Effect of the organophosphorous insecticide, chlorpyrifos (Dursban), on growth, fecundity and mortality of Biomphalaria alexandrina and on the production of Schistosoma mansoni cercariae in the snail.

Exposure of Biomphalaria alexandrina to sublethal concentrations (0.125, 0.25 and 0.05 ppm) of the organophosphorous insecticide, chlorpyrifos (Dursban), induced a reduction in egg production and egg hatchability. Exposure of Schistosoma mansoni miracidia to the insecticide (60 min, 0.50 ppm) prior to infection of B. alexandrina did not affect the subsequent production of cercariae. However, exposure of S. mansoni-infected snails to the insecticide until day 55, from day 20 to day 62 and from day 35 to 62 following infection resulted in blockage of cercarial shedding. Cercarial shedding commenced in some snails when the treatment stopped. Exposure to the insecticide in concentrations of 0.125 and 0.25 ppm during the first 20 days following infection did not affect the subsequent production of cercariae, but exposure to 0.5 ppm during the first 20 days affected markedly the production of cercariae due to a high snail mortality. The findings indicate that the cercaria is the target stage for the activity of chlorpyrifos on the intramolluscan larval development. It is suggested that S. mansoni cercarial production in B. alexandrina may be a useful system for monitoring the effect of low concentrations of pesticides on the aquatic environment, and that the ability by chemical means to interrupt the cercarial production might be a useful tool in further analyses of important aspects of the snail/parasite relationship.     

Molluscicidal activity and new flavonoids from Egyptian Iris germanica L. (var. alba)

The molluscicidal activity of leaf and rhizome extracts of Iris germanica L. (var. alba) against Biomphalaria alexandrina snails was evaluated and the rhizome extracts were found to be the most potent. Activity-guided fractionation revealed that the chloroform extract showed the highest molluscicidal activity (LC90 = 1.26 mg/l) among the tested extracts of the rhizomes. Fraction B prepared from the chloroform extract was the most potent molluscicide (LC90 = 0.96 mg/l) in addition, it showed a significant heart rate reduction in the snail after a 6- to 24-h exposure period. It also displayed a significant level of cercaricidal potential in a time-concentration relationship pattern. Chromatographic fractionation and purification of fraction B resulted in the isolation of two novel compounds: 5,2'-dihydroxy-3-methoxy-6,7-methylenedioxyflavone and 5,7,2'-trihydroxy-6-methoxyflavanone. Their structures were established by one- and two-dimensional NMR methods and mass spectrometry.     

Synthesis and evaluation of molluscicidal and larvicidal activities of some novel enaminones derived from 4-hydroxyquinolinones: part IX

A series of 10 3-(hetarylaminomethylene)quinolinediones, 12 3-(substituted aminopropenoyl)-4-hydroxyquinolinones, and 10 3-(substituted aminomethylene-5-oxo-pyrazolinyl)-4-hydroxyquinolinones were synthesized as novel enaminones derived from 3-(un)substituted 4-hydroxyquinolin-2(1H)-ones in 72-94% yields and assayed for their molluscicidal activities against Biomphalaria alexandrina and Lymnaea natalensis snails. Some of the tested enaminones presented high molluscicidal activities (LC(50)20ppm). The new compounds showed more potency against hatchability of B. alexandrina egg masses, the infection rate and prepatent period of the snails. In addition, these derivatives revealed potential larvicidal effects (100% mortality) on both miracidia and cercariae of Schistosoma mansoni at reduced exposure time. The selected active derivatives were examined against Daphnia magna and their nontoxic effect at all sublethal, lethal, and higher concentrations suggests that these compounds can play an important role as molluscicides and larvicides with environmental safe properties.     

