Plasmodium Berghei-infected Mice Research Articles

In vivo antimalarial efficacy of Artemisia afra powder suspensions.

A global death toll of 608,000 in 2022 and emerging parasite resistance to artemisinin, the mainstay of antimalarial chemotherapy derived from the Chinese herb Artemisia annua, urge the development of novel antimalarials. A clinical trial has found high antimalarial potency for aqueous extracts of A. annua as well as its African counterpart Artemisia afra, which contains only trace amounts of artemisinin. The artemisinin-independent antimalarial activity of A. afra points to the existence of other antimalarials present in the plant. However, the publication was retracted due to ethical and methodological concerns in the trial, so the only evidence for antimalarial activity of A. afra is built on in vitro studies reporting efficacy only in the microgram per milliliter range. Our study aims to shed more light on the controversy around the antimalarial activity of A. afra by assessing its efficacy in mice. In particular, we are testing the hypothesis that A. afra contains a pro-drug that is inactive in vitro but active in vivo after metabolization by the mammalian host. Plasmodium berghei-infected mice were treated once or thrice (on three consecutive days) with various doses of A. afra, A. annua, or pure artemisinin. Aqueous powder suspensions of A. annua but not A. afra showed antimalarial activity in mice. Our experiments conducted in mice do not support the pro-drug hypothesis.

Mechanism of antimalarial action and mitigation of infection-mediated mitochondrial dysfunction by phyto-constituents of Andrographis paniculata ((Burm f.) Wall. ex Nees) in Plasmodium berghei-infected mice

Ethnopharmacological relevanceAndrographis paniculata (AP) ((Burm f.) Wall. ex Nees) is a medicinal plant, documented for its folkloric use in the treatment of malaria. AimThis study was designed to determine the potency of extract and fractions of A. paniculata (AP) as a curative, both for susceptible and resistant malaria and to also determine the plant's mechanism of action. This study was also designed to determine whether AP extract and its most potent fraction will mitigate infection-mediated mitochondrial dysfunction, and to assess the phytochemical constituents of the most potent fraction. Materials and methodsn-Hexane, dichloromethane, ethylacetate and methanol were used to partition the methanol extract of A. paniculata. Graded doses of these extract and fractions were used to treat mice infected with chloroquine-sensitive strain of P. berghei in a curative model. The most potent fraction was used to treat mice infected with resistant (ANKA strain) P. berghei. Inhibition of hemozoin formation, reversal of mitochondrial dysfunction and antiinflammatory potentials were determined. A combination of ultraperformance liquid chromatography-quadrupole time of flight-mass spectrometry and nuclear magnetic resonance spectroscopy were used for chemical analysis. ResultsMicroscopy revealed that the dichloromethane fraction decreased the parasite burden the most, and inhibition of the hemozoin formation is one of its mechanisms of action. The dichloromethane fraction reversed parasite-induced mitochondrial pore opening in the host, enzyme-dependent ATP hydrolysis and peroxidation of host mitochondrial membrane phospholipids as well as its antiinflammatory potentials. The UPLC-qTOF-MS report and NMR fingerprints of the dichloromethane fraction of A. paniculata yielded fourteen compounds of which sibiricinone C was identified from the plant for the first time. ConclusionFractions of A. paniculata possess antiplasmodial effects with the dichloromethane fraction having the highest potency. The potent effect of this fraction may be attributed to the phytochemicals present because it contains terpenes implicated with antimalarial and antiinflammatory activities.

Antimalarial Efficacy of Ethanol Extract of Bridelia micrantha Stem Bark against Plasmodium berghei-Infected Mice.

The spread of drug resistance is a significant issue, particularly in endemic countries with limited resources. The aim of this study was to evaluate antimalarial and antioxidant activity of B. micrantha in order to justify its use in traditional medicine. Evaluation of the in vivo antimalarial activity of B. micrantha was carried out according to the model of the suppressive and curative test of Peters' over 4 days in infected Swiss albino mice. Antioxidant parameters and stress were measured after intraperitoneal administration of 1 × 107 infected red blood cells. At doses of 150 mg/kg, 300 mg/kg, and 600 mg/kg, administration of B. micrantha substantially produced suppression of P. berghei infection by 67.75%, 73.46%, and 78.99%, respectively, while 84.64% of the untreated group (1% DMSO) had suppression from chloroquine. The curative test significantly decreased the levels of parasitaemia and death in the treated groups. Furthermore, after B. micrantha extract was given to infected mice, a noteworthy increase in total protein, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) was observed. On the other hand, hepatic catalase (CAT) and superoxide dismutase (SOD) productions were considerably greater than that of the healthy control. Mice had considerably lower levels of nonenzymatic antioxidant markers such as glutathione, NO, and MDA showing that the liver was protected. The infected groups responded favorably to the ethanol extract of B. micrantha. This result justifies investigation for its use in Cameroon.

