Biodemography of the Mediterranean fruit fly: aging, longevity and adaptation in the wild

The purpose of this paper is to summarize recent research on longevity, aging and adaptation in wild medfly populations and in a close relative of the medfly. The key findings include a new life table identity that relates age structure and the distribution of deaths in stationary populations, seasonal variation in the post-capture longevity of trapped medflies of unknown age, greater longevity of once-wild (wild-caught) adult medflies relative to never-wild (laboratory-emerged) individuals, differences in age specificity of different medfly field capture methods, large variation in the sex-specific longevity of six medfly global biotypes (e.g. Kenya; Brazil; Greece), and the extraordinary longevity of the natal fruit fly — a sister species of the medfly. The discussion contains a listing of discoveries derived from this recent research that appear to be unique to the investigations on medfly aging in the wild. It is suggested that studies of aging in wild populations of Drosophila melanogaster have the potential to exploit this model organism in an entirely new aging research domain and thus complement the already deep literature on aging in this species.     

Tissue- and sex-specific regulation of CYP19A1 expression in the Atlantic croaker (Micropogonias undulatus)

To better define the tissue- and sex-specific roles of aromatase in fishes, we have isolated a CYP19A1 cDNA sequence from a well-developed model of teleost reproduction, the Atlantic croaker (Micropogonias undulatus). This cDNA encodes a protein which has high identity (57-90%) to known CYP19A1 proteins and segregates with teleost CYP19A1 proteins in molecular phylogenetic analysis. In both sexes, the gene encoding Atlantic croaker CYP19A1 is expressed primarily in gonadal tissue, but also in the brain and other tissues at much lower levels, as determined relative to ribosomal 18S RNA expression by real-time quantitative RT-PCR. In females, the highest levels of CYP19A1 mRNA are found in the developing ovary compared to spawning, regressing and resting ovaries. In contrast, testicular CYP19A1 expression is lowest in developing testes and increases in spawning and regressing testes, although there were no statistically significant differences between stages. Brain CYP19A1 mRNA levels are lower in animals with developing gonads compared to spawning fish. In vitro treatment with human chorionic gonadotropin (10 IU/ml) for 6 or 24h increases CYP19A1 mRNA approximately 16- and 43-fold, respectively, in isolated Atlantic croaker ovarian follicles, but has no effect on CYP19A1 mRNA in testicular or brain minces. Six hour in vitro treatment with sex steroids (estradiol, testosterone or 17,20 beta,21-trihydroxy-4-pregnen-3-one; 290 nM) does not alter CYP19A1 mRNA in ovary, testis or brain. The regulation of CYP19A1 in the Atlantic croaker therefore differs in a tissue- and sex-specific manner.     

The effects of aging on stem cell behavior in Drosophila

Throughout life, adult stem cells play essential roles in maintaining tissue and organ function by providing a reservoir of cells for homeostasis and regeneration. A decline in stem cell number or activity may, therefore, lead to compromised organ and tissue function that is characteristic of aging. Drosophila has emerged as an ideal system for studying the relationship between stem cells and aging, as it has a short lifespan, tissues that are maintained by adult stem cells, and conserved pathways known to regulate aging. In this review, we highlight recent findings describing intrinsic and extrinsic age-related changes that affect the behavior of Drosophila germline and intestinal stem cells. We also discuss whether pathways affecting lifespan can act autonomously or non-autonomously in stem cells during aging.     

Testing the rate-of-living/oxidative damage theory of aging in the nematode model Caenorhabditis elegans

The integration of the rate-of-living and oxidative damage theory of aging predicts that lifespan extension is linked to low energy metabolism, low ROS production rates, low molecular damage and a slow aging rate. In the long-lived Caenorhabditis elegans Ins/IGF-1 mutant daf-2(e1370), low carbonylation levels and postponed morphological decline comply with the latter two of these predictions. However, metabolic rates in daf-2(e1370) refute the rate-of-living theory. The apparent contradiction between increased ROS generation and long lifespan in daf-2(e1370) is reconciled by an enhanced stress defense, acknowledging oxidative damage as a probable cause of aging.     

