties in the function of the cibarial pump. The role of the platelet plugs and the changes in blood viscosity at the R. prolixus feeding site are discussed. The method introduced in the present study to analyze the images brings new insights into the interaction between hematophagous vectors and their hosts, reinforcing the importance of insect saliva throughout the feeding process.     

Life cycle and reproductive parameters of Clerada apicicornis signoret (Hemiptera: Lygaeidae) under laboratory conditions

The life cycle of Clerada apicicornis was determined under laboratory conditions. Mean development times in days were: egg 27.2, nymph I 12.5, nymph II 12, nymph III 13.4, nymph IV 16.4, nymph V 26. The life expectancy of adults ranged from 117 to 317 days (mean 196 days). Based on a cohort of 29 females of C. apicicornis, a horizontal life table was constructed. The following predictive parameters were obtained: net rate of reproduction (Ro = 48.31), intrinsic rate of population increase (r(m) = 0.153), generation time (Tc = 28.20 weeks), and finite rate of population increment (lambda = 1.16). The reproductive value (Vx) for each age class of the cohort females was calculated. The following observed parameters were calculated after mortality in each stage: net rate of reproduction (R'o=13.4), intrinsic rate of population increase (r c' =0.09 ), and finite rate of population increment (lambda' =1.1). The generation time (Tc' =27.4) was estimated using the methods of Laughlin and Bengstron. A vertical life table was elaborated and mortality was described for one generation of the cohort.     

A missing metabolic pathway in the cattle tick Boophilus microplus.

Heme proteins are involved in a wide variety of biological reactions, including respiration, oxygen transport and oxygen metabolism [1]. The heme prosthetic group is synthesized in almost all living organisms except for a few pathogenic bacteria and trypanosomatids that use blood as food [2] [3]. There is a general belief that all nucleated animal cells synthesize heme [1] [4]. However, blood-feeding arthropods ingest enormous amounts of vertebrate blood in a single meal and the heme pathway has not been studied in these animals. We have examined heme synthesis in two hematophagous arthropods - the blood-sucking bug Rhodnius prolixus and the cattle tick Boophilus microplus. We show that R. prolixus makes heme and has a fully operative heme biosynthetic pathway, while B. microplus does not. To our knowledge, this is the first report of an animal that does not synthesize its own heme and relies solely on the recovery of heme present in the diet. Because of the inability of Boophilus to synthesize heme and its ability to deal efficiently with large amounts of free heme, we propose this organism as a good model for studying heme transport and reutilization in animal cells.     

Morphometric analysis of Rhodnius prolixus Stal (Hemiptera:Reduviidae) midgut cells during blood digestion

Post-feeding ultrastructural modifications to the midgut cells of Rhodnius prolixus are quantified using morphometry. Changes in relative and absolute volumes and/or surface areas are demonstrated for the whole cells, nuclei, mitochondria, rough endoplasmic reticulum, lysosomes, Golgi apparatus, storage vesicles, glycogen, microvilli, and basal labyrinth, before and during blood digestion. These parameters are separately determined for cells from each of the three midgut regions, and are correlated against previously published cycles of digestive enzyme activities. The results support the proposed division of the midgut of R. prolixus into three functional regions: the anterior midgut or crop is the site of water transport immediately after feeding, and of lipid and glycogen storage. No protein digestion occurs in this region. The anterior intestine is the site of most proteinase synthesis and secretion, although limited absorption and nutrient storage also occurs. The posterior intestine is responsible for some secretory activity, but is also implicated as the most important region for absorption of digested nutrients and for carbohydrate absorption and storage.     

Antidiuretic mechanism in Rhodnius prolixus (Stal, 1859) (Hemiptera, Reduviidae)

An antidiuretic mechanism is proposed for Rhodnius prolixus, whose activity is manifested when the diuretic phase has terminated and is maintained until the insect has fed again. Presumably this mechanism acts at the level of the proximal rectal sphincter and is inhibited by ingestion, mechanical distension of the gut and central disinhibition by decapitation. It is suggested that the antidiuretic activity is maintained by the nervous system and is modified when there is distention of the gut or when the nervous signal is interrupted by decapitation. It is demonstrated that the excretion of urine in R. prolixus is not controlled exclusively by the diuretic hormone, but rather that factors not linked to the haemolymph maintain the insects in a diuretic or nondiuretic state.     

