Resistance of Biomphalaria alexandrina to Schistosoma mansoni and Bulinus truncatus to Schistosoma haematobium Correlates with Unsaturated Fatty Acid Levels in the Snail Soft Tissue.

Only a fraction of the Biomphalaria and Bulinus snail community shows patent infection with schistosomes despite continuous exposure to the parasite, indicating that a substantial proportion of snails may resist infection. Accordingly, exterminating the schistosome intermediate snail hosts in transmission foci in habitats that may extend to kilometres is cost-prohibitive and damaging to the ecological equilibrium and quality of water and may be superfluous. It may be more cost effective with risk less ecological damage to focus on discovering the parameters governing snail susceptibility and resistance to schistosome infection. Therefore, laboratory bred Biomphalaria alexandrina and Bulinus truncatus snails were exposed to miracidia of laboratory-maintained Schistosoma mansoni and S. haematobium, respectively. Snails were examined for presence or lack of infection association with soft tissue and hemolymph content of proteins, cholesterol, and triglycerides, evaluated using standard biochemical techniques and palmitic, oleic, linoleic, and arachidonic acid, assayed by ultraperformance liquid chromatography-tandem mass spectrometry. Successful schistosome infection of B. alexandrina and B. truncatus consistently and reproducibly correlated with snails showing highly significant (up to P < 0.0001) decrease in soft tissue and hemolymph content of the monounsaturated fatty acid, oleic acid, and the polyunsaturated fatty acids, linoleic, and arachidonic acids as compared to naïve snails. Snails that resisted twice infection had soft tissue content of oleic, linoleic, and arachidonic acid similar to naïve counterparts. High levels of soft tissue and hemolymph oleic, linoleic, and arachidonic acid content appear to interfere with schistosome development in snails. Diet manipulation directed to eliciting excessive increase of polyunsaturated fatty acids in snails may protect them from infection and interrupt disease transmission in a simple and effective manner.

Biological activity of artemisinin-naphthoquine phosphate on Schistosoma haematobium stages and the vector Bulinus truncatus.

BACKGROUND: Schistosoma haematobium infection is a major public health problem in most of Africa and the Middle East and praziquantel remains the only drug used for schistosomiasis control, therefore emergence of drug resistance is unavoidable. The antimalarial artemisinin-naphthoquine phosphate combination (co-ArNp) was recently documented to have promising effects on Schistosoma mansoni and its snail host. METHODS: We conducted this in vitro study to assess the bioactivity of co-ArNp on S. haematobium and its snail vector Bulinus truncatus. RESULTS: Treatment of S. haematobium worms with 1 µg/ml co-ArNp for 24 h reduced worm motility, while 20 µg/ml resulted in 25-100% mortality of adult flukes within 48-72 h. Incubation of S. haematobium miracidia and cercariae with the molluscicidal co-ArNp (50% lethal concentration 7.5 µg/ml) killed all the free larval stages within 40 and 15 min, respectively. Also, exposure of B. truncatus adult snails to 20 ppm of the combined regimen caused a mortality rate of 100% within 24 h. CONCLUSIONS: Co-ArNp therapy has also shown encouraging activity against the other major human schistosome, S. haematobium, as well as its vector.

ECHINOCHASMUS, NEW SPECIES (TREMATODA: ECHINOSTOMATIDAE) FROM EGYPT.

In this study a new species of Echinochasnius (Dietz, 1909) is recorded for the first time in Egypt. Life cycle of one of gymnocephalous cercariae procured from Melanoides tuberculata snail was successfully completed in the laboratory. A total of 407 Melanoides tuberculata snails were collected from Mansouriya Canal, Giza Governorate. They were individually exposed to artificial light to determine natural infection with trematode larvae, Seven snails were found infected with gymnocephalous cercariae (infection index of 1.71). These cercariae were used to infect Gambusia affinis fish as second intermediate host. The infected gills were given to clean laboratory bred Rattus norvegicus as experimental final host. Adult worms were obtained ten days post-infection from rats' intestine identified as Echinochasmus after accurate comparson with standard keys. The diagnostic morphology of developmental stages were given.

Spotlight on the in vitro effect of artemisinin-naphthoquine phosphate on Schistosoma mansoni and its snail host Biomphalaria alexandrina.