Reproductive maturation of the tropical clawed frog: Xenopus tropicalis

The tropical clawed frog, Xenopus tropicalis, is a relatively new model species being used in developmental biology and amphibian toxicology studies. In order to increase our understanding of reproductive maturation and the role of steroid hormones in X. tropicalis, we collected baseline reproductive data in this species from metamorphosis to adulthood. One cohort of frogs was maintained for 42 weeks post-metamorphosis (PM) with endpoints representative of important reproductive parameters collected at 1- or 2-week intervals. These endpoints were then correlated to titers of either estradiol or testosterone. Male frogs exhibited nuptial pads, starting at 8 weeks (PM) when measureable concentrations of circulating testosterone (5.3 ng/mL plasma) first appeared. Testosterone concentrations remained above this level at all later time points, but were highly variable among individuals. Testes sizes in males reached their peak at 22 weeks PM (21 mg) with sperm counts peaking at the same time (25 million sperm/male). In females, estradiol becomes elevated in the blood at 16 weeks PM (1.5 ng/mL plasma) which corresponds with the presences of vitellogenin (4.4 mg/mL plasma), vitellogenic oocytes in the ovary, ovarian growth, and oviduct growth. Vitellogenic oocytes increased in number up to 15,000 per female at 30 weeks PM and accounted for 75% of the total number of oocytes present in the ovary. The ovary and oviducts continued to grow in mass until 30 weeks PM at which point they had reached sizes of 3.6 g and 0.8 g, respectively. These data indicate that male and female X. tropicalis reach reproductive maturation at 22 and 30 weeks PM, respectively. Results from this study are valuable for the design of amphibian toxicology assays and increase our understanding of the reproductive biology of this relatively new model species.     

Comparison of the effects of extracts from three Vitex plant species on Anopheles gambiae s.s. (Diptera: Culicidae) larvae

Acetone and methanol extracts of different parts of three Vitex species (leaves and stem bark of Vitex trifolia, leaves, stem bark and root bark of Vitex schiliebenii and stem and root bark of Vitex payos) were evaluated for their potential to control Anopheles gambiae Giles s.s. larvae (Diptera: Culicidae). The extracts gave different levels and rate of mortality of the larvae. Some (methanol extract of V. trifolia leaves, acetone extracts of stem bark and leaves of V. schiliebenii, acetone extract of root bark of V. payos) caused 100% mortality at 100 ppm in 72 h, with those of V. schiliebenii and V. payos showing faster rate of mortality (LT₅₀ = 8 h) than that of V. trifolia (LT₅₀ = 14 h). At lower doses of these extracts (≤50 ppm), most of the larvae failed to transform to normal pupae but gave larval–pupal intermediates between 4 and 14 days of exposure. Some pupated normally but the adults that emerged appeared to be weak and died within 48 h. Extracts of the stem bark of V. payos showed interesting effects on the larvae. Initially, the larvae were relatively hyperactive compared to those in control treatments. Later, the ones that did not transform to larval–pupal intermediates became stretched and inactive and died and floated in clusters on the surface. These observations suggest some interesting growth-disrupting constituents in the plants, with possible application in the practical control of mosquito larvae in aquatic ecosystems.     

A further test of the equation of lifespan by C. elegans: effective activation energy for aging and lifespan

We previously proposed a rate theory of chemical reaction as well as a lifespan equation derived by a stochastic fluctuation theory. Both were applied to biodemographic data by C. elegans to quantitatively explain that respiratory activity declines exponentially with age and that it has a physiological decline rate and a finite value (threshold) in advanced age. In this work, using the poikilothermic nature of Caenorhabditis elegans, we demonstrate the further validity of the rate theory of chemical reaction as well as the lifespan equation by changing two methods. First, to test the appropriateness of the lifespan equation from another aspect, lifespan assays were conducted by varying the time interval of observation employing the egl-1 mutant. The results indicate that, as the time interval is reduced, mortality rates gradually approach the force of mortality expected from the fitting equation of the survival curve. Second, based on the dependence of lifespan on the temperature of the culture, the physiological decline rate, and the onset of biodemographic aging, we show that the effective activation energy or energy barrier for aging and lifespan may be closely related to the standard free-energy change of ATP or ADP for a wild type and some lifespan-related mutants of C. elegans.     

