Synthesis of Carvacrol-Loaded Invasomes Nanoparticles Improved Acaricide Efficacy, Cuticle Invasion and Inhibition of Acetylcholinestrase against Hard Ticks.

Carvacrol is a monoterpenoid phenol found in many essential oils that has antibacterial, antifungal and antiparasitic activities. Drug loaded-invasome systems are used to deliver drugs utilizing nanoparticles to improve bioavailability, efficacy, and drug release duration. As a result, the present study developed carvacrol-loaded invasomes and evaluated their acaricidal effect against Rhipicephalus annulatus (cattle tick) and Rhipicephalus sanguineus (dog tick). Carvacrol loaded-invasome (CLI) was prepared and characterized using UV/Vis spectrophotometer, zeta potential measurements, Scanning Transmission Electron Microscopy (STEM), Fourier Transform Infrared (FT-IR) Spectroscopy, and Differential Scanning Calorimetry Analysis. CLI (5%) induced significant mortality (100%) in R. annulatus adult ticks with LC50 of 2.60%, whereas the LC50 of pure carvacrol was 4.30%. Carvacrol and CLI were shown to have a significant larvicidal action on both tick species, with LC50s of 0.24 and 0.21% against R. annulatus and 0.27 and 0.23% against R. sanguineus, respectively. Carvacrol and CLI (5%) induced significant repellent activities for 24 h against R. annulatus and R. sanguineus, as evidenced by the rod method and the petri-dish selective area choice method, respectively. High-performance liquid chromatography (HPLC) demonstrated that the CLI form had 3.86 times the permeability of pure carvacrol. Moreover, carvacrol and CLI inhibited acetylcholinesterase activity and decreased glutathione and malonedealdehyde levels in the treated ticks. In conclusion, invasomes significantly improved adulticidal and repellency activities of carvacrol against both tick species.

Anticoccidial and Antioxidant Activities of Moringa oleifera Leaf Extract on Murine Intestinal Eimeriosis.

BACKGROUND: Coccidiosis is an intestinal disease caused by protozoan parasites of the genus Eimeria and responsible for considerable economic loss in the livestock and poultry industries. Resistance to the current anticoccidial drugs is now a major challenge to efforts to control the disease, and this has stimulated the search for new compounds as alternative treatments. In this context, plant extracts have emerged as an alternative and complementary approach to control coccidiosis. In the present study, an ethanol extract of Moringa oleifera leaves was screened for its anticoccidial activity against Eimeria papillata infection in mice. METHODS: To this end, albino mice were allocated into three groups: the first group was the non-infected control; the second and third groups were infected with 103 E. papillata oocysts. Of these, the second group was kept as an infected control; while the third group was gavaged with 100 µl of moringa leaf extract (MLE) at a dose of 400 mg MLE/kg, once daily, for five days. RESULTS: MLE significantly suppressed oocyst excretion in faeces, and histological study of the jejunum showed a significant decrease in the number of parasitic stages, with significant improvement in the numbers of goblet cells. Furthermore, the expression of MUC2 gene was upregulated in the treated mice compared with infected, which further supports the anticoccidial potential of MLE. Moreover, our study evidenced that MLE reduced oxidative damage by decreasing TBARS and iNOS expression, and increasing the GSH and GPX levels. Also, treatment with MLE promoted the expression of Bcl-2 and ultimately, inhibited the apoptosis of host cells in the treated mice. CONCLUSION: Our data indicate that MLE has anticoccidial, anti-oxidant and anti-apoptotic activities in mice infected with Eimeria papillata.

Phyllodistomum vaili(Plagiorchiida: Gorgoderidae) infecting Parupeneus rubescens (Perciformes: Mullidae): morphology and phylogeny.

