<Emphasis Type="Italic">Eimeria biarmicus</Emphasis> sp.n. (Apicomplexa: Eimeriidae) infecting falcons from the genus <Emphasis Type="Italic">Falco</Emphasis> in Saudi Arabia

The oocysts of Eimeria biarmicus sp. n. were described from the feces of the lanner falcon, Falco biarmicus, collected from the falcon market in Riyadh City, Saudi Arabia. The prevalence of infection was 5% (2/40). The majority of the oocysts examined had completed sporulation within 84 h at 24 ± 2°C. Sporulated oocysts are ovoid in shape, measuring 22.4 × 17.9 (20.5–24.7 × 15.8–18.5) μm; shape index (L/W) is 1.25 (1.14–1.36) μm. The oocyst wall is smooth and bi-layered. Micropyle and oocyst residuum are absent. A polar granule is present, consisting of 2–4 globules. Sporocysts are ovoid, 10.1 × 6.1 (9.4–11.2 × 5.4–6.8) μm; with a smooth single-layered wall and a minute Stieda body, but there is no substieda body. The sporocyst residuum consists of numerous small granules. Sporozoites are comma shaped, each contains two refractile bodies. E. biarmicus sp. n. is the second eimerian species described from F. biarmicus.     

<Emphasis Type="Italic">Myxobolus lubati</Emphasis> n. sp. (Myxosporea: Myxobolidae), a new parasite of haffara seabream <Emphasis Type="Italic">Rhabdosargus haffara</Emphasis> (Forsskal, 1775), Red Sea,Egypt: a light and transmission electron microscopy

A new myxosporean parasite, Myxobolus lubati n. sp., was described from the wall of the intestine of haffara seabream Rhabdosargus haffara (Forsskal 1775), Red Sea, Egypt. Macroscopic plasmodia of about 300 μm diameter were located in the circular muscle layer of the intestine. The spores were ovoid and sometimes ellipsoid and measured 9.8 × 7.2 μm. The shell wall of the spore was thickened at the posterior end and marked with 5–7 sutural markings. Polar capsules were equal and pyriform with three polar filament turns situated in the posterior half of the polar capsule. Polar capsules measured 4.2 × 1.6 μm. Histological evaluation of the infection revealed a slight distention of the intestinal layer of muscularis. Ultrastructure of the plasmodial wall and sporogenesis of the present species followed the usual pattern valid for most studied myxosporean species.     

<Emphasis Type="Italic">Sarcocystis</Emphasis> sp. from the goldeneye (<Emphasis Type="Italic">Bucephala clangula</Emphasis>) and the mallard (<Emphasis Type="Italic">Anas platyrhynchos</Emphasis>): cyst morphology and ribosomal DNA analysis

By light microscopy, cysts of Sarcocystis sp. (cyst type I) from the goldeneye (Bucephala clangula) seemed filamentous with a smooth and thin (<1 μm) cyst wall. Ultrastructurally, the cyst wall surface was irregular with minute undulations of the primary cyst wall. These sarcocysts had type-1 cyst wall. Cystozoites were banana-shaped and measured 7.0–8.5 μm in length. By light microscopy, cysts of Sarcocystis sp. (cyst type II) from the mallard (Anas platyrhynchos) were ribbon-shaped, very long, and thin. On the surface of the wall (up to 1.5 μm), they had palisade-like villar protrusions closely crowded together. Electron micrographs showed villar protrusions (up to 1.3 μm in length) different in size and shape. The latter had short microprojections especially obvious in the oblique sections. Cystozoites were slightly bent with blunt ends, broader at one end, and measured 13.0–16.1 × 1.8–2.5 μm. Phylogenetic analysis based on the comparison of partial 28S rRNR gene sequences of Sarcocystis sp. (cyst type II) derived from the mallard, Sarcocystis sp. (cyst type I) and Sarcocystis sp. (cyst type III) both derived from the white-fronted goose (Anser albifrons) suggested that these sequences belonged to separate Sarcocystis species.     

