Infected erythrocyte choline carrier inhibitors: exploring some potentialities inside Plasmodium phospholipid metabolism for eventual resistance acquisition

We have developed a model for designing antimalarial drugs based on interference with an essential metabolism developed by Plasmodium during its intraerythrocytic cycle, phospholipid (PL) metabolism. The most promising drug interference is choline transporter blockage, which provides Plasmodium with a supply of precursor for synthesis of phosphatidylcholine (PC), the major PL of infected erythrocytes. Choline entry is a limiting step in this metabolic pathway and occurs by a facilitated-diffusion system involving an asymmetric carrier operating according to a cyclic model. Choline transport in the erythrocytes is not sodium dependent nor stereospecific as demonstrated using stereoisomers of alpha and beta methylcholine. These last two characteristics along with distinct effects of nitrogen substitution on transport rate demonstrate that choline transport in the infected erythrocyte possesses characteristics quite distinct from that of the nervous system. This indicates a possible discrimination between the antimalarial activity (inhibition of choline transport in the infected erythrocyte) and a possible toxic effect through inhibition of choline entry in synaptosomes. Apart from the de novo pathway of choline, PC can be synthesized by N-methylation from phosphatidylethanolamine (PE). There is a de novo pathway for PE biosynthesis from ethanolamine in infected cells but phosphatidylserine (PS) decarboxylation also occurs. In addition, PE can be directly and abundantly synthesized from serine decarboxylation into ethanolamine, a pathway which is absent from the host. The variety of the pathways that exist for the biosynthesis of one given PL led us to investigate whether an equilibrium can occur between all PL metabolic pathways. Indeed, if alternative (compensative) pathway(s) can operate after blockage of the de novo PC biosynthesis pathway this would indicate a potential mechanism for resistance acquisition. Up until now, there is no evidence of such a compensative process occurring in Plasmodium-infected erythrocytes under physiological conditions. Besides, the discovery of a highly parasite-specific pathway (serine decarboxylation and the presence of PS synthase) constitutes a very attractive and promising target, which could be attacked if resistances are built up against choline analogs. Indeed, potential inhibitions of the serine decarboxylase pathway could be very useful in acting instead of, or in surgery with, choline analogs.

Effect of Niclosamide (Bayluscide WP 70 (R)), Anacardium occidentale hexane extract and Euphorbia splendens latex on behavior of Biomphalaria glabrata (Say, 1818), under laboratory conditions

The repellent effect of the molluscicides Niclosamide (Bayluscide WP 70 (R)), Anacardium occidentale and the latex of Euphorbia splendens on Biomphalaria glabrata was observed through the investigation of the occurrence of escape behavior among molluscs that were exposed to dosages lower than the LD 50. The total number of individuals out of water among the surviving snails in the control group provided a "Natural Escape Index". The comparison between this total and the total number of surviving snails in each group exposed to the different dosages of the molluscicides after 24 hr provided the "Molluscicide Escape Index" and the detection of a "Repellency Range" to these snails. The escape indexes for Niclosamide, A. occidentale and E. splendens were 10, 6.22 and 6.44 respectively. Repellency occurred at the following concentration ranges: 0.01, 0.02 and 0.03 ppm Bayluscide, 0.1, 0.2 and 0.3 ppm A. occidentale and 0.05, 0.10, 0.15 and 0.20 ppm E. splendens. The Natural Escape Index obtained in the control group was zero.

Larval and pupal periods of Peckia chrysostoma and Adiscochaeta ingens (Diptera: Sarcophagidae) reared under laboratory conditions

Peckia chrysostoma obtained mean viability of 97.0±2.4% for larvae and of 96.9±2.5% for pupae (total viability of 94.0±3.7%). Adiscochaeta ingens obtained mean viability of 93.0±7.5% for larvae and of 92.8±7.6% for pupae (total viability of 86.0±7.3%). P. chrysostoma obtained mean larval period of 185±4 hr at 18ºC, of 94±2 hr at 27ºC and of 88±2 hr at room temperature (range of 23ºC and 29ºC). A. ingens obtained mean larval period of 169±1 hr at 18ºC, of 77±1 hr at 27ºC and of 84±2hr at room temperature. P. chrysostoma obtained mean pupal period of 23.5±1.3 days at 18ºC, of 12.5±0.7 days at 27ºC and of 15.5±0.7 days at room temperature. A. ingens obtained mean pupal period of 33.0±2.2 days at 18ºC, of 16.0±1.0 days at 27ºC and of 19.0±1.0 days at room temperature.

