Blood-sucking Bug Research Articles

SIFamide Influences Feeding in the Chagas Disease Vector, Rhodnius prolixus.

SIFamides are a family of highly conserved neuropeptides in arthropods, and in insects are mainly expressed in four medial neurons in the pars intercerebralis of the brain. Although SIFamide has been shown to influence sexual behavior, feeding, and sleep regulation in holometabolous insects such as Drosophila melanogaster, little is known about its role in hemimetabolous insects, including the blood-sucking bug, Rhodnius prolixus. In this study, we confirm the nucleotide sequence for R. prolixus SIFamide (Rhopr-SIFa) and find characteristic phenotypic expression of SIFamide in four cells of the pars intercerebralis in the brain. In addition to extensive SIFa projections throughout the entire central nervous system, SIFamidergic processes also enter into the corpus cardiacum, and project along the dorsal vessel, suggestive of Rhopr-SIFa acting as a neurohormone. Physiologically, Rhopr-SIFamide induces dose-dependent increases in heartbeat frequency in vitro suggesting the presence of peripheral receptors, and thereby indicating Rhopr-SIFa is released to act upon peripheral targets. We also explore the function of Rhopr-SIFa in R. prolixus, specifically in relation to feeding, since R. prolixus is a blood-gorging insect and a vector for Chagas disease. The intensity of SIFamide-like staining in the neurons in the brain is diminished 2 h following feeding, and restocking of those cells is finished 24 h later, indicating Rhopr-SIFa may be released at feeding. The results of temporal qPCR analysis were consistent with the immunohistochemical findings, showing an increase in Rhopr-SIFa transcript expression in the brain 2 h after feeding. We also observed enhanced feeding (size of meal) in insects injected with Rhopr-SIFa whereas insects with RNAi-mediated knockdown of the Rhopr-SIFa transcript consumed a significantly smaller blood meal relative to controls. These data suggest that the four SIFamidergic neurons and associated arborizations may play an important function in the neuronal circuitry controlling R. prolixus feeding, with Rhopr-SIFa acting as a central and peripheral neuromodulator/neurohormone.

Juvenile hormone III skipped bisepoxide, not its stereoisomers, as a juvenile hormone of the bean bug Riptortus pedestris

Juvenile hormone (JH) plays a pivotal role in many aspects of insect physiology. Although its presence was first reported in a blood-sucking bug belonging to the suborder Heteroptera (true bugs), JH species in the group has long been controversial. Although some recent studies proposed a putative JH molecular species in several Heteropteran species, it is not conclusive because physicochemical analyses were insufficient in most cases. Here, we studied this issue with an ultraperformance liquid chromatography–tandem mass spectrometer (UPLC-MS/MS) equipped with C18 and chiral columns in the bean bug Riptortus pedestris (Heteroptera, Alydidae), in which the JH species has long been controversial. Although a recent study describes JHSB3 as the major JH of this species, that finding was not conclusive because its chirality has not been clarified. In the present study, we detected methyl (2R,3S,10R)-2,3;10,11-bisepoxyfarnesoate, commonly named juvenile hormone III skipped bisepoxide (JHSB3), in the culture media of the corpora cardiaca-corpus allatum (CC-CA) complex and in the hemolymph of this species by a chiral ultraperformance liquid chromatography- tandem mass spectrometer (UPLC-MS/MS). Other JHSB3 stereoisomers were not detected. Topical application of JHSB3 effectively averted diapause. These results indicate that JHSB3 is the major JH of R. pedestris. The present study further revealed that JHSB3 and its (2R,3S,10S) isomer are more potent than (2S,3R,10R) and (2S,3R,10S) isomers, which suggests that there is a significance to the configuration of the 2,3-epoxide moiety in JH action. We further found a supplemental significance to the configuration of the 10-position.

Blood as fuel: the metabolic cost of pedestrian locomotion in Rhodnius prolixus.