Sex reversal effects on Caiman latirostris exposed to environmentally relevant doses of the xenoestrogen bisphenol A

Exposure to environmental contaminants known as endocrine disruptors (EDs) alters the development and function of reproductive organs in several species. Bisphenol A (BPA) is an estrogenic chemical that leaches from dental materials and plastic food and beverage containers. BPA has been found in sewage, surface and drinking water, and therefore poses a potentially significant risk for human and wildlife. Prenatal exposure of rodents to environmentally relevant doses of BPA alters the development of the reproductive organs of male and female offspring. Species with temperature dependent sex determination (TSD) could act as sentinels of ecosystem health by providing sensitive biomarkers of endocrine disruptor's effects. We selected Caiman latirostris as an animal model to study endocrine disruption caused by BPA. The aim of this study was to determine whether exposure in ovum to BPA could cause estrogen-like effects on the reproductive system of C. latirostris. Sex determination and gonadal histoarchitecture were the endpoints evaluated after in ovum exposure to different doses of BPA and 17beta-estradiol (E(2)). We confirmed that C. latirostris is a species with TSD and additionally demonstrated that BPA causes estrogen-like developmental effects by reversing gonadal sex and altering gonadal histoarchitecture. Differences in responses to BPA and E(2) in our in vivo system were on the order of 100-fold. In contrast published in vitro studies have reported differences on the order of 10,000x or more. These results support the utility of C. latirostris, a species in which sex determination is temperature dependent, as a tool in assessing estrogenic activity in vivo and as a sentinel to monitor EDs in aquatic environment.     

Malaria transmission dynamics in Niono, Mali: the effect of the irrigation systems

The type of water management and drainage system could be a potential reason for variation in malaria transmission in rice cultivation areas. To investigate this we have compared the population dynamics of Anopheles mosquitoes (Diptera, Culicidae) in rice plots with controlled and uncontrolled water depth, i.e. casiers and hors-casiers, respectively in the Office du Niger, Mali. We also compared malaria transmission in areas with mixed and casiers plots. Larval collection was performed fortnightly with the standard WHO dipping technique. Adult Anopheles were collected both by pyrethrum spray and landing catches. During the dry season rice cultivation cycle, the larval density in the hors-casier was significantly higher than in the casier plots. The larval peak in the casier plots was considerably smaller than the one in the hors-casier. During the rainy season, no significant difference was observed between the two plot types. However, larval densities begin to rise approximately one month earlier in the casier then in the hors-casier plots, and continued to increase trough the rice development phases until the grain filling/maturation phase, declining thereafter. In contrast, in the hors-casier rice plots larval density increased throughout the rice development. This difference was not significantly reflected in the adult vector density and man biting rate. However, high relative frequencies of Anopheles funestus, survival and entomological inoculation rates of An. gambiae s.l. were observed in the mixed plot sector.     

The role of sex ratio on spawning performance and on the free and conjugated sex steroids released into the water by common dentex (Dentex dentex) broodstock

