Pharmacological Efficacy of Ethanol Leaf Extract of Justicia secunda in Swiss Albino Mice Experimentally Infected with Plasmodium berghei

Pharmacological Efficacy of Ethanol Leaf Extract of Justicia secunda in Swiss Albino Mice Experimentally Infected with Plasmodium berghei | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Pharmacological Efficacy of Ethanol Leaf Extract of Justicia secunda in Swiss Albino Mice Experimentally Infected with Plasmodium berghei Nasiru Ademola ADEYEMI, Sonnie ONIYE, Peter Ibrahim REKWORT, Abdullateef YUSUF, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3879871/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 26 Aug, 2024 Read the published version in International Journal of Applied Biology and Pharmaceutical Technology → Version 1 posted You are reading this latest preprint version Abstract Background Plasmodium parasite that causes malaria has developed resistance to most antimalarial medications and this has prompted research on natural products. The plant, Justicia secunda , is domesticated in the tropical region of Africa, and is used for the treatment of anaemia and other debilities. We evaluated the antiplasmodial and immunomodulatory activity of ethanol leaf extract of J. secunda in mice experimentally infected with Plasmodium berghei . Result Ethanol leaf extract of Justicia secunda contains steroids, tannins, flavonoids, terpenoids, phenols, glycosides and carbohydrate as well as thirty (30) compounds with numerous pharmacological activities. The LD 50 was above 5,000mg/kg with no mortality. A dose-dependent percentage suppression of P . berghei was observed in the curative test for the three doses respectively (53.13%, 60.71%, 71.21%). The suppressive test for ethanol leaf extract of J. secunda also showed a similar trend (52.2%, 62.16%, 75.18%). The ethanol leaf extract of J . secunda effectively prevented anaemia, reduced the level of ALT enzyme and also increased the level of TNF-α and IL-10 in the plasma of P. berghei -infected mice. It also significantly reduced the oxidative stress and vascular congestion in the liver of mice infected with P . berghei . Conclusion This study revealed that ethanol leaf extract of J. secunda could be utilized to treat infection caused by Plasmodium parasite and also ameliorate the pathogenesis of the disease. Antiplasmodial Phytochemicals Immunomodulatory Histopathology Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1 Background Malaria is more prevalent in African nations than it is worldwide. According to the World Malaria Report 2020; Nigeria (31.3%), the Democratic Republic of Congo (12.6%), the United Republic of Tanzania (4.1%) and Niger (3.9%) account for more than half of all malaria death [ 1 ]. Although, sociopolitical issues are connected to the causes and consequences of malaria disease [ 2 ]; explicitly, the major contributing factor to the prevalence of malaria is the lack of mosquito protection and appropriate malaria treatment [ 3 ]. Malaria is an infectious disease affecting humans and animals [ 4 ]. The etiologic agent of the disease is the parasitic protozoans of the genus Plasmodium , and the sole biological vector is the female Anopheles mosquitoes [ 5 ]. The asymptomatic human is an unwitting parasite reservoir, allowing continuous transmission [ 6 ]. The initial clinical symptom of this illness leads to headaches, fatigue, soreness in the joints and muscles, vomiting, and anorexia. Acidemia, hypoglycemia, anaemia, pulmonary oedema, coma and renal failure are common signs of acute malaria [ 7 ]. Antimalarial drug resistance occurs in the malaria parasite due to ineffective chemoprophylaxis [ 8 ], fake and irregular pharmaceutical administration leading to genetic mutation [ 9 ]. Therefore, searching for a new anti-malarial compound is a continuous process, and numerous studies on African medicinal plants with anti-malarial potentials have been conducted over time. However, there is a dearth of practical information regarding the pharmacological efficacy of J. secunda as an antimalarial agent. Justicia secunda is taxonomically classified in the family Acanthaceae and is notably utilized for treating anaemia in tropical regions of Africa [ 10 ]. It is commonly known as 'Christ blood' and 'Blood herb' in Nigeria. The herbaceous plant has green leaves and pink inflorescence (Fig. 1 ) [ 11 ]. According to Irinmwinuwa and Afonne [ 12 ], the ethanol leaf extract of J. secunda contains saponin (9.2%), tannins (9.0), flavonoids (7.0%) and alkaloid (2.4%). It has also been established that J . secunda has hematinic effect in rats with anaemia induced by phenylhydrazine [ 13 ]. Likewise, it exhibited anti-inflammatory and antioxidant activity in an in vitro study using heat-induced bovine serum albumin (BSA) denaturation and erythrocyte membrane stabilization assay [ 14 ]. This study evaluates the phytochemical constituent and lethal dose of the ethanol leaf extract of J. secunda , as well as the effect of the extract on the parasitaemia level, hematological profile, pro-inflammatory (TNF-α) and anti-inflammatory (IL-10) cytokines, liver biochemicals, and histopathology of the liver in Plasmodium berghei NK65-infected mice. 2 Methods 2.1 Collection of Plant Sample Matured leaves were harvested from stands of J. secunda plants from three different sites in Lusada, Ado Odo Ota Local Government Area (LGA) Ogun State, Nigeria. 