Evaluation of Andrographis paniculata-supplemented Diet on the reproductive indices of Plasmodium berghei-infected mice

Ethnopharmacological relevanceThe King of Bitters (Andrographis paniculata) is a plant used to cure a wide range of infectious diseases which includes malaria, fever and others. However, there is a paucity of scientific evidence of its effect on male reproductive indices during malaria treatment. Aim of the studyThe aim of this study is to evaluate the effect of supplemented diet on antiplasmodial, hematological and male reproductive indices in mice infected with Plasmodium berghei. Materials and methodsAqueous extract of A. paniculata (King of Bitters, KGB) was prepared and the total phenol and flavonoid contents were determined. Forty-two mice, weighing 20–25 g, were distributed into 7 groups consisting of 6 mice each. The mice were innoculated with strain NK65 Plasmodium berghei (Chloroquine, CQ sensitive) and the parasitemia suppression was assessed. The mice were fed with the dietary supplementation of KGB at varying inclusions (2.5%, 5%, 7.5%, and 10%) and administered 10 mg/kg CQ (which served as the positive control) for 5 consecutive days after infection was established. The reactive malondialdeahyde (MDA), antioxidant [superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH)] and the hematological (hemoglobin, packed cell volume and red blood cell) parameters in the infected mice were determined. The reproductive indices (serum testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), sperm count, sperm motility, and sperm viability) and testis histopathology were also assessed. ResultThe result revealed that KGB had a total phenol content of 32.55 mgGAE/g and total flavonoid content of 19.71 mgQUE/g. The infected mice treated with the dietary supplementation of KGB showed significantly decreased (p < 0.05) parasitaemia and MDA levels. Furthermore, the 7.5% dietary inclusion showed significant improvement in the antioxidant, hematological and reproductive indices as well as the restoration of testis morphology as seen in the histopathology plate of the infected mice treated with KGB. Hence, this study suggests that the KGB- supplemented diet (7.5%) may be a potential alternative and complementary therapy in the treatment of malaria infection and reproductive disorders.

Evaluation of the antimalarial activity of SAM13-2HCl with morpholine amide (SKM13 derivative) against antimalarial drug-resistant Plasmodium falciparum and Plasmodium berghei infected ICR mice.

Antimalarial drugs are an urgently need and crucial tool in the campaign against malaria, which can threaten public health. In this study, we examined the cytotoxicity of the 9 antimalarial compounds chemically synthesized using SKM13-2HCl. Except for SKM13-2HCl, the 5 newly synthesized compounds had a 50% cytotoxic concentration (CC50) &gt; 100 µM, indicating that they would be less cytotoxic than SKM13-2HCl. Among the 5 compounds, only SAM13-2HCl outperformed SKM13-2HCl for antimalarial activity, showing a 3- and 1.3-fold greater selective index (SI) (CC50/IC50) than SKM13-2HCl in vitro against both chloroquine-sensitive (3D7) and chloroquine -resistant (K1) Plasmodium falciparum strains, respectively. Thus, the presence of morpholine amide may help to effectively suppress human-infectious P. falciparum parasites. However, the antimalarial activity of SAM13-2HCl was inferior to that of the SKM13-2HCl template compound in the P. berghei NK65-infected mouse model, possibly because SAM13-2HCl had a lower polarity and less efficient pharmacokinetics than SKM13-2HCl. SAM13-2HCl was more toxic in the rodent model. Consequently, SAM13-2HCl containing morpholine was selected from screening a combination of pharmacologically significant structures as being the most effective in vitro against human-infectious P. falciparum but was less efficient in vivo in a P. berghei-infected animal model when compared with SKM13-2HCl. Therefore, SAM13-2HCl containing morpholine could be considered a promising compound to treat chloroquine-resistant P. falciparum infections, although further optimization is crucial to maintain antimalarial activity while reducing toxicity in animals.

