Evaluation of anthelminthic and antiprotozoal activity of myrrh (Commiphora myrrha) methanolic extract.

To treat and control parasitic infections, traditional medical remedies using plant products are utilized as antiparasitic agents rather than standard synthetic chemicals due to drug resistance. Myrrh, a resinous exudate of Commiphora myrrha (Burseraceae), is a powerful antioxidant with a variety of medicinal uses. This study aimed to investigate the effect of the myrrh methanolic extract (MyE) of three concentrations (100, 50, and 25 mg/ml) on the sporulation of oocysts and as an anthelminthic effector via in vitro study. Characterization of the plant was done by Fourier-transform infrared spectroscopy (FT-IR). The earthworm, Eisenia fetida, is used as a model worm to evaluate the anthelminthic activity of MyE. Eimeria labbeana-like oocysts are used as a model protozoan parasite in anticoccidial assays. The sporulation and inhibition (%) of E. labbeana-like were assessed by MyE compared to other chemical substances. FT-IR revealed the presence of twelve active compounds. Our results showed that paralysis and death of earthworms at MyE (100 mg/ml) were 7.88 ± 0.37 and 9.24 ± 0.60 min, respectively, which is more potency when compared to mebendazole (reference drug). In all treated worms, microscopic examinations revealed obvious surface architecture abnormality. This study shows that MyE affects oocysts sporulation in a dose-dependent manner. At 24 and 36 hr, a high concentration of MyE (100 mg/ml) inhibits sporulation by 90.95 and 87.17 %. At 36 hr, other concentrations of MyE (50 and 25 mg/ml), as well as amprolium, DettolTM, and phenol inhibits oocyst sporulation by 40.17 %, 29.34 %, 45.09 %, 85.11 %, and 61.58 %, respectively. According to our research, the MyE extract had powerful anthelmintic and anticoccidial properties.

Detection of Raillietina saudiae from the domestic pigeon in Saudi Arabia through 18S and 28S rDNA genes.

Raillietina saudiae is a well-studied avian gastrointestinal parasite belonging to the family Davaineidae and is the most prevalent cyclophyllid tapeworm infecting pigeon in Saudi Arabia. The present study considered as a complementary analysis of Al-Quraishy et al. (2019; Parasitol Int 71, 59-72) with molecular studies for two ribosomal DNA genes employed for precise recognition of this Raillietina species. The annotated partial 18S and 28S rDNA gene regions were found to be 888 and 900 bp long that utilized further to elucidate their genetic relationships at species level using maximum likelihood method. The query sequence of R. saudiae is well aligned and placed within the Davaineidae family, with the same clade of all species of Raillietina that well separated from other cyclophyllidean cestodes especially taeniid and hymenolepid species. Sequence data recorded the monophyly of Raillietina species. The current phylogeny supports the usage of the partial 18S and 28S rDNA genes as reliable markers for phylogenetic reconstructions.

Molecular phylogeny of Sarcocystis fayeri (Apicomplexa: Sarcocystidae) from the domestic horse Equus caballus based on 18S rRNA gene sequences and its prevalence.

Sarcocystosis is a parasitic disease caused by an intracellular protozoan parasite Sarcocystis belonging to the phylum Apicomplexa. These parasites have a requisite two-host life cycle. Recently, there are many Sarcocystis species that identified morphologically. In the present study, diaphragmatic muscle samples from the domestic horse (Equus caballus) were examined for Sarcocystis infection. The natural infection with sarcocysts was recorded to be 62·5% for only microcysts in the infected muscles. Molecular analysis using the 18S rRNA gene was conducted to swiftly and accurately identify the recovered species. Studies on the expression of the 18S rRNA gene have confirmed that the present parasite isolates belong to the Sarcocystis genus. The sequence data showed significant identities (>80%) with archived gene sequences from species within the Sarcocystidae family, and a dendrogram showing the phylogenetic relationship was constructed. The most closely related species were the previously described Sarcocystis fayeri and Sarcocystis bertrami. The current data showed that the present species was identified as S. fayeri and deposited in GenBank (accession number MF614956.1). This study highlights the importance of the genetic data in the exact taxonomy within sarcocystid species.

