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.

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.

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.

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.

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.

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.

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.

Testosterone response of hepatic gene expression in female mice having acquired testosterone-unresponsive immunity to Plasmodium chabaudi malaria.

Blood-stage malaria of Plasmodium chabaudi is characterized by its responsiveness to testosterone (T): T suppresses development of protective immunity, whereas once acquired immunity is T-unresponsive. Here, we have analyzed the liver, a T target and lymphoid organ with anti-malaria activity, for its T-responsiveness of gene expression in immune mice. Using Affymetrix microarray technology, in combination with quantitative RT-PCR, we have identified (i) T-unresponsive expression of newly acquired mRNAs encoding diverse sequences of IgG- and IgM-antibodies, (ii) 24 genes whose expression has become T-unresponsive including those encoding the T(H)2 response promoting EHMT2 and the erythrocyte membrane protein band 7.2 STOM, (iii) T-unresponsive expression of mRNAs for the cytokines IL-1ß, IL-6, TNFα, and IFNγ, as well as iNOS, which are even not inducible by infection, and (iv) 35 genes retaining their T-responsiveness, which include those encoding the infection-inducible acute phase proteins SAA1, SAA2, and ORM2 as well as those of liver metabolism which encode the T-downregulated female-prevalent enzymes CYP2B9, CYP2B13, CYP3A41, CYP7A1, and SULT2A2 and the T-upregulated male-prevalent enzymes CYP2D9, CYP7B1, UGT2B1, HSD3B2, HSD3B5, respectively. The mRNA of the latter T-metabolizing enzyme is even 5-fold increased by T, suggesting a decrease in the effective T concentrations in the liver of immune mice. Collectively, our data suggest that the liver, which has acquired a selective T-unresponsiveness of gene expression, contributes to the acquired T-unresponsive, antibody-mediated protective immunity to blood-stage malaria of P. chabaudi.

Anticoccidial and antiinflammatory activity of garlic in murine Eimeria papillata infections.

Coccidiosis with the protozoan parasite Eimeria as the infectious agent causes enormous economic losses, particularly in poultry farms. Here, we investigated the effects of garlic on the outcome of coccidiosis caused by Eimeria papillata in male Balb/c mice. The data showed that mice infected with E. papillata revealed an output of 3260 ± 680 oocysts per gram faeces on day 4 p.i.. This output is significantly decreased to 1820 ± 415 oocysts in garlic-treated mice. Infection also induced inflammation and injury of the liver. This was evidenced (i) as increases in inflammatory cellular infiltrations, dilated sinusoids, and vacuolated hepatocytes, (ii) as increased mRNA levels of inducible nitric oxide synthase (iNOS) and of the cytokines interferon gamma (IFN-γ), and interleukin-6 (IL-6), (iii) as increased plasma levels of alanine and aspartate aminotransferases, alkaline phosphatase, γ-glutamyl transferase and total bilirubin, (iv) as increased production of nitric oxide derived products (nitrite/nitrate) and malondialdehyde, and (v) as lowered glutathione levels and decreased activities of catalase and superoxide dismutase, respectively. All these infection-induced parameters were significantly less altered during garlic treatment. In particular, garlic counteracted the E. papillata-induced loss of glutathione and the activities of catalase and superoxide dismutase. Our data indicated that garlic treatment significantly attenuated inflammation and injury of the liver induced by E. papillata infections.

Hepatic tissue damage induced in Meriones ungliculatus due to infection with Babesia divergens-infected erythrocytes.

Babesia divergens is an intraerythrocytic parasite which is capable of infecting a wide range of vertebrates causing huge economic losses. Histopathological, hematological and biochemical changes during B. divergens infection in female Meriones ungliculatus were reported. Animals were challenged with 5 × 10(6) B. divergens-infected erythrocytes. Parasitemia were maximum at day 5 postinfection where all gerbils died. Infection of gerbils with Babesia induced a significant decrease in erythrocytic count as well as the hemoglobin concentration and hematocrit percentage but leucocytes were increased significantly when compared to uninfected gerbils. Liver enzymes aspartate aminotransferase (AST) and aniline aminotransferase (ALT) were significantly increased while albumin and total bilirubin were significantly decreased at day 5 postinfection with B. divergens-infected erythrocytes. Histopathological scores of inflammation after infection of gerbils were done using Ischak's activity index and indicated that the liver was severely affected. In conclusion, the study indicated that the course of infection by B. divergens-induced alternations in hematology, biochemistry and histopathology of the hepatic tissue.