Malaria and schistosomiasis are the two most important parasitic diseases in the tropics and sub-tropics with geographic overlap. Efforts have been made for developing new schistosomicidal drugs, or testing existing drugs originally used for non-related diseases. The antimalarial artemisinin-naphthoquine phosphate combination (CO-ArNp) was recently reported to be a promising novel antischistosomal therapy with potent in vivo activity against Schistosoma mansoni. In this work, we report the in vitro dose- and time-response effect of CO-ArNp against the Egyptian strain of S. mansoni, and its snail host, Biomphalaria alexandrina. Incubation of adult S. mansoni with CO-ArNp at 40 or 20 µg/ml for 48 or 72 h killed all worms. Exposure of S. mansoni miracidia and cercariae to the molluscicidal LC50 of CO-ArNp (16.8 µg/ml) resulted in 100% mortality of the free larval stages within 90 and 15 min, respectively. Moreover, incubation of adult B. alexandrina snails with this drug combination killed all snails at 40 µg/ml within 24h. Scanning electron microscope revealed marked morphological and tegumental alterations on the different stages of the parasite and its snail soft tissue. Our study highlights the schistosomicidal and molluscicidal effects of artemisinin-naphthoquine phosphate. No doubt more studies are needed to clarify its potential value to control schistosomiasis.

Description of two new cercariae (an echinostome cercaria and a xiphidiocercaria) procured from Biomphalaria pfeifferi (Krauss) from Nigeria.

During parasitological examination of Biomphalaria pfeifferi snails obtained from Niger state (Nigeria), 2 new types of cercariae were found. They are identified to the level of referring to the major group and described here for the first time. They were examined viable and stained with vital stains as well as fixed in 70% alcohol. They were drawn with a camera lucida and photographed. They are identified as an echinostome cercaria and a xiphidiocercaria. The echinostome is characterized by having a ventral sucker almost double in size the oral one. It has a semicircular structure located beyond the oral sucker. Three pairs of penetration glands are found at the anterior portion of the body. The number of collar spines is relatively large (44-46). The flame cellsare 17 x 2 in number. Two main lateral excretory ducts extend anteriorly, form two typical echinostome loops then pass posteriorly to open together in a diverticulated excretory vesicle. Its tail is relatively long and flattened with 3 fin folds. The tail (640 µm) is longer than the body (475 µm). The xiphidiocercaria belongs to the "ornatae" group. It is relatively small (180.5 x 110 µm) with a long stylet (30 µm). Its oral sucker is one and half times the size of the ventral sucker. Two excretory ducts extend posteriorly in both sides and become dilated and unite to open in a circular excretoryvesicle. Tail is slender shorter than the body and has a dorso-ventral fin fold.

Experimental evaluation of Candonocypris novaezelandiae (Crustacea: Ostracoda) in the biocontrol of Schistosomiasis mansoni transmission.

OBJECTIVE: To test Candonocypris novaezelandiae (Baird) (C. novaezelandiae), sub-class Ostracoda, obtained from the Nile, Egypt for its predatory activity on snail, Biomphalaria alexandrina (B. alexandrina), intermediate host of Schistosoma mansoni (S. mansoni) and on the free-living larval stages of this parasite (miracidia and cercariae). METHODS: The predatory activity of C. novaezelandiae was determined on B. alexandrina snail (several densities of eggs, newly hatched and juveniles). This activity was also determined on S. mansoni miracidia and cercariae using different volumes of water and different numbers of larvae. C. novaezelandiae was also tested for its effect on infection of snails and on the cercarial production. RESULTS: C. novaezelandiae was found to feed on the eggs, newly hatched and juvenile snails, but with significant reduction in the consumption in the presence of other diet like the blue green algae (Nostoc muscorum). This ostracod also showed considerable predatory activity on the free-living larval stages of S. mansoni which was affected by certain environmental factors such as volume of water, density of C. novaezelandiae and number of larvae of the parasite. CONCLUSIONS: The presence of this ostracod in the aquatic habitat led to significant reduction of snail population, infection rate of snails with schistosme miracidia as well as of cercarial production from the infected snails. This may suggest that introducing C. novaezelandiae into the habitat at schistosome risky sites could suppress the transmission of the disease.

Schistosomicidal and molluscicidal activities of aminoalkylamino substituted neo- and norneocryptolepine derivatives.

