Cytotoxic and Antioxidative Effects of Geranium Oil and Ascorbic Acid Coloaded in Niosomes against MCF-7 Breast Cancer Cells.

Geranium oil (GO) has antiproliferative, antiangiogenic, and anti-inflammatory properties. Ascorbic acid (AA) is reported to inhibit the formation of reactive oxygen species, sensitize cancer cells, and induce apoptosis. In this context, AA, GO, and AA-GO were loaded into niosomal nanovesicles to ameliorate the physicochemical properties of GO and improve its cytotoxic effects using the thin-film hydration technique. The prepared nanovesicles had a spherical shape with average diameters ranging from 200 to 300 nm and exhibited outstanding surface negative charges, high entrapment efficiencies, and a controlled sustained release over 72 h. Entrapping AA and GO in niosomes resulted in a lower IC50 value than free AA and GO when tested on MCF-7 breast cancer cells. In addition, flow cytometry analysis showed higher apoptotic cells in the late apoptotic stage upon treating the MCF-7 breast cancer cells with AA-GO niosomal vesicles compared to treatments with free AA, free GO, and AA or GO loaded into niosomal nanovesicles. Assessing the antioxidant effect of the free drugs and loaded niosomal nanovesicles showed enhanced antioxidant activity of AA-GO niosomal vesicles. These findings suggest the AA-GO niosomal vesicles as a potential treatment strategy against breast cancer, possibly through scavenging free radicals.

Ceratonia siliqua pods (Carob) methanol extract alleviates doxorubicin-induced nephrotoxicity via antioxidant, anti-inflammatory and anti-apoptotic pathways in rats.

Doxorubicin (DOX) is an anti-neoplastic therapy, but its use is limited by its deleterious toxic effects including nephrotoxicity and cardiotoxicity. This work aimed at assessing the potential protective effect of Ceratonia siliqua methanol extract (CME) on DOX-induced nephrotoxicity in 5 groups of Wistar rats. Nephrotoxicity was induced experimentally by intraperitoneal (IP) injection of DOX (15 mg/kg). DOX increased serum creatinine, urea, sodium, and potassium levels. It elevated MDA levels in the renal tissue but decreased the concentration of GSH and the activity of GST, CAT, and SOD. Meanwhile, it decreased the level of immunomodulatory anti-inflammatory mediators: IL-10 and TGF-ß, as well as the activity of MPO but increased the level of IL-6, TNF-α, and caspase-3 in the renal tissue. DOX has upregulated COX-2, caspase-9, and Bax gene expression and downregulated the Bcl-2 gene expression. Immunolabeling of renal tubular epithelium in DOX-intoxicated rats was moderate to strong against Bax, COX-2, and NF-kß and weak against Bcl-2. Treatment with CME significantly restored the levels of kidney function parameters and the levels of oxidative stress markers. It stimulated the production of IL-10 and TGF-ß and decreased the level of IL-6 and TNF-α. CME reverted the gene expression of COX-2, caspase-9, and Bax. Microscopically, CME alleviated the DOX-induced renal damage. Phytochemical analysis revealed the presence of 26 compounds in the CME. No signs of acute toxicity were recorded by CME up to 4000 mg/kg b. wt. orally into mice. Finally, CME could effectively alleviate the deleterious effects of DOX on the kidney. The safety of carob extract encourages its use in the preparation of valuable therapeutic agents.

New NSAID Conjugates as Potent and Selective COX-2 Inhibitors: Synthesis, Molecular Modeling and Biological Investigation.

New sets of ibuprofen and indomethacin conjugates comprising triazolyl heterocycle were synthesized via click chemistry, adopting an optimized protocol through the molecular hybridization approach affording the targeted agents in good yields. The new non-steroidal anti-inflammatory drug (NSAID) conjugates were designed and synthesized and could be considered as potential drug candidates for the treatment of pain and inflammation. The anti-inflammatory properties were investigated for all the synthesized conjugates. Among 14 synthesized conjugates, four (5a, 5b, 5d, and 5e) were found to have significant anti-inflammatory properties potency 117.6%, 116.5%, 93.8%, and 109.1% in comparison to reference drugs ibuprofen (97.2%) and indomethacin (100%) in the rat paw edema carrageenan test without any ulcerogenic liability. The suppression effect of cytokines IL-6, TNF-α, and iNOS in addition to NO in the LPS-induced RAW264.7 cells supports the promising anti-inflammatory properties observed in the ibuprofen conjugates. In addition, several conjugates showed promising peripheral and central analgesic activity. The selectivity index (SI) of compound 5a (23.096) indicates the significant efficacy and selectivity for COX-2 over COX-1. Molecular modeling (docking and QSAR) studies described the observed biological properties.