Access to Artemisinin-Triazole Antimalarials via Organo-Click Reaction: High In Vitro/In Vivo Activity against Multi-Drug-Resistant Malaria Parasites.

Malaria is one of the most widespread diseases worldwide. Besides a growing number of people potentially threatened by malaria, the consistent emergence of resistance against established antimalarial pharmaceuticals leads to an urge toward new antimalarial drugs. Hybridization of two chemically diverse compounds into a new bioactive product is a successful concept to improve the properties of a hybrid drug relative to the parent compounds and also to overcome multidrug resistance. 1,2,3-Triazoles are a significant pharmacophore system among nitrogen-containing heterocycles with various applications, such as antiviral, antimalarial, antibacterial, and anticancer agents. Several marketed drugs possess these versatile moieties, which are used in a wide range of medical indications. While the synthesis of hybrid compounds containing a 1,2,3-triazole unit was described using Cu- and Ru-catalyzed azide-alkyne cycloaddition, an alternative metal-free pathway has never been reported for the synthesis of antimalarial hybrids. However, a metal-free pathway is a green method that allows toxic and expensive metals to be replaced with an organocatalyst. Herein, we present the synthesis of new artemisinin-triazole antimalarial hybrids via a facile Ramachary-Bressy-Wang organocatalyzed azide-carbonyl [3 + 2] cycloaddition (organo-click) reaction. The prepared new hybrid compounds are highly potent in vitro against chloroquine (CQ)-resistant and multi-drug-resistant Plasmodium falciparum strains (IC50 (Dd2) down to 2.1 nM; IC50 (K1) down to 1.8 nM) compared to CQ (IC50 (Dd2) = 165.3 nM; IC50 (K1) = 302.8 nM). Moreover, the most potent hybrid drug was more efficacious in suppressing parasitemia and extending animal survival in Plasmodium berghei-infected mice (up to 100% animal survival and up to 40 days of survival time) than the reference drug artemisinin, illustrating the potential of the hybridization concept as an alternative and powerful drug-discovery approach.

Effect of black cumin-enriched acha-based cookies on antioxidant status and cholinergic enzymes in the brain of Plasmodium berghei-infected diabetic mice

Effect of black cumin-enriched acha-based cookies on antioxidant status and cholinergic enzymes in the brain of Plasmodium berghei-infected diabetic mice

SAR Guided Synthesis of Potent, Selective and Artemisinin Synergistic New Imidazole Derivatives with Promising In Vitro Antiplasmodial and In Vivo Antimalarial Activities

AbstractGuided by Structural Activity Relationship (SAR), a series of synthetic tri‐ and tetra‐substituted imidazoles were studied for in vitro antiplasmodial activities against erythrocytic stages of chloroquine sensitive and resistant strains of Plasmodium falciparum using SYBR green I assay. This led to the identification of three potent (IC50 μM) derivatives : Im2 (0.75), Im5 (0.75) and Im22 (0.5). These three lead molecules present unique structural features with a common molecular architecture. Thus, Im2 and Im22 have phenyl rings at positions 4 and 5 of imidazole ring along with n‐hexyl at N3 position, and differ in the C2 substituent (3‐cyclopentyloxy‐4‐methoxy phenyl for Im2) versus naphthyl for Im22. In contrast, positions 4 and 5 in Im5 have pyridyl rings, with unsubstituted N3. Further, curative antimalarial studies with Im2 and Im5 in a Plasmodium berghei (ANKA) infected mouse model of malaria, revealed an increase in average survival time to 28.6±4.2 and 27.4±4 days respectively, as compared to 20.2±2.3 days for untreated control mice. Results of in vitro combination (Im22+Artemisinin) with ΣFIC50: 0.34–0.79, indicated promising synergy and prospects for the use of Im22 as a new constituent in Artemisinin combination therapy (ACT).