2.2 Preparation of Plant Extract Cold maceration technique was adopted for the extraction by simply soaking five hundred grams (500g) of the powdered leaves in five (5) litres of eighty percent (80%) ethanol for three days, and the bottle containing the mixtures was agitated at intervals. The liquid extract was filtered into a clean container using a piece of chiffon material after 72 hours. The filtrate was poured into an evaporating dish and placed on a water bath to obtain a solid and more concentrated form of the extract [15]. 2.3 Evaluation of Phytochemical Constituent of Plant Extract A standard method [16] was adopted in the phytochemical screening of the extract. The Gas Chromatography-Mass Spectrometry (GC-MS) analysis was conducted using a 7890B GC System (Agilent Technologies, USA) coupled with a 5977A Mass Selective Detector (Agilent Technologies, USA). The leaf extract of J. secunda was diluted in methanol, and 2μL of the mixture was injected into the GC-MS machine using a micro-syringe. The experimental parameters for the GC-MS system were: initial oven temperature: 70 o C, Equilibration Time: 1 min, Max Temperature: 325 ̊C, Slow Fan: Disabled, Oven Program: 5°C/min On 250°C for 1 min, #1 then 30°C/min to 300°C for 0 min, Post Run: 50 °C. Cryo: Off, Front SS Inlet He – Mode: Split. Heater: On 250°C, Pressure: On 11.089psi, Total Flow on 19.204mL/min, Septum Purge Flow: On 3 mL/min, Gas Saver: off, Purge flow to Split vent: on 15mL/min at 0.75min. Holdup time: 1.2386min, Flowrate: 1.0mL/minute 2.4 Maintenance of Experimental Animals Seventy-five (75) male Swiss albino mice with body weight between 20 and 30g were obtained for this study. Chicken grower’s marsh and water was provided at all time for the experimental mice [17]. The overall maintenance of the experimental animals was in accordance with the procedures approved by the ethical committee of the institution where the experiment was conducted. 2.5 Determination of Lethal Dose of Plant Extract The median lethal dose of the ethanol leaf extract of J. secunda was determined per the Up and Down method described by the Organization for Economic Co-orporation and Development [18]. 2.6 Evaluation of Antiplasmodial Activity of Plant Extract The infected experimental mice were inoculated intraperitoneally with 0.2mL of blood from donor mouse infected with P. berghei NK65 (30% parasitaemia) and normal saline (1:9) containing 10 5 P. berghei parasitized erythrocytes. The seven-day curative test was evaluated seventy-two (72) hours after inoculation and confirmation of parasitemia in thirty (30) mice, and they were randomly assigned into six (6) groups. The suppressive test was conducted, and the inception was precisely three (3) hours after the infection of mice for a four-day duration using thirty (30) mice. These tests were performed using the method of Onyegeme-Okerentaa et al . [19], with a slight modification. Groups 1–3 were the control groups treated with 1 mL of normal saline (the negative control), 25 mg/kg of chloroquine (the positive control), and the uninfected and untreated groups (the normal control), respectively. Groups 4-6 were given 500 mg/kg, 1000 mg/kg, and 1,500 mg/kg of the crude extract based on the LD 50 for the ethanol leaf extract of J. secunda determined in this study. The oral treatment was done once a day. 2.7 Collection of Blood and Liver Samples The animals were sacrificed after inhaling vapour from ketamine-damped cotton wool at the end of the experiment. The blood of experimental animals was collected in EDTA bottles for haematological and biochemical analysis through cardiac aspiration. A portion of the liver was placed in cold phosphate buffer for antioxidant and MDA analysis. A small portion of liver of mice was preserved in phosphate-buffered formalin. 2.8 Determination of Parasitaemia and Percentage Chemosuppression The number of parasitized erythrocytes in ten (10) slides prepared with thin blood smears was counted, and the average was computed to give the parasitemia of each mouse [20]. Percentage parasitaemia and chemosuppression was calculated as: Parasitaemia (%) = (Number of parasitized RBC/Total number of RBC) x 100 A= mean % parasitaemia in negative control B= mean % parasitaemia in treated group 2.9 Assessment of Haematological Parameters The Red Blood Cell (RBC), White Blood Cell (WBC), Packed Cell Volume (PCV) and Haemoglobin (HGB) concentration were analyzed using a Biobase Bk6100 haematology analyzer [21] in blood samples collected in Ethylene Diamine Tetracetic Acid (EDTA) bottles. 2.10 Assessment of Plasma and Liver Biochemicals The liver homogenate was used to evaluate the levels of Malondialdehyde (MDA), Superoxide dismutase (SOD), Catalase (CAT) and reduced Glutathione (GSH) using a Sigma Aldrich Assay kit. Blood in EDTA bottles was used to check the concentrations of Alanine aminotransferase (Randox kit), Interleukin-10 and Tumour Necrosis Factor-α using ELISA kits by FineTest. 