The effect of exposure to MoO3-NP and common bean fertilized by MoO3-NPs on biochemical, hematological, and histopathological parameters in rats.

Nanotechnologies has been used to introduce several beneficial tools in the agricultural field. Herein, the effect of molybdenum oxide nanoparticles (MoO3-NPs) was investigated by evaluating the hematological, biochemical, and histopathological parameters in rats orally exposed to MoO3-NPs or fed common beans (CB) fertilized by MoO3-NPs. In the first study, 18 rats were randomly divided into 3 groups: G1 (control group) was given water orally, while G2 and G3 were administered 10 and 40 ppm MoO3-NPs by oral gavage tube, respectively. There was a significant increase in the levels of alanine aminotransferase (ALT), albumin, and total protein; however, there was a a significant decrease in body weight change (BWC), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatinine, creatine kinase-MB (CK-MB), thyroid-stimulating hormone (TSH), free triiodothyronine (FT3), and testosterone levels in G3 compared to G1. In the second study, 24 rats were divided into 4 groups: the control (C) group was fed a balanced diet, and three groups were fed on a balanced diet plus 10% CB that was fertilized with 0, 10, and 40 ppm MoO3-NPs, resulting in nCB, CB10, and CB40 groups, respectively. This revealed a significant increase in BWC and total food intake (TFI) but a significant decrease in relative kidney weight in all the CB groups compared to the control group. In CB10 and CB40 groups ALT, LDH, TSH, FT3, and testosterone levels were significantly lower than the respective levels in the control group. We concluded that high doses of MoO3-NPs caused more side effects than low doses in both experiments.

Development of Isatin-Based Schiff Bases Targeting VEGFR-2 Inhibition: Synthesis, Characterization, Antiproliferative Properties, and QSAR Studies.

Three sets of isatin-based Schiff bases were synthesized utilizing the molecular hybridization approach. Some of the synthesized Schiff bases show significant to moderate antiproliferative properties against MCF7 (breast), HCT116 (colon), and PaCa2 (pancreatic) cancer cell lines with potency compared to reference drugs 5-fluorouracil (5-FU) and Sunitinib. Among all, compound 17 f (3-((1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-4-yl)imino)-1-((1-(2-methoxyphenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-methylindolin-2-one) exhibits promising antiproliferative properties against the MCF7 cancer cell line with 2.1-fold more potency than Sunitinib. However, among all the synthesized compounds, three (5-methylisatin derivatives) were the most effective against HCT116 in comparison to 5-FU. Compound 17 f exhibited the highest anti-angiogenic effect on the vasculature as it significantly reduced BV from 43 mm to 2 mm in comparison to 5.7 mm for Sunitinib and flow cytometry supports the arrest of the cell cycle at G1/S phases. In addition, compound 17 f also showed high VEGFR-2 inhibition properties against breast cancer cell lines. Robust 2D-QSAR studies supported the biological data.

PEGylated Chitosan Nanoparticles Encapsulating Ascorbic Acid and Oxaliplatin Exhibit Dramatic Apoptotic Effects against Breast Cancer Cells.