CONTEXT: The cryptolepines originate from the roots of the climbing shrub Cryptolepis sanguinolenta (Lindi) Schitr(Periplocaeae) which is used in Central and West Africa in traditional medicine for the treatment of malaria. OBJECTIVES: Evaluation for the first time of a series of chloro- and aminoalkylamino derivatives of neo- and norneocryptolepines for potential schistosomicidal and molluscicidal activities. MATERIALS AND METHODS: A series of chloro- and aminoalkylamino substituted neo- and norneocryptolepine derivatives were synthesized. They were tested in vitro against viable Schistosoma mansoni Sambon mature worms in culturemedium with fetal serum and antibiotics and in dechlorinated water against the snail vector Biomphalaria alexandrina Ehrenberg. Active compounds were further subjected to determination of their IC50 values. RESULTS: Results showed that six neocryptolepine and two norneocryptolepine derivatives had in vitro schistosomicidal activity on Egyptian and Puerto Rican strains of S. mansoni. The most effective derivative (2-chloro-5-methyl-N-(2-morpholin-4-ethyl)-5H-indolo[2,3b]quinoline-11-amine) has IC50 and IC90 1.26 and 4.05 µM and 3.54 and 6.83 µM with the Egyptian and Puerto Rican strains of Schistosoma, respectively. All eight derivatives showed molluscicidal activity against the vector snail B. alexandrina. The most active compound (2-chloro-11-(4-methylpiperazin-1-yl)-6H-indolo[2,3-b] quinoline) has LC50 0.6 and LC90 3.9 ppm after 24 h. DISCUSSION AND CONCLUSIONS: The findings demonstrate that introducing chloro- and aminoalkylamino side chain initiated both schistosomicidal and molluscicidal activities in these derivatives. The structure­activity relationship of this series of compounds is discussed.

Contribution to in vitro screening of Egyptian plants for schistosomicidal activity.

CONTEXT: This study is a continuation of our previous work in which a bioassay screening of 346 methanol extracts from 281 Egyptian plant species was carried out for in vitro schistosomicidal activity. OBJECTIVE: Another 309 methanol extracts from 278 plant species were subjected to the bioassay screening using the same technique on viable Schistosoma mansoni Sambon (Schistosomatidae) mature worms in specialized culture medium (Roswell Park Memorial Institute medium 1640) in a trial to discover a source for a schistosomiasis drug from Egyptian flora. MATERIAL AND METHODS: The methanol plant extracts were tested in vitro against viable S. mansoni mature worms in culture medium. Viability of worms was examined after exposure to 100 µg/ml of the extract in the medium for 24 h. Negative (dimethyl sulfoxide) and positive (praziquantel) controls were simultaneously used. Extracts showing schistosomicidal activity were further subjected to determination of their (Lethal concentration) LC50 and LC90 values. RESULTS: Confirmed in vitro antischistosomal activity was found in 42 extracts. Of these, 14 plant species possessed considerably high antischistosomal activity (LC50 ≤ 15 µg/ml), viz. Callistemon viminalis (Soland. Ex Gaertn) Cheel, C. rigidus R.Br., C. speciosus (Sims.) DC, C. citrinus Stapf, Eucalyptus citriodora Hook, E. rostrata Dehnh., Eugenia edulis Vell, E. javanica Lam syn. Syzygium samarangense (Blume) Merril, Melaleuca leucadendron (L.) L., M. stypheloides Sm. (all belong to Myrtaceae), Cryptostegia grandiflora R.Br. (Asclepiadaceae), Zilla spinosa (L.) Prantl (Cruciferae), Ficus trijuja L. (Moraceae) and Fagonia mollis Delile (Zygophylacae). DISCUSSION AND CONCLUSION: These species may represent additional natural sources of bioactive material that deserve further investigation for drug discovery against schistosomiasis.

Bioactivity of miltefosine against aquatic stages of Schistosoma mansoni, Schistosoma haematobium and their snail hosts, supported by scanning electron microscopy.

BACKGROUND: Miltefosine, which is the first oral drug licensed for the treatment of leishmaniasis, was recently reported to be a promising lead compound for the synthesis of novel antischistosomal derivatives with potent activity in vivo against different developmental stages of Schistosoma mansoni. In this paper an in vitro study was carried out to investigate whether it has a biocidal activity against the aquatic stages of Schistosoma mansoni and its snail intermediate host, Biomphalaria alexandrina , thus being also a molluscicide. Additionally, to see whether miltefosine can have a broad spectrum antischistosomal activity, a similar in vitro study was carried out on the adult stage of Schistosoma haematobium, the second major human species, its larval stages and snail intermediate host, Bulinus truncutes. This was checked by scanning electron microscopy. RESULTS: Miltefosine proved to have in vitro ovicidal, schistolarvicidal and lethal activity on adult worms of both Schistosoma species and has considerable molluscicidal activity on their snail hosts. Scanning electron microscopy revealed several morphological changes on the different stages of the parasite and on the soft body of the snail, which further strengthens the current evidence of miltefosine's activity. This is the first report of mollusicidal activity of miltefosine and its in vitro schistosomicidal activity against S.haematobium. CONCLUSIONS: This study highlights miltefosine not only as a potential promising lead compound for the synthesis of novel broad spectrum schistosomicidal derivatives, but also for molluscicidals.