Synthesis, antileishmanial, antimalarial evaluation and molecular docking study of some hydrazine-coupled pyrazole derivatives

Pyrazole-bearing compounds are known for their diverse pharmacological effects including potent antileishmanial and antimalarial activities. Herein, some hydrazine-coupled pyrazoles were successfully synthesized and their structures were verified by employing elemental microanalysis, FTIR, and 1H NMR techniques. The in vitro antileishmanial and in vivo antimalarial activities of the synthesized pyrazole derivatives (9–15) were evaluated against Leishmania aethiopica clinical isolate and Plasmodium berghei infected mice, respectively. The result revealed that compound 13 displayed superior antipromastigote activity (IC50 = 0.018) that was 174- and 2.6-fold more active than the standard drugs miltefosine (IC50 = 3.130) and amphotericin B deoxycholate (IC50 = 0.047). The molecular docking study conducted on Lm-PTR1, complexed with Trimethoprim was acquired from the Protein Data Bank (PDB ID:2bfm), justified the better antileishmanial activity of compound 13. Furthermore, the target compounds 14 and 15 elicited better inhibition effects against Plasmodium berghei with 70.2% and 90.4% suppression, respectively. In conclusion, the hydrazine-coupled pyrazole derivatives may be considered potential pharmacophores for the preparation of safe and effective antileishmanial and antimalarial agents.

Evaluation of the antimalarial properties of Solanum incanum L. leaf extract fractions and its ability to downregulate delta aminolevulinate dehydratase to prevent the establishment of malaria infection

Ethnopharmacological relevanceSolanum incanum L. is commonly used in traditional herbal medicine (THM) in Kenya for treating various ailments. Recent developments in disease treatment have introduced the concept of host-directed therapy (HDT). This approach involves targeting factors within the host cell that can impede the growth or replication of a pathogen. One such host factor is delta aminolevulinate dehydratase (δ-ALAD), the second enzyme in the heme biosynthesis pathway utilized by Plasmodium for growth. Studies using mice models have shown an increase in δ-ALAD expression during Plasmodium berghei infection. Another plant in the Solanum genus, S. guaranticum, has been found to inhibit δ-ALAD in red blood cells in vitro and in the brain in vivo. Is it possible that the bioactive compounds in S. incanum extracts could also be effective in HDT for malaria treatment? Aim of studyTo better assess the effectiveness of S. incanum leaf extracts as a curative and prophylaxis in malaria parasite infection, and to test the plant’s ability to decrease δ-ALAD expression. Materials and methodsThe leaves of S. incanum were collected, dried, and pulverized before being subjected to a successive extraction protocol to obtain crude, hexane, ethyl acetate, and aqueous extract fractions. Phytochemical analysis was conducted on all extract fractions, followed by GC-MS analysis of the fraction with the most potent antimalarial activity. An acute toxicity study was also performed on the extracted fractions. The potency of the extract fractions as curative and prophylactic antimalarial was then evaluated in THM using Plasmodium berghei-infected mice at a dose of 100 mg/kg. The extract fraction with the highest activity was further evaluated at varying doses and its effect on δ-ALAD was measured using RT-qPCR. The percentage of parasitemia and chemosuppression, and mean survival time were used as indices of activity. ResultsPhytochemical analysis revealed that the ethyl acetate and aqueous extract fractions contained high terpenoids, flavonoids, and phenols levels. However, alkaloids were only present in moderate quantities in the aqueous extract, and quinones were found in high levels only in the crude extract. Additionally, all extract fractions contained saponins in high levels but lacked tannins. While the plant extracts were found to be non-toxic, they did not exhibit curative antimalarial activity. However, all extract fractions showed prophylactic antimalarial activity, with the ethyl acetate extract having the highest percentage of chemosuppression even at doses of 250 and 1000 mg/kg. In the negative control, the expression of δ-ALAD was 5.4-fold, but this was significantly reduced to 2.3-fold when mice were treated with 250 mg/kg of the ethyl acetate fraction. GC-MS analysis of the ethyl acetate fraction revealed high percentages of 2-methyloctacosane, tetracosane, and decane. ConclusionThe fractions extracted from S. incanum leaves have been found to possess only antimalarial prophylactic properties, with the ethyl acetate extract fraction showing the most effective results. The activity of this fraction may be attributed to its ability to decrease the expression of δ-ALAD, as it contains an alkane compound implicated with enzyme-inhibitory activity.

Antimalarial Properties of Sida cordifolia L. Leaf Extract in Mice: Survivability Depends Less on Parasitaemia Suppression.

