Molecular and phylogenetic analysis of a type K1 strain Trypanosoma evansi isolate from Nigerian cattle: An evaluation of the therapeutic effects of compounds from Brassica oleracea on the histopathology of infected wister rats.

Background: Understanding the pathogenesis of animal trypanosomiasis can be improved by studying the genetics of bovine trypanosomes. Pathogenic animal trypanosomes are a major impediment to livestock production, with negative economic consequences spreading beyond Sub-Saharan Africa to subtropical regions of Northern Africa, Southeast Asia, and Central and South America. An atypical K1 strain of Trypanosoma evansi (T. evansi) isolates from infected cattle in Nigeria was analyzed. The therapeutic effect of phenolic-rich compounds on the histopathology of wistar rats infected with the K1 strain was studied. Methods: The K1 strain T. evansi was analyzed molecularly using PCR and sequence analysis of the Spacer-1 ribosomal RNA gene. To assess the evolutionary relationship, this was phylogenetically compared to other species studied in different parts of the world. Thirty adult male wistar rats were divided into six groups of five each. Animals in group A served as the standard control (not infected). Group B animals were infected but not treated. Group C animals were infected and given 3.5 mg/kg body weight of the standard drug diminazene aceturate. Animals in groups D, E, and F were infected and treated with phenolic-rich compounds isolated from Brassica oleracea (B. oleracea) at concentrations of 100, 200, and 400 mg/kg body weight, respectively. The phytochemicals were extracted using standard analytical procedures, and GCMS analysis revealed the presence of phenolic-rich compounds. The animals were given 0.2 mg/ml trypanosome intraperitoneally, diluted with normal saline. The vital organs of the animals were harvested and histologically examined. Results: The nested PCR amplification of the trypanosome's ITS-1 region revealed a DNA amplicon of 627 base pairs. The rRNA nucleotide sequence was deposited in GenBank under the accession number MN462960. Basic Local Alignment search of the obtained ITS-1 rRNA sequences revealed that the K1 strain trypanosome and other strains from different regions have an evolutionary relationship. The phenolic-rich compounds had protective effects on the organs of infected animals, resulting in a decrease in parasitemia levels. They have anti-trypanosome activities at the minimum and maximum effective doses of 200 and 400 mg/kg body weight, respectively. Conclusions: The K1 strain T. evansi was isolated from naturally infected cattle in this study. The results indicate that phenolic-rich compounds have anti trypanosoma activities capable of healing organ damage caused by trypanosomiasis.

Evaluation of the toxicity of crude aqueous Pleurotus tuber-regium (Fr.) Singer extract in mice.

OBJECTIVES: The study aimed to determine the phytochemical profile and safety of the aqueous extract of Pleurotus tuber regium using laboratory animal model. METHODS: The pulverized sample was macerated with water and allowed to stand for 24 h, then filtered. Preliminary qualitative phytochemical screening was carried out on the crude extract and LD50 determination was done using standard procedures. Haematological and serum biochemical parameters to assess the kidney and liver functions were carried out using standard methods. RESULTS: The phytochemical screening revealed the presence of alkaloids, cardiac glycosides, saponins, phenolic compounds, tannins, steroids, carbohydrates, flavonoids and terpenoids. However, anthraquinones were observed to be absent in the extracts. The LD50 of the extract was found to be greater than 5,000 mg/kg when administered orally with no physical signs of toxicity and death over the 14-day period. There was no significant (p>0.05) increase in the level of serum alanine aminotransferase, aspartate transaminase, alkaline phosphatase, blood urea nitrogen and creatinine (p<0.05) when compared to the control group. There was a significant (p<0.05) increase in the total white blood cells of the mice administered with P. tuber-regium. CONCLUSIONS: This study showed that P. tuber-regium (Fr.) Singer does not possess toxic principles, hence, the wide safety margin observed in the mice.

Effects of crude methanol extract of Adansonia digitata fruit pulp on Naja nigricollis venom-induce toxicity in Wistar rats.