Light and electron microscopic studies on <Emphasis Type="Italic">Myxobolus egyptica</Emphasis> sp. nov. (Myxozoa,Myxosporea), infecting the hornlip mullet <Emphasis Type="Italic">Oedalechilus labiosus</Emphasis> from the Red Sea

A new myxosporean, Myxobolus egyptica sp. nov., was described from the gills of the hornlip mullet Oedalechilus labiosus, collected from the Red Sea at Al-Quseir city, Egypt. The prevalence of infection was 12/72 (16.66%). Myxobolus egyptica was identified on the basis of spore morphometry, histology and transmission electron microscopy. It was distinguished from all previously reported Myxobolus spp. by its shape, dimensions of the mature spore 10.0 ± 0.6 (9.5–10.5) μm in length, 8.5 ± 0.4 (8.0–9.0) μm in width and 8.7 ± 0.5 (8.4–9.2) μm in thickness, polar capsules, locality and host. The parasite formed intrafilamental cyst-like plasmodia. These plasmodia caused curling and atrophy of the gill lamellae. The ultrastructural analysis revealed a double-unit plasmodial membrane which was in direct contact with the host cells and had numerous vesicles. Some mitochondria were found below this membrane. The disporic pansporoblast was earliest recognizable stage of sporogenesis. Advanced developmental stages of spores and mature spores were reported.     

Biometrical and genetical characterization of large <Emphasis Type="Italic">Babesia Ovis</Emphasis> in Iran

One species of Babesia was identified on the blood smear of 20 different naturally infected sheep in the Northwest of Iran. It was polymorphic, including double pyriform with acute or obtuse angle, single pyriform, and ring form. The size of typical paired pyriforms with acute angle was 2.7 × 0.4 μm (n = 10) and with obtuse angle was 3.5 × 0.6 μm (n = 10). Although the morphological and biometrical parameters resembled the Babesia motasi, the results of seminested polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism using primers specific for small subunit of 18S rRNA confirmed this species as Babesia ovis. Furthermore, the sequence analysis of hypervariable region of small subunit of 18S rRNA revealed the corresponding sequences for B. ovis as well. Experimental infection of healthy lambs with the morphological larger B. ovis showed a milder clinical signs compared to the small one.     

Morphological and molecular biological characterization of <Emphasis Type="Italic">Pleistophora aegyptiaca</Emphasis> sp. nov. infecting the Red Sea fish <Emphasis Type="Italic">Saurida tumbil</Emphasis>

One hundred three out of 225 (45.8%) of the Red Sea fish Saurida tumbil were infected with microsporidian parasites. The infection was recorded as tumor-like masses (whitish macroscopic cysts) or xenomas often up to 2 cm in diameter and embedded in the peritoneal cavity. Generally, the infection was increased during winter 63.8% (86 out of 135) and fall to 18.9% (17 out of 90) in summer. Light microscopic study revealed that xenomas were encapsulated by a fibrous layer encircling numerous sporophorous vesicles filled with mature spores measuring 1.7 ± 0.6 (1.5–2.7 μm) × 1.5 ± 0.3 μm (1.2–1.8 μm) in size. Ultrastructural microscopic study showed the presence of smooth membranes of the sarcoplasmic reticulum forming a thick, amorphous coat surrounding various developmental stages of the parasite. The various recognizable stages of the parasite were uninuclear, binucleated, and multinucleated meronts followed by detachment of the plasmalemma of the sporont from the sporophorous vesicle producing sporoblasts. Mature spores consist of a spore coat and spore contents. The spore contents consist of the uninucleated sporoplasm and a posterior vacuole located at the posterior end. The polar tube consists of a straight shaft and a coiled region (26–32 coils) arranged in many rows along the inside periphery of the spore. The polaroplast consisted of an anterior region of closely and loosely packed membranes. Molecular analysis based on the small subunit rDNA gene was performed to determine the phylogenetic position of the present species. The percentage identity between this species and a range of other microsporidia predominantly from aquatic hosts demonstrated a high degree of similarity (>92%) with eight Pleistophora species. Comparison of the nucleotide sequences and divergence showed that the sequence of the present microsporidium was most similar to that of Pleistophora anguillarum (99.8% identity) differing in 13 nucleotide positions. So, the present species was recorded and phylogenetically positioned as a new species of Pleistophora.     