Knowledge of the Arabian Gulf fish's parasite fauna is very poor. Until recently, only scattered reports from different locations are known for ecto- and endoparasites. Therefore, the present study aimed to investigate the digenean species that infects one of the most economically fish species in the Arabian Gulf, the rosy goatfish Parupeneus rubescens . One plagiorchiid species has been described, belonging to the Gorgoderidae family, and has been named as Phyllodistomum vaili Ho, Bray, Cutmore, Ward & Cribb, 2014 based on its morphological and morphometric characteristics. In order to accurately classify and characterize this plagiorchiid species, molecular analysis was carried out using both nuclear 18S and 28S rRNA gene regions and revealed that the present plagiorchiid species was associated with other species belonging to the Gorgoderidae family and deeply embedded in the Phyllodistomum genus, closely related to the previously described P. vaili (gb| KF013187.1, KF013173.1). The present study therefore revealed that the species Phyllodistomum is the first account as endoparasites from the rosy goatfish inhabiting the Arabian Gulf.

Enhancement of Anti-allergic Effect of Diethylcarbamazine Citrate in Asthmatic Mouse Model: Testing of Anti-drug Antibodies and Quercetin.

Diethylcarbamazine citrate (DEC) is known as an effective treatment for bronchial asthma because of its ability to reduce eosinophil trafficking to the lung tissue. The current study aimed to potentiate the anti-allergic effect of the drug by passive immunization of the asthmatic model with anti-DEC antibody or prior treatment with quercetin (Qur). Eight mice groups were categorized into control, the model of lung asthma, treated with DEC, passively immunized with anti(α)-bovine serum albumin Ab, anti-DEC Ab, prior exposure to 10, 20, or 40 mg Qur/Kg. b.wt. Both eosinophil peroxidase (EPO) and eotaxin2 in the lung tissues were performed. Serum levels of cytokines, bronchoalveolar lavage fluid  (BALF) IgE, rabbit anti-bovine serum albumin (anti-BSA), and DEC IgG in lung tissue homogenates were assayed by ELISA. Regarding the effect of anti-DEC Ab and Qur on DEC-induced recovery of histopathological alterations showed that the Ova group had peri-bronchial hyperplasia, mononuclear leukocyte infiltration, thickening in the wall of alveoli, and congested blood vessels. However, the reduction of inflammatory cells and thickened alveolar walls was dependent on the Qur dose. Qur40 enhanced the anti-allergic effect of DEC. Moreover, the present data revealed high levels of Th2 cytokines (IL-4 and IL-5) and IgE in the Ova group. An increased leukocyte infiltration/thickening of the alveolar wall and lung tissue EPO/eotaxin2 were also observed. Qur-40 could show an enhancement effect on DEC for the reduction of IL-4, IL-5, IgE, EPO, and eotaxin 2. Consequently, the IFN-γ/IL-4 ratio was increased. Qur at 40 mg/Kg could be recommended to enhance the DEC effect suggesting a novel approach for treatment.

Phyllodistomum vaili(Plagiorchiida: Gorgoderidae) infecting Parupeneus rubescens (Perciformes: Mullidae): morphology and phylogeny / Phyllodistomum vaili (Plagiorchiida: Gorgoderidae) infectando Parupeneus rubescens (Perciformes: Mullidae): morfologia e filogenia

Abstract Knowledge of the Arabian Gulf fish's parasite fauna is very poor. Until recently, only scattered reports from different locations are known for ecto- and endoparasites. Therefore, the present study aimed to investigate the digenean species that infects one of the most economically fish species in the Arabian Gulf, the rosy goatfish Parupeneus rubescens . One plagiorchiid species has been described, belonging to the Gorgoderidae family, and has been named as Phyllodistomum vaili Ho, Bray, Cutmore, Ward & Cribb, 2014 based on its morphological and morphometric characteristics. In order to accurately classify and characterize this plagiorchiid species, molecular analysis was carried out using both nuclear 18S and 28S rRNA gene regions and revealed that the present plagiorchiid species was associated with other species belonging to the Gorgoderidae family and deeply embedded in the Phyllodistomum genus, closely related to the previously described P. vaili (gb- KF013187.1, KF013173.1). The present study therefore revealed that the species Phyllodistomum is the first account as endoparasites from the rosy goatfish inhabiting the Arabian Gulf.