OBJECTIVES: This study evaluated the effects of Crude Methanol Extract of Adansonia digitata Fruit Pulp on Naja nigricollis Venom-Induce Toxicity in Wistar rats. METHODS: A. digitata was extracted using 70% methanol and median lethal dose (LD50) of both the extract and venom were determined using the up-and-down method. Sixty Wistar rats were randomly assigned into 10 groups of 6 rats each and were administered with normal saline, venom only, venom + antivenom, 125 mg/kg, 250 mg/kg and 500 mg/kg crude methanol extract (CME) pre-envenomation, 125 mg/kg, 250 mg/kg and 500 mg/kg CME post-envenomation, and venom + antivenom + 250 mg/kg CME, respectively. Blood samples were collected 8-h post-envenomation in EDTA and plain sample bottles. Erythrocyte osmotic fragility (EOF) test was carried out on the EDTA blood samples while serum was harvested and used for Malondialdehyde (MDA) and Superoxide Dismutase (SOD) assays. RESULTS: LD50 of the CME and venom was >5,000 mg/kg and 0.889 mg/kg, respectively. N. nigricollis-induced oxidative stress was evident in group B through increased % haemolysis, MDA and lowered SOD activities. The groups treated with antivenom only, 250 mg/kg CME post-envenomation and antivenom +250 mg/kg CME significantly (p<0.05) reduced EOF, MDA values and increased SOD. The CME revealed better ameliorative effect than protective via inhibition of EOF, MDA values and increased SOD activity. CONCLUSIONS: The CME when administered singly showed more ameliorative properties and the combination of CME with antivenom for protection was not as effective as when compared to single administration.

Comparative effect of dietary supplements on the performance and severity of experimental Eimeria tenella infection in broiler chickens.

The effect of the dietary supplements, sugarcane molasses (prebiotic), Antox® (probiotic) and Enflorax® (synbiotic), on gut health, performance and severity of infection with Eimeria tenella in broiler chickens was evaluated in this study. Ninety-day-old broiler chicks were divided into six groups (A, B, C, D, E and F). Groups B to F were challenged with Eimeria tenella (2.0 × 104 oocysts/chick). Groups C, D and E were supplemented from day old with prebiotic, probiotic and synbiotic, respectively, while F was administered amprolium. Groups A and B served as negative and positive controls, respectively. Feed intake and performance parameters were assessed weekly for each group. After infection clinical signs, morbidity and mortality rates were monitored alongside oocyst output, gross and microscopic caecal lesions. Infected chickens exhibited clinical signs 4 days post infection (dpi) with 100% morbidity in all infected groups. Infected groups showed significant (P < 0.05) drop in feed intake and weight gain from 3 to 5 weeks of age. Feed conversion ratio was highest in B but lower in the supplemented groups. Oocyst output in faeces were significantly lower (P < 0.05) in the supplemented groups compared with B. Macroscopic lesion scores 7 dpi were significantly lower in the supplemented groups compared with B, though group F had the lowest mean score. Histopathological examination of caeca tissues showed milder lesions in the supplemented groups. In conclusion, the supplements prebiotic, probiotic and synbiotic ameliorated the consequences of caecal coccidiosis in broiler chickens and therefore recommended for use in broiler production.

Comparative effects of artemether and in combination with diminazene aceturate in the treatment of experimental Trypanosoma brucei brucei infection in Wistar rats.

This study assessed the effects of artemether and in combination with diminazene aceturate on parasitaemia, weight, haematology and pathology induced by experimentally Trypanosoma brucei brucei infection in Wistar rats. Fifty adult rats comprising 25 each of males and females were assigned into 5 groups of ten rats (five males and five females). Rats in group I was uninfected while groups II-V were infected with T b brucei. Groups II were untreated; III administered diminazene aceturate once; IV and V administered artemether only and in combination with diminazene aceturate respectively for 5 days. Parasitaemia was determined daily, blood was collected for haematology and weight obtained every four days for a period of 32 days. At 24 days post-treatment, rats were humanely euthanized and organs harvested for pathological examination. Results revealed parasitaemia at day 4 post-infection, significant (p < 0.05) decrease in weight, erythrogram and leucogram in all infected rats. Following treatment, there was significant (p < 0.05) decrease in parasitaemia, increased weight gain and improved haematology. Pathological examination revealed significantly (p < 0.05) decreased gross and histopathological lesions in treated groups compared to group II. In conclusion, artemether and in combination with diminazene aceturate produced antitrypanosomal effects against experimental trypanosomosis.