Active searching for vertebrate blood is a necessary activity for haematophagous insects, and it can be assumed that this search should also be costly in terms of energetic expenditure. Whether by swimming, walking, running or flying, active movement requires energy, increasing metabolic rate relative to resting situations. We analysed the respiratory pattern and energetic cost of pedestrian locomotion in the blood-sucking bug Rhodnius prolixus using flow-through respirometry, by measuring carbon dioxide emission and water loss before, during and after walking. We observed an increase in the metabolic rate during walking as compared with resting of up to 1.7-fold in male R. prolixus and 1.5-fold in females, as well as a change in their respiratory pattern, which switched from cyclic during rest to continuous when the insects started to walk, remaining in this condition during locomotion and for several minutes after stopping. Walking induced a significant loss of mass in both males and females. This can be explained by an increase in both metabolic rate and water loss during walking. These data constitute the first metabolic measures of active haematophagous insects and provide the first insights into the energetic expenditure associated with the active search for blood in this group.

The Octopamine Receptor Is a Possible Target for Eugenol-Induced Hyperactivity in the Blood-Sucking Bug Triatoma infestans (Hemiptera: Reduviidae).

Eugenol is a major component of the essential oils in cloves and other aromatic plants. In insects, it produces toxic effects and repellency, and there is evidence that its site of action is the octopamine receptor. The objective of the present study was to explore whether the octopamine receptor is involved in the hyperactivity produced by eugenol in the blood-sucking bug Triatoma infestans (Klug). This insect is the main vector of Chagas disease in Latin America. Four treatments were topically applied on third instar nymphs: 1) octopamine, 2) eugenol, 3) phentolamine hydrochloride (an antagonist of the octopamine receptor) followed by octopamine, and 4) phentolamine hydrochloride followed by eugenol. Both octopamine and eugenol hyperactivated the nymphs. However, pretreatment with phentolamine hydrochloride inhibited the hyperactivating effect of both compounds. These results are in agreement with previous works on Drosophila melanogaster (Meigen) (Diptera: Drosophilidae) and the American cockroach. They suggest that the octopamine receptor is a possible site of action for eugenol.

Eugenol-hyperactivated nymphs of Triatoma infestans become intoxicated faster than non-hyperactivated nymphs when exposed to a permethrin-treated surface

BackgroundEugenol is a botanical monoterpene that hyperactivates the blood-sucking bug Triatoma infestans, and permethrin is a pyrethroid with a strong triatomicide effect. In the present work, we tested the hypothesis that eugenol-hyperactivated nymphs of T. infestans pick up more insecticide, and then become intoxicated faster, than non-hyperactivated nymphs when exposed to a permethrin-treated surface.ResultsValues of knockdown time 50% (KT50) for third-instar T. infestans exposed to a paper impregnated with permethrin were obtained under the following situations: (a.i.) immediately after topical application of eugenol (KT50: 66.75 min for acetone pre-treated controls, and 46.27 min for eugenol pre-treated nymphs); (a.ii.) 30 min after topical application of eugenol (KT50: 66.79 min for controls, and 66.79 min for eugenol pre-treated nymphs); (b) simultaneously with exposure to eugenol vapors (KT50: 51.90 min for controls, and 39.5 min for nymphs exposed to an eugenol-treated filter paper); and (c) immediately after an injection of eugenol (on average, controls were knocked down after 63.00 min, whereas nymphs injected with eugenol were knocked down after 65.30 min). In other experimental series, the distance traveled (DT) by nymphs exposed to eugenol was quantified in the same situations previously described, but without exposure to permethrin. In (a.i.), the DT in interval 0–30 min after topical application of eugenol was 487.00 (control) and 1127.50 (eugenol) cm; in (a.ii.), the DT in the interval 31–60 min after topical application was 336.75 (control) and 256.75 (eugenol) cm; in (b), DT was 939.08 (control) and 1048.53 (eugenol) cm; and in (c), it was 589.20 (control) and 700.00 (eugenol) cm. The KT50 values for permethrin decreased significantly in situations (a.i.) and (b), and eugenol only produced a significant hyperactivity in the same situations. Finally, the amount of permethrin picked up by non-hyperactivated and hyperactivated nymphs exposed to a film of permethrin was quantified by gas chromatography. Non-hyperactivated nymphs picked up 0.34 μg/insect of permethrin, while hyperactivated nymphs picked up 0.65 μg/insect.ConclusionResults support the hypothesis that eugenol-hyperactivated nymphs of T. infestans pick up more insecticide, and then become intoxicated faster, than non-hyperactivated nymphs when exposed to a permethrin-treated surface.