Interaction of photoperiod and nutritive state in female reproduction of Lymnaea stagnalis

Summary

A study was made of the interaction of the photoperiod and the availability of food in influencing egg laying of the freshwater snail Lymnaea stagnalis. At a long day photoperiod (LD = 16 h light/8 h dark) egg laying of fed snails had increased compared with that of fed animals kept at a medium day photoperiod (MD = 12 h light/12 h dark). In MD snails oviposition ceased within a week of the beginning of a starvation period. This is most probably due to a reduction in the activities of the neuroendocrine caudodorsal cells which secrete an ovulation hormone. In contrast, in starved LD snails a low rate of ovipository activity continued, indicating that a lowered frequency of caudodorsal cell release cycles occurred under these conditions. The decreased mean size (number of eggs) of the egg masses in starved LD snails indicates that the activities of the endocrine dorsal bodies, which control vitellogenesis and synthetic activities in the female accessory sex organs, had decreased.

All MD snails survived, but nearly all LD snails died during the course of the experiment. Determinations of the mantle glycogen stores of LD snails suggest that the high mortality of LD snails is due to exhaustion of the animal's energy reserves.     

The Effects of Isolation,Grouping and Population Density on Fecundity of Bulinus truncatus

Summary

Bulinus truncatus is the intermediate host of Schistosoma haematobium, a trematode parasite causing bladder bilharziasis in man. In the struggle against this disease, control of snail populations is an important goal and expansion of our knowledge of the reproductive activity of Bulinus is relevant in this regard.

In this study the effects of various factors on the fecundity of Bulinus were investigated in specimens kept under continuous water refreshment conditions. The results are considered to be more indicative of the natural situation than results obtained under the current use of discontinuous water refreshment.

Isolated snails produce considerably higher numbers of egg masses and eggs than grouped snails. The effect of grouping on isolated snails, and the reverse situation, is very rapid, being fully effective within one week. The increase in fecundity of isolated snails is most probably caused by the absence of copulatory activity, since transfer of semen is known to curtail egg laying in freshwater pulmonate snails. Egg mass production was shown to decrease with increasing population density. This may be caused either by the difference in space available or by the possible difference in copulatory activity. Isolated snails showed a faster body growth than grouped snails.

The possible role of hormones involved in establishing the above effects and the consequences of the results for snail control programs are discussed.     

The effects of starvation and refeeding on egg laying and the synthetic activity of the albumen gland in Bulinus truncatus, a snail vector of urinary schistosomiasis

1. The effects of starvation (for 1, 2, 3, 6, 9 and 12 days, respectively) and refeeding (12 days starvation and 1, 2 and 3 days refeeding, respectively) on egg laying and albumen gland activity in the freshwater snail, Bulinus truncatus were studied. 2. The egg laying of starving snails rapidly decreased and ceased by day 6 of starvation. Egg laying was restored 24 hr after refeeding. The recorded decrease in albumen gland wet weight was proportional to the starvation periods. The DNA contents of the glands of the different experimental groups was not statistically different from the controls. 3. Albumen gland synthetic activity expressed as 14C-glucose incorporation into galactogen/microgram DNA and 3H-amino acids into total protein was determined. The glands showed an abrupt decrease in synthetic activity after 1 day of starvation and gradually decreased further until days 9-12. The decrease in activity of the glands was more rapid than that of egg laying. Upon refeeding, the activity of the glands recovered rapidly, simultaneous with the increase in wet weight and egg laying. 4. In conclusion, there is a correlation between egg laying and the in vitro activity of albumen glands. The results show a short-term effect of starvation on the fecundity of the snails. Such studies could be useful in field studies as well as snail control by applying molluscicides under optimal conditions.     