Proximate Analysis and Safety Profile of Farmed Edible Bird's Nest in Malaysia and Its Effect on Cancer Cells.

Edible bird's nest (EBN) which is solidified swiftlet's saliva contains high nutritional value. It is widely consumed in countries like Malaysia, Indonesia, and Thailand. However, previous proximate analysis of Malaysia EBN was not representative of all the regions in Malaysia. In recent years, safety issues such as high nitrate and nitrite contents, presence of heavy metal, adulteration, fungal infection, and cancer cell stimulation were associated with EBN. Hence, this study aimed to determine the proximate analysis, safety profile during normal weather and hazy periods, and its effect on cancer cells stimulation in Malaysia-farmed EBN. Seven raw cleaned EBN samples were sourced from 6 different regions in Malaysia. Proximate analysis and safety profile were performed using official AOCA methods and Malaysian Standard. High protein (53.03-56.37%) and carbohydrate content (27.97-31.68%) with an acceptable level of moisture (10.8-14.04%) and ash (2.22-3.38%) were reported. A good safety profile was obtained with low nitrite and nitrate levels, with undetectable heavy metals and no significant growth of pathogenic microorganism except mould. Epidermal growth factor was detected but below the quantification level with the chicken EGF ELISA kit. The microculture tetrazolium (MTT) assay was performed for growth stimulation assessment comparing human EGF and EBN. There was no significant cell growth observed in cancer cells after EBN treatment. In conclusion, EBN Malaysia has a good nutritional profile, free of heavy metals, and an acceptable level of nitrate, nitrite, and microorganism profile except for mould contents. Furthermore, the in vitro study indicated that EBN was not associated with cancer cell growth.

A standardised Andrographis paniculata Burm. Nees aqueous extract prevents Lipopolysaccharide-induced cognitive deficits through suppression of inflammatory cytokines and oxidative stress mediators.

Substantial evidence has shown that most cases of memory impairment are associated with increased neuroinflammation and oxidative stress. In this study, the potential of a standardised Andrographis paniculata aqueous extract (APAE) to reverse neuroinflammation and cognitive impairment induced by lipopolysaccharide (LPS) was examined in vivo. Rats were treated with APAE (50, 100, 200, and 400 mg·kg-1, p.o.) for 7 consecutive days prior to LPS (1 mg·kg-1, i.p.)-induced neuroinflammation and cognitive impairment. Spatial learning and memory were evaluated using the Morris water maze (MWM) test, while neuroinflammation and oxidative stress were assessed through the measurement of specific mediators, namely, tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1ß, superoxide dismutase (SOD), catalase (CAT), antioxidant glutathione (GSH), reactive oxygen species (ROS), and thiobarbituric acid reactive substance (TBARS). Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) were also evaluated. LPS caused significant memory deficits in the 2-day MWM protocol, whereas pretreatment with standardised APAE dose-dependently improved performance in the MWM test. APAE treatment also blocked the LPS-induced hippocampal increase in the concentration and expression of proinflammatory cytokines (TNF-α, IL-1ß, and IL-6) and production of ROS and TBARS and enhanced the activities of AChE and BChE. Furthermore, APAE enhanced the decrease in the levels and expression of hippocampal antioxidant enzymes (SOD and CAT) following LPS-induced neuroinflammation and cognitive deficit. The findings from these studies suggested that standardised APAE improved memory and had potent neuroprotective effects against LPS-induced neurotoxicity.

Characterization of thymoquinone/hydroxypropyl-ß-cyclodextrin inclusion complex: Application to anti-allergy properties.