Human Antimicrobial Peptide Isolated From Triatoma infestans Haemolymph, Trypanosoma cruzi-Transmitting Vector.

The importance of antimicrobial peptides (AMPs) in relation to the survival of invertebrates is well known. The source and the mode of action on the insects' immune system of these molecules have been described from different perspectives. Insects produce their own AMPs as well as obtain these molecules from various sources, for example by absorption through the intestinal tract, as previously described for Boophilus microplus. Blood-sucking barber bug Triatoma infestans attracts social, economic and medical interest owing to its role in the transmission of Chagas disease. Despite new studies, descriptions of AMPs from this insect have remained elusive. Thus, the aims of this work were to characterize the antimicrobial potential of human fibrinopeptide A (FbPA) obtained from the T. infestans haemolymph and identify its natural source. Therefore, FbPA was isolated from the T. infestans haemolymph through liquid chromatography and identified by mass spectrometry. This peptide exhibited antimicrobial activity against Micrococcus luteus. Native FbPA from human blood and the synthetic FbPA also exhibited antimicrobial activity. The synthetic FbPA was conjugated with fluorescein isothiocyanate and offered to the insects. The haemolymph collected after 72 h exhibited fluorescence at the same wavelength as fluorescein isothiocyanate. Our experiments show that beyond intrinsic AMP production, T. infestans is able to co-opt molecules via internalization and may use them as AMPs for protection.

Females’ sequential mating decisions depend on both the quality of the courting male and the quality of the potential mates in a blood-sucking bug

This study investigates whether female sequential decisions on accepting or rejecting a courting male are based exclusively on the quality of the male present or are affected by the quality of potential mates. We used blood-sucking bugs Rhodnius prolixus as experimental subjects as females’ exhibit stereotyped discrimination behavior towards undesirable males, which allows sequential testing of a female’s preference for each male. We expect higher levels of female rejection when the benefit of finding a better quality male overrides the cost of rejecting a mating opportunity. Male quality was manipulated through changes in the feeding regime prior to the experiment because in this species nutritional condition affects males’ reproductive success. Females were simultaneously exposed to both a focal male currently available and a background male potentially available. The quality of each focal and background male could be either low or high and all the possible combinations were tested for each pair (low/high, high/low, high/high, low/low). We found that female rejection behavior depends not only on the quality of the focal male but also on the quality of the background male. This behavioral flexibility may allow females to maximize the benefits of their mating decisions. Using an experimental approach, we study how females of a blood-sucking bug assess male quality and decide whether to accept or reject a mating opportunity in a sequential choice paradigm. Male quality (high/low) and male availability (current/potential) were manipulated. We show that female rejection depends on both the quality of the current and potential mates. Female rejection increases when better quality males are potentially available. This behavioral flexibility may allow females to maximize the benefits of mating decisions.

A paradoxical thrombogenic mutation in factor II at the target site of arthropod bleeding toxin

Loss-of-function mutations in coagulation cascade proteins lead to bleeding diasthesis. In contrast, gain-of-function mutations in these proteins, which are exceptionally rare, lead to hereditary thrombosis. This is best exemplified by Factor V (i.e., Factor V Leiden) and Factor II (i.e., p.Arg596Leu). Here, we report a family with hereditary thrombosis. The proposita presented with cerebral venous thrombosis accompanied by infarction at the age of 12 years. Despite anticoagulation therapy with oral warfarin, she later developed deep venous thrombosis in her hepatic portal veins at the age of 27 years. A medical exome analysis identified a de novo heterozygous mutation p.Tyr434His in Factor II, which was segregated within the family. A retrospective molecular diagnosis was made using a preserved surgical specimen from the proposita's mother, who had died 10 years earlier. The p.Tyr434 residue, which was substituted in the presently reported family, was located in “exosite I” of thrombin, a critical recognition site for fibrinogen, protein C, and thrombin aptamer. Therefore, the mutant thrombin likely exerted its thrombophilic effect by altering the affinity of thrombin to downstream substrates. Furthermore, the “exosite I” domain of thrombin was inhibited by the arthropod bleeding toxin Triabin (a protein discovered from the saliva of the blood-sucking triatomine bug Triatoma pallidipennis). Our observation that patients with a p.Tyr434His mutation exhibited recurrent thrombosis provides the first proof-of-concept in humans that a pharmacologic agent targeting “exosite I” could be an effective means of specifically modulating the thrombogenic-side of the coagulation system via the inhibition of factor II.