Resumo O conhecimento da fauna de parasitas dos peixes do Golfo Árabe é escasso. Atualmente, apenas relatórios dispersos de diferentes locais são conhecidos para ecto e endoparasitas. Portanto, o presente estudo teve como objetivo investigar as especies digenéticas que infectam uma das espécies economicamente mais importantes do Golfo Arábico, o peixe-cabra rosado Parupeneus rubescens . Uma espécie de plagiorquídeo foi descrita, pertencente à família Gorgoderidae e foi denominada Phyllodistomum vaili Ho, Bray, Cutmore, Ward & Cribb, 2014, com base em suas propriedades morfológicas e morfométricas. A fim de classificar e caracterizar com precisão essa espécie de plagiorquídeo, a análise molecular foi realizada usando as regiões nucleares do gene 18S e 28S rRNA, revelando que a atual espécie de plagiorchídeo estava associada a outras espécies pertencentes à família Gorgoderidae e, profundamente incorporada ao gênero Phyllodistomum , intimamente relacionado ao P. vaili descrito anteriormente (gb - KF013187.1, KF013173.1). O presente estudo revelou, portanto, que a espécie Phyllodistomum vailli é o primeiro relato como endoparasita do peixe-cabra rosado que habita o Golfo Arábico.

Heat shock protein 70 of filarial parasite Setaria equina: Cloning, expression, and analysis of binding with diethylcarbamazine citrate.

Setaria equina heat shock protein (SeqHSP) 70 gene was characterized, cloned and expressed to recombinant protein (rSeqHSP70). The protein was tested for binding with an anti-filarial drug "diethylcarbamazine citrate (DEC)" by equilibrium dialysis method. Molecular docking was also used to determine the binding sites and residues of binding with DEC. The mice were immunized with the protein alone or bound to DEC. Serum IFN-γ levels in the immunized group with protein-drug complex were significantly higher (P < 0.05) than the protein-immunized group. Mouse anti-SeqHSP70 polyclonal IgG recognized 2 bands at 70 and 75 kDa in S. equina adult worm and human cancer cell lines (HepG2 and MCF-7) extracts. The proliferation assay for mice splenocytes revealed a potentiation and down-regulating effects in non-immunized and immunized groups, respectively with the drug-protein complex. The proliferation and IFN-γ assays for purified human NK cells indicated a potentiating effect of the drug-protein complex (DEC concentration is 50 µM) comparable to the protein. DEC at lower concentration (25 mM) could also show a significant increase (P < 0.05) in IFN-γ. From the results, DEC was postulated to induce conformational changes in the protein exposing more epitopes for NK cell binding and activation.

Antitumoral effect of Ocoxin in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is becoming one of the most prevalent types of cancer worldwide. The most efficient types of treatment at present include surgical resection and liver transplantation, but these treatments may only be used in a small percentage of patients. In order to identify novel therapeutic strategies for this disease, the present study explored the potential antitumoral effect of Ocoxin® oral solution (OOS) in HCC. OOS inhibited the proliferation of HCC cell lines in a time- and dose-dependent manner, being more efficient when used in combination with sorafenib, a standard of care treatment for patients diagnosed with advanced-stage disease. Mechanistic studies indicated that the effect of OOS was due to the induction of cell cycle arrest rather than the stimulation of apoptotic cell death. The cell cycle was slowed down in all phases in the HCC cell lines treated with OOS. Finally, when tested in animal models of HCC, OOS reduced tumor progression through the induction of necrosis in xenograft tumor models. Considering the poor prognosis and high resistance to antitumor treatments of HCC, the antiproliferative action of OOS, particularly in combination with sorafenib, provides the opportunity to investigate the effect of combined therapy in a clinical setting.

Morphological re-description and phylogenetic relationship of five myxosporean species of the family Myxobolidae infecting Nile tilapia.