Comparative studies of the anti-leishmanial activity of three <Emphasis Type="Italic">Crotalus durissus</Emphasis> ssp. venoms

In this study, we compared the anti-leishmanial activity of three crotalic venoms (Crotalus durissus terrificus–Cdt, Crotalus durissus cascavella–Cdca, and Crotalus durissus collilineatus–Cdcol). Different concentrations of each venom incubated with Leishmania (Leishmania) amazonensis promastigotes were used. Cdt venom exhibited a higher anti-leishmanial activity (Inhibitory concentration-IC50-value of 4.70 ± 1.72 μg/ml) in comparison with that of Cdca venom (IC50 value of 9.41 ± 1.21 μg/ml), while Cdcol venom increased parasite numbers in 50% at a concentration of 44.30 ± 2.18 μg/ml. In addition, this venom showed a low anti-leishmanial activity in higher concentrations (IC50 value of 281.00 ± 9.50 μg/ml). The main fractions of Cdca venom were isolated and assayed under similar conditions used for assessing crude venom. The most active fractions were gyroxin and crotamine that had IC50 values of 3.80 ± 0.52 μg/ml and 19.95 ± 4.21 μg/ml, respectively. Convulxin also inhibited parasite growth rate, although this effect was not dose-dependent. Crotoxin was the least effective fraction with an IC50 value of 99.80 ± 2.21 μg/ml. None of the protein fractions presented cytotoxic effects against J774 cells in culture. In vivo assays using BALB/c mice revealed that crotoxin and crotamine were the main toxic fractions. In conclusion, C. durissus cascavella venom has three main fractions with anti-leishmanial activity. These results open new possibilities to find proteins that might be used as possible agents against cutaneous leishmaniasis.     

A novel herbal formulation against dengue vector mosquitoes <Emphasis Type="Italic">Aedes aegypti</Emphasis> and <Emphasis Type="Italic">Aedes albopictus</Emphasis>

The objective of this study was to develop a herbal formulation to control dengue vector mosquitoes. PONNEEM, a novel herbal formulation prepared using the oils of neem (Azadirachta indica), karanj (Pongamia glabra) and their extracts, was tested for larvicidal, ovicidal and oviposition deterrent activities against Aedes aegypti and Aedes albopictus at 1, 0.5, 0.3 and 0.1 ppm concentrations. Cent percent larvicidal and ovicidal activities were observed at 0.1 ppm in the two mosquito species under laboratory and sunlight-exposed conditions up to 12 months from the date of manufacture. Oviposition deterrent activity of 69.97% and 71.05% was observed at 1 ppm concentration of PONNEEM against A. aegypti and A. albopictus, respectively. Reduction in enzyme levels for α-esterase was 0.089 ± 0.008 and 0.099 ± 0.140 μg napthol produced/min/mg larval protein; for β-esterase, it was 0.004 ± 0.009 and 0.001 ± 0.028 μg napthol produced/min/mg larval protein; for glutathione S-transferase, it was 10.4814 ± 0.23 and 11.4811 ± 0.21 μmol/min/mg larval protein and for total protein, it was 0.177 ± 0.010 and 0.008 ± 0.005 mg/individual larva in treated groups of A. aegypti and A. albopictus, respectively. The nontarget organisms such as Gambusia affinis and Diplonychus indicus were not affected. No mortality was observed in control. PONNEEM can be used effectively for the management of human vector mosquitoes.     

Two new <Emphasis Type="Italic">Isospora</Emphasis> species from Brazilian tanager (<Emphasis Type="Italic">Ramphocelus bresilius dorsalis</Emphasis>) of South America

Two new coccidian (Protozoa: Apicomplexa: Eimeriidae) species from the Brazilian tanager Ramphocelus bresilius dorsalis are reported in the current study. Isospora cadimi n. sp. oocysts are spheroidal to sub-spheroidal, 24.2 × 22.9 μm, with a smooth and bi-layered wall, ∼1.1 μm. Micropyle, oocyst residuum, and polar granule are absent. Sporocysts are broadly ovoidal, 16.9 × 11.6 μm. Stieda and substieda bodies are present. Sporocyst residuum is present and sporozoites have refractile body and nucleus. Isospora navarroi n. sp. oocysts are spheroidal to sub-spheroidal, 21.4 × 20.6 μm, with a smooth and bi-layered wall, ∼1.1 μm. Micropyle, oocyst residuum, and polar granule are absent. Sporocysts are ellipsoidal, 16.1 × 10.2 μm. Stieda and substieda bodies are present. Sporocyst residuum is present and sporozoites have a robust posterior refractile body.     