2.11 Liver Histopathological Studies The liver of mice preserved in phosphate-buffered formalin was prepared for histological study using standard histotechnique and viewed with a microscope at 400 × magnification. The photomicrographs were taken using a digital camera [22]. 2.12 Data Analyses One-way Analysis of Variance (ANOVA) was used to determine statistical differences in the antiplasmodial activity, haematological and biochemical parameters in the in vivo suppressive and curative test. Tukey HSD test was used to rank the difference between significant parameters at p <0.05. Data were represented in tables and summarized in form of mean ± standard error. Statistical analyses were done using 'R' statistical package (version 4.1.2) for windows. 3 Results 3.1 Phytochemical Characterization and Acute Toxicity The phytochemical screening detected eight different classes of phytochemicals (Table 1 ), and the GC-MS technique confirmed the presence of thirty different compounds, as shown in Table 2 . Treatment of mice with limit dose of ethanol leaf extract of J. secunda (5000mg/kg) caused no mortality and signs of overtoxication. This suggests that the LD 50 of the extract is greater than 5000mg/kg. Table 1 Phytochemical screening of ethanol leaf extract of Justicia secunda S/No Phytoconstituent Inference 1. Alkaloids + 2. Cardiac glycosides + 3. Saponin - 4. Phenolic compounds + 5. Tannins + 6. Steroids + 7. Carbohydrates + 8. Flavonoids + 9. Terpenoids + 10 Anthraquinones - Keys: (+) = Present, (-) = Absent Table 2 GC-MS profile of ethanol leaf extract of Justicia secunda Peak Retention Time (Minute) Library I.D Peak area (%) Chemical Abstract Service Number 1 17.4652 17-Pentatriacontene 0.1691 006971-40-0 2 17.64 Oxalic acid, cyclobutyl heptadecyl ester 0.2012 1000309-70-7 3 20.0166 Hexadecanoic acid, methyl ester 2.6316 000112-39-0 4 21.4568 3-Chloropropionic acid, heptadecyl ester 0.2485 1000283-05-1 5 23.0506 9,12-Octadecadienoic acid, (linoleic) methyl ester, (E,E)- 1.1048 002566-97-4 6 23.2025 cis-13-Octadecenoic acid, methyl ester 6.3996 1000333-58-3 7 23.7984 Methyl stearate 0.8041 000112-61-8 8 25.1498 Carbonic acid, but-3-en-1-yl hexadecyl ester 0.1945 1000383-23-7 9 25.279 Aspidospermidin-17-ol, 1-acetyl-19,21-epoxy-15,16-dimethoxy- 0.2787 002122-26-1 10 28.5241 8-Hexadecenal, 14-methyl-, (Z)- 1.1543 060609-53-2 11 26.5546 Cyclododecane 1.5526 000294-62-2 12 26.7618 9,12-Octadecadienal 0.6438 026537-70-2 13 26.8969 p-Menth-8(10)-en-9-ol, cis- 0.1951 015714-13-3 14 26.9881 Oxirane, [(dodecyloxy)methyl]- 0.7356 002461-18-9 15 27.3173 Oleic Acid 2.1944 000112-80-1 16 27.7978 22-Stigmasten-3-one 3.3243 004736-95-2 17 28.2861 (S)(+)-Z-13-Methyl-11-pentadecen-1-ol acetate 3.9133 1000130-84-8 18 28.5241 8-Hexadecenal, 14-methyl-, (Z)- 1.2379 060609-53-2 19 28.9265 3-Cyclohexylthiolane,S,S-dioxide 1.317 071053-08-2 20 28.9874 cis-7, cis-11-Hexadecadien-1-yl acetate 1.3175 052207-99-5 21 29.3867 cis-9-Hexadecenal 0.7726 056219-04-6 22 29.8609 9-Octadecenoic acid (Z)-, 2,3-dihydroxypropyl ester 3.1331 000111-03-5 23 30.1094 Z-8-Methyl-9-tetradecenoic acid 3.1701 1000130-84-5 24 30.2394 1-Nonadecene 1.2528 018435-45-5 25 30.394 Cyclododecanol, 1-ethenyl- 5.5624 006244-49-1 26 30.9115 Cyclododecane, ethyl- 3.3424 028981-49-9 27 31.2385 2-Methyl-Z,Z-3,13-octadecadienol 3.9236 1000130-90-5 28 31.5878 E-9-Tetradecenal 0.9144 1000131-35-7 29 32.575 β-Sitosterol 19.7234 000083-46-5 30 34.706 Squalene 4.1521 000111-02-4 3.2 Antiplasmodial Activity of the Crude Extract The crude extract was tested for curative and suppressive antiplasmodial activity as presented in Table 3 and the result was expressed in the form of percentage parasitaemia and percentage chemosuppression for the infected controls and the treatment groups. The extract significantly ( p < 0.05) reduced parasitaemia level in infected mice for both curative and suppressive tests compared with the negative control. A dose-dependent chemosuppression was observed for the experimental groups, which was lower than the standard drug with 100% chemosuppression. Table 3 Antiplasmodial activity of ethanol leaf extract of Justicia secunda against P . berghei in mice Curative test Suppressive test Experimental groups % Parasitaemia % Chemo-suppression % Parasitaemia % Chemo-suppression C- 44.8 ± 1.61 a 0.00 41.5 ± 2.40 a 0.00 C+ 0.00 c 100 0.00 c 100 NC 0.00 0.00 0.00 0.00 T1 21.0 ± 1.29 b 53.13 19.8 ± 3.70 b 52.2 T2 17.6 ± 0.959 b 60.71 15.7 ± 1.46 b 62.16 T3 13.9 ± 2.81 b 71.21 10.3 ± 2.67 b 75.18 p -value 0.00 0.00 NOTE: Data are expressed as mean ± SEM, n=5 Statistical analysis performed amongst Negative control (C-), Positive control (C+), Normal control (NC), 500mg/kg(T1), 1000mg/kg(T2) and 1,500mg/kg(T3) dose of ethanol leaf extract of J . secunda , means with different superscript within a column are statistically significant at p <0.05 3.3 Haematological Profile of Experimental Mice The haematological profile of mice measured at the end of the experiment shows that the extract significantly ( p < 0.05) prevented the reduction in Packed Cell Volume (PCV), Red Blood Cell (RBC) and Haemoglobin (HGB) for both curative and suppressive test as shown in Fig. 2 . The group of mice that received chloroquine (4.26 ± 0.189) had a statistically ( p < 0.05) reduced White Blood Cell (WBC) than and the negative control (5.76 ± 0.356) and experimental groups treated for seven days (4.8 ± 0.362, 4.72 ± 0.252, 4.76 ± 0.218). 