Chagas Disease Recommendations for Solid-Organ Transplant Recipients and Donors.

Chagas Disease Recommendations for Solid-Organ Transplant Recipients and Donors.

Chagas disease study using satellite image processing: A Bolivian case

Remote sensing is the technology that has enabled us to obtain information about the Earth's surface without directly contacting it. For this reason, currently, the Bolivian state has considered a list of interesting applications of remote sensing in the country, including the following: biodiversity and environment monitoring, mining and geology, epidemiology, agriculture, water resources and land use planning. The use of satellite images has become a great tool for epidemiology because with this technological advance we can determine the environment in which transmission occurs, the distribution of the disease and its evolution over time. In that context, one of the important diseases related to public health in Bolivia is Chagas disease, also known as South American Trypanosomiasis.Chagas is caused by a blood-sucking bug or Vinchuca, which causes serious intestinal and heart long term problems and affects 33.4% of the Bolivian population. This disease affects mostly humble people, so the Bolivian state invests millions of dollars to acquire medicine and distribute it for free.Due to the above reasons, the present research aims to analyze some areas of Bolivia using satellite images for developing an epidemiology study. The primary objective is to understand the environment in which the transmission of the disease happens, and the climatic conditions under which occurs, observe the behavior of the blood-sucking bug, identify in which months occur higher outbreaks, in which months the bug leaves its eggs, and under which weather conditions this happens. All this information would be contrasted with information extracted from the satellite images and data from the Ministry of Health, and the Institute of Meteorology in Bolivia. All this data will allow us to have a more integrated understanding of this disease and promote new possibilities to prevent and control it.

Toxicity, repellency and flushing out in Triatoma infestans (Hemiptera: Reduviidae) exposed to the repellents DEET and IR3535.

DEET and IR3535 are insect repellents present worldwide in commercial products; their efficacy has been mainly evaluated in mosquitoes. This study compares the toxicological effects and the behavioral responses induced by both repellents on the blood-sucking bug Triatoma infestans Klug (Hemiptera: Reduviidae), one of the main vectors of Chagas disease. When applied topically, the Median Lethal Dose (72 h) for DEET was 220.8 µg/insect. Using IR3535, topical application of 500 µg/insect killed no nymphs. The minimum concentration that produced repellency was the same for both compounds: 1,15 µg/cm2. The effect of a mixture DEET:IR3535 1:1 was similar to that of their pure components. Flushing out was assessed in a chamber with a shelter containing groups of ten nymphs. The repellents were aerosolized on the shelter and the number of insects leaving it was recorded for 60 min. During that time, 0.006 g/m3 of the positive control tetramethrin flushed out 76.7% of the nymphs, while 1.76 g/m3 of DEET or IR3535 flushed out 30 and 0%, respectively. The concentrations required for both compounds to produce toxicity or flushing out are too high to have any practical applications. However, they showed a promising repellency. Additional research should be done to evaluate their possible use for personal protection against T. infestans bites.