The relationship between genetic variability and the susceptibility of Biomphalaria alexandrina snails to Schistosoma mansoni infection

In the present study, Biomphalaria snails collected from five Egyptian governorates (Giza, Fayoum, Kafr El-Sheikh, Ismailia and Damietta), as well as reference control Biomphalaria alexandrina snails from the Schistosome Biological Supply Center (SBSC) (Theodor Bilharz Research Institute, Egypt), were subjected to species-specific polymerase chain reaction (PCR) assays to identify the collected species. All of the collected snails were found to be B. alexandrina and there was no evidence of the presence of Biomphalaria glabrata. Randomly amplified polymorphic DNA (RAPD)-PCR assays showed different fingerprints with varying numbers of bands for the first generation (F?) of B. alexandrina snail populations (SBSC, Giza, Fayoum, Kafr El-Sheikh, Ismailia and Damietta). The primer OPA-1 produced the highest level of polymorphism and amplified the greatest number of specific bands. The estimated similarity coefficients among the B. alexandrina populations based on the RAPD-PCR profiles ranged from 0.56 (between SBSC and Ismailia snails) to 0.72 (between Ismailia and Kafr El-Sheikh snails). Experimental infection of the F? of progeny from the collected snails with Schistosoma mansoni (SBSC strain) showed variable susceptibility rates ranging from 15% in the Fayoum snail group to 50.3% in SBSC snails. A negative correlation was observed between the infection rates in the different snail groups and the distances separating their corresponding governorates from the parasite source. The infection rates of the snail groups and their similarity coefficients with SBSC B. alexandrina snails were positively correlated. The variations in the rates of infection of different B. alexandrina groups with S. mansoni, as well as the differences in the similarity coefficients among these snails, are dependent not only on the geographical distribution of the snails and the parasite, but also on the genetic variability of the snails. Introduction of this variability into endemic areas may reduce the ability of the parasite to infect local hosts and consequently reduce schistosomiasis epidemiology.     

Sexual dimorphism of egg size in the European Blackbird Turdus merula

Recently, a number of studies have shown that female birds are able to control the sex of their progeny at the stage of the gamete. There is also some evidence that females adjust their investment in offspring depending on the sex of the embryo during egg formation. Differential maternal investment to the eggs depending on their sex is usually interpreted as an adaptive strategy, by which females can increase competitive abilities of the smaller sex, or preferentially invest towards the sex with the potentially higher fitness returns. Here, we studied variation in egg size in relation to embryo sex and laying order in the European Blackbird Turdus merula. We found male and female eggs to differ in size, with larger eggs containing male embryos, as well as a significant interaction between embryo sex and laying order. This interaction resulted from the fact that egg size increased with the laying sequence among eggs bearing females but did not change with laying order among eggs bearing males. There was no relationship between offspring sex and the laying sequence within a clutch. We suggest that sexual dimorphism in egg size recorded in the European Blackbird may reflect favouritism of the sex which may give higher fitness returns.     

The effect of temperature,darkness, starvation and various food types on growth,survival and reproduction of Helisoma duryi,Biomphalaria alexandrina and Bulinus truncatus (Gastropoda: Planorbidae)

Helisoma duryi has been proposed as a biological control agent in schistosomiasis due to its superiority in laboratory competition experiments with various species of the intermediate host snails. Therefore it was considered important to evaluate the response of this snail species and the intermediate host species, Biomphalaria alexandrina and Bulinus truncatus, to various physical, chemical and biological factors under laboratory conditions in order to obtain information on the similarities in the ecological niches of these species. The factors considered in the present paper are: temperature, darkness, starvation and food. All three species had optimal growth and egg laying at 26–28 °C. Only H. duryi survived for a longer period at 33°C and it was capable of starting egg laying at this temperature although the onset was delayed. However, low temperature (18°C) caused a relatively larger decrease in egg laying of H. duryi than in the other two species. Growth and egg laying was reduced for H. duryi and B. truncatus kept under darkness and B. alexandrina could not tolerate maintenance under darkness. A few days of starvation of juvenile snails had no effect on later growth and egg laying capacity of the survivors, although mortality in B. truncatus was increased. B. alexandrina had a lower tolerance to starvation than the other two species. Egg laying of snails fed only one of the three laboratory food types decreased for all three species in the order: Vov-vov (dog food in dry pellets), Tetramin (fish food) and lettuce. Combinations of lettuce and one or more proteinaceous food types gave optimal growth and egg laying for all three species.     