Plasmodium chabaudi-infected mice spleen response to synthesized silver nanoparticles from Indigofera oblongifolia extract.

Malaria is a worldwide serious-threatening infectious disease caused by Plasmodium and the parasite resistance to antimalarial drugs has confirmed a significant obstacle to novel therapeutic antimalarial drugs. In this article, we assessed the antioxidant and anti-inflammatory activity of nanoparticles prepared from Indigofera oblongifolia extract (AgNPs) against the infection with Plasmodium chabaudi caused in mice spleen. AgNPs could significantly suppress the parasitaemia caused by the parasite to approximately 98% on day 7 postinfection with P. chabaudi and could improve the histopathological induced spleen damage. Also, AgNPs were able to increase the capsule thickness of the infected mice spleen. In addition, the AgNPs functioned as an antioxidant agent that affects the change in glutathione, nitric oxide and catalase levels in the spleen. Moreover spleen IL1ß, IL-6 and TNF-α-mRNA expression was regulated by AgNPs administration to the infected mice. These results indicated the anti-oxidant and the anti-inflammatory protective role of AgNPs against P. chabaudi-induced spleen injury.

Ultrastructure and phylogeny of the parasite Henneguya carolina sp. nov. (Myxozoa), from the marine fish Trachinotus carolinus in Brazil.

Microscopic and molecular procedures are used to describe a new myxosporean species, Henneguya carolina sp. nov., found infecting the intestine of the marine teleost fish Trachinotus carolinus on the southern Atlantic coast of Brazil. Spherical to ellipsoid cysts, measuring up to ~750 µm, display synchronous development. Mature myxospores are ellipsoidal with a bifurcated caudal process. Myxospore body length, width, and thickness are 12.7 ± 0.8 (12.0-13.4) µm, 8.8 ± 0.6 (7.5-9.6) µm, and 5.8 ± 0.4 (5.0-6.4) µm, respectively; 2 equal caudal processes are 16.8 ± 1.1 (15.9-18.0) µm long, and the total myxospore length is 29.4 ± 0.8 (28.4-30.4) µm. Two pyriform polar capsules measure 5.0 ± 0.5 (4.6-5.6) × 2.4 ± 0.4 (1.9-2.9) µm, and each contains a polar filament forming 3 to 4 coils. Sporoplasm is binucleated and presents a spherical vacuole surrounded by numerous globular sporoplasmosomes. Molecular analysis of the small subunit rRNA gene by maximum parsimony, neighbor joining, and maximum likelihood reveals the parasite clustering together with other myxobolids that are histozoic in marine fish of the order Perciformes, thereby strengthening the contention that the host phylogenetic relationships and aquatic environment are the strongest evolutionary signal for myxosporeans of the family Myxobolidae.

In vitro cultivation model for Heterosporis saurida (Microsporidia) isolated from lizardfish, Saurida undosquamis (Richardson).

Heterosporis saurida is a microsporidian that infects lizardfish, Saurida undosquamis (Richardson, 1848), in the Arabian Sea. Spores were isolated from infected lizardfish and used to infect derived fish cell lines: common carp brain (CCB), epithelioma papulosum cyprinid (EPC), fathead minnow epithelial (FHM), rainbow trout gonad (RTG), bluegill fry (BF-2) and chinook salmon embryo (CHSE). Non-fish cell lines were also tested that include: insect (SF-9), rabbit (RK-13) and African green monkey (Vero E6). No growth of H. saurida was observed in any fish cell line, SF-9 or Vero E6 cell lines. H. saurida spores grew only in RK-13 cell line and were detected by immunofluorescence. Developmental stages of H. saurida were seen in RK-13 cells by light and transmission electron microscopy, and species identification was confirmed by sequencing. This study demonstrated that H. saurida was able to proliferate in the mammalian RK-13 cell line, which thus represents an in vitro model for conducting molecular genetics and cell-pathogen interaction studies of Heterosporis.