Tick-borne encephalitis viral titers of engorged adult Ixodes persulcatus ticks

The results of titration of adult virus infected ticks of Ixodes persulcatus are given. The ticks were collected in various natural nidi of tick-borne encephalitis and individually fed on laboratory animals. The results of titration of ticks of the same species primarily infected on laboratory animals during virusemia are given as well. Changes of the virus titer in engorged ticks were observed till the egg production. A role of the presence of virus in engorged ticks for the process of the agent circulation in natural nidi is discussed.     

Predators of the tickAmblyomma variegatum (Acari: Ixodidae) in Guadeloupe,French West indies

Some vertebrate species in Guadeloupe are predators of free or parasitic stages ofAmblyomma variegatum (Fabricius). Among birds, 1.9% of the 421 identified animals found in the stomachs of grackles (Quiscalus lugubris), 1.6% of the 364 animals found in the stomachs of free-ranging chickens, and 0.3% of the 4642 animals found in the stomach of cattle egrets (Bubulcus ibis) wereA. variegatum ticks. The most efficient predator of ticks was the tropical fire ant,Solenopsis geminata, which was observed to only attack engorged stages of ticks. An average of 8% of the 564 individual ticks or batch of ticks released on the ground, mainly in a grass environment, were attacked by this ant species. Mice (Mus musculus) and mongooses (Herpestes auropunctatus) feed on engorged nymphs and female ticks. None of the 15 strains of entomophagous nematodes, generaSteinernema andHeterorhabditis, experimentally put in contact with engorged larvae and nymphs were able to parasitize them. In the laboratory, the insectsMegaselia scalaris (Diptera) andTineola sp. (Lepidoptera) were occasionally found to feed on engorged and unfed ticks, respectively.     

Biological effect of 1-dodecanol in teneral and post-teneral Rhodnius prolixus and Triatoma infestans (Hemiptera: Reduviidae)

Topical application of 1-dodecanol was significantly more toxic against teneral first nymphs (1-3 h old) than post-teneral first nymphs (24 h old). The lethal dose ratios were 711,500 for Rhodnius prolixus and 3613 for Triatoma infestans. No significative difference between LD50 was found when 1-dodecanol was injected in recently hatched adult R. prolixus (1-4 h old) nor in older adults (24 h old). These values were similar to those calculated for deltamethrin (an effective triatomicide), showing that 1-dodecanol had no insecticidal properties when it was applied by injection. Topical application of high dose of 1-dodecanol (1 microg/i) on teneral first nymphs of R. prolixus, produced an interruption of the darkening process of the cuticle, and probably in the development of its physiological properties.     

氯虫苯甲酰胺对黑肩绿盲蝽实验种群的影响

采取稻茎浸渍法测定了氯虫苯甲酰胺对黑肩绿盲蝽毒力,利用生命表技术研究了氯虫苯甲酰胺对黑肩绿盲蝽实验种群的影响,为协调水稻害虫的化学防治和生物防治提供参考。结果表明,氯虫苯甲酰胺对黑肩绿盲蝽若虫LC50和LC10分别为83.5mg/L和61.3 mg/L,氯虫苯甲酰胺对黑肩绿盲蝽成虫LC50和LC10分别为64.3 mg/L和39.0 mg/L。氯虫苯甲酰胺对黑肩绿盲蝽若虫和成虫的LC10分别大于和接近于大田使用剂量40mg/L。以大田使用剂量40 mg/L氯虫苯甲酰胺稻茎浸渍法处理黑肩绿盲蝽3龄若虫后,其产卵期、寿命和产卵量降低了4.3 d、3.0 d和22.0粒,与对照相比差异显著,表明氯虫苯甲酰胺对当代成虫寿命与生殖力影响较大;药剂处理后次代种群的成虫前期延长了2.3 d;存活率、平均日产卵量跟对照相比明显降低;种群净增值率、周限增长率、内禀增长率跟对照相比明显降低,分别为21.0(对照63.3)、18.8(对照19.2)、0.16(对照0.22),而种群加倍时间延长为4.3(对照3.2);这些结果表明,在40 mg/L浓度下,氯虫苯甲酰胺能够降低黑肩绿盲蝽种群的增长。     

The ability of Rhodnius prolixus (Hemiptera; Reduviidae) to approach a thermal source solely by its infrared radiation

The ability of the haematophagous bug Rhodnius prolixus to approach a pure IR source only by its long-wave infrared radiation (IR) was investigated. To exclude any heated air from reaching the bug a cooled IR-transmitting window was placed between the IR-source (a Peltierelement heated to 35 degrees C) and the bug. Starved bugs were tested under invisible short-wave IR illumination (lambda<1 μm) in order to exclude also any visual cues. The number of bugs approaching the IR-source was significantly increased compared to the controls in which the IR-source was turned off (chi(2)-test, P<0.01). Our results show that Rhodnius prolixus can use infrared stimuli to find a host.     