<Emphasis Type="Italic">Kudoa unicapsula</Emphasis> n. sp. (Myxosporea: Kudoidae) a parasite of the Mediterranean mullets <Emphasis Type="Italic">Liza ramada</Emphasis> and <Emphasis Type="Italic">L. aurata</Emphasis> (Teleostei: Mugilidae)

A new multivalvulid myxozoan parasite, Kudoa unicapsula n. sp., is described from the intestinal mesentery, intestine and pyloric caeca of the thin-lipped grey mullet Liza ramada (Risso 1826) and the golden grey mullet L. aurata (Risso, 1810) from the Mediterranean coastal waters of Spain. It is characterized by the presence of elongated, rice corn-like white cysts of 0.47–0.56 × 0.18–0.38 mm, filled with tetracapsulate, slightly asymmetric spores, rectangular in apical view and tear-shaped in lateral view with four polar capsules of considerably different size and slightly unequal spore valves with rounded edges, overlapping each other on the apex of the spore. One large polar capsule includes a polar filament coiled in two to three turns, and the other three polar capsules, which are very small, posses only a rudimental filament. Both light and electron microscopy data showed that this species differs from all previously described Kudoa spp. with unequal polar capsules. The molecular analysis based on 18S and 28S ribosomal deoxyribonucleic acid DNA sequence data of K. unicapsula n. sp. indicates a close relationship and thus phylogenetic clustering together with K. trifolia, a myxozoan from the same host and the same geographical location.     

A new species of <Emphasis Type="Italic">Falcaustra</Emphasis> (Nematoda,Kathlaniidae) from <Emphasis Type="Italic">Cnemaspis</Emphasis> aff. <Emphasis Type="Italic">tro pidogaster</Emphasis> (Squamata,Gekkonidae) from Sri Lanka

Falcaustra desilvai sp. nov. (Ascaridida, Kathlaniidae) from the large intestine of Cnemaspis aff. tropidogaster (Squamata, Gekkonidae) is described and illustrated. Falcaustra desilvai represents the 4th nematode species from Sri Lanka to be assigned to the genus and is distinguished from other Sri Lankan species by the distribution pattern of caudal papillae (12 precloacal, 2 adcloacal, 10 postcloacal, and 1 median), length of spicules (956–1046 μm) and absence of a pseudosucker.     

Infestation status of gnathiid isopod juveniles parasitic on Dusky grouper (<Emphasis Type="Italic">Epinephelus marginatus</Emphasis>) from the northeast Mediterranean Sea

This is the first detailed documented record of Gnathiid isopod praniza larvae infestating dusky grouper, (Epinephelus marginatus Lowe 1834) in the northeast Mediterranean Sea (36°36′N–36°07′E, 35°52′N–36°25′E). Fish were sampled monthly from Iskenderun Bay during a 3-year period from 2000 to 2003 [N = 468, W ± SD (range) = 503.69 ± 342.35 g (177–2,832 g), TL ± SD (range) = 32.39 ± 9.22 cm (16.1–67.0 cm), W _total = 0.213L _total ^2.19, r _total ² = 0.85]. Juveniles of the Gnathia sp. were only extracted from the epithelium of the buccal cavity. The monthly and seasonal patterns in infestation rates (mean prevalence, P = 27.35% and mean intensity, MI ± SD = 21.35 ± 16.19), and the relationship between length–weight and infested/non-infested fish were calculated. This study suggests that gnathiid parasite has no effect on the growth and general health condition of infested fish, although high intensities were observed in fish.     