Ultrastructure of novel thrombocytes in the dog snapper Lutjanus jocu.

Myxosporean cysts containing spores of Henneguya sp. were observed in the gills of the dog snapper Lutjanus jocu. Adjacent to the cysts were capillaries, allowing observation of peripheral blood cells. Numerous white blood cells displaying uncommon cytoplasmic projections were observed amongst the erythrocytes. Their morphology allowed them to be identified as thrombocytes (TCs). Each TC displayed 18-26 cytoplasmic projections, most of which were in close proximity to erythrocytes. At their apical end, each cytoplasmic projection presented an ellipsoidal vacuole (c. 0·6 µm × 0·3 µm) from which a secretory tubule, 0·3-0·4 µm long and c. 120 nm in total diameter, extended towards the periphery of the TC plasmalemma and fused with the cellular membrane. From this opening, contents of vacuoles were apparently released into the lumen of the capillaries. Other vacuoles with similar features, and containing an electron-lucent matrix, were observed in the cytoplasm of the TC. This is the first description of fish TC with these ultrastructural features and organization, which suggest that they perform a secretory function.

Tissue metal distribution and risk assessment for important fish species from Saudi Arabia.

The concentrations of nine heavy metals (Cr, Cd, Pb, Ni, Cu, Zn, Mn, As and Fe) in the muscle tissue of four fish species (Oreochromis niloticus, Clarias gariepinus, Poecilia latipinna and Aphanius dispar dispar) collected from Wadi Hanifah were detected in two different seasons. The concentrations of the studied heavy metals, except for Cd, Pb, Ni and Cu in A. d. dispar and P. latipinna, were found to be below the safe limits recommended by various authorities and thus provide an indication of the extent of pollution of these metals. The present study also demonstrates that Zn and Cr were the most and least accumulated metals, respectively, in the studied fish muscle tissues. From the standpoint of human health, this study suggests that there is a possible health risk to consumers due to the current consumption rate of A. d. dispar and P. latipinna in Saudi Arabia.

Caryospora cherrughi sp. n. (Apicomplexa: Eimeriidae) infecting Falco cherrug in Saudi Arabia.

The oocysts of Caryospora cherrughi sp. n. were described from the feces of the saker falcon, Falco cherrug, collected from the falcon market in Riyadh City, Saudi Arabia. The prevalence of infection was 21.4 % (15/70). The majority of the oocysts examined had completed sporulation within 64-84 h at 24 ± 2 °C. Sporulated oocysts are ovoid in shape, measuring 32.1 × 29.3 (30.7-34.5 × 27.7-29.8) µm; shape index (L/W) is 1.1 (1.04-1.27) µm. The oocyst wall is smooth and bilayered. Micropyle, polar granule, and oocyst residuum are absent. Sporocysts are ellipsoidal, measuring 24.1 × 19.6 (23.0-25.5 × 18.7-20.5) µm; with a smooth single-layered wall, but there is no Stieda body. The sporocyst residuum is present as numerous small granules. Sporozoites are banana-shaped, blunt at one end, and slightly tapered at the other, each with a large single refractile body. C. cherrughi sp. n. is the third caryosporian species described from F. cherrug.

Ultrastructural and molecular studies of Microgemma carolinus n. sp. (Microsporidia), a parasite of the fish Trachinotus carolinus (Carangidae) in Southern Brazil.

A new species of Microsporidia Microgemma carolinus n. sp. found in the marine teleost Trachinotus carolinus collected in Florianópolis, Brazil was described based on light, ultrastructural and phylogenetic studies. This parasite developed in the liver forming whitish xenomas that contained different developmental stages with monokaryotic nuclei. The periphery of the xenoma presented some vacuolization and possessed several small projections in the membrane. The mature spores, measuring 3·8 ± 0·4 µm in length and 2·4 ± 0·4 µm in width, were slightly pyriform to ellipsoidal and had rounded ends. The polaroplast was bipartite and the isofilar polar filament was coiled with 8 - 9 turns in a single or double row at the posterior end of the spore. The nucleus was voluminous and in a central position, measuring ~0·9 µm in diameter. A large posterior vacuole appeared as a pale area, occupying about a third of the spore length. The SSU rRNA gene was sequenced and analysed using maximum parsimony, maximum likelihood and neighbour-joining methods. This study allowed us to conclude that this was a new species of the genus Microgemma, being the first description of this genus from among South America fauna.