Role of superoxide and reactive nitrogen intermediates in Rhodnius prolixus (Reduviidae)/Trypanosoma rangeli interactions.

This study compares aspects of the superoxide, nitric oxide and prophenoloxidase pathways in Rhodnius prolixus hemolymph, measured in parallel, in response to Trypanosoma rangeli inoculation. Responses to two strains of T. rangeli, and two developmental forms, were studied, and the results obtained were correlated with the ability of the parasites to survive, multiply, and complete their life cycles in the hemolymph of the host. T. rangeli H14 strain parasites, which fail to complete their life cycle in Rhodnius by invading the salivary glands, stimulated high levels of superoxide and prophenoloxidase activity, which peaked 24 h after inoculation. Simultaneously, the concentration of hemolymph nitrites and nitrates increased, indicative of nitric oxide activity, but parasite numbers remained low. T. rangeli Choachi strain parasite inoculation also stimulated superoxide and prophenoloxidase activity, which, though significantly lower than the equivalent responses to the H14 strain, also peaked at 24 h. However, nitrate and nitrite levels in Choachi strain-inoculated hemolymph remained low, and this parasite strain multiplied rapidly, especially following peak superoxide activity, and eventually invaded the salivary glands for transmission to a vertebrate host. In both strains, short form epimastigotes stimulated greater superoxide and prophenoloxidase responses than long form epimastigotes. Injection of the NADPH oxidase inhibitor N-ethylmaleimide or the inducible nitric oxide synthase inhibitor S-methyl isothiourea sulfate caused significantly higher insect mortalities in groups of R. prolixus inoculated with either parasite strain compared with those of uninfected control insects. This indicates that both NADPH oxidase and nitric oxide synthase activity may be involved in the immune response of R. prolixus to infection by T. rangeli. Finally, Western blotting of R. prolixus hemocyte lysates revealed the presence of a protein immunologically related to the human NADPH oxidase complex, the initiator enzyme of the respiratory burst.     

Epidemiology of Chagas disease in Guatemala: infection rate of Triatoma dimidiata,Triatoma nitida and Rhodnius prolixus (Hemiptera,Reduviidae) with Trypanosoma cruzi and Trypanosoma rangeli (Kinetoplastida,Trypanosomatidae)

A five-year domiciliary collection in the 22 departments of Guatemala showed that out of 4,128 triatomines collected, 1,675 were Triatoma dimidiata (Latreille, 1811), 2,344 were Rhodnius prolixus Stal 1859, and only 109 were T. nitida Usinger 1939. The Chagas disease parasite, Trypanosoma cruzi, was found in all three species. Their natural infection rates were similar in the first two species (20.6%; 19.1%) and slightly lower in T. nitida(13.8%). However there was no significant difference in the infection rates in the three species (p = 0.131). T. dimidiata males have higher infection rates than females (p = 0.030), whereas for R. prolixus there is no difference in infection rates between males and females (p = 0.114). The sex ratios for all three species were significantly skewed. More males than females were found inside houses for T. dimidiata (p < 0.0001) and T. nitida (p = 0.011); a different pattern was seen for R. prolixus (p = 0.037) where more females were found. Sex ratio is proposed as an index to show the mobility of T. dimidiata in different populations. T. dimidiata is widely distributed in the country, and is also the main vector in at least ten departments, but R. prolixus with higher vectorial capacity is an important vector in at least two departments.     

Functional consequences of the human leptin receptor (LEPR) Q223R transversion

Perturbations in the functional integrity of the leptin axis are obvious candidates for mediation of altered adiposity. In a large number of genetic association studies in humans, the nonconservative LEPR Q223R allele has been inconsistently associated with adiposity. Subtle, long-term effects of such genetic variants can be obscured by effects of the environment and other confounders that render definitive inferences difficult to reach. We directly assessed the biological effects of this variant in 129P3/J mice segregating for the humanized Lepr allele at codon 223. No effects of this allele were detected on body weight, composition, or energy expenditure in animals fed diets of varying fat content over periods as long as 235 days. In vitro, Q223R did not affect leptin signaling as reflected by activation of STAT3. We conclude that Q223R is unlikely to play a significant role in regulation of human adiposity. This approach to vetting of human allelic variation might be more widely used.     