3.4 Blood Plasma Biochemicals of Experimental Mice The immunomodulatory and hepatoprotective potentials of the plant extract was assessed by analysing the biochemical components of plasma such as TNF-α, IL-10 and ALT. The outcome of this biochemical assay for the therapeutic and inhibitory test is presented in Fig. 3 . The infected mice treated with ethanol leaf extract of J. secunda had significantly higher ( p < 0.05) concentrations of TNF-α and IL-10 compared to negative control for both curative and suppressive test. Furthermore, the plasma concentration of ALT in negative control was significantly higher ( p < 0.05) than groups that received plant extract treatment. 3.5 Antioxidant and Lipid Peroxidation Status of the Liver Figure 4 shows the liver’s antioxidant and lipid peroxidation profile of experimental mice for the curative and suppressive test. The groups treated with plant extract exhibited a significant ( p < 0.05) increase in the bioactivity of antioxidants, including SOD, CAT, and GSH, compared to the negative control. The SOD concentration was, however, significantly ( p < 0.05) lower in the extract-treated group (3.32 ± 0.447u/ml, 3.48 ± 0.526u/ml, 3.47 ± 0.579u/ml) and negative control (3.38 ± 0.481u/ml) than in the normal control (7.26 ± 0.462u/ml) and positive control (6.84 ± 0.331u/ml). In addition, the hepatic malondialdehyde (MDA) concentrations were significantly ( p < 0.05) reduced in treatment groups compared to the negative control. 3.6 Liver Histology of Experimental Mice The liver of negative control shows mass vascular congestion when compared to liver sections of other groups for the curative test (Fig. 5 ). The positive control and normal control have normal architecture of a liver photomicrograph, while the 500mg/kg, 1000mg/kg and 1,500mg/kg have a moderate hepatic necrosis, moderate vacuolation and slight Kupffer cell hyperplasia respectively. Comparing the histological sections of the treatment groups to those of the negative control group for the suppressive test (Fig. 6 ), the treatment groups showed slight vacuolation and necrosis (500 mg/kg), slight Kupffer cell hyperplasia (1,000mg/kg and 1,500 mg/kg) and the negative control had slight vascular congestion. Also, the liver photomicrographs of the positive and normal control exhibit normal features in their hepatic histology. 4 Discussion Eight (8) different secondary metabolites were present in the ethanol leaf extract of J. secunda namely: alkaloids, cardiac glycosides, phenolic compounds, tannins, steroids, carbohydrates, flavonoids and terpenoids. In addition, several phytochemical compounds with antioxidant, anti-inflammatory and anti-plasmodial activity were detected in the extract. For instance, β-sitosterol has anti-inflammatory properties [ 23 ], Squalene has antiparasitic and anti-inflammatory properties [ 24 ], 2-Methyl-Z,Z-3,13-octadecadienol has antimicrobial properties [ 25 ], 17-Pentatriacontene has anti-inflammatory properties [ 26 ], hexadacenoic acid methyl ester has anti-inflammatory properties [ 27 ], 9,12-Octadecanoic (linoleic) acid methyl ester has anti-plasmodial and anti-inflammatory [ 28 ], Methyl stearate has anti-inflammatory properties [ 24 ], cis-13-Octadecenoic acid, methyl ester has anti-inflammatory activity [ 29 ], 16-dimethoxy-aspidospermidin-17-ol has anti-plasmodial activity [ 30 ], 8-Hexadecenal 14-methyl-, (Z)- has antioxidant activity [ 31 ], Oleic acid has antioxidant activity [ 32 ], cis-9-Hexadecenal has anti-inflammatory properties [ 33 ] and cyclododecane, ethyl has antioxidant activity [ 34 ]. These myriads of phytochemicals may have conferred on this plant its popularity and wide use in African traditional medicine [ 35 ]. The phytochemical screening contrasts with the findings of a study conducted on a similar extract of J. secunda leaves collected from a farm in south-east Nigeria that is devoid of steroids and glycosides [ 36 ]; however, hexadecenoic acid methyl ester and octadecanoic acid methyl ester was also detected in extract of J. secunda [ 37 ]. The difference in phytochemical constituents in both studies could be attributed to ecological variation [ 38 ]. According to Hodge and Sterner's [ 39 ] toxicity scale, this plant extract is practically non-toxic. A study [ 40 ] that processed the leaves of J . secunda for three weeks and macerated them in ethanol for 24 hours reported an LD 50 of 3,800 mg/kg body weight in rats due to the cyanide content of the extract. It is very likely that the selection of mature leaves and the longer processing duration of the extract for this present study may be reasons for the reduced toxicity observed [ 41 ]. One of the major foci of this study is to include Justicia