Identifying Current and Missing Knowledge in the Control of Pyrethroid-Resistant Triatoma infestans, Vector of Chagas Disease

Triatomines are blood-sucking bugs that occur mainly in Latin America. They are vectors of Trypanosoma cruzi, the parasite that causes Chagas disease. Chemical control of Chagas disease´s vectors by using pyrethroid insecticides has been highly successful for the elimination of domestic infestation and consequently the reduction of the vector transmission. However, at the beginning of the 2000s a decrease in the effectiveness of the chemical control of triatomines was detected in several areas from Argentina and Bolivia, particularly in the Gran Chaco eco-region.During the last 15 years, several studies demonstrated the evolution of insecticide resistance in Triatoma infestans and established the presence of different toxicological profiles, the autosomal inherence of resistance, the biological costs of deltamethrin resistance, the expression of deltamethrin resistance thorough the embryonic development, and the main mechanisms of resistance (target-site insensitivity and metabolic detoxification of insecticides).The emergence of pyrethroid resistance coupled with the usual difficulties in sustaining adequate rates of insecticide applications emphasize the need of incorporating other tools for integrated vector and disease control, such as the proposal of the organo-phosphorus insecticide fenitrothion as an alternative chemical strategy for the management of the resistance because it was effective against pyrethroid-resistant populations in laboratory and semi-field trials.New studies on the current situation of presence and spread of resistant populations of triatomines and the acceptance of the use of alternative insecticides are critical requirements in the implementation of strategies for the management of resistance and for the rational design of campaigns oriented to reducing the vector transmission of Chagas’ disease.

Detection of a Potential New Bartonella Species "Candidatus Bartonella rondoniensis" in Human Biting Kissing Bugs (Reduviidae; Triatominae).

BackgroundAmong the Reduviidae family, triatomines are giant blood-sucking bugs. They are well known in Central and South America where they transmit Trypanosoma cruzi to mammals, including humans, through their feces. This parasitic protozoan is the causative agent of Chagas disease, a major public health issue in endemic areas. Because of the medical and economic impact of Chagas disease, the presence of other arthropod-borne pathogens in triatomines was rarely investigated.Methodology/Principal findingsIn this study, seven triatomines species involved in the transmission of T. cruzi were molecularly screened for the presence of known pathogens generally associated with arthropods, such as Rickettsia, Bartonella, Anaplasmataceae, Borrelia species and Coxiella burnetii. Of all included triatomine species, only Eratyrus mucronatus specimens tested positive for Bartonella species for 56% of tested samples. A new genotype of Bartonella spp. was detected in 13/23 Eratyrus mucronatus specimens, an important vector of T. cruzi to humans. This bacterium was further characterized by sequencing fragments of the ftsZ, gltA and rpoB genes. Depending on the targeted gene, this agent shares 84% to 91% of identity with B. bacilliformis, the agent of Carrion’s disease, a deadly sandfly-borne infectious disease endemic in South America. It is also closely related to animal pathogens such as B. bovis and B. chomelii.ConclusionsAs E. mucronatus is an invasive species that occasionally feeds on humans, the presence of potentially pathogenic Bartonella-infected bugs could present another risk for human health, along with the T. cruzi issue.

Sleeping habits affect access to host by Chagas disease vector Triatoma dimidiata.

BackgroundChagas disease, caused by the parasite Trypanosoma cruzi, is mainly transmitted by blood-sucking bugs called triatomines. In the Yucatán Peninsula, Mexico, the main vector of T. cruzi is Triatoma dimidiata. While this species may colonize houses in other regions, it is mostly intrusive in Yucatán: it generally lives in sylvan and peridomestic areas, and frequently enters inside homes, likely attracted by potential vertebrate hosts, without establishing colonies. Bugs collected inside homes have a low nutritional status, suggesting that they cannot efficiently feed inside these houses. We hypothesized that this low nutritional status and limited colonization may be associated, at least in part, with the local practice in Mayan communities to sleep in hammocks instead of beds, as this sleeping habit could be an obstacle for triatomines to easily reach human hosts, particularly for nymphal instars which are unable to fly.MethodsWe used an experimental chamber in which we placed a miniature bed in one side and a miniature hammock on the other side. After placing a mouse enclosed in a small cage on the bed and another one in the hammock as baits, T. dimidiata bugs were released in the chamber and their activity was video recorded during the night.Results T. dimidiata adults and nymphs were able to reach the mouse in bed significantly more often than the mouse in hammock (Binomial test, P < 0.0001). Moreover, females reached the mice twice as often as did males. Most of the adult bugs reached the mouse in bed by walking, while they reached the mouse in hammock by flying. Nymphs presented a host-seeking index ten times lower than adult bugs and were also able, on a few occasions (4/132 released bugs), to reach the mouse in hammock.ConclusionsWe conclude that sleeping in hammocks, as done in rural Yucatán, makes human hosts less accessible to the bugs. This, combined with other factors (e.g. absence of domestic animals sleeping inside houses), may explain, at least in part, the low nutritional status of bugs collected inside homes and the limited colonization of houses by T. dimidiata in the region. Nevertheless, while this sleeping habit limits contact with the bugs, it does not confer complete protection as adult bugs as well as some nymphs were still able to reach the host in hammock in our study.