Thymoquinone is an effective phytochemical compound in the treatment of various diseases. However, its practical administration has been limited due to poor aqueous solubility and bioavailability. In this work, we developed a novel inclusion complex of thymoquinone and hydroxypropyl-ß-cyclodextrin that features improved solubility and bioactivity. The drug solubility was markedly accelerated in the increasing ratio of hydroxypropyl-ß-cyclodextrin to thymoquinone amount. The formation of the thymoquinone/hydroxypropyl-ß-cyclodextrin inclusion complex was evidenced using X-ray diffraction, differential scanning calorimetry, thermal gravimetric analysis, Fourier transform infrared, scanning electron microscopy and nuclear magnetic resonance. The release behavior of the complex, as well as of their mixtures, was examined in artificial gastric (pH 1.2) and intestinal (pH 6.8) dissolution media. The formulated complex released the drug rapidly at the initial stage, followed by a slow release. Thermodynamic parameters ΔH, ΔS and ΔG were calculated with temperatures ranging from 20 to 45 °C to evaluate the complexation process. The activity of the inclusion complex was evaluated on IgE-mediated allergic response in rat basophilic leukemia (RBL-2H3) cells by monitoring key allergic mediators. The results revealed that compared with free thymoquinone, the inclusion complex more strongly inhibited the release of histamine, tumor necrosis factor-α, and interleukin-4, and was not cytotoxic at the tested thymoquinone concentrations (0.125-4 µg/mL) indicating the inclusion complex possibly had better antiallergic effects. Our finding suggested that the inclusion complex achieved prolonged action and reduced side-effect of thymoquinone.

Beneficial effects of TQRF and TQ nano- and conventional emulsions on memory deficit, lipid peroxidation, total antioxidant status, antioxidants genes expression and soluble Aß levels in high fat-cholesterol diet-induced rats.

The study determined the effect of thymoquinone rich fraction (TQRF) and thymoquinone (TQ) in the forms of nano- and conventional emulsions on learning and memory, lipid peroxidation, total antioxidant status, antioxidants genes expression and soluble ß-amyloid (Aß) levels in rats fed with a high fat-cholesterol diet (HFCD). The TQRF was extracted from Nigella sativa seeds using a supercritical fluid extraction system and prepared into nanoemulsion, which later named as TQRF nanoemulsion (TQRFNE). Meanwhile, TQ was acquired commercially and prepared into thymoquinone nanoemulsion (TQNE). The TQRF and TQ conventional emulsions (CE), named as TQRFCE and TQCE, respectively were studied for comparison. Statin (simvastatin) and non-statin (probucol) cholesterol-lowering agents, and a mild-to-severe Alzheimer's disease drug (donepezil) were served as control drugs. The Sprague Dawley rats were fed with HFCD for 6 months, and treated with the intervention groups via oral gavage daily for the last 3 months. As a result, HFCD-fed rats exhibited hypercholesterolaemia, accompanied by memory deficit, increment of lipid peroxidation and soluble Aß levels, decrement of total antioxidant status and down-regulation of antioxidants genes expression levels. TQRFNE demonstrated comparable effects to the other intervention groups and control drugs in serum biomarkers as well as in the learning and memory test. Somehow, TQRFNE was more prominent than those intervention groups and control drugs in brain biomarkers concomitant to gene and protein expression levels. Supplementation of TQRFNE into an HFCD thus could ameliorate memory deficit, lipid peroxidation and soluble Aß levels as well as improving the total antioxidant status and antioxidants genes expression levels.

Nanoemulsion-based Parenteral Drug Delivery System of Carbamazepine: Preparation, Characterization, Stability Evaluation and Blood-Brain Pharmacokinetics.

Carbamzepine (CBZ) was encapsulated in a parenteral oil-in-water nanoemulsion, in an attempt to improve its bioavailability. The particle size, polydispersity index and zeta potential were measured using dynamic light scattering. Other parameters such as pH, osmolality, viscosity, drug loading efficiency and entrapment efficiency were also recorded. Transmission electron microscopy revealed that emulsion droplets were almost spherical in shape and in the nano-range. The in vitro release profile was best characterized by Higuchi's equation. The parenteral nanoemulsion of CBZ showed significantly higher AUC0→5, AUC0→∞, AUMC0→5, AUMC0→∞, Cmax and lower clearance than that of CBZ solution in plasma. Additionally, parenteral nanoemulsion of CBZ showed significantly higher AUC0→∞, AUMC0→∞ and Cmaxthan that of CBZ solution in brain. The parenteral nanoemulsion of CBZ could therefore use as a carrier, worth exploring further for brain targeting.