Sida cordifolia has been used to treat malaria in Ghana albeit without scientific evidence of antimalarial activity and safety. This work aimed to assess the antimalarial properties and acute toxicity of the aqueous leaf extract of S. cordifolia in murine models. Aqueous extract of the plant was analysed for both suppressive and curative antimalarial properties in chloroquine-sensitive ANKA strains of rodent Plasmodium berghei-infected mice. Acute toxicity evaluation was performed in rats according to the OECD 425 guidelines. The extract displayed antiplasmodial activity in vivo with ED50 of 117.49 ± 15.22 mg/kg and 144.84 ± 18.17 mg/kg in suppressive and curative studies, respectively. The highest % parasitaemia suppression exerted was 76.90 ± 0.64% and 61.50 ± 0.97%, respectively, in the suppressive and curative studies. Survival of infected mice treated with the extract was significantly prolonged. This was dependent on the dose of the extract but imperfectly related to the % parasitaemia suppression. Related antimalarial parameters including percentage hematocrit, changes in body weight, and temperature of experimental mice indicated alleviation of malarial symptoms of treated animals. The extract did not show toxicity in rats. Sida cordifolia L. has antimalarial properties, and was safe. It suppressed parasitaemia in both suppressive and curative studies, was not toxic to animals and prolonged the life of infected animals under treatment. This, therefore, justifies the traditional use of S. cordifolia for the treatment of malaria in Ghana.

Diverse mitochondrial effects, antiplasmodial and anti-inflammatory potentials of Costus afer (Ker Gawl), Nauclea latifolia (Sm) and Sphenocentrum jollyanum (Pierre) in mice infected with Plasmodium berghei

ObjectivesThis study reported the antiplasmodial, anti-inflammatory and mito-protrective effects of Costus afer (CA), Nauclea latifolia (NA) and Sphenocentrum jollyanum (SJ) methanol extracts in Plasmodium berghei-infected mice. MethodsAir-dried CA, NA and SJ were extracted with methanol. Antiplasmodial activity of these extracts were monitored using chloroquine-sensitive and resistant strains of Plasmodium berghei. Heme and hemozoin contents, interleukins and C-reactive protein as well as mitochondrial permeability transition (mPT) pore opening, lipid peroxidation (mLPO) and F0F1 ATPase activity were determined by spectrophotometry. Phytochemical constituents were determined using UPLC-QTOF-MS and NMR spectroscopy. Results and ConclusionsCA, NL and SJ decreased percentage parasitemia to 0.25 ± 0.07; 0.30 ± 0.14 and 0.25 ± 0.07% relative to control (8.60 ± 0.15%) in the chloroquine-sensitive model and to 0.40 ± 0.14; 0.30 ± 0.14 and 0.45 ± 0.07, respectively as against 10.88 ± 0.26% of the infected control in the chloroquine-resistant model. In chloroquine-resistant model, NL decreased mLPO (0.41 ± 0.04) F0F1 ATPase (0.15 ± 0.02 mmol pi/mg protein /min) while CA enhanced mPT pore opening at 100 mg/kg,and SJ (50 mg/kg) reversed parasite-induced mPT pore opening (1.66 vs 9.4 folds). The NL increased heme, decreased hemozoin, IL-6, CRP, TNF-α, while SJ dose-dependently increased IL-10. UPLC-QTOF-MS analysis showed that coumaric acid, divaricatinic acid, diocin and aferiosides A and C were present in CA, 3-caffeoylquinic acid, 18, 19-dihydroangustine, jatrorrhizine, 17-epinaucleidinal, strictosamide and quinovic acid 3-O-rhamnoside in NL and quinic acid, jatrorrhizine and mabioside B in SJ. While the three medicinal plants have varying antimalarial effects, their decoction will be better for a synergistic purpose.

In Vitro and in Vivo Antimalarial Activity, Cytotoxicity and Phytochemical HRMS2 Profile of Plants from the Western Pará State, Brazilian Amazonia.