This paper reports the results of an experiment to determine whether sex ratio has any effect on the spawning efficiency of common dentex, Dentex dentex, broodstock. The paper also reports preliminary data on diurnal changes in water concentrations of steroids (measured on two separate occasions) in the same groups of fish. These latter observations were made to determine whether, using a non-invasive procedure, it was possible to relate hormonal changes in the fish to the "success of spawning." Mature fish were placed in broodstock tanks at three different sex ratios (Group A, 4 female:1 male; Group B, 1 female:1 male; and Group C, 1 female:4 male) and regularly monitored for egg production. At 18 days after the start of spawning, water samples were removed from all three tanks at 130 min intervals over a period of 24 h. At the end of spawning, water samples were removed at 4 h intervals. Free and conjugated steroids were extracted from the water and measured by radioimmunoassay. The highest number of eggs released, percentage of viable eggs, number of spawnings, spawning index, and relative fecundity were observed in Group B (1 female:1 male). The sex steroids testosterone (T), 11-ketotestosterone (11-KT), 17,20beta-P, and 17,20beta, 21-trihydroxy-pregn-4-en-3-one could be readily detected in water from all tanks. Concentrations of the conjugated forms (sulphate and glucuronide) were higher than those of the free form--with the exception of sulphated T and 11-KT. The concentrations of glucuronidated 17,20beta-P, sulphated 17,20beta-P, and free T appeared to be associated with the number of females in the tank and glucuronidated 11-KT concentrations with the number of males. In the first set of water samples (taken during the spawning period) there was also evidence for a cycle in steroid release with an acrophase during the afternoon and early evening, a few hours before the actual act of spawning. We were not able to show any association between steroid concentrations and success of spawning. However, since the data were derived from a "snapshot" of just a single day in what was a prolonged spawning period, this was perhaps not surprising. The results from the present study will help us to design experiments that are more able to answer this question.     

New administration model of trans-chalcone biodegradable polymers for the treatment of experimental leishmaniasis

The present study was designed to investigate a new administration model and the antileishmanial activity of a semi-synthetic chalcone, benzylideneacetophenone (trans-chalcone). The antileishmanial activity of this product was first tested in vitro against promastigotes of L. braziliensis, L. tropica, L. infantum and L. amazonensis. An in vivo experiment was carried out using subcutaneous administration of trans-chalcone and implants of synthetic biodegradable polymers, polylactic acid (PLA) and polylactic/glycolic acid (PLGA). This compound showed potent inhibitory effects on the growth of all Leishmania strains examinated. Subcutaneous administration of trans-chalcone at a single dose of 4 mg/kg of body weight reduced lesion development in mice infected with L. amazonensis. A similar inhibition of the lesion growth in mice treated with trans-chalcone and pentamidine was observed. PLA and PGLA implants of trans-chalcone at 4 mg/kg were administered to mice infected with L. amazonensis. PLGA implants induced a highest reduction in the lesion size (31.25%) than PLA implants (10.75%). Treatment in vitro with trans-chalcone at IC50, completely inhibited the pathogenicity of this parasite in vivo. The development of this model provides a new practical technique for delivering drugs and can be useful for experimental leishmaniasis treatment.     

First report of the activity of predatory fungi on Angiostrongylus cantonensis (Nematoda: Angiostrongylidae) first-stage larvae

The nematode Angiostrongylus cantonensis causes eosinophilic meningoencephalitis in humans and thus alternative methods of control should be studied. The objective of this work was to evaluate the predatory capacity of eight fungal isolates of the species Duddingtonia flagrans (AC001, CG768 and CG722), Monacrosporium thaumasium (NF34), M. sinense (SF53) and Arthrobotrys robusta (I31), A. cladodes (CG719) and A. conoides (I40) on first-stage larvae (L₁) of A. cantonensis under laboratory conditions. The treated groups contained 1000 conidia of the fungal isolates and 1000 A. cantonensis L₁ in Petri dishes containing 2% water-agar medium (2% WA). The control group (without fungi) contained only 1000 A. cantonensis L₁ in 2% WA. Evidence of predation was observed at the end of 7 days. Percentage reductions in L₁ were: AC001, 82.8%; CG768, 71.0%; CG722, 72.8%; NF34, 86.7%; SF53, 89.7%; I40, 48.3%; CG719, 84.7%; and I31, 80.4%. No significant difference was observed (p > 0.01) between the actions of the isolates used; however, a difference was noted (p < 0.01) in relation to the control group. The results of the present work, confirm previous reports of the effectiveness of the fungi D. flagrans, M. thaumasium, M. sinense and A. robusta in controlling larvae of potentially zoonotic nematodes, this being the first report on A. cantonensis L₁.     