This study aims to design a pH-responsive dual-loaded nanosystem based on PEGylated chitosan nanoparticles loaded with ascorbic acid (AA) and oxaliplatin (OX) for the effective treatment of breast cancer. In this regard, non-PEGylated and PEGylated chitosan nanoparticles (CS NPs) loaded with either ascorbic acid (AA), oxaliplatin (OX), or dual-loaded with AA-OX were fabricated using the ionotropic gelation method. The hydrodynamic diameters of the fabricated AA/CS NPs, OX/CS NPs, and AA-OX/CS NPs were 157.20 ± 2.40, 188.10 ± 9.70, and 261.10 ± 9.19 nm, respectively. While the hydrodynamic diameters of the designed AA/PEG-CS NPs, OX/PEG-CS NPs, and AA-OX/PEG-CS NPs were 152.20 ± 2.40, 156.60 ± 4.82, and 176.00 ± 4.21 nm, respectively. The ζ-potential of the prepared nanoparticles demonstrated high positive surface charges of +22.02 ± 1.50, +22.58 ± 1.85 and +40.4 ± 2.71 mV for AA/CS NPs, OX/CS NPs, and AA-OX/CS NPs, respectively. The ζ-potential of the PEGylated CS NPs was reduced owing to the shielding of the positive charges by the PEG chains. Additionally, all the prepared nanoparticles exhibited high entrapment efficiencies (EE%) and spherical-shaped morphology. The chemical features of the prepared nanoparticles were investigated using Fourier transform infrared (FTIR) spectroscopy. Release studies showed the capability of the prepared non-PEGylated and PEGylated chitosan NPs to release their cargo in the acidic environment of cancer tissue (pH 5.5). Furthermore, the AA/CS NPs, AA/PEG-CS NPs, OX/CS NPs, OX/PEG-CS NPs, AA-OX/CS NPs and AA-OX/PEG-CS NPs exhibited remarkable cytotoxic activities against breast adenocarcinoma (MCF-7) cells with IC50 values of 44.87 ± 11.49, 23.3 ± 3.73, 23.88 ± 6.29, 17.98 ± 3.99, 18.69 ± 2.22, and 7.5 ± 0.69 µg/mL, respectively; as compared to free AA and OX (IC50 of 150.80 ± 26.50 and 147.70 ± 63.91 µg/mL, respectively). Additionally, treatment of MCF-7 cells with IC50 concentrations of AA, AA/CS NPs, AA/PEG-CS NPs, OX, OX/CS NPs, OX/PEG-CS NPs, AA-OX/CS NPs or AA-OX/PEG-CS NPs increased the percentages of early apoptotic cells to 5.28%, 9.53%, 11.20%, 5.27%, 13.80%, 8.43%, 2.32%, and 10.10%, respectively, and increased the percentages of late apoptotic cells to 0.98%, 0.37%, 2.41%, 2.06%, 0.97%, 9.66%, 56%, and 81.50%, respectively. These results clearly indicate that PEGylation enhances the apoptotic effect of AA and OX alone, in addition to potentiating the apoptotic effect of AA and OX when combined on MCF-7 cells. In conclusion, PEGylated chitosan nanoparticles encapsulating AA, OX, or AA and OX represent an effective formula for induction of apoptosis in MCF-7 cells.

Current perspective on veterinary drug and chemical residues in food of animal origin.

The marked increase in the demand for animal protein of high quality necessitates protecting animals from infectious diseases. This requires increasing the use of veterinary therapeutics. The overuse and misuse of veterinary products can cause a risk to human health either as short-term or long-term health problems. However, the biggest problem is the emergence of resistant strains of bacteria or parasites. This is in addition to economic losses due to the discarding of polluted milk or condemnation of affected carcasses. This paper discusses three key points: possible sources of drug and chemical residues, human health problems, and the possible method of control and prevention of veterinary drug residues in animal products.

Design and synthesis of ibuprofen-quinoline conjugates as potential anti-inflammatory and analgesic drug candidates.

A new set of ibuprofen-quinoline conjugates comprising quinolinyl heterocycle and ibuprofen moieties linked by an alkyl chain were synthesized in good yields utilizing an optimized reaction procedure in a molecular hybridization approach to overcome the drawbacks of the current non-steroidal anti-inflammatory drugs. The synthesized conjugates were screened for their anti-inflammatory, and ulcerogenic properties. Several conjugates were found to have significant anti-inflammatory properties in the carrageenan-induced rat paw edema test without showing any ulcerogenic liability. In addition, most conjugates showed promising peripheral analgesic activity in the acetic acid-induced writhing test as well as central analgesic properties in the in vivo hot plate test. The most promising conjugates were the unsubstituted and 6-substituted fluoro- and chloro-derivatives of 2-(trifluoromethyl)quinoline linked to ibuprofen by a propyl chain. Their anti-inflammatory activity was evaluated against LPS-stimulated inflammatory reactions in RAW264.7 mouse macrophages. In this regard, it was found that most of the conjugates were able to significantly reduce the release and production of nitric oxide in the LPS-stimulated macrophages. The secretion and expression of the pro-inflammatory cytokines IL-6, TNF-α, and inducible nitric oxide synthase (iNOS) were also significantly suppressed.