Ethnopharmacology and botanical taxonomy are valid criteria used to selecting plants for antimalarial bioprospection purposes. Based on these two criteria, ethanol extracts of 11 plants from Santarém City vicinities, Western Pará State, Brazilian Amazonia, had their in vitro antiplasmodial activity against chloroquine-resistant Plasmodium falciparum (W2 clone) assessed by the PfLDH method, whereas their cytotoxicity to HepG2-A16 cells was assessed through MTT assay. Acmella oleracea, Siparuna krukovii and Trema micrantha extracts disclosed the highest rate of parasite growth inhibition (90 %) in screening tests. In vivo antimalarial assays were conducted with these extracts against Plasmodium berghei (NK 65 strain) infected mice. Inhibition rate of parasite multiplication ranged from 41.4 % to 60.9 % at the lowest extract dose (25 mg/kg). HPLC-ESI-HRMS2 analyses allowed the putative identification of alkylamides, fatty acids, flavonoid glycosides and alkaloids in ethanol extracts deriving from these three plant species. Results pointed towards A. oleracea flowers ethanol extract as the most promising potential candidate to preclinical studies aiming the development of antimalarial phytomedicine.

Antiplasmodial Profiling of Mangifera indica's Herbal Formulation and Its Ability to Ameliorate Hematological Derangements in Plasmodium berghei-Infected Mice

Background Malaria caused by Plasmodium falciparum with an average prevalence of 75% in Nigeria, can be treated. Chemical-based medications, one of the methods used to treat it, frequently causes hematological abnormalities in the blood tissues. However, the ethnobotanical and scientific usage of Mangifera indica for the management of malaria in Nigeria necessitated this study on the antiplasmodial evaluation, the hematological profiling of Plasmodium berghei-infected mice treated with Mangifera indica's herbal formulation as well as the isolation and characterization of bioactive compounds from the plants. Method The plant's coarse leaves (3 kg) and stem bark (3.2 g) were weighed, divided into groups of leaves (250.71 g), stem bark (509.34 g), leaves/stem bark (1:1) (101.24/101.24 g), leaves/stem bark (1:2) (164.39/328.78 g), and leaves/stem bark (2:1) (218.50/109.25 g), macerated in 3 L of ethanol for 72 h, filtered, and concentrated. For each extract, the following weights and yields were recorded: 31.3 g leaves (15.2%), 43.8 g stem bark (8.7%), 34.8 g leaves/stem bark (1:1) (17.2%), 29.2 g leaves/stem bark (1:2) (5.92%), and 26.1 g leaves/stem bark (2:1) (7.96%). According to established methods, the herbal formulation was employed for hematological testing, GC-MS analysis, and in vivo antiplasmodial evaluation based on standard protocols. Results The leaves: Stem bark (1:2) extract (the best therapeutic response) of 98.92% was better than ACT (98.63%). GCMS analysis revealed predominantly mangiferin and esters of linoleic acids that could have enhanced erythropoiesis, mitigated infections, as well as boosted platelet aggregation in P berghei-infected mice as observed from the hematological assay. Conclusion Therefore, the extract of leaves: stem bark (1:2) had the best antiplasmodial therapeutic response and hematoprotective activity in P berghei-infected mice.

Therapeutic Potentials of Azadirachta indica and Ocimum gratissimum, and their Synagistic Effects in Treatment of Plasmodium berghei Infected Albino Mice

Antimalarial resistance is a major challenge for effective control of malaria. This triggered the need to monitor the efficacy of Azadirachta indica and Ocimum gratissimum in the disease management. The present study compared antimalarial activities of the leaf extracts of the individual plants and evaluated their combined effects on malaria infected mice. The leaves of the different plants were extracted with absolute ethanol (BDL 95%) for the test. Clean albino mice were experimentally infected intraperitoneally with chloroquine-sensitive Plasmodium berghei Nk65 strain. Parasitaemia level was determined before parasite inoculation and at 24 hours post treatment period. Efficacy of the leaf extracts was tested on the infected mice using Peter’s 4-days suppressive and curative tests, and secondary biological assessment procedures.The lethal median dose (LD50) recorded for neem and clove basil leaf extracts were 31.62 and 1246.9 mg/kg body weight, respectively. Infected mice treated with leaf extracts of the plants and their combinations produced significant dose- dependent activity against the parasite (P&lt;0.05). Highest reduction of parasitaemia was observed on day 4. Maximum parasitaemia reduction (78.65%) was attained with 30mg/kg of the combination of the extracts on the 7th day. These observations indicate better anti-malarial activity of the combination therapy as compared with the individual extracts of the plants, and also show their good antimalarial potentials.