secunda in the record of antimalarial study, and the crude extract has a percentage chemosuppression of 71.21% (1,500 mg/kg) and 75.18% (1,500 mg/kg) in the suppressive and curative tests, respectively. The result is less than the antimalarial activity of the ethanol leaf extract of Justicia carnea (800 mg/kg dose) with a percentage chemosuppression of 82% [ 42 ]. The efficacy of plant extracts in treating malaria disease depends on several factors, such as the Plasmodium species, the dosage, the duration of treatment, and the host immune response [ 43 ]. Experimental research on 16-dimethoxy-aspidospermidin-17-ol [ 31 ] and linoleic acid-methyl esters [ 29 ] suggest that these phytochemicals inhibit the growth of Plasmodium parasites by binding with the Fab-I enzyme responsible for fatty acid biosynthesis. This is to create awareness that J. secunda could be an alternative source of antimalarial compounds. The ethanol leaf extract of J. secunda prevented PCV, HGB, and RBC reduction in mice, but there was an increase in WBC in the curative test. A different study observed similar trend for these haematological parameters with a normal WBC in curative and suppressive tests on the antioxidant and antimalarial activity of the ethanol stem bark extract of Terminalia macroptera in P. berghei- infected mice [ 44 ]. The haematinic properties of the extract in this study may be due to the presence of flavonoid compounds that have been reported to increase iron absorption and deposition in tissues while reducing iron excretion [ 45 ]. In addition, many antimalarial herbal preparations may exert their anti-infective activity not only by affecting the parasite directly but may also stimulate the defensive system of the host through many other mechanisms [ 46 ]. The assessment of modulatory and hepatoprotective potentials reveals that TNF-α and IL-10 increased and the concentrations of ALT reduced in treatment groups. This outcome corroborates the TNF and IL-10-increasing potentials observed in P. berghei- infected mice treated with the ethyl acetate leaf extract of Sonchus arvensis [ 47 ] and the hepatoprotective nature of J. secunda in reducing plasma ALT in rats administered carbon tetrachloride [ 48 ]. Infection and immunological reactions to the extract may be reasons for the increased level of TNF-α in treated mice. The anti-inflammatory activity of β-sitosterol was harnessed in reducing inflammation in Zebra fish [ 49 ] and the presence of this phytochemical in the extract utilized for this research may be the reason for the observed pharmacological effect. This is an indication that J. secunda may be utilized for treatment associated with pathogenesis of malaria infection. The liver homogenate of the experimental groups sustained an increased bioactivity of antioxidants (SOD, GSH, and CAT). Yet, a reduced level of SOD was observed in mice treated days after infection. The lipid peroxidation biomarker (MDA) is equally reduced in mice treated with crude extract. Hepatic antioxidant and MDA concentrations follow the same pattern as those of experimental mice treated with an extract of Croton membranaceus but differ with an increased SOD level [ 50 ]. The infection also caused a reduction in hepatic SOD in mice infected with P. berghei and treated with a stem-bark extract of Terminalia macroptera [ 51 ]. Justicia secunda exhibited antioxidant properties in this study, which may be attributed to components of the extract with the relevant pharmacological activity [ 49 , 52 , 53 ]. The ethanol leaf extract of J. secunda reduced vascular congestion in the liver of mice, as the photomicrograph of the treatment groups only shows sections with Kupffer cell hyperplasia, vacuolation, and hepatic necrosis. Ibukunoluwa [ 54 ] also reported similar histopathology in the antiplasmodial activity of polyherbal mixtures. The slight changes seen in the liver when used to test the efficacy of the plant extract may be due to the induced infection and the activation of immune cells in the organs. Ayawa et al. [ 55 ] also advanced cellular damage by immunological reactions as a reason for histopathological changes in the liver of mice. Pure compounds should be isolated from J . secunda extract to conduct trials on their pharmacological activity in relation to malaria infection and this could potentially uncover another effective antimalarial compound. 