The effect of different proportions of males and females over the Chrysomya albiceps (Wiedemann 1819) (Diptera, Calliphoridae) biotic potential and longevity under laboratory conditions

Chrysomya albiceps specimens were derived from colonies kept under laboratory conditions. The oviposition period, total number of eggs-mass and the weight of the eggs-mass (average/female) presented significant differences between colonies regarding the sexual ratio of 1male/1female (situation I), when compared to the other ratios (1male/3female, situation II), (1male/5female, situation III), (3male/1female, situation IV) and (5 male/1female, situation V). It was ascertained that the increase in the proportion of females, resulted in higher weight and greater number of ovipositions and lenghtening of the period of oviposition, leads to a decrease in their lifespan.

Dynamics of experimental populations of native and introduced blowflies (Diptera: Calliphoridae): mathematical modelling and the transition from asymptotic equilibrium to bounded oscillations

The equilibrium dynamics of native and introduced blowflies is modelled using a density-dependent model of population growth that takes into account important features of the life-history in these flies. A theoretical analysis indicates that the product of maximum fecundity and survival is the primary determinant of the dynamics. Cochliomyia macellaria, a blowfly native to the Americas and the introduced Chrysomya megacephala and Chrysomya putoria, differ in their dynamics in that the first species shows a damping oscillatory behavior leading to a one-point equilibrium, whereas in the last two species population numbers show a two-point limit cycle. Simulations showed that variation in fecundity has a marked effect on the dynamics and indicates the possibility of transitions from one-point equilibrium to bounded oscillations and aperiodic behavior. Variation in survival has much less influence on the dynamics.

Temperature requirements of Chrysomya albiceps (Wiedemann, 1819) (Diptera, Calliphoridae) under laboratory conditions

Chrysomya albiceps specimens were obtained from colonies established with larvae and adults collected at the Federal Rural University in Rio de Janeiro, Seropédica, State of Rio de Janeiro. The larval stage of C. albiceps was allowed to develop in climatic chambers at temperatures of 18, 22, 27 and 32ºC, and the pupal stage was allowed to develop at 22, 27 and 32ºC (60 ± 10% RH and 14 hr photoperiod). The duration and viability of the larval stage of C. albiceps at 18, 22, 27 and 32ºC were 21.30, 10.61, 5.0 and 4.0 days and 76.5, 88.5, 98.5 and 99.5%, respectively, with mean mature larval weights of 45.16, 81.86, 84.35 and 70.53 mg, respectively. Mean duration and viability of the pupal stage at 22, 27 and 32ºC were 9.36, 4.7 and 3.0 days and 93.8, 100 and 100%, respectively. The basal temperature for the larval and pupal stage and for the larval and adult phase were 15.04, 17.39 and 15.38ºC, corresponding to 65.67, 44.15 and 114.23 DD.

Physical Factors Influencing the Oviposition of Lutzomyia migonei (Diptera: Psychodidae) in Laboratory Conditions

To determine the influence of physical factors on oviposition of Lutzomyia migonei (Diptera: Psychodidae) under laboratory conditions, two sets of experiments were performed. The first test was to determine the influence of the size of pots on oviposition. Gravid flies were placed individually or in groups in different oviposition pots. The number of eggs laid, oviposition time and survival of gravid females were observed. In the second experiment, the influence of irregular surfaces on oviposition was studied. The results suggested that physical space was not an important factor in the oviposition behavior of L. migonei and that the flies showed a preference to oviposit on irregular horizontal surfaces in response to thigmotropic behavior

Vital Statistics of Panstrongylus geniculatus (Latreille 1811) (Hemiptera: Reduviidae) under Experimental Conditions