3-Alkenyl-2-oxindoles: Synthesis, antiproliferative and antiviral properties against SARS-CoV-2.

Sets of 3-alkenyl-2-oxindoles (6,10,13) were synthesized in a facile synthetic pathway through acid dehydration (EtOH/HCl) of the corresponding 3-hydroxy-2-oxoindolines (5,9,12). Single crystal (10a,c) and powder (12a,26f) X-ray studies supported the structures. Compounds 6c and 10b are the most effective agents synthesized (about 3.4, 3.3 folds, respectively) against PaCa2 (pancreatic) cancer cell line relative to the standard reference used (Sunitinib). Additionally, compound 10b reveals antiproliferative properties against MCF7 (breast) cancer cell with IC50 close to that of Sunitinib. CAM testing reveals that compounds 6 and 10 demonstrated qualitative and quantitative decreases in blood vessel count and diameter with efficacy comparable to that of Sunitinib, supporting their anti-angiogenic properties. Kinase inhibitory properties support their multi-targeted inhibitory activities against VEGFR-2 and c-kit in similar behavior to that of Sunitinib. Cell cycle analysis studies utilizing MCF7 exhibit that compound 6b arrests the cell cycle at G1/S phase while, 10b reveals accumulation of the tested cell at S phase. Compounds 6a and 10b reveal potent antiviral properties against SARS-CoV-2 with high selectivity index relative to the standards (hydroxychloroquine, chloroquine). Safe profile of the potent synthesized agents, against normal cells (VERO-E6, RPE1), support the possible development of better hits based on the attained observations.

A Naturally Derived Carrier for Photodynamic Treatment of Squamous Cell Carcinoma: In Vitro and In Vivo Models.

Photodynamic therapy (PDT) is a non-invasive treatment strategy that includes the combination of three components-a photosensitizer, a light source, and tissue oxygen. PDT can be used for the treatment of skin diseases such as squamous cell carcinoma. The photosensitizer used in this study is the naturally derived chlorophyll derivative chlorin e6 (Ce6), which was encapsulated in ultradeformable ethosomes. Singlet oxygen production by Ce6 upon laser light irradiation was not significantly affected by encapsulation into ethosomes. PDT of squamous cell carcinoma cells treated with Ce6 ethosomes triggered increased mitochondrial superoxide levels and increased caspase 3/7 activity, resulting in concentration- and light-dose-dependent cytotoxicity. Ce6 ethosomes showed good penetration into 3D squamous cell carcinoma spheroids, which upon laser light irradiation exhibited reduced size, proliferation, and viability. The PDT effect of Ce6 ethosomes was specific and showed higher cytotoxicity against squamous cell carcinoma spheroids compared to normal skin fibroblast spheroids. In addition, PDT treatment of squamous cell carcinoma xenografts grown on chorioallantoic membranes of chick eggs (CAM) exhibited reduced expression of Ki-67 proliferation marker and increased terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining, indicating reduced proliferation and activation of apoptosis, respectively. The results demonstrate that Ce6-loaded ethosomes represent a convenient formulation for photodynamic treatment of squamous cell carcinoma.

Evaluation of the curative effect of Balanites aegyptiaca fruits ethanolic extract on Haemonchosis experimentally induced in Egyptian Baladi goats: phytoanalytical, parasitological and hematological studies.