5 Conclusion In conclusion, the ethanol leaf extract of J . secunda contains phytochemicals with numerous pharmacological activities, and is non-toxic at 5,000mg/kg. It exhibits a dose-dependent percentage of chemosuppression of P . berghei at 500mg/kg, 1,000mg/kg and 1,500mg/kg. The extract prevented the reduction of the studied haematological parameters such as RBC, HGB and PCV. The concentrations of TNF-α and IL-10 increased and ALT concentrations reduced in the plasma of mice treated with extract. Evaluation of liver biochemicals revealed that MDA concentration was reduced with increased bioactivity of the studied antioxidants for the infected mice treated with the plant extract. Vascular congestion was also reduced in the liver histology of mice infected and treated with the plant extract but shows increased infiltration of immune cells and necrosis. This present study infers that ethanol leaf extract of J. secunda contains phytochemicals that have been tested individually to treat malaria and inflammation, which can be isolated for treating malarial pathogenesis. Abbreviations ALT Alanine aminotransferase MDA Malondialdehyde SOD Superoxide dismutase CAT Catalase GSH Reduced Glutathione IL-10 Interleukin-10 TNF-α Tumour Necrosis Factor-alpha HGB Haemoglobin PCV Packed Cell Volume RBC Red Blood Cell WBC White Blood Cell Declarations Ethical approval Maintenance and procedures performed involving animals in this study follow the ethical standards of Ahmadu Bello University, Zaria Committee on Animal Use and Care (ABUCAUC) with the approval number ABUCAUC/2023/055. Consent for publication Not applicable Availability of data and materials All data generated or analyzed in this study are included in this article. Competing interest None Funding None Authors’ contributions N.A.A, S.J.O, P.I.R, A.Y., and Y.A.W conceptualized the study, conducted the investigation and formal analysis. N.A.A and M.H. prepared figures and tables. All authors reviewed the manuscript. Acknowledgements A sincere gratitude to the Department of Pharmacology, Ahmadu Bello University (ABU), Zaria for providing the animal house and Swiss albino mice used for this research, Department of Pharmacognosy and Drug Development ABU, Zaria for providing the facility to achieve the extraction of the plant and the phytochemical screening, the Multi-User Science Research Laboratory ABU, Zaria for the Gas Chromatography-Mass Spectrometric analysis of the plant extract and the Teaching laboratory, Department of Human Anatomy ABU, Zaria for the successful investigation of histological, haematological and biochemical aspect of this work. We also appreciate Mr. S. Namadi from the Department of Botany, ABU, Zaria for the authentication of the plant (voucher number: ABU06885). A special thank you to the laboratory staff of these departments for their assistance in successfully conducting this experiment. References World Health Organization. (2022). 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Ayoola AA, Ekunseitan DA, Muhammad SB, Oguntoye MA, Adejola YA (2020) Phytochemicals analysis and gc-ms determination of ethanolic extracts of Azadirachta indica and Mangifera indica stem bark and their biological potentials. The Pac J Sci Technol , 21(1), 219-22. Kane NF, Kyama MC, Nganga JK, Hassanali A, Diallo M, Kimani FT (2023) Expression of the Fab enzymes (Fab I and Fab Z) from Plasmodium falciparum after exposure to Artemisia afra plant extracts and drugs screening. J Parasit Dis, 47(1), 46-58. Revathi N, Dhanaraj T (2019) Evaluation of bioactive phytochemicals in leaves extract of dodonaea angustifolia using gas chromatography and mass spectroscopic technique. J Pharmacogn Phytochem, 8(3), 4406-4409. Alabi K, Lajide L, Owolabi B (2018) Biological activity of oleic acid and its primary amide: experimental and computational studies. J Chem Soc Niger, 43(2). 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Asian Pac J Trop Biomed, 10(11), 479. Dhawan D, Gupta J (2017) Research article comparison of different solvents for phytochemical extraction potential from Datura metel plant leaves. Int J Biol Chem Sci, 11(1), 17-22. Hodge A, Sterner B (2005) Toxicity classes. In: Canadian Center for Occupational Health and Safety. http://www.ccohs.ca/oshanswers/chemicals/id50.htm Onochie AU, Oli AH, Oli AN, Ezeigwe OC, Nwaka AC, Okani CO, Okam PC, Ihekwereme CP, Okoyeh JN (2020). The pharmacobiochemical effects of ethanol extract of Justicia secunda Vahl leaves in Rattus norvegicus . J Exp Pharmacol , 423-437. Panter KE (2018) Cyanogenic glycoside–containing plants. In R. C. Gupta (Ed.), Veterinary Toxicology (3rd ed., pp. 935-940). Academic Press. doi.org/10.1016/B978-0-12-811410-0.00064-7 Chidi AS, Mattew WO, Benjamin AA, Peter UA, Uche NC (2018) The modulatory activity of Justicia carnea in Plasmodium infected mice. Trends J Sci Res , 3(4), 151-160 White NJ (2017) Malaria parasite clearance. Malar J, 16(1), 88. Haidara M, Haddad M, Denou A, Marti G, Bourgeade-Delmas S, Sanogo R, Bourdy G, Aubouy A (2018) In vivo validation of anti-malarial activity of crude extracts of Terminalia macroptera , a Malian medicinal plant. Malar J, 17(1), 1-10. Carneiro MR, Sallum LO, Martins JL, Peixoto JD, Napolitano HB, Rosseto LP (2023) Overview of the Justicia genus: insights into its chemical diversity and biological potential. Molecules, 28(3), 1190. Arrey TP, Okalebo FA, Ayong LS, Agbor GA, Guantai AN (2014) Anti-malarial activity of a polyherbal product (Nefang) during early and established Plasmodium infection in rodent models. Malar J, 13 , 1-11. Wahyuni DK, Wacharasindhu S, Bankeeree W, Wahyuningsih S P, Ekasari W, Purnobasuki H, Punnapayak H, Prasongsuk S (2023) In vitro and in vivo antiplasmodial activities of leaf extracts from Sonchus arvensis L. BMC Complement Med, 23(1), 1-12. Anyasor GN, Moses N, Kale O (2020) Hepatoprotective and hematological