Impact of the age of Biomphalaria alexandrina snails on Schistosoma mansoni transmission: modulation of the genetic outcome and the internal defence system of the snail

Of the approximately 34 identified Biomphalaria species,Biomphalaria alexandrina represents the intermediate host of Schistosoma mansoni in Egypt. Using parasitological and SOD1 enzyme assay, this study aimed to elucidate the impact of the age of B. alexandrina snails on their genetic variability and internal defence against S. mansoni infection. Susceptible and resistant snails were reared individually for self-reproduction; four subgroups of their progeny were used in experiment. The young susceptible subgroup showed the highest infection rate, the shortest pre-patent period, the highest total cercarial production, the highest mortality rate and the lowest SOD1 activity. Among the young and adult susceptible subgroups, 8% and 26% were found to be resistant, indicating the inheritance of resistance alleles from parents. The adult resistant subgroup, however, contained only resistant snails and showed the highest enzyme activity. The complex interaction between snail age, genetic background and internal defence resulted in great variability in compatibility patterns, with the highest significant difference between young susceptible and adult resistant snails. The results demonstrate that resistance alleles function to a greater degree in adults, with higher SOD1 activity and provide potential implications for Biomphalaria control. The identification of the most susceptible snail age enables determination of the best timing for applying molluscicides. Moreover, adult resistant snails could be beneficial in biological snail control.     

Dephenolization of industrial wastewaters catalyzed by polyphenol oxidase

A new enzymatic method for the removal of phenols from industrial aqueous effluents has been developed. The method uses the enzyme polyphenol oxidase which oxidizes phenols to the corresponding o-quinones; the latter then undergo a nonenzymatic polymerization to form water-insoluble aggregates. Therefore, the enzyme in effect precipitates phenols from water. Polyphenol oxidase has been found to nearly completely dephenolize solutions of phenol in the concentration range from 0.01 to 1.0 g/L. The enzymatic treatment is effective over a wide range of pH and temperature; a crude preparation of polyphenol oxidase (mushroom extract) is as effective as a purified, commercially obtained version. In addition to phenol itself, polyphenol oxidase is capable of precipitating from water a number of substituted phenols (cresols, chlorophenols, naphthol, etc.). Also, even pollutants which are unreactive towards polyphenol oxidase can be enzymatically coprecipitated with phenol. The polyphenol oxidase treatment has been successfully used to dephenolize two different real industrial waste-water samples, from a plant producing triarylphosphates and from a coke plant. The advantage of the polyphenol oxidase dephenolization over the peroxidase-catalyzed one previously elaborated by the authors is that the former enzyme uses molecular oxygen instead of costly hydrogen peroxide (used by peroxidase) as an oxidant.     

Porcine testicular extract inhibits T cell proliferation by blocking cell cycle transition from G1 phase to S phase

Since T cells express diverse sex steroid hormone receptors, they might be a good model to evaluate the effects of sex steroid hormones on immune modulation. Porcine testicular extract contains several sex steroid hormones and may be useful to study the effects of sex steroid hormones during T cell activation. We have examined the effects of the porcine testicular extract on T cell activation: proliferation and secretion of cytokines (IL-2 and IFN-γ) by activated T cells were severely decreased after treatment with porcine testicular extract. The extract produced an immunosuppressive effect and inhibited the proliferation of activated T cells by blocking the cell cycle transition from the G(1) phase to S phase. These effects were mediated by a decrease in the expression of cyclin D1 and cyclin E and constitutive expression of p27(KIP1) after T cell activation.     

Desmettianosides A and B, bisdesmosidic furostanol saponins with molluscicidal activity from Yucca desmettiana

Bioactivity-guided separation of the aqueous methanolic extract of Yucca desmettiana leaves, which in a preliminary screening exhibited significant molluscicidal activity, led to the isolation and structure elucidation of two new steroidal saponins (1 and 2). The structures of desmettianosides A and B, identified as bisdesmosidic furostanol glycosides with six and five sugar units, respectively, were established by detailed spectroscopic analyses of their NMR and MS data. Compounds 1 and 2 exhibited high molluscicidal activity against Biomphalaria alexandrina snails with LC100 values of 6 and 11 mg/L, respectively.     