Geographical Variation of Deltamethrin Susceptibility of Triatoma infestans (Hemiptera: Reduviidae) in Argentina With Emphasis on a Resistant Focus in the Gran Chaco.

Chagas disease is one of the most important parasitic infections in Latin America. The main vector of the protozoan Trypanosoma cruzi in America is Triatoma infestans, a blood-sucking triatomine bug who is widely distributed in the Gran Chaco ecoregion. Control programs in endemic countries are focused in the elimination of triatomine vectors with pyrethroid insecticides. However, chemical control has failed in the Gran Chaco over the last two decades because of several factors. Previous studies have reported the evolution of different levels of resistance to deltamethrin in Tri. infestans Recently, very high resistance has been found in the central area of the Argentine Gran Chaco. However, the origin and the extension of this remarkably resistant focus remain unknown. The aim of this study was to evaluate the geographical variation of deltamethrin susceptibility of Tri. infestans in different endemic provinces of Argentina, with emphasis in the center of the Argentine Gran Chaco ecoregion where this main vector has not been reduced. Populations of Mendoza, San Juan, Santiago del Estero, and Tucumán provinces were all susceptible. Resistant populations were only detected in the province of Chaco, where a mosaic resistant focus was described at the Güemes Department. It was characterized into three pyrethroid resistance categories: susceptible, low, and highly resistant populations. We found the populations with the highest resistance levels to deltamethrin, with resistant ratios over 1000.

Symbiont-mediated RNA interference in insects.

RNA interference (RNAi) methods for insects are often limited by problems with double-stranded (ds) RNA delivery, which restricts reverse genetics studies and the development of RNAi-based biocides. We therefore delegated to insect symbiotic bacteria the task of: (i) constitutive dsRNA synthesis and (ii) trauma-free delivery. RNaseIII-deficient, dsRNA-expressing bacterial strains were created from the symbionts of two very diverse pest species: a long-lived blood-sucking bug, Rhodnius prolixus, and a short-lived globally invasive polyphagous agricultural pest, western flower thrips (Frankliniella occidentalis). When ingested, the manipulated bacteria colonized the insects, successfully competed with the wild-type microflora, and sustainably mediated systemic knockdown phenotypes that were horizontally transmissible. This represents a significant advance in the ability to deliver RNAi, potentially to a large range of non-model insects.

Molecular Individual-Based Approach on Triatoma brasiliensis: Inferences on Triatomine Foci, Trypanosoma cruzi Natural Infection Prevalence, Parasite Diversity and Feeding Sources.

We used an individual-based molecular multisource approach to assess the epidemiological importance of Triatoma brasiliensis collected in distinct sites and ecotopes in Rio Grande do Norte State, Brazil. In the semi-arid zones of Brazil, this blood sucking bug is the most important vector of Trypanosoma cruzi—the parasite that causes Chagas disease. First, cytochrome b (cytb) and microsatellite markers were used for inferences on the genetic structure of five populations (108 bugs). Second, we determined the natural T. cruzi infection prevalence and parasite diversity in 126 bugs by amplifying a mini-exon gene from triatomine gut contents. Third, we identified the natural feeding sources of 60 T. brasiliensis by using the blood meal content via vertebrate cytb analysis. Demographic inferences based on cytb variation indicated expansion events in some sylvatic and domiciliary populations. Microsatellite results indicated gene flow between sylvatic and anthropic (domiciliary and peridomiciliary) populations, which threatens vector control efforts because sylvatic population are uncontrollable. A high natural T. cruzi infection prevalence (52–71%) and two parasite lineages were found for the sylvatic foci, in which 68% of bugs had fed on Kerodon rupestris (Rodentia: Caviidae), highlighting it as a potential reservoir. For peridomiciliary bugs, Galea spixii (Rodentia: Caviidae) was the main mammal feeding source, which may reinforce previous concerns about the potential of this animal to link the sylvatic and domiciliary T. cruzi cycles.