A quantitative framework for a multi-group model of Schistosomiasis japonicum transmission dynamics and control in Sichuan,China

A quantitative framework is presented for the site-specific characterization of schistosomiasis transmission with the object of developing local control strategies. Central to the framework is a worm-burden model using ordinary differential equations of disease transmission in risk groups defined by residence and occupation. The model incorporates temperature- and precipitation-dependent seasonality of infectious stages, snail population dynamics, and seasonal patterns of human water contact specific to the local agricultural setting. The model's parameters are separated into two main subsets, those associated with the general biology of the parasite and its life cycle in the human and the snail and those associated with directly measurable features of disease status in the local population or relevant aspects of the local environment. In this regard, the model is structured and parameterized to take maximum advantage of data that can be collected in rural China by conventional methods. For example, it includes a statistical model for egg excretion to the environment by each risk group which is based on local population surveys of the prevalence and intensity of infection. The second element of the framework of analysis relates to the strategy for parameter estimation and calibration to local conditions. We propose a Bayesian approach in which parameter estimates are refined over time by methods employing extensive computer simulations. An early analysis of data collected between 1987 and 1989 in endemic villages near Xichang City in southwestern Sichuan provides encouragement that parametric uncertainty can be reduced to levels adequate to explore effective control strategies.     

Feeding behavior of triatomines from the southwestern United States: An update on potential risk for transmission of Chagas disease

Chagas disease is an emerging infectious disease in North America due to the immigration of individuals from endemic areas. The parasite has been transmitted to patients in non-endemic areas by blood transfusion and organ donation. Only six autochthonous cases have been described in humans in the United States yet the parasite is widespread among native mammals and resident triatomines are competent vectors. We attempted to determine if common southwestern triatomines, Triatoma protracta and Triatoma rubida have the potential to amplify the disease among human residents of the Southwest. The defecation patterns of the bugs were studied while feeding upon immobilized mice. Wild-caught adult male and female triatomines were observed feeding one to three times for a total of 71 observed feedings. T. rubida (15 bugs) appeared to be more aggressive, beginning feeding shortly after being placed in proximity to the host (within 2.3 min) whereas Triatoma protracta (12 bugs) was more deliberate, beginning feeding, on average, at 4 min. There were 40 observations of T. rubida, which fed for 27.9 ± 13.6 min, whereas T. protracta fed for 22.8 ± 7.5 min (n = 31). Bugs were weighed pre- and post-feeding and T. rubida females ingested > T. protracta females > T. rubida males > T. protracta males. Weight gain did not correspond to the feeding duration. Defecation occurred on 42% of the feedings (30 of 71), and no bugs defecated on the host. The majority of the defecations occurred within 1 min of feeding, usually at the time of repletion. A large proportion of defecations occurred after the bugs left the vicinity of the host. All bugs and at least one fecal smear from each feeding bug were tested for Trypanosoma cruzi and 21% of T. protracta were positive by PCR (4 bugs and 1 feces). No T. rubida tested positive for T. cruzi. The bugs’ defecation pattern is similar to that reported >50 years ago. Furthermore, there is no indication that they are becoming more domesticated in the desert Southwest. Thus, based on our observations, we do not believe that T. protracta and T. rubida pose an imminent risk for transmission of Chagas disease to residents of the southwestern United States.     

The effects of the fungus Metarhizium anisopliae var. acridum on different stages of Lutzomyia longipalpis (Diptera: Psychodidae)

The control of Visceral Leishmaniasis (VL) vector is often based on the application of chemical residual insecticide. However, this strategy has not been effective. The continuing search for an appropriate vector control may include the use of biological control. This study evaluates the effects of the fungus Metarhizium anisopliae var. acridum on Lutzomyia longipalpis. Five concentrations of the fungus were utilized, 1 × 10⁴ to 1 × 10⁸ conidia/ml, accompanied by controls. The unhatched eggs, larvae and dead adults previously exposed to fungi were sown to reisolate the fungi and analysis of parameters of growth. The fungus was subsequently identified by PCR and DNA sequencing. M. anisopliae var. acridum reduced egg hatching by 40%. The mortality of infected larvae was significant. The longevity of infected adults was lower than that of negative controls. The effects of fungal infection on the hatching of eggs laid by infected females were also significant. With respect to fungal growth parameters post-infection, only vegetative growth was not significantly higher than that of the fungi before infection. The revalidation of the identification of the reisolated fungus was confirmed post-passage only from adult insects. In terms of larvae mortality and the fecundity of infected females, the results were significant, proving that the main vector species of VL is susceptible to infection by this entomopathogenic fungus in the adult stage.     