Offspring sex is not related to maternal allocation of yolk steroids in the lizard Bassiana duperreyi (Scincidae)

The eggs of birds and reptiles contain detectable levels of several steroid hormones, and experimental application of such steroids can reverse genetically determined sex of the offspring. However, any causal influence of maternally derived yolk steroids on sex determination in birds and reptiles remains controversial. We measured yolk hormones (dihydrotestosterone, testosterone, and 17 beta-estradiol) in newly laid eggs of the montane scincid lizard Bassiana duperreyi. This species is well suited to such an analysis because (1) offspring sex is influenced by incubation temperatures and egg size as well as by sex chromosomes, suggesting that yolk hormones might somehow be involved in the complex pathways of sex determination, and (2) experimental application of either estradiol or fadrozole to such eggs strongly influences offspring sex. We obtained yolk by biopsy, before incubating the eggs at a temperature that produces a 50:50 sex ratio. Yolk steroid levels varied over a threefold range between eggs from different clutches, but there were no significant differences in yolk steroids, or in relative composition of steroids, between eggs destined to become male versus female. Further, yolk steroid concentrations were not significantly related to egg size. Thus, yolk steroid hormones do not appear to play a critical role in sex determination for B. duperreyi.     

Inhibition of lactate dehydrogenase isoenzyme associated with anaerobic respiration in schistosomiasis intermediate host snails

Lactate dehydrogenase isoenzyme (LD5) which is associated with anaerobic respiration was inhibited to a certain degree in Biomphalaria alexandrina snails, the intermediate host for Schistosoma mansoni. Urea and thiourea were used as inhibitors. The effect of LD5 inhibition on the mortality rate of infected Biomphalaria alexandrina snails and on the susceptibility of the snails to the trematode infection was also studied.     

Trichobilharzia ocellata: physiological characterization of giant growth, glycogen depletion, and absence of reproductive activity in the intermediate snail host, Lymnaea stagnalis

Giant growth, depletion of energy stores, and inhibition of reproductive activity are striking effects of many trematode parasites on their intermediate snail hosts. Two hypotheses have been put forward to explain these phenomena: (1) host and parasite compete for energy rich and other essential nutrients, with the parasite as the winner, and (2) the parasite intervenes in the endocrine control of reproduction of the snail. These hypotheses were tested in the present study with the Trichobilharzia ocellata/Lymnaea stagnalis association. The snails were infected at a juvenile stage, and release of cercariae started on Day 55 after exposure. It was shown that enhanced growth of infected snails is not paralleled by a greater increase in dry weight, but hemolymph volume does increase, being 35% greater than in the noninfected controls. Control snails, on the other hand, showed an increase in the percentage body dry weight during sexual maturation. The conclusion is that infected snails retain an essentially juvenile body structure. In control snails, glycogen was depleted from the mantle store at the start of egg laying but the onset of cercariae production marked a severe glycogen depletion from the headfoot and the mantle in infected snails, being nearly complete on Day 68 after exposure. The hemolymph glucose concentration was only slightly lower in infected than in control snails and it did not change (in both groups) during glycogen mobilization. This suggests that glycogen mobilization does not result from the snail and the parasite competing directly for metabolites within the hemolymph. Infection inhibited the maturation of the accessory sex organs: there was no increase in the relative wet weights nor in the amounts of DNA and secretion products in the albumin and prostate glands. Infected snails did not lay eggs. It is presumed that the parasite produces one or more agents which intervene in the action of the gonadotrophic hormones. The release of these agents commences at an early stage of infection.     