Studies on Trypanosoma rangeli Tejera, 1920. VII--Its effect on the survival of infected triatomine bugs

The pathological effects of Trypanosoma rangeli on Rhodnius prolixus and R. robustus, and the relation of mortality to infection, were studied under laboratory conditions. Frequent observations revealed that when the first instar nymphs of R. prolixus and R. robustus were infected with T. rangeli, survival of the bugs during the stages of development to the adult stage decreased. This decrease was statistically significant when compared with uninfected control-bugs, indicating that T. rangeli is pathogenic for both species of triatomine. In R. prolixus the most affected nymphal stages were the first, second and fifth instars, where a higher mortality was also observed. In R. robustus a progressive increase of the mortality from the first to fifth instars, was observed. The pathogenicity of T. rangeli as measured by overall mortality was the same in R. prolixus and R. robustus. The possible pathogenic mechanism of T. rangeli in triatomine-bugs and its epidemiological implications, are discussed.     

Biology and defensive adaptations in Rodatus major (Coleoptera: Coccinellidae) and its prey, Monophlebulus pilosior (Hemiptera: Margarodidae)

The biology of the Australian coccinellid Rodatus major (Blackburn) and its prey, the hemipteran margarodid Monophlebulus pilosior (Maskell), are described. New predators and a new parasite of M. pilosior are recorded. Rodatus major makes use of elaborate and distinctive defensive adaptations and behaviour to protect itself from predators. They include cryptic coloration, aposematic behaviour, production of wax by larvae, production of a large, thick protective covering concealing the pupa and resembling an M. pilosior ovisac, and reflex bleeding. It is the only known coccinellid species in which both prepupa and pupa are hidden by a protective shroud of wax threads. Rodatus major has a specialized feeding behaviour. Its potential as a biological control agent is assessed. It is only the second Australian margarodid-feeding coccinellid to be studied, Rodolia cardinalis (Mulsant) being the first.     

Vital statistics of Rhodnius neivai Lent, 1953 (Hemiptera: Reduviidae) under experimental conditions

A statistical evaluation of the population dynamics of R. neivai is based on six cohorts experiments conducted under controlled laboratory conditions. Two blood sources were offered to animals: rabbit and hen. Egg hatching, nymphal development time and mortality, adult longevity and age-specific mortality, female age-specific fecundity and fertility were determined. In addition, some population parameters were evaluated, such as: life expectancy, intrinsic rate of natural increase, net reproduction rate, finite rate of increase, reproductive value and stable age distribution. Life cycle was longer in the animals fed on rabbit, nymphal survival was slightly higher in the individuals fed on hen. Age of first reproduction was lower in the insects fed on hen, but reproductive output and total number of reproductive weeks were greater in the cohorts fed on rabbit. Intrinsic and finite rate of increase were greater in the animals fed on hen. Generation time was slightly greater in the cohorts fed on rabbit. Net reproduction rate was similar on both blood sources, although it was slightly bigger in the individuals fed on rabbit. Reproductive value in the insects fed on rabbit was twice as much as the registered in the animals fed on hen.     

Feeding behaviour of morphologically similar Rhodnius species: influence of mechanical characteristics and salivary function

Despite their morphological similarities, very similar Rhodnius species (R. prolixus, R. robustus, R. nasutus and R. neglectus) displayed a distinct feeding behaviour when fed on artificial feeder, pigeon or mouse. On pigeon hosts, these species showed distinct groups in terms of cumulative probing time - quicker species (R. prolixus and R. neglectus) followed by R. nasutus and finally a much slower species (R. robustus). On mouse hosts, R. nasutus showed quicker probing time compared to the other three species. Moreover, R. prolixus displayed quicker probing time compared to R. robustus and R. neglectus. Except for R. nasutus, the mean total ingestion rate tended to have different values between feeding sources (artificial feeder>pigeon>mouse). The volume ingested by each cibarial pump contraction and maximum frequency obtained using the artificial feeder are expected to be related to intrinsic mechanical characteristics of the insect feeding apparatus. However, probing time and the modulation of cibarial pump frequency on live hosts may be related to salivary function. R. prolixus showed high mechanical and salivary efficiency, achieving high values of total ingestion rate when fed on artificial feeder or either of the hosts. Comparative analysis suggests that species which possess higher total ingestion rates tend to achieve higher nutritional status, allowing them to reach higher densities.