A new microsporidian parasite, Heterosporis saurida n. sp. (Microsporidia) infecting the lizardfish, Saurida undosquamis from the Arabian Gulf, Saudi Arabia: ultrastructure and phylogeny.

A new microsporidian that infects the lizardfish Saurida undosquamis (Richardson, 1848) that are caught in the Arabian Gulf in Saudi Arabia is described here. This parasite invades the skeletal muscle of the abdominal cavity forming white, cyst-like structures containing numerous spores. The prevalence of the infection was 32·1% (135/420). The spores were oval to pyriform in shape and measured approximately 3·3 µm×2·0 µm. The developing spores were found within parasitophorous vacuoles. In mature spores, the polar filament was arranged into 5 coils in a row. Molecular analysis of the rRNA genes, including the ITS region, and phylogenetic analyses using maximum parsimony, maximum likelihood, and Bayesian inference were performed. The ultrastructural characteristics and phylogenetic analyses support the recognition of a new species, herein named Heterosporis saurida n. sp.

Eimeria biarmicus sp.n. (Apicomplexa: Eimeriidae) infecting falcons from the genus Falco 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.

Morphological and ultrastructural description of Pleistophora dammami sp. n. infecting the intestinal wall of Saurida undosquamis from the Arabian Gulf, Saudi Arabia.

Pleistophora dammami sp. n. is described from Saurida undosquamis from the Arabian Gulf in Saudi Arabia. Infection appeared as whitish cysts in the intestinal wall. Cysts ranged in size from 1 to 4 mm. The prevalence of the infection across both fish sexes was 17.5% (24/420). Two kinds of spores were recognized, microspores and macrospores, and each were ovoid in shape. The microspores measured ~2.5 × 2.0 µm in size, while the macrospores measured ~6.0 × 3.0 µm. Ultrastructurally, the parasite did not form xenoma but it formed cysts surrounded by thick cyst wall. All stages of development as meronts, sporonts, sporoblast and spores occurred in the cytoplasm of the host cells within sporophorous vesicles. The stages of development occurred asynchronously and thus all stages were randomly distributed within the cysts. Meronts were elliptical and multinucleated, with unpaired nuclei which constantly divided giving rise to new sporonts. During the transition to sporonts, the border of the meronts increased in thickness to form dense discontinuous cell coat. Later, the sporont divided into sporoblast cells which gradually differentiated the typical organelles of the spores. In mature spores, the polar filament was arranged in 20-24 coils in two rows either side of the posterior vacuole. All ultrastructural and morphological criteria indicate that the described species belongs to the genus Pleistophora.

Acroeimeria lineri (McAllister, Upton, Freed, 1988) Paperna and Landsberg, 1989 in Mediterranean Geckos (Hemidactylus turcicus): Oocyst Morphometrics, Endogenous Developmental Stages, and Molecular Sequencing Support its Placement into Acroeimeria.