This study aimed to screen phytochemical components and antioxidant activity of Balanites aegyptiaca ethanolic extract (BAF-EE) as well as to evaluate its curative effect on experimentally induced haemonchosis in goats. Phytochemical constitutes of BAF-EE were screened and identified using Gas Chromatography-mass Spectrometry (GC-MS) analysis and antioxidant effect was determined. Infective third larval stage (L3) of Haemonchus contortus (H. contortus) were obtained by culturing feces of goat harboring monospecific infection of the parasite. Twelve male goats were randomly divided into four groups (n = 3) as: G1 (infected-untreated) which served as control positive, G2 (infected-BAF-EE treated), G3 (infected-albendazole treated) and G4 (uninfected-BAF-EE treated) that served as control negative. Experimental infection was conducted with a single oral dose of 10,000 L3 at 0-time, whereas treatment with BAF-EE and albendazole were given at a single oral dose of 9 g and 5 mg/kg BW, respectively in the 5th week post infection (PI). Egg count per gram of feces (EPG) was conducted once a week and blood samples were drawn on zero time, 3rd week PI and then biweekly for 9 weeks, for conduction of hemogram. At the end of the experiment, all animals were slaughtered and adult worms in their abomasa were counted. GC-MS analysis confirmed 28 compounds in the extract which revealed presence of saponins, flavonoids, terpenoids, phenolics and alkaloids, and exhibited in vitro antioxidant activity. Clinical signs observed on the infected animals were signs of anemia, which were gradually disappeared post treatment (PT). A maximum reduction in EPG (88.10%) and worm burden (94.66%) was recorded on 4th week PT due to efficacy of BAF-EE in contrast to 98.29% and 96.95% efficacy of albendazole. All infected groups showed a significant decrease in hemoglobin (Hb) and packed cell volume (PCV) and presence of microcytic hypochromic anemia compared with G4. Goats treated with B. aegyptiaca and albendazole, exhibited significant increase in Hb and PCV 2 weeks PT and anemia changed to be normocytic hypochromic or microcytic normochromic in G2 and G3, respectively. Total white blood cells (WBCs) were elevated significantly in all infected groups which attributed to increase in lymphocytes, monocytes and eosinophils on expense of neutrophils. After treatments, WBCs and previously mentioned cells tended to decease. This study demonstrated that BAF-EE has anthelmintic effect against H. contortus and can improve hemogram and health condition of infected goats.

Ethosomes and lipid-coated chitosan nanocarriers for skin delivery of a chlorophyll derivative: A potential treatment of squamous cell carcinoma by photodynamic therapy.

Photodynamic therapy (PDT) is a localized treatment strategy used for skin cancers such as squamous cell carcinoma (SCC), the second most common form of skin cancer. PDT combines a photosensitizer, laser source and tissue oxygen. In this study, the selected photosensitizer, ferrous chlorophyllin (Fe-CHL) was loaded in ethosomes and lipid coated chitosan (PC/CHI) nanocarriers to enhance skin delivery of Fe-CHL for potential PDT of squamous carcinoma. The nanocarrier formulations were characterized and studied for their skin retention and penetration depth of Fe-CHL across mouse skin ex vivo using high performance liquid chromatography and confocal microscopy. Confocal microscope images of mouse skin showed deeper penetration of ethosomes down to the dermis when compared to PC/CHI that was confined to the epidermis, although they showed no significant difference in skin retention. Immunohistochemistry (IHC) staining with ki67 and TUNEL show maintained skin structure and no cytotoxic effects of the nanocarrier gel formulations before laser exposure to mouse skin. The nanocarriers were also studied for their PDT effect against human SCC monolayer and three-dimensional (3-D) spheroids. When compared to ethosomes, PC/CHI showed higher cytotoxicity in MTT assay and live confocal microscopy showed cell disintegration after laser exposure. For 3-D spheroids, PC/CHI also showed higher cytotoxicity using acid phosphatase assay and a decrease in spheroid size was observed using light microscopy. In conclusion, both types of nanocarriers can be used for their potential treatment of SCC using PDT depending on the tumour localization in the skin.

Beneficial effects of antioxidants in improving health conditions of sheep infected with foot-and-mouth disease.

The effect of multinutrient antioxidant treatment on sheep naturally infected with FMD virus was investigated in terms of general health conditions, serum proteins profile, and antioxidant/oxidant parameters. Twenty diseased sheep were divided into 4 equal groups (n = 5) and underwent certain therapeutic protocols for 8 weeks as follows: GI, infected not treated group; GII, infected and treated with the ideal and usual line of treatment against FMD virus infection; GIII, infected animals supplemented orally zinc methionine at a dose of 5 g/head/day and vitamin E with selenium-enriched yeast at the same dose level; GIV, infected animals received both the ideal treatment and antioxidants. The animals under experiment were clinically evaluated. Blood samples were obtained for the comet assay and biochemical examination at zero time and at the 8th week after treatment. Results revealed that DNA damage reduced in both GIII and GIV groups which received antioxidants. In the GI group, the activity of SOD and GPx and the level of total antioxidant capacity (TAC) markedly decreased. However, in both GIII and GIV groups treated with multinutrient antioxidants, GPx and TAC values significantly increased after treatment in comparison with the values of the same groups before treatment. After treatment with multinutrient antioxidants, α1-, ß-, and γ-globulins levels markedly increased in GII and GIII groups while α2-globulin level decreased. The improvement in healing of clinical signs and general health conditions was clear in the GIV group. Finally, FMD infection in sheep was found to be associated with oxidative stress. The use of antioxidants as therapeutic approaches recovers and improves general health conditions and performance of affected animals.