Two new <Emphasis Type="Italic">Isospora</Emphasis> species from the saffron finch, <Emphasis Type="Italic">Sicalis flaveola</Emphasis> in Brazil

Two new coccidian species (Protozoa, Apicomplexa, Eimeriidae) are reported from the saffron finch Sicalis flaveola Linnaeus, 1766, a very common species in South America. Isospora cetasiensis sp. nov. oocysts are subspherical to ellipsoidal, 23.1 × 21.6 μm, with smooth, bilayered wall, ∼1.0 μm. Micropyle, oocyst residuum and polar granule are absent. Sporocysts are ovoidal, 15.1 × 10.9 μm. Stieda body is knob-like and substieda body is rounded. Sporocyst residuum is composed of many scattered granules and spherules of different sizes. Sporozoites are vermiform with one refractile body and a nucleus. Isospora sicalisi sp. nov. oocysts are subspherical to ellipsoidal, 27.5 × 25.2 μm, with a smooth, bilayered wall, ∼1.1 μm. Micropyle, oocyst residuum and polar granule are absent. Sporocysts are ellipsoidal, 17.2 × 11.7 μm. Stieda body is knob-like and substieda body is trapezoidal. Sporocyst residuum is composed of scattered granules and spherules of different sizes. Sporozoites are vermiform with one refractile body and a nucleus.     

<Emphasis Type="Italic">Sarcocystis</Emphasis> sp. from the herring gull (<Emphasis Type="Italic">Larus argentatus</Emphasis>) identity to <Emphasis Type="Italic">Sarcocystis wobeseri</Emphasis> based on cyst morphology and DNA results

Having studied 11 herring gulls (Larus argentatus) Sarcocystis cysts were found in neck and leg muscles of 4 birds. One type of sarcocysts (cyst type I) that have a thin (∼1.0 μm), smooth, or slightly wavy cyst wall without clearly visible protrusions and small (6.0–8.0 μm) lancet- or banana-shaped cystozoites was identified by the light microscopy. Sarcocysts extracted from one herring gull were used for electron microscopy and DNA analysis. Ultrastructurally, Sarcocystis sp. from the herring gull had the same tissue cyst wall type-1 as S. calchasi, S. columbae, and S. wobeseri parasitizing in birds. According to first internal transcribed spacer (ITS-1) region, 18S rRNA and 28S rRNA gene sequences, Sarcocystis sp. from the herring gull belongs to S. wobeseri. Nevertheless, without evidences of cross-transmission experiment sarcocysts extracted from herring gull at present time are named as S. wobeseri-like.     

A new xenoma-forming microsporidium infecting intestinal tract of Atlantic bluefin tuna (<Emphasis Type="Italic">Thunnus thynnus</Emphasis>)

An undescribed species of Microsporidia Balbiani, 1882 was isolated from the muscularis mucosa of the intestinal mucosa of the reared Atlantic bluefin tuna (Thunnus thynnus). Transmission electron microscopy showed that the pyriform unikaryotic microsporidium, measuring 3.6 μm ± 0.08 in length and 1.8 μm ± 0.04 in width (inferred from TEM sections), had two layers of 14–17 coils of polar filament and a robust manubrium of the filament. No developmental stages were observed in xenoma. Phylogenetic analysis of the small subunit rRNA showed closest similarity with Kabatana spp. (88.4%), clustering the microsporidium together in a sister group with K. takedai and Kabatana sp. (JI-2008)/K. newberryi clade. While recognized Kabatana members show ovoid, rounded to pyriform spore, lack of sporophorous vesicle or xenoma, 3–10 coils of polar filament in 1–2 rows and are localized within cytoplasm of trunk muscle, the new species is markedly pyriform, xenoma forming, with many filament coils in 2 rows and parasitizing smooth intestinal muscle. Since morphological features were not typically congruent with any of Kabatana spp. so far described and molecular clustering indicated paraphyletic position within Kabatana clade, we suggest assigning described species to collective group Microsporidia, as Microsporidium milevae sp. nov., until more evidence permits potential formation of the new genus.     