Culture-dependent and culture-independent characterization of microorganisms associated with Aedes aegypti (Diptera: Culicidae) (L.) and dynamics of bacterial colonization in the midgut

In this work we show that the lumen of Aedes aegypti midgut is highly colonized by bacteria that were identified by culture-dependent and culture-independent methods. rDNA sequences obtained were compared with those from GenBank and the main bacterial genera identified were: Serratia, Klebsiella, Asaia, Bacillus, Enterococcus, Enterobacter,Kluyvera and Pantoea. All genera were identified in midgut except Enterobacter that was observed only in eggs. Asaia and Pantoea were also identified in eggs and ovary, respectively. In addition two yeast genera were observed in A. aegypti: Pichia isolated from midgut and Candida identified in midgut and ovary. The genus Serratia was dominant in all isolation assays representing 54.5% of the total of microorganisms. Thirty-nine and 24 bacterial clones were successfully obtained from midguts 24 and 48 h after blood feeding (ABF), respectively. The majority of clones obtained were from Serratia sp. (48.7% and 50% for 24 and 48 h ABF, respectively). Light microscopy showed that bacteria were located preferentially in the posterior midgut, around the blood meal and associated with peritrophic matrix. Scanning electron microscopy images showed a high number of bacteria in midgut during blood digestion and the peak of bacterial enumeration was reached 48 h ABF, stage in which lumen was almost totally occupied by bacteria that were also interacting with epithelial microvilli. Our results show the dynamics of microbial colonization and their distribution in midgut during blood digestion.     

Use of sentinel snails for the detection of Schistosoma haematobium transmission on Zanzibar and observations on transmission patterns

Urogenital schistosomiasis is an important public health issue in Zanzibar. Transmission of the parasite to the human population is related to the distribution of the intermediate snail host, Bulinus globosus. We measured B. globosus abundance and Schistosoma haematobium prevalence within snails in a series of naturally occurring populations and compared prevalence detected by observing cercarial shedding for patent infections, and by PCR using DraI repeat. A total of 2146 B. globosus were collected throughout the study period from 2003 to 2007; of these 85 (3.96%) were shedding cercariae. The levels of infection detected by PCR were consistently higher (40–100 percent). Levels of exposure to miracidia in the field were measured using sentinel snails. B. globosus (a susceptible host) and B. nasutus (a non-susceptible host) were placed in cages at transmission sites for 72 h to observe rates of penetration by miracidia. Both B. globosus and B. nasutus tested positive for S. haematobium by PCR (up to 24% infected) indicating frequent contamination of the waterbodies with S. haematobium miracidia. The use of sentinel snails coupled with PCR diagnostics could be a sensitive tool for mapping and monitoring transmission of schistosomiasis in areas of low prevalence.     

Contrasting effects of Trichinella spiralis and Trichuris muris antigens on the infection by Leishmania infantum in BALB/c mice

The interaction of Trichinella spiralis and Trichuris muris derived antigens with the infection by Leishmania infantum was investigated in BALB/c mice. Infection with 10(6) promastigotes of L. infantum did not induce relevant serum antibody (IgG subclasses), nor cytokine (IFN-gamma, IL-4) responses despite that mice could partially control the infection. Immunization with T. spiralis activated a moderate IgG1 and secondarily an IgG2a anti-leishmanial response whereas immunization with T. muris elicited only a weak and late activation of IgG1 anti-leishmanial response. Immunization with T. muris caused an elevation of serum IFN-gamma levels which was drastically reinforced by the L. infantum infection, and that was accompanied by almost complete parasitological cure of infected mice. Immunization with T. spiralis induced an elevation of serum IL-4 levels but this response was greatly (about 60%) neutralized by the infection with L. infantum, and this was associated to exacerbation of the infection.