Between June 2016 and June 2019, we surveyed 62 Mediterranean geckos, Hemidactylus turcicus, from Abu Rawash, Giza, Egypt, for the presence of endoparasites. In June 2016, we found 3 individuals to be infected with Eimeria lineri. We studied the morphology and inner structures of its sporulated oocysts, and the locations of its intestinal endogenous stages. We also extracted genomic DNA from these sporulated oocysts and successfully sequenced a 632-bp fragment of the 18S rRNA gene. Phylogenetic analyses using this partial sequence allowed us to support previous studies that assigned E. lineri to the genus Acroeimeria. Our consensus sequence was used to query similar 18S rDNA sequences from GenBank, and 14 sequences were selected. The phylogenetic analysis inferred by maximum likelihood and Bayesian inference methods gave similar results, as both separated the sequences into 2 clades: (1) a monophyletic group of Goussia species (from fish); and (2) a strongly supported clade that separated 4 Choleoeimeria species from a polyphyletic group of species that clustered A. lineri with 3 other Acroeimeria species and 3 Eimeria species from lizards, including Eimeria tiliquae from Tiliqua rugosa (Gray, 1825), Eimeria tokayae from Gecko gecko (L., 1758), and Eimeria eutropidis from Eutropis macularia (Blyth, 1853). Our study supports the placement of E. lineri into the Acroeimeria and contributes additional life history information toward understanding the evolutionary origin of the Eimeria-like species that have sporocysts without Stieda bodies in their oocysts and that infect saurian reptiles. We also support the concept that several traits (morphological, endogenous, and gene sequences) are both necessary and important for authors to include when making generic reassignments within the eimeriid coccidia.

Genebank accession numbers of sequences of Culicoides species vectors of bluetongue virus in Germany.

The paper offers the genebank accession numbers of Culicoides obsoletus, Culicoides scoticus and Culicoides pulicaris sequences (ITS 1, ITS 2, 18S rRNA) that had been shown to be vectors of the bluetongue virus serotype 8, which was introduced in 2006 into Germany and spread until 2009 all over Central Europe, including parts of England. The numbers are FN 263292 until FN 263323.

Infection with Plasmodium chabaudi diminishes plasma immune complexes and ameliorates the histopathological alterations in different organs of female BWF1 lupus mice.

OBJECTIVE: To determine the effect of Plasmodium chabaudi infection on the plasma level of circulating immune complexes (CICs), haemoglobin (Hb) content, urine profile, and histological features of female BWF1 mice, the murine model of systemic lupus erythematosus (SLE). MATERIALS AND METHODS: A total of 30 female BWF1 lupus mice were randomly divided into three groups as follows: group (I) control group (P. chabaudi uninfected); group (II) lupus mice infected with live P. chabaudi; group (III) lupus mice infected with irradiated P. chabaudi. Urine samples were daily collected from the second week-post infection. Mice from the three groups were killed at day 14 post-infection and heparinized blood was collected for further haemoglobin contents and plasma analysis. Paraffin-embedded kidney, liver, lung, heart, brain, ovary and skin tissues were stained with Hematoxylin and Eosin (H&E) and examined under light microscope. RESULTS: Our results reveal that infection of lupus mice with live P. chabaudi was associated with an increase in urinary Hb and a decrease in plasma Hb and CIC levels. Interestingly, infection of lupus mice with live P. chabaudi ameliorates the histopathological alterations mediated by lupus disease in kidney tissues. Although no parasite sequestration was observed in any of the investigated organs, P. chabaudi pigment deposition was observed in the liver of both live and irradiated P. chabaudi infected groups. CONCLUSIONS: This study in lupus prone BWF1 mice indicated that gamma-irradiated P. chabaudi infection has the desired lupus ameliorating effect without negative effects of malaria which assist the understanding of different responses to plasmodium sp. infection in human lupus patients.

Development of a novel in vitro method for drug development for fish; application to test efficacy of antimicrosporidian compounds.

Few drugs are approved for treating diseases caused by parasites in minor species such as fish. This is due, in part, to the expense of drug development and to the comparatively small market. In vivo effectiveness trials for antiparasitic drugs are costly, time consuming and require ethics approval, therefore an in vitro screening approach is a cost-effective alternative to finding promising drug candidates. We developed an in vitro testing system to test antimicrosporidial compounds against a microsporidian pathogen Heterosporis saurida. Five antiparasitic compounds, albendazole, fumagillin, TNP-70, nitazoxanide and lufenuron, were assayed for antimicrosporidial activity. All compounds reduced the number of H saurida spores in infected cells when applied at a concentration that did not appear to be toxic to the host cells. Albendazole inhibited replication of H saurida by >60 per cent, fumagillin and its analogue TNP-470 inhibited H saurida >80 per cent, nitazoxanide and lufenuron inhibited growth >70 per cent. The data suggest that both fumagillin and its analogous TNP-70 hold the best promise as therapeutic agents against H saurida. The ability to use fish cell cultures to assess drugs against H saurida demonstrates an approach that may be helpful to evaluate other drugs on different microsporidia and host cells.