The intestinal nematode <Emphasis Type="Italic">Trichuris arvicolae</Emphasis> affects the fecundity of its host,the common vole <Emphasis Type="Italic">Microtus arvalis</Emphasis>

Parasites have detrimental effects on host fitness. Consequently, they play a major role for host population dynamics. In this study, we investigated experimentally the impact of the nematode Trichuris arvicolae on the reproduction of its host, the common vole Microtus arvalis. Wild common voles were trapped in east of France and reared in standardized conditions before being experimentally infected. Infection with Trichuris arvicolae did not affect host consumption of food or water. Parasitized females gave birth to slightly less pups (mean 3.36 ± 0.38) than unparasitized females (mean 3.60 ± 0.40). Controlling for natal litter size using analysis of covariance (ANCOVA), T. arvicolae infection had a significant effect on the individual mass at birth, with pups from parasitized females having significantly lower mass (2.11 g ± 0.01) than pups from unparasitized females (2.20 g ± 0.01). Other measures of host reproductive outputs (time to first reproduction, mass of pups at weaning, litter survival) were not affected by maternal parasite infection. We discuss how these changes in M. arvalis reproductive investments associated with T. arvicolae infection must now be investigated in the context of physiological trade-offs.     

The comparative morphology of <Emphasis Type="Italic">Marshallagia marshalli</Emphasis> and <Emphasis Type="Italic">Ostertagia occidentali</Emphasis>s (Nematoda: Strongylida,Trichostrongylidae) by scanning electron microscopy

The systematics of the Ostertagiinae is unsettled with no agreement on how many genera and species are present in cattle and sheep. Ten species of Ostertagiinae are commonly parasitic in cattle and sheep. In the global fauna, six of 13 ostertagiine genera are endemic to Iran. The life cycle of Ostertaginae is direct and ingested third-stage larvae after exsheatment in the rumen, penetrate the gastric glands in the abomasal mucosa where two parasitic moults occur before the L5 emerges from the gland. In the present work, Marshallagia marshalli and Ostertagia occidentalis, collected from the abomasums of sheep from Mashhad, Iran, is described. The association of light and scanning electron microscopy (SEM) allowed a detailed analysis of the morphology and ultrastructure of these nematodes. The male body length of M. marshalli and O. occidentalis were 9.3–10.20 and 9.60–10.50 mm, respectively. The female body length of M. marshalli and O. occidentalis were 10.10–15.30 and 10.4–15.70 mm, respectively. One of cervical papillae is seen 333 and 250 μm from the anterior end of male and female body surface in O. occidentalis and 287.5 and 200 μm from the anterior end of male and female body surface in M. marshalli, respectively. The size of cervical papillae is 13.3 μm in male and 10 μm in female in O. occidentalis and 9.33 μm in male and 8.57 μm in female in M. marshalli. Some other taxonomic features of M. marshalli and O. occidentalis, such as details of cephalic region, the system of longitudinal and surface cuticular ridges (synlophe), the orientation of rays of the copulatory bursa, localization of vulva, morphology of vulvar flap, and posterior end of females are also documented by SEM.     

Characterization of <Emphasis Type="Italic">Taenia solium</Emphasis> cysticerci microsomal glutathione <Emphasis Type="Italic">S</Emphasis>-transferase activity

Glutathione S-transferase activity has been shown to be associated with the microsomal fraction of Taenia solium. Electron microscopy and subcellular enzyme markers indicate the purity of the microsomal fraction that contains the glutathione S-transferase activity. T. solium microsomes were solubilized under conditions used to solubilize integral microsomal proteins. This procedure proved necessary to obtain enzymatic activity. To characterize this parasite enzyme activity, several substrates and inhibitors were used. The optimum activity for microsomal glutathione S-transferase was found to be pH 6.6, with a specific enzyme activity of 0.9, 0.1, 0.067, 0.03, and 0.05 μmol min⁻¹ mg⁻¹ with the substrates 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene, 4-hydroxynonenal, 2,4-hexadienal, and trans-2-nonenal, respectively. No activity of glutathione peroxidase was observed. T. solium microsomes had an _app K _{m (GSH)} = 0.161 μM, _app K _{m (CDNB)} = 14.5 μM, and _app V _max of 0.15 and 27.9 μmol min⁻¹ mg⁻¹ for GSH and CDNB, respectively. T. solium microsomes were inhibited by several glutathione S-transferase enzyme inhibitors, and it was possible to establish a simple inhibition system as well as corresponding K _i ’s for each inhibitor. These results indicate that the T. solium microsomal glutathione S-transferase is different from the parasite cytoplasmic enzymes that catalyze similar reactions.     