Antimalarial Effects of Ethanolic Leaf Extracts of Azadirachta indica and Ocimum gratissimum, and Their Histologic Effects on Some Organs (Liver, Kidney and Heart) of Plasmodium berghei Infected Albino Mice

Azadirachta indica (Neem) and Ociumum gratissimum (clove Basil) have long been employed locally for the management of malaria. The present study compared antimalaria activities of the ethanolic leaf extracts of the individual plants, and assessed their combined effects on some organs of malaria-infected mice, at the Parasitology and Histopathology units, Federal Medical Centre (FMC), Owerri, from January to March, 2021. The leaves of the different plants were extracted with absolute ethanol (BDL 95%) for the test. Clean albino mice were experimentally infected intraperitoneally with chloroquine-sensitive Plasmodium berghei NK65 strain. Parasitaemia level was determined before parasite inoculation and at 24 hours post treatment period. Histopathological study on the liver, kidney, and heart was carried out using the Paraffin Setions method. Extracts of the leaves were administered orally, while chloroquine administration was intramuscular. The efficacy of the leaf extracts was tested on the P. berghei infected albino mice using the 4-day curative test. The lethal median dose (LD50) recorded for neem and clove basil leaf extracts were 31.62 and 1246.9 mg/kg body weight, respectively. Significant activity against the parasite was produced by infected mice treated with extracts of A. indica and O. gratissimum, and their combinations throughout the treatment period (P&lt;.05). Highest reduction of parasitaemia was observed on day 4. Maximum parasitaemia reduction (78.65%) was attained with 30mg/kg of the combined extracts on the 7th day. Mild pathological lesions were observed in mice treated with A. indica leaf extract. These observations indicate better anti-malaria activity of the combination therapy as compared with the individual extracts of A. indica and O. grassimum, and indicate good antimalarial and protective roles of the plant extracts on the parasitized mice at large, as it slows down development of resistance.

Comparative Study of Pharmaceutical, Herbal Medicine, and Low-Value Alternating Electromagnetic Radiation as Anti-Malaria Treatment

Malaria is a significant global health burden, particularly in regions with limited healthcare resources. Pharmaceutical interventions, specifically artemisinin-based combination therapies (ACTs), have long been the mainstay of malaria treatment due to their proven efficacy in eliminating Plasmodium parasites. Herbal medicine has also gained attention as a potential alternative or adjunctive therapy. Nevertheless, the emergence of drug-resistant strains, notably in Southeast Asia and Africa, poses a considerable challenge to the effectiveness of these pharmaceutical treatments. Using low-value alternating electromagnetic field (EMF) radiation as an anti-malaria treatment is a novel approach that requires further investigation to determine its therapeutic potential. This research studies the comparative analysis of the effect of pharmaceutical intervention, herbal medicine, and low-value alternating EMF as anti-malaria treatments. The research methodology involves a systematic literature review of relevant scientific databases, design, construction, and administration of 20mT &amp; 30mT electromagnetic field, Lumartem and Thitonia Diversifolia (Jume 12) to plasmodium berghei-infected mice for four days consecutively at the same time interval. Results revealed that all treatments were significantly effective. Lumartem is maximally effective from the third day, while Tihonia Diversifolia was maximally effective on the second day. EMF 10mT was maximally effective on the third day, while EMF 20mT did not fully align with the non-infected mice trend, but it is likely to align if treatment continues. The results of this comparative study will contribute to the body of knowledge regarding the effectiveness of pharmaceutical, herbal medicine, and low-value alternating electromagnetic field treatments for malaria.

FORMULATION AND CHARACTERIZATION OF GDL-BASED ARTESUNATE SOLID LIPID NANOPARTICLE

Objective: The present research aimed to prepare and characterize glyceryl dilaurate (GDL) containing solid lipid nanoparticles (SLN) with tween 80 and lecithin as an emulsifier in which the artesunate drug was loaded. Methods: SLNs were synthesized by solvent emulsification–diffusion technique. The formulation was characterized for size, size distribution, zeta potential, shape and morphology, and DSC. In vitro drug release studies were performed at pH 5.0 and pH 7.4 to mimic in vivo conditions. Hemolytic studies and In vitro, antiplasmodial activities were carried out. Plasmodium berghei (NK65 resistant strain) infected mice were used to test the in vivo antimalarial efficacy of SLN. Results: SLN exhibited 327 nm average sizes with 0.164 PDI and-23.4 mV zeta potential. TEM images revealed a spherical structure. The entrapment efficiency of the ART was calculated as 85.68%. In vitro, drug release studies showed that entrapped drug was released in a weakly acidic environment (83.45% for ART). Hemolytic studies revealed that ART-GDL conjugate was stable and safe for parenteral delivery. IC50 value of the GDL-based ART-SLNs was calculated to be 0.32 µM. Furthermore, the GDL-based ART-SLNs resulted in enhanced parasite killing in P. berghei-infected mice and improved survivability as compared to free ART administration. Conclusion: The present research allows safe and effective intravenous administration of artesunate. Thus GDL-Based ART-SLNs could be a potential drug delivery system for antimalarial therapy.