Hepatoprotective and immunomodulatory effects of copper-nicotinate complex against fatty liver in rat model.

AIM: The current study was designed to evaluate the potential hepatoprotective and immunomodulatory effects of copper-nicotinate complex (CNC) against methionine- and choline-deficient diet (MCDD)-induced fatty liver in rats. MATERIALS AND METHODS: Forty male Wistar rats were randomly allocated into one of four equal-sized groups (G1-G4). The G1 group was fed a balanced diet and kept under normal conditions; the G2 group received CNC orally at a dose of 0.043 mg/kg body weight, 3 times/week for 4 weeks, and a balanced diet; the G3 group was fed an MCDD for 4 weeks; and the G4 group was fed an MCDD and administered CNC at the same dose and route as G2. Blood samples were collected for the determination of serum enzyme activity. After 4 weeks of treatment, liver specimens were collected for the evaluation of the oxidative/antioxidative markers, cytokine gene expression, and histopathological examination. RESULTS: CNC improved MCDD-induced liver dysfunctions by recovering serum alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyl transferase activities to their normal levels. The glutathione (GSH) level and superoxide dismutase (SOD) activity significantly decreased, while lipid peroxidation (as reflected by malondialdehyde [MDA]) markedly increased in the liver tissue of the MCDD group. After cotreatment with MCDD and CNC, the GSH level and SOD activity markedly increased and the MDA level significantly decreased to return to normal levels. After cotreatment with MCDD and CNC, significant downregulation of the mRNA expression of hepatic interleukin (IL)-1ß, IL-4, macrophage inflammatory protein-1α, and monocyte chemoattractant protein-1 genes was found. Moreover, CNC reduced fatty liver complications by reducing the number of hepatic vacuolations, degenerative changes in the hepatocytes, and hemorrhage. CONCLUSION: CNC has the potential to limit tissue injury and possibly prevent the progression to severe liver disease caused by an MCDD.

Effect of Egyptian propolis on cryptosporidiosis in immunosuppressed rats with special emphasis on oocysts shedding, leukogram, protein profile and ileum histopathology.

OBJECTIVES: To investigate the activity of Egyptian propolis extracts (ethanol and water) on cryptosporidiosis in experimentally infected dexamethasone-immunosuppressed rats. METHODS: A total of 180 male rats (190-220) g BWt were randomly divided into 9 equal groups (G1-G9). Groups of rats were kept as (G1): normal control, (G2-G9): immunosuppressed with dexamethasone and (G3-G9): infected with Cryptosporidium oocysts. Rats from (G4-G9) were given orally ethanol and water extract of propolis (at a dose of 50 mg/kg BWt) and nitazoxanide (standard anti-cryptosporidial drug at a dose of 100 mg/kg BWt) to infected rats with different regimes. Faecal pellets were collected from all groups to monitor oocysts shedding from the 2nd to the 15th day post infection. At the end of the experiment, blood was collected from all groups for determination of leukogram and serum proteins. Ileum specimens were also examined histopathologically. RESULTS: The highest reduction of oocysts shedding in faecal samples was 88% in rats prophylactically treated with propolis ethanol extract at the 4th dpi, and in rats prophylactically treated with water extract of propolis, was 91% at the 6th dpi. There was a marked increase in neutrophils count and α2- and ß-globulins levels in infected rats treated with both extracts, while a significant decrease was detected in lymphocytes compared to the infected non treated group. ß-Globulin level markedly increased in the rats administered nitazoxanide. Histopathological changes were observed in the ileum of rats infected with Cryptosporidium. CONCLUSIONS: Egyptian propolis extracts have an activity on cryptosporidiosis in rats. Moreover, propolis modulated the immunity in dexamethasone-immunosuppressed rats.

Use of allicin as feed additive to enhance vaccination capacity of Clostridium perfringens toxoid in rabbits.