effects of justicia secunda vahl leaves on carbon tetrachloride induced toxicity in rats. Biotech Histochem, 95 (5), 349-359. Zhang P, Liu N, Xue M, Zhang M, Liu W, Xu C, Fan Y, Meng Y, Zhang Q, Zhou Y (2023) Anti-Inflammatory and antioxidant properties of β-sitosterol in copper sulfate-induced inflammation in zebrafish. Antioxidants , 12(2), 391. doi.org/10.3390/antiox12020391 Afriyie D, Ameyaw EO, Acheampong F, Appiah-Opong R (2021) In vitro and in vivo antioxidant properties of stem extracts of Croton membranaceus . J Med Plant Stud, 9 (4), 1-8. Sidiki NN, Nadia NA, Cedric Y, Guy-Armand GN, Sandra TN, Kevin TD, Azizi MA, Payne VK (2023) Antimalarial and antioxidant activities of ethanolic stem bark extract of Terminalia macroptera in swiss albino mice infected with Plasmodium berghei . J Parasit Res, 2023 Zhang P, Liu N, Xue M, Zhang M, Xiao Z, Xu C, Fan Y, Liu W, Qiu J, Zhang Q (2023) Anti-Inflammatory and antioxidant properties of squalene in copper sulfate-induced inflammation in zebrafish ( Danio rerio ). Int J Mol Sci, 24(10), 8518. Wei CC, Yen PL, Chang ST, Cheng PL, Lo YC, Liao VHC (2016) Antioxidative activities of both oleic acid and Camellia tenuifolia seed oil are regulated by the transcription factor DAF-16/FOXO in Caenorhabditis elegans . PloS one, 11(6), e0157195. Ibukunoluwa MR (2017) In vivo anti-plasmodial activity and histopathological analysis of water and ethanol extracts of a polyherbal antimalarial recipe. J Pharmacogn Phytother, 9(6), 87-100. Ayawa NG, Ramon-Yusuf SB, Wada YA, Oniye SJ, Shehu DM (2021) Toxicity study and anti-trypanosomal activities of aqueous and methanol whole plant extracts of Brillantaisia owariensis on Trypanosoma brucei -induced infection in BALB/c mice. Clin Phytoscience , 7(1), 39. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3879871","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":268248641,"identity":"488efa81-f724-4c3d-aca3-2a1c8e388e65","order_by":0,"name":"Nasiru Ademola 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University","correspondingAuthor":false,"prefix":"","firstName":"Yunusa","middleName":"","lastName":"WADA","suffix":""},{"id":268248647,"identity":"94ecc965-0b72-460f-aea9-91354fb39340","order_by":5,"name":"Muhammad HUSSAINI","email":"","orcid":"","institution":"Ahmadu Bello University","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"HUSSAINI","suffix":""}],"badges":[],"createdAt":"2024-01-19 21:14:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3879871/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3879871/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.26502/ijabpt.202130","type":"published","date":"2024-08-27T00:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":50091751,"identity":"2a2c9965-ef46-4ed6-b9ae-0d8abcfbca9e","added_by":"auto","created_at":"2024-01-24 12:00:49","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":376408,"visible":true,"origin":"","legend":"\u003cp\u003eAerial part of \u003cem\u003eJusticia\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3879871/v1/52624c1d1dc2bfdfbdbf8cd5.png"},{"id":50091753,"identity":"aed4758f-fd85-4e54-8445-92915517f768","added_by":"auto","created_at":"2024-01-24 12:00:49","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":24417,"visible":true,"origin":"","legend":"\u003cp\u003eHaematological profile of experimental mice treated with ethanol leaf extract of \u003cem\u003eJusticia\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis performed amongst negative control (C-), positive control (C+), normal control (NC), 500mg/kg(T1), 1000mg/kg(T2) and 1500mg/kg(T3) dose of ethanol leaf extract of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e respectively, groups with different letters on bars are statistically significant at \u003cem\u003ep\u003c/em\u003e\u0026lt;0.05, each point is the mean ± SEM (Standard error of mean) n=5.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3879871/v1/e86ad039e96e66840efbb551.png"},{"id":50091749,"identity":"8fdbf0a2-b0a2-4c50-bbc3-360b8ca00f14","added_by":"auto","created_at":"2024-01-24 12:00:49","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":15194,"visible":true,"origin":"","legend":"\u003cp\u003ePlasma biochemical composition of mice treated with ethanol leaf extract of \u003cem\u003eJusticia\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis performed amongst negative control (C-), positive control (C+), normal control (NC), 500mg/kg(T1), 1000mg/kg (T2), 1500mg/kg (T3) dose of ethanol leaf extract of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e, groups with different letters on bars are statistically significant \u003cem\u003ep\u003c/em\u003e\u0026lt;0.05, each point is the mean ± SEM (Standard error of mean) n=5.