Sex and immunity in the yellow dung fly Scathophaga stercoraria

The immune system is of increasing interest to evolutionary biologists. Immunity may trade-off against other fitness components, with recent work suggesting reproduction in particular impinges on immune defence. There may also be sex differences in the immune system. Additionally, while life history traits typically have low heritability, little is known about additive genetic variance of immunity. An insect's major defence against multicellular pathogens is to encapsulate the invader. Phenoloxidase (PO) is a key enzyme in the cascade resulting in the melanized capsule, and is often used to estimate resistance to an immune insult. We examined the effects of copulation, egg laying, sex and age on PO in Scathophaga stercoraria. We also measured the heritability of PO activity. The sexes differed in haemolymph PO activity and PO was significantly affected by age, but not by copulation or egg laying. There was significant heritable variation for haemolymph PO.     

Energy allocation patterns in Biomphalaria alexandrina snails in response to cadmium exposure and Schistosoma mansoni infection

This study was aimed to investigate the effects of both parasitism and environmental stress on the growth, reproduction, and survival of Biomphalaria alexandrina snails. Resource allocation strategies may be influenced by both biotic and abiotic factors. Using the planorbid snail B. alexandrina and Schistosoma mansoni, this hypothesis was examined by raising snails fed the same diet under two stressors (infection and Cd exposure). The snails divided into four groups, uninfected, infected, Cd-exposed uninfected, and Cd-exposed infected snails. Egg production, growth, and survival of the snails were monitored over a 9-week period postinfection. Inhibition of snail reproductive activity by parasitism results in increased snail growth in the first week postinfection, termed gigantism, during which the snail is hypothesized to allocate excess energy normally used for reproduction to somatic growth. Infection status and Cd exposure had significant effects on snail growth and reproduction. The infected and Cd-exposed infected snails exhibiting reduced survival relative to snails of other treatments. It was found that parasite development influenced by Cd exposure. Results of this study suggest that energy allocation patterns are context-dependent in B. alexandrina snails, influenced by infection and Cd exposure.     

Antimicrobial activities of Gloriosa superba Linn (Colchicaceae) extracts

The methanol extract of the rhizomes of Gloriosa superba Linn (Colchicaceae) and its subsequent fractions in different solvent systems were screened for antibacterial and antifungal activities. Excellent antifungal sensitivity was expressed by the n-butanol fraction against Candida albicans and Candida glaberata (up to 90%) and against Trichophyton longifusus (78%) followed by the chloroform fraction against Microsporum canis (80%). In the antibacterial bioassay, the crude extract and subsequent fractions showed mild to moderate antibacterial activities. Chloroform fraction displayed highest antibacterial sensitivity against Staphylococcus aureous (88%) followed by the crude extract (59%). The total phenol content of the crude extract and fractions of the plant expressed no significant correlation with the antimicrobial activities.     

Behavioural and electrophysiological responses of grape berry moth (Lep., Tortricidae) to selected plant extracts

Abstract:  Four non-host plant extracts from Bifora radians , Arctium lappa , Humulus lupulus and Xanthium strumarium , were tested against adult grape berry moths, Paralobesia viteana , for their potential as repellents or oviposition deterrents. Responses were compared with those elicited by the major component of the P. viteana sex pheromone. Moths of both sexes exhibited varying electroantennogram (EAG) responses from 1.7 to 2.4 mV to volatile odours from plant extracts, with the greatest response to extracts of H. lupulus (2.4 mV). A multi-arm olfactometer was used to assay the behavioural response of moths to the same treatments. Male moths responded significantly to H. lupulus extract, although their strongest response was to the sex pheromone (30.0%). Female moths showed no behavioural response to the sex pheromone, attracting only 6.7% of moths, but they were attracted to extracts of H. lupulus , X. strumarium and A. lappa , with 25.0%, 21.7% and 15.0% of the released insects responding to these treatments, respectively. Choice tests were conducted to compare oviposition by P. viteana on untreated grapes and on grapes treated with one of the extracts. Despite the attraction in the olfactometer assays, H. lupulus extract significantly reduced egg laying; only 12.5% of the total deposited eggs were laid on berries treated with this extract. The extract of B. radians provided complete protection against oviposition.