The present study assessed the efficacy of Clostridium perfringens (C. perfringens) toxoid and/or allicin - as feed additive - in rabbits for preventing or minimizing the severity of infection with locally isolated strain of C. perfringens type A. Serum biochemical, immunological and pathological investigations were also done. One hundred rabbits of 6 weeks of age were divided into five equal groups (G1-G5). G1 were kept as normal control. G2 was allocated for C. perfringens type A infection. G3 was vaccinated with C. perfringens toxoid at zero time and then with a booster dose at the 3rd week of the experimental period. G4 was treated with allicin 20% added to the ration (200mg/kg ration) all over the experimental period. G5 was vaccinated with C. perfringens toxoid at the zero time then with a booster dose at the 3rd week of the experiment period, and treated with allicin 20% from the zero time till the end of the experiment. At the 4th week, G2, G3, G4 and G5 were challenged orally (5 ml) and subcutaneously (2 ml) with 24h cooked meat broth containing 1 × 10(7) colony-forming units/ml of C. perfringens type A strain. Blood and tissue samples were collected from all groups po st-vaccination then post-challenge for biochemical analysis, serum neutralization test and histopathological examinations. Results revealed that rabbits treated with both allicin and toxoid vaccine demonstrated high level of antitoxin titre post-challenge, improved liver and kidney functions, and reduced morbidity and mortality rates and the severity of histopathological changes associated with challenge of rabbits with C. perfringens type A strain. In conclusion, vaccination of rabbits with C. perfringens toxoid combined with allicin 20% gave better protection, enhanced immune response and had no adverse effects on the general health conditions against C. perfringens type A infection compared to rabbits vaccinated with C. perfringens toxoid only.

Tegumental Effects of Methanolic Extract of Balanites aegyptiaca Fruits on Adult Paramphistomum microbothrium (Fischoeder 1901) under Laboratory Conditions.

BACKGROUND: Weak efficacy of different fasciolicidal compounds used for treatment of paramphistomosis has drawn the attention of many authors to alternative drugs. The purpose of this study was to assess, for the first time, the effect of the methanolic extract of Balanites aegyptiaca fruits (BAE) on adult Paramphistomum microbothrium. METHODS: The effect of BAE on adult P. microbothrium after 24 h incubating the parasites in RPMI 1640 culture medium containing 10, 50, 100 and 200 µg/ml BAE was determined by light and scanning electron microscopic studies. RESULTS: Differences in response to BAE action were concentration dependent.The major target organ that was highly affected was the tegument. Maximum anthelmintic activity was found with a dose of 200 µg/ml BAE, at which distinct damage to the whole body surface of the trematodes was very much distinct. Shape and structure of both suckers were deformed due to BAE. This damage would undoubtedly disrupt many of the physiological processes associated with the tegument. Besides, the damage of the tegumental folds of the acetabular region might disrupt its function in drawing the rumen wall tissue of the host into the acetabular cavity. CONCLUSION: the use of methanolic extract of B. aegyptiaca fruits offers a new dimension and potential for control of such a neglected infectious disease in ruminants, at a time when paramphistomosis has emerged as an important cause of productivity loss.

Serological and molecular diagnostic surveys combined with examining hematological profiles suggests increased levels of infection and hematological response of cattle to babesiosis infections compared to native buffaloes in Egypt.

BACKGROUND: Babesiosis threatens the development of the cattle and buffaloes industries in Egypt and improved control is needed. The main objectives of this study are surveying the presence of bovine babesiosis in distinct selected bovine and buffalo populations in Egypt using novel molecular and previously validated serological methods, while also comparing the occurrence of hematological alterations among Babesia infected cattle and buffalos. METHODS: A total of 253 and 81 blood samples from apparently healthy cattle and buffaloes, respectively, were randomly collected from diverse locations in Egypt. All samples were tested for Babesia bovis and B. bigemina infection using blood film examination, competitive ELISA (cELISA) and PCR. Novel semi-nested and nested PCR assays for the detection of B. bovis and B. bigemina respectively, were developed and used to analyze DNA extracted from bovine and buffalo samples. Hematological profiles were studied using a hematological analyzer. RESULTS: Blood films examination revealed 13.8% and 7.4% Babesia infection rates in cattle and buffaloes, respectively. However, in cattle, the cELISA detected 32.8%, 21.3% and 10.7% infection rates with B. bigemina, B. bovis and mixed infection, respectively. In addition, cELISA identified 22.2%, 22.2% and 6.2% infection rates with B. bigemina, B. bovis and mixed infection, respectively in buffaloes. The semi-nested PCR assay showed that 15% of the tested samples were positive for B. bovis in cattle, but just 3% in buffaloes. Infections with B. bigemina were also found in cattle (32.4%), but not in buffaloes upon nested PCR analysis. Sequencing analysis confirmed the identity of the PCR amplicons and showed that Egyptian genotypes of B. bigemina and B. bovis highly resemble sequences previously deposited in GenBank. Hemograms performed on the sampled animals revealed macrocytic hypochromic anemia associated with reduced platelet counts in infected cattle with babesiosis. In addition, marked increases in total leukocyte and granulocytic counts and decreases in lymphocytic counts were found in infected cattle. In contrast, no such hematological anomalies were found in presumably Babesia-infected buffaloes. CONCLUSIONS: Frequent occurrence of babesiosis among apparently healthy bovines in Egypt, suggests the need for appropriately designed prevalence studies in this country. Infected bovine, but not buffalo, populations often present hematological disorders compatible with intravascular hemolysis and thrombocytopenia.