Antileishmanial potential of a marine sponge, <Emphasis Type="Italic">Haliclona exigua</Emphasis> (Kirkpatrick) against experimental visceral leishmaniasis

In this study, we are reporting antileishmanial activity of a marine sponge Haliclona exigua, belonging to phylum Porifera. The crude methanol extract and its three fractions were tested both in vitro and in vivo. The crude extract exerted almost complete inhibition of promastigotes at 50 μg/ml and 76.4 ± 6.5% inhibition of intracellular amastigotes at 100 μg/ml concentration with IC₅₀ values of 18.6 μg/ml and 47.2 μg/ml, respectively. When administered to Leishmania donovani infected hamsters at a dose of 500 mg/kg × 5, p.o., it resulted in 72.2 ± 10.4% inhibition of intracellular amastigotes. At a lower dose (250 mg/kg), it exhibited 43.9 ± 5.1% inhibition. Among the fractions, highest antileishmanial activity both in vitro (>90%) and in vivo (60.9 ± 18.3%) was observed in n-butanol (soluble) fraction with IC₅₀ values of 8.2 μg/ml and 31.2 μg/ml against promastigotes and intracellular amastigotes, respectively. Hexane fraction also showed comparatively good activity against both the stages of parasites in vitro but was moderately active in leishmania-infected hamsters. Chloroform fraction resulted in 45 ± 10.2% inhibition in vivo at a dose of 500 mg/kg × 5, p.o., whereas it was inactive in vitro. n-Butanol (insoluble) fraction was inactive both in vitro and in vivo. Araguspongin C, an alkaloid isolated from n-butanol (soluble) fraction exhibited moderate inhibition of promastigotes and intracellular amastigotes at 100 μg/ml but showed weak antileishmanial action in vivo. Our findings indicate that this marine sponge has the potential to provide new lead toward development of an effective antileishmanial agent and, hence, calls for more exhaustive studies for exploiting the vast world of marine resources to combat the scourge of several parasitic diseases.     

Serum protein alterations in dogs naturally infected with <Emphasis Type="Italic">Toxoplasma gondii</Emphasis>

We conducted this study to describe the serum electrophoretic pattern in dogs associated with the infection of Toxoplasma gondii (T. gondii). The serum protein pattern of 25 dogs with confirmed T. gondii infection and 15 clinically healthy dogs were evaluated using native polyacrylamide gel electrophoresis. Albumin, alpha-1 globulin, alpha-2 globulin, beta globulin, and gamma globulin bands were seen from the serum electrophoresis of infected and healthy dogs. Compared to the control group, significant decreases in the mean percentages of albumin (from 46.1 ± 7.2 to 40.8 ± 4.5%, P < 0.05), alpha-1 globulin (from 3.9 ± 0.4 to 0.8 ± 0.2%, P < 0.001), alpha-2 globulin (from 9.0 ± 0.4 to 8.3 ± 0.8%, P < 0.01), and beta globulin (from 18.4 ± 1.2 to 12.1 ± 0.6%, P < 0.001) in the infected group were determined. In contrast, gamma globulin fraction was significantly higher in infected dogs (38.1 ± 4.6%) than in control dogs (22.7 ± 7.2%; P < 0.001). Moreover, significant correlations were determined between the percentages of the albumin and gamma globulin fractions and liver enzyme tests including aspartate aminotransferase and alanine aminotransferase in infected dogs; however, no correlation was observed for the other protein fractions. In conclusion, marked alterations in serum protein pattern associated with strong modifications of serum protein concentrations are in accordance with the hepatic injury as affirmed by liver enzyme tests that were demonstrated in the canine toxoplasmosis. These findings showed that serum protein electrophoresis can be used in the diagnosis and prognosis of canine toxoplasmosis as a supplementary analysis in combination with serological, clinical, and laboratory findings of this disease.