Morphological and molecular characterization of a new myxozoan species (Myxosporea) infecting the gall bladder of Raja clavata (Chondrichthyes), from the Portuguese Atlantic Coast.

Microscopic and molecular procedures are used to describe a new myxosporean species, Chloromyxum clavatum n. sp., infecting the cartilaginous fish Raja clavata Linnaeus, 1758 (Chondrichthyes: Rajidae), collected from the northwest Atlantic coast of Portugal. Young plasmodia and mature spores were found floating free in the gall bladder of R. clavata . Spores were spherical to subspherical with a pointed anterior end, measuring14.4 ± 0.5 µm (n = 25) in length, 11.9 ± 0.5 µm (n = 25) in width, and 9.4 ± 0.5 µm (n = 15) in thickness. The spore's wall was composed of 2 equally sized valves, each displaying 6-8 elevated surface ridges and a bundle of several tapering caudal filaments attached to the basal portion. Spores contained 4 pyriform equally sized polar capsules (5.5 ± 0.4 µm × 2.9 ± 0.5 µm) (n = 25), each possessing an obliquely arranged isofilar polar filament coiled in 7-8 coils. Morphological data, host specificity, tissue tropism, and molecular analysis of the SSU rDNA gene identify this parasite as a new species of Chloromyxum. Neighbor-joining and maximum likelihood further reveal the parasite clustering with other species of Chloromyxum infecting the gall bladder of marine cartilaginous fish to form a clade positioned at the base of the freshwater clade, therefore constituting an exception to the major division of the class Myxosporea into the freshwater and marine clades, while supporting the existence of a correlation between tissue tropism and myxosporean phylogeny.

Hepatic oxidative stress in Mongolian gerbils experimentally infected with Babesia divergens.

The present study aimed to investigate oxidative stress, DNA damage, and histopathological alterations in hepatic tissues of Mongolian gerbils experimentally infected with Babesia divergens. It was found that parasitaemia reached approximately 77% at day 5 post-infection. The liver became dark-brown and extremely friable, and hepatic sinusoids were dilated and contained macrophages and parasite-containing erythrocytes. Infection also induced inflammation and injury of the liver. This was illustrated by (1) an increase in inflammatory cellular infiltrations, (2) a decrease in total antioxidant capacity, as indicated by lowered glutathione and catalase levels, (3) increased production of nitric oxide-derived products (nitrite/nitrate) and malondialdehyde, and (4) increased lactic acid dehydrogenase activity and protein carbonyl content in the liver. Infection also interfered with the normal cell cycle of the hepatic tissue, as indicated by a significant increase in the percentage of liver cells at G0/G1 from approximately 86.2% to 97.5% and in S phases from 0.28% to 2.2%. Collectively, the present data suggest that B. divergens infection could induce cell-cycle alteration following oxidative stress and DNA damage in hepatic tissue. Further work is required to investigate the mechanism by which this hepatic tissue damage takes place.

A new species of Isospora (Apicomplexa: Eimeriidae) infecting the Baiuch rock gecko, Bunopus tuberculatus, in Saudi Arabia.

Isospora bors n. sp. (Apicomplexa: Eimeriidae) is described from 6 of 30 (20%) Baiuch rock gecko Bunopus tuberculatus Blanford in Saudi Arabia. Sporulated oocysts are subspheroidal to spheroidal, 18 × 16 (17-20 × 15-17) µm, with a bilayered, smooth, yellow-orange wall, without striae or micropyle. Polar body and oocyst residuum, both absent. Sporocysts are ovoidal, 10 × 7 (9-11 × 6-9) µm, with a Stieda body and sporocyst residuum. Endogenous stages developed in the cytoplasm of epithelial cells of the small intestine and above the host cell nucleus.