Evaluation of the Efficacy of Artemether/Lumefantrine/Doxycycline Combination against Plasmodium berghei in Mice

Background: Malaria is common in regions like sub-Saharan Africa and South East Asia, where it results in millions of mortalities per year. Resistance to currently available antimalarials further compounds the problem. Drug combination has been demonstrated to be effective in overcoming resistance to antimalarials. This study aimed to assess the efficacy of the combination of Artemether/Lumefantrine (AL) and Doxycycline (DX) against Plasmodium berghei infection in mice. Methods: Thirty Adult Swiss albino mice were inoculated with the chloroquine (CQ)-sensitive P. berghei NK65 strain. The curative test involved oral administration of any of DX, AL, AL/DX, or Chloroquine (CQ) to mice with established infections, while the suppressive test was performed by treating mice shortly after parasite inoculation. Parasitemia levels were monitored using thin smears stained with Giemsa solution. Mean Survival Time (MST) was determined for each group. Results: In the curative test, DX, AL, and AL/DX exhibited parasitemia inhibitions of 81.4%, 94.5%, and 93.2%, respectively, while CQ achieved 93.5% inhibition. MST was significantly extended by DX, AL, and AL/DX treatments compared to negative control (NC). In the suppressive test, parasitemia inhibitions were 79.9%, 96.1%, 95.5%, and 94.9% for DX, AL, AL/DX, and CQ, respectively. The administration of DX, AL, and AL/DX also significantly prolonged MST compared to NC. Conclusion: The study demonstrates that the drug combination, particularly AL/DX, exhibits significant antimalarial effects in both curative and suppressive tests. The combination treatments led to substantial reductions in parasitemia levels and extended mean survival time, indicating their potential as effective antimalarial strategies. These findings highlight the promising role of drug combinations in addressing malaria infections and suggest avenues for further investigation and development of combination therapies.

Prophylactic effects of probiotic bacterium Latilactobacillus sakei on haematological parameters and cytokine profile of mice infected with Plasmodium berghei ANKA during early malaria infection

Malaria is a deadly parasitic disease caused a by protozoan parasite of the genus plasmodium. The challenges facing by chemotherapy and vector control couple with the lack of vaccine against malaria necessitate an urgent need for the development of alternative treatment regimens to combat this disease. One possible antimalarial treatment regimen is the use of probiotic bacteria as dietary supplements. Traditionally fermented milk is a rich source of probiotic bacteria that up to date, very few studies have been carried out on their immunoprotective effects against early malaria infection in mice. This study sought to assess the prophylactic activities of a probiotic bacterium Latilactobacillus sakei on malaria and inflammation in Plasmodium berghei infected mice. The probiotic bacterium was isolated from the Fulani's traditionally fermented milk and identified using the sequencing of the 16S r RNA gene. The repository activity of L. sakei on malaria was assessed using the method described by Peters with slight modification. Eighty-four BALB/c mice were randomly divided into two sets of seven groups of six mice each. One set received orally different doses of L. sakei Chloroquine and Sulfadoxine/Pyrimethamine for seven days before infection while the other set received for fourteen days before infection with 0.1 mL of 107Plasmodium berghei. Parasitaemia density, haematological parameters and inflammatory cytokines profile were evaluated. Data were presented as Mean ± SEM and analysed using SPSS version 20.0. The results of this study revealed that L. sakei significantly (p < 0.05) reduced in dose dependent manner parasite load, body weight loss and reduction of body temperature in all the treated mice when compare to untreated mice. Leukocytopenia, thrombocytosis and inflammation were also found to be significantly (p < 0.05) prevented in treated mice as compared to untreated mice.This study suggested that L sakei possesses immunomodulation and protective effects on early malaria infection in Plasmodium berghei mice.