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-3879871/v1/393c2faa9fbae394eabcb9f5.png"},{"id":50092157,"identity":"51f5caf9-1003-4195-bdd8-19e13deaaad4","added_by":"auto","created_at":"2024-01-24 12:08:49","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":60967,"visible":true,"origin":"","legend":"\u003cp\u003eAntioxidant and lipid peroxidation status of the liver for mice treated with ethanol leaf extract of \u003cem\u003eJusticia\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis performed amongst negative control (C-), positive control (C+), normal control (NC), 500mg/kg(T1), 1000mg/kg (T2), 1500mg/kg (T3) dose of ethanol leaf extract of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e, groups with different letters on bars are statistically significant \u003cem\u003ep\u003c/em\u003e\u0026lt;0.05, each point is the mean ± SEM (Standard error of mean) n=5.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-3879871/v1/5adbd8052a613c672211c1e7.png"},{"id":50091754,"identity":"73baa6d8-af95-4314-a5fc-d0fbba50af64","added_by":"auto","created_at":"2024-01-24 12:00:50","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":284195,"visible":true,"origin":"","legend":"\u003cp\u003eLiver Photomicrograph of mice treated with ethanol leaf extract of \u003cem\u003eJusticia\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e(Curative test)- 400 × magnification\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKEYS:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNegative control (C-), positive control (C+), normal control (NC), 500mg/kg(T1), 1000mg/kg (T2), 1500mg/kg (T3) dose of ethanol leaf extract of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e, MVC= Mass Vascular Congestion, NF= Normal Feature, VC= Vacuolation, MHN= Moderate Hepatic Necrosis, MVC= Mass Vascular Congestion, SKH= Slight Kupffer cell Hyperplasia,\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-3879871/v1/8a541b12ae5796667f189a25.png"},{"id":50092158,"identity":"dd26189e-ed0b-4ada-93fa-2ec0b41ee00c","added_by":"auto","created_at":"2024-01-24 12:08:49","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":291941,"visible":true,"origin":"","legend":"\u003cp\u003eLiver Photomicrograph of mice treated with ethanol leaf extract of \u003cem\u003eJusticia\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e(Suppressive test)- 400 × magnification\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKEYS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNegative control (C-), positive control (C+), normal control (NC), 500mg/kg(T1), 1000mg/kg (T2), 1500mg/kg (T3) dose of ethanol leaf extract of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e, SVC= Slight Vascular Congestion, NF= Normal Feature, VC= Vacuolation, SKH= Slight Kupffer cell Hyperplasia.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-3879871/v1/8d76fe4e0f7426bbfe05e95b.png"},{"id":84307811,"identity":"0e7de3a4-1c64-4e91-bd69-7d487e0eac29","added_by":"auto","created_at":"2025-06-10 11:43:12","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2435349,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3879871/v1/af47edf5-a34a-41d1-958a-ff21ba250986.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Pharmacological Efficacy of Ethanol Leaf Extract of Justicia secunda in Swiss Albino Mice Experimentally Infected with Plasmodium berghei","fulltext":[{"header":"1 Background","content":"\u003cp\u003eMalaria is more prevalent in African nations than it is worldwide. According to the World Malaria Report 2020; Nigeria (31.3%), the Democratic Republic of Congo (12.6%), the United Republic of Tanzania (4.1%) and Niger (3.9%) account for more than half of all malaria death [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Although, sociopolitical issues are connected to the causes and consequences of malaria disease [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]; explicitly, the major contributing factor to the prevalence of malaria is the lack of mosquito protection and appropriate malaria treatment [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMalaria is an infectious disease affecting humans and animals [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The etiologic agent of the disease is the parasitic protozoans of the genus \u003cem\u003ePlasmodium\u003c/em\u003e, and the sole biological vector is the female \u003cem\u003eAnopheles\u003c/em\u003e mosquitoes [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The asymptomatic human is an unwitting parasite reservoir, allowing continuous transmission [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. The initial clinical symptom of this illness leads to headaches, fatigue, soreness in the joints and muscles, vomiting, and anorexia. Acidemia, hypoglycemia, anaemia, pulmonary oedema, coma and renal failure are common signs of acute malaria [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAntimalarial drug resistance occurs in the malaria parasite due to ineffective chemoprophylaxis [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], fake and irregular pharmaceutical administration leading to genetic mutation [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Therefore, searching for a new anti-malarial compound is a continuous process, and numerous studies on African medicinal plants with anti-malarial potentials have been conducted over time. However, there is a dearth of practical information regarding the pharmacological efficacy of \u003cem\u003eJ. secunda\u003c/em\u003e as an antimalarial