Freshwater fish have a major economic and nutritional importance worldwide. Myxosporeans are highly dangerous parasites that infect different fish species, causing severe damage to a large number of economically important species, especially in aquaculture. We conducted a survey of myxosporean parasites infecting Nile tilapia Oreochromis niloticus (Perciformes: Cichlidae) collected from different localities along the River Nile passing through Giza province, Egypt. Out of 100 fish specimens collected, 45 were found to be naturally infected with these parasites in the region of the trunk kidney. Light microscopic examination revealed the presence of 5 distinct myxosporean species belonging to 2 different genera, viz. Myxobolus and Triangula, belonging to the family Myxobolidae; all 5 species have been previously described. Morphological characteristics, host specificity and geographical distribution, tissue tropism, and molecular analysis of the partial sequence of small subunit ribosomal DNA gene revealed that the recovered myxosporean species described herein were genetically distinct from other myxozoan species but had 95% sequence similarity to M. cerebralis. Also, phylogenetic analysis placed the present myxosporean species in the freshwater Myxobolus clade, which is a sister group of freshwater Myxobolus/Henneguya species.

Immunomodulatory effect of diethylcarbamazine citrate plus filarial excretory-secretory product on rat hepatocarcinogenesis.

Diethylcarbamazine citrate (DEC) had a significance in anti-filarial chemotherapy, while excretory-secretory product (ES) is released from adult filarial females. The target of the current study was to examine the immunomodulatory effect of DEC, Setaria equina ES or a combination of them on rat hepatocellular carcinoma (HCC) induced by diethylnitrosamine (DEN). In vitro effect of combined DEC and ES or ES alone on lipopolysaccharide (LPS)-stimulated rat peripheral blood mononuclear cells (PBMCs) was tested through IFN-γ assay in culture supernatants. In addition, single or repeated doses of DEC, ES or DEC+ES have been applied in white albino rats to test the effect on HCC. Levels of IFN-γ and anti-ES IgG antibodies in rat serum were assayed using ELISA. Hemolytic complement activity (CH50) was determined in serum while the concentration of nitric oxide (NO) was assayed in liver tissue. The infiltration of NK cells as well as the expression of MHC Iproliferating cell nuclear antigen (PCNA), inducible NO synthase (iNOS), Bcl2 and p53 were determined using immunohistochemistry. There was a dose-dependent increase in IFN-γ after in vitro exposure to DEC+ES. Repeated ES doses increased NO concentration (p<0.05) and expression of iNOS but reduced CH50 (p<0.001), while repeated DEC+ES doses could increase anti-ES IgG (p<0.01), IFN-γ level (p<0.05) and NK cell infiltration. The same treatments could also reduce the expression of MHC I expression, PCNA, Bcl2 and p53. This study has shown immunomodulatory and protective effects of DEC+ES repeated doses on rat HCC.

Effect of diethylcarbamazine citrate and Setaria equina excretory-secretory material on rat hepatocellular carcinoma.

Diethylcarbamazine citrate (DEC) has been known for its efficacy to eradicate bancroftian filariasis in Egypt and other countries in the world. One of the known effects was to decrease the level of circulating filarial antigen in the patient's serum. The target of this study was to examine the effect of DEC, excretory-secretory (ES) material from the filarial parasite Setaria equina or a combination of both on the status of oxidative stress and pathogenesis of rat hepatocellular carcinoma (HCC) induced by diethylnitrosamine and 2-acetylaminofluorene. This could be tested in vitro using nitroblue tetrazolium reduction test for measuring the level of superoxide anion (O2(•-)) released from rat peritoneal macrophages. For in vivo test, a single dose before induction of carcinogenesis or continually repeated doses with DEC, ES or DEC + ES was used. Exposure of macrophages to ES could lead to a significant decrease (p < 0.01) in O2(•-) release, while DEC (200 µM) could modulate such effect with significant increase (p < 0.05). Pathogenesis of liver cancer and treatment were evaluated using histological investigation, level of antioxidant and liver function enzymes. Repeated ES doses could increase the activity of antioxidant enzymes, especially the catalase enzyme and show a protective effect on liver architecture. DEC could modulate the later effects when combined with ES. No significant effect on the liver function enzymes after treatment was observed. Nuclear factor κB was found to be localized only in the cytoplasm after single and repeated treatments with ES. This study could indicate the effect of S. equina ES as antioxidant against rat HCC, while DEC could modulate such effect when combined with it.