Vector identification and clinical, hematological, biochemical, and parasitological characteristics of camel (Camelus dromedarius) theileriosis in Egypt.

The objectives of the present study were to identify a possible tick vector and to determine the prevalence of camel theileriosis in Egypt using blood smears stained with Giemsa's stain and PCR assay. Hemogram and serum biochemical constituents were also investigated. A total of 243 camels, aged 3-5 years, were examined. The results revealed that 75 (30.86 %) camels were infected with Theileria spp. of Giemsa-stained blood smears. Hyalomma dromedarii was identified as the carrier tick of Theileria spp. Multinucleated sporoblast and free sporozoite were observed in the salivary gland smears from collecting ticks. PCR result revealed that Theileria annulata was the most abundant in camels (60 %) followed by Theileria spp. (10 %). Macrocytic hypochromic anemia was recorded in the infected camels with T. annulata. Leukocytosis, neutrophilia, eosinophilia, and lymphopenia were also observed in the infected group. In the serum of infected camels, total proteins, albumin, ß-globulin, and A/G ratio were significantly decreased (P < 0.05); however, total globulins and α- and γ-globulins were markedly increased (P < 0.05). The activity of aspartate aminotransferase and the levels of glucose, creatinine, and high-density lipoprotein cholesterol were markedly increased (P < 0.05) in the infected group. In contrast, triglycerides and total cholesterol concentrations were significant decreased (P < 0.001) in the infected group. In conclusion, a high prevalence of camel theileriosis was recorded in apparently healthy camels. H. dromedarii commonly infested these camels and were found infected with the transmissible forms of Theileria, indicating a role in transmission. Camels infected with T. annulata induced alterations in the cellular and biochemical constituents.

Protective effect of Egyptian propolis against rabbit pasteurellosis.

The present study was conducted to study the protective effect of ethanolic extract of propolis given subcutaneously (S/C) either alone or in combination with inactivated formalized Pasteurella multocida (P. multocida) vaccine in rabbits challenged with virulent P. multocida strain. Twenty-eight New-Zealand rabbits, 6-8 weeks old and not vaccinated against pasteurellosis, were randomly divided into four equal groups. Group (1) was kept as nonvaccinated control. Group (2) was injected S/C with propolis. Group (3) was vaccinated (S/C) with P. multocida vaccine only. Group (4) was injected with vaccine mixed with propolis as adjuvant. Groups (2, 3, and 4) received the same doses of propolis and vaccine after 4 weeks as a booster dose. The experiment continued for six weeks during which clinical signs, body weight, and mortality rate were recorded. Blood samples were collected every 2 weeks of treatment for evaluating the erythrogram and biochemical parameters. At the end of six weeks, all groups were subjected to challenge with a virulent strain of P. multocida. Two weeks later, tissue specimens were collected from different organs for histopathological investigation. Results showed that before challenge all rabbits of different groups were apparently healthy and had good appetite. After challenge, control group (1) showed acute form of the disease, 100% mortality rate, and severe histopathological changes. Rabbits of groups (2 and 3) showed less severe clinical signs, mortality rate, and histopathological changes than control. Rabbits of group (4) were apparently healthy with normal histological picture. In conclusion, an ethanolic extract of propolis injected alone or combined with formalized inactivated P. multocida vaccine improved general health conditions, liver and kidney functions in addition to reduction of the severity of adverse clinical signs, mortality rates, and histopathological changes associated with challenge of rabbits with P. multocida strain.