Nitric oxide heme interactions in nitrophorin from Cimex lectularius

The nitrophorin from the bedbug Cimex lectularius (cNP) is a nitric oxide (NO) carrying protein. Like the nitrophorins (rNPs) from the kissing bug Rhodnius prolixus, cNP forms a stable heme Fe(III)-NO complex, where the NO can be stored reversibly for a long period of time. In both cases, the NPs are found in the salivary glands of blood-sucking bugs. The insects use the nitrophorins to transport the NO to the victim’s tissues, resulting in vasodilation and reduced blood coagulation. However, the structure of cNP is significantly different to those of the rNPs from Rhodnius prolixus. Furthermore, the cNP can bind a second NO molecule to the proximal heme cysteine when present at higher concentrations. High field Mossbauer spectroscopy on 57Fe enriched cNP complexed with NO shows reduction of the heme iron and formation of a ferrous nitric oxide (Fe(II)-NO) complex. Density functional theory calculations reproduce the experimental Mossbauer parameters and confirm this observation.

Female choosiness and mating opportunities in the blood-sucking bug Rhodnius prolixus

We investigated the relationship between female choosiness and mating opportunities in the blood sucking bug Rhodnius prolixus. Rhodnius prolixus females exhibit active discrimination behaviour to male mating attempts which delays or prevents copulation. Female rejection behaviours can be beneficial if mating opportunities are readily available and the pool of males varies in their quality. Thus, the benefits of finding a better quality male may override the cost of rejecting a mating opportunity. Since the availability of mating opportunities is affected by the ratio of sexually active males to females, we randomly assigned focal pairs to arenas with a sex ratio biased toward males, females or without other individuals. More females exhibited rejection behaviour when conspecifics were present, however, no differences were found when the sex ratio was biased toward either males or females. We discuss possible explanations for these results and hypothesize about the adaptive function of female rejection behaviour.

Cytogenetic Examination of South American Tapirs, Tapirus Terrestris (Perissodactyla, Tapiridae), from the Wroclaw Zoological Garden

Abstract Cytogenetic Examination of South American Tapirs, Tapirus terrestris (Perissodactyla, Tapiridae) from the Wroclaw Zoological Garden. Kosowska, B., Strzała, T., Moska, M., Ratajszczak, R., Dobosz, T. - Seven lowland tapirs (Tapirus terrestris) from Wrocław ZOO (three females and four males), differing from each other with exterior and sexual behaviour were verified with cytogenetic analysis in order to check their taxonomic status. Cytogenetic analysis was done using two alternative methods of blood collection: 1) conventionally with venepuncture, and 2) with blood sucking bugs from the Reduviidae family. Lymphocytes capable of growing were obtained only with conventional method of blood sampling. Karyotypes and karyograms of all analyzed tapirs were created using classical cytogenetic methods of chromosomes staining. All possessed karyograms had diploid chromosome number equal 80 (2n = 80). Homologous chromosomes did not differ between each other with quantity, size, centromeres location, length of arms, G bands and all were classified as proper karyograms of Tapirus terrestris species representatives. The X chromosomes as well as the first pair of chromosomes (both metacentric), were the largest among all analyzed, respectively. All remaining 38 pairs of chromosomes were acrocentric with Y chromosome as the smallest one (in males’ karyograms). Blood collected with blood sucking bugs proved to be unsuitable for cell culture. None of the seven established cultures was effective as lymphocytes obtained with this method did not show growth potential in prepared media. Thus, blood collected from the tapirs via Dipetalogaster maxima species did not show usefulness for cytogenetic studies due to the inability of cells to proliferation, even after a relatively short period of time elapsed since the blood sampling (1 to 2 hours).