A statistical evaluation of the population dynamics of Panstrongylus geniculatus is based on a cohort experiment conducted under controlled laboratory conditions. Animals were fed on hen every 15 days. Egg incubation took 21 days; mean duration of 1st, 2nd, 3rd, 4th, and 5th instar nymphs was 25, 30, 58, 62, and 67 days, respectively; mean nymphal development time was 39 weeks and adult longevity was 72 weeks. Females reproduced during 30 weeks, producing an average of 61.6 eggs for female on its lifetime; the average number of eggs/female/week was 2.1. Total number of eggs produced by the cohort was 1379. Average hatch for the cohort was 88.9%; it was not affected by age of the mother. Age specific survival and reproduction tables were constructed. The following population parameters were evaluated, generation time was 36.1 weeks; net reproduction rate was 89.4; intrinsic rate of natural increase was 0.125; instantaneous birth and death rates were 0.163 and 0.039 respectively; finite rate of increase was 1.13; total reproductive value was 1196 and stable age distribution was 31.2% eggs, 64.7% nymphs and 4.1% adults. Finally the population characteristics of P. geniculatus lead to the conclusion that this species is a K strategist.

Population Biology of Rhodnius domesticus Neiva & Pinto, 1923 (Hemiptera: Reduviidae) under Laboratory Conditions

The entire life cycle of Rhodnius domesticus, fed weekly on mice, was studied under controlled conditions. Aspects related to hatching, life time, mortality, feeding behaviour and fecundity for each stage of the insect life-cycle were evaluated. The hatching rate observed in 100 eggs was 57% and the mean time of hatching was 15.6 days. Forty-six nymphs (80.7%) completed the cycle and the mean time from NI to adult was 93.8 days. The average span in days for each stage was 12.4 for NI, 9.8 for NII, 14.2 for NIII, 16.8 for NIV and 25.0 for NV. The number of bloodmeals in each nymphal stage varied from 1 to 3. The mortality rate was 12.3% for NI, 3.5% for NII and 1.7% for NIII and NV nymphs. The mean number of eggs laid per female in a 9-month period was 333.1. Average adult survival rates were 287.6 +133 and 328 +73 days for males and females respectively.

Life Cycle and Reproductive Patterns of Triatoma rubrofasciata (De Geer, 1773) (Hemiptera: Reduviidae), under Laboratory Conditions

The life cycle and reproductive patterns of Triatoma rubrofasciata were studied along with laboratory conditions for the establishment of a prolific colony. The insects were divided into four groups: two of them were maintained at room temperature (20.5°C to 33°C and 85% ± 5% of relative humidity), the other two in a climatic chamber (CC) (temperature: 29°C, humidity: 80% ± 5%). The groups were fed weekly or fortnightly on Swiss mice. The females from the group kept in the CC and fed weekly had longer life span, as well as a higher number of eggs, fertile eggs and hatchings; the group kept in the CC and fed fortnightly had a shorter life span for the 1st, 2nd and 3rd instars and a lower mortality rate for all instars. It was concluded that a constant high temperature (CC at 29°C) is the most suitable condition for the maintenance of a colony of T. rubrofasciata regardless of the interval between repasts.

Resistance to Starvation of Triatoma rubrofasciata (De Geer, 1773) under Laboratory Conditions (Hemiptera: Reduviidae: Triatominae)

The present work aims at learning the period of resistance to starvation (molting/death) of Triatoma rubrofasciata in different stages of development and the respective loss of weight until death. Eggs of specimens from the greater area of the city of São Luis in the State of Maranhão, Brazil, yielded approximately 300 nymphs. These nymphs were placed in labelled Borrel glasses, in which they were weekly fed on rats (Rattus norvegicus), until reaching the stage to be observed. The experiments were conducted in a climatic chamber regulated at 29 ± 1° C, 70% relative humidity and 12 hr photoperiod. The resistance to starvation increased according to the stage of development, except for adult bugs, whose results were similar to the 3rd stage nymphs. In all these development stages there was an abrupt loss of weight in the first week, followed by a gradual loss until death. Comparing this work with those of other authors, it was observed that T. rubrofasciata is among the less resistant triatomine species.