Fibroblasts in the human vocal fold mucosa: an ultrastructural study of different age groups.

INTRODUCTION: An investigation was carried out to determine the morphological characteristics of fibroblasts in two portions of the vocal fold (VF) mucosa, the macula flava (MF) and Reinke's space (RS), of three different age groups: newborns, adults and geriatrics. METHODS: Normal human VF obtained from autopsy cases were included in this study: four from mature newborns; four from middle-aged adults; and four from geriatric cases. Fibroblasts in RS and MF were investigated by transmission electron microscopy. RESULTS: The fibroblasts of the MF in both adults and newborns tended to be stellate in shape, with a small nucleus/cytoplasm (N/C) ratio and a well-developed rough endoplasmic reticulum (rER) and Golgi apparatus (GA). Most of the fibroblasts present in RS were oval in newborns and spindle-shaped in adults, with a large N/C ratio and less developed rER and GA. The majority of fibroblasts of the geriatric MF were stellate in shape; while in geriatric RS, the majority of fibroblasts were spindle-shaped with an N/C ratio of 0.5 to 2.0 as in the case of adults. However, the development of rER and GA was less marked in geriatrics than in adults. CONCLUSION: Histological changes of fibroblasts in the VF mucosa are one of the important causes of the change in voice quality with ageing. Furthermore, geriatric changes in the vocal ligament can be attributed to the activities and the presence of ageing processes in fibroblasts of geriatric VF mucosa.

Structural alterations of the glomerular wall and vessels in early stages of diabetes mellitus: (light and transmission electron microscopic study).

OBJECTIVE: The capillary changes at the initial stage of diabetes may show an angioarchitecture clearly different from those of later stages and/or very severe glomerular change. However, the onset of alterations in the early phases is unclear. This study attempts to determine the structural alterations of the glomerular wall and vessels in the early stage of diabetes. MATERIAL AND METHODS: Twenty-five adult rats were used in this study. They were divided into two groups: the first group of five was used as a control. The second group of 20 (the experimental group) was injected intraperitoneally by a single dose of streptozotocin to induce hyperglycemia. Rats were sacrificed after ten days, two months, and four months. Five rats at two months of age with hyperglycemia were treated with insulin for eight weeks. Renal tissues were prepared by routine technique for light and transmission electron microscopic evaluation. RESULTS: By light microscopy after ten days of induced hyperglycemia, there were no structural modifications detected either in renal glomerular fine vessels or in the glomerular basement membrane of the glomerular capillaries. After two months, there was a moderate glomerular enlargement and dilatation of glomerular capillaries, afferent, and efferent arterioles. After four months, glomerular basement membrane thickening was the only structural alteration observed. Recovery of the glomerular alterations was observed after two months of treatment with insulin. CONCLUSION: In early stages of diabetes mellitus in rats, there was an increase in the diameter of glomerular vessels. In later stages of the disease, the reverse was seen, but insulin treatment had a positive role in reversing these changes in the study subjects.

Piroxicam-Induced Hepatic and Renal Histopathological Changes in Mice

Piroxicam is a non-steroidal anti-inflammatory drug widely used in rheumatic diseases. The aim of this study was to investigate Piroxicam-induced histopathological changes in livers and kidneys of male albino mice.Methods: Animals were classified into a control group and 4 treated groups. Piroxicam was injected intraperitoneally using 0.3 mg/kg every day for four weeks. Each week a group of mice was sacrificed. Liver and kidneys were obtained for histological and histochemical examination. Animals were classified into a control group and 4 treated groups. Piroxicam was injected intraperitoneally using 0.3 mg/kg every day for four weeks. Each week a group of mice was sacrificed. Liver and kidneys were obtained for histological and histochemical examination.Results: Liver sections appeared with inflammatory cellular infiltration; vacuolated hepatocytes; dilated sinusoids; and increased number of Kupffer cells. Kidney sections appeared with some cellular inflammations. The glomeruli were shrunk resulting in widening of the urinary space. Oedema and vacuolations were noticed in the tubular cells. There was a positive correlation between these pathological changes and the increased treatment periods. Histochemical staining revealed that glycogen and protein contents had decreased in the hepatocytes. This depletion worsened gradually in liver cells after two; three; and four weeks. Similar depletion of the glycogen content was observed in kidney tissue. However; protein content appeared to be slightly decreased in the kidney tubules and glomeruli. Incensement of coarse chromatin in the nuclei of hepatocytes; Kupffer cells and most inflammatory cells were detected by Fuelgen method. Kidney tissues appeared with a severe decrease in coarse chromatin in the nuclei.Liver sections appeared with inflammatory cellular infiltration; vacuolated hepatocytes; dilated sinusoids; and increased number of Kupffer cells. Kidney sections appeared with some cellular inflammations. The glomeruli were shrunk resulting in widening of the urinary space. Oedema and vacuolations were noticed in the tubular cells. There was a positive correlation between these pathological changes and the increased treatment periods. Histochemical staining revealed that glycogen and protein contents had decreased in the hepatocytes. This depletion worsened gradually in liver cells after two; three; and four weeks. Similar depletion of the glycogen content was observed in kidney tissue. However; protein content appeared to be slightly decreased in the kidney tubules and glomeruli. Incensement of coarse chromatin in the nuclei of hepatocytes; Kupffer cells and most inflammatory cells were detected by Fuelgen method. Kidney tissues appeared with a severe decrease in coarse chromatin in the nuclei.Conclusion: Piroxicam has a time-dependent toxic effect on both liver and kidney tissues

Structural Alterations of the Glomerular Wall And Vessels in Early Stages of Diabetes Mellitus: Light and Transmission Electron Microscopic Study

Objective: The capillary changes at the initial stage of diabetes may show an angioarchitecture clearly different from those of later stages and;/or very severe glomerular change. However; the onset of alterations in the early phases is unclear. This study attempts to determine the functional and structural alterations of the glomerular wall and vesicles in the early stage of diabetes. Material and Methods: Twenty-five adult rats were used in this study. They were divided into two groups: the first group of five was used as a control .The second group of 20 (the experimental group) was injected intraperitoneally by a single dose of streptozotocin to induce hyperglycemia. Rats were sacrificed after ten days; two months; and four months.Five rats at two months of age with hyperglycemia were treated with insulin for eight weeks. Renal tissues were prepared by routine technique for light and transmission electron microscopic evaluation

Deletion of LTbetaR augments male susceptibility to Plasmodium chabaudi.

SUMMARY Disruption of the lymphotoxin beta receptor (LTbetaR) gene has been shown to result in enhanced resistance of female mice to blood-stage Plasmodium chabaudi malaria. Here, we investigate the effect of LTbetaR deletion on host defence of males. In contrast to females, male LTbetaR(-/-) mice do not exhibit any increase in resistance. Conversely, they are even more susceptible than wild-type C57BL/6 mice, which becomes evident after lowering circulating levels of testosterone by castration, which makes C57BL/6 males resistant, whereas LTbetaR(-/-) remain susceptible. Gene-expression analysis using cDNA arrays revealed no differences in immunological responses in spleen of malaria-resistant female and malaria-susceptible castrated male LTbetaR(-/-) mice. In the liver, however, expression levels of plasminogen activator inhibitor PAI1, chemokine CXCL10, dual specificity phosphatase DUSP1, and hydroxysteroid-specific sulfotransferases Sult2a1/2 were decreased 6- to 85-fold in susceptible castrated male LTbetaR(-/-) mice in comparison to resistant female LTbetaR(-/-) mice at maximal parasitaemia, as evidenced by Northern blot analyses. The present data support our previous view that the liver is involved in the combat against malarial blood stages and that down-regulation of the genes DUSP1 and Sult2a1/2 signals dysregulation of protective liver responses, thus possibly contributing to male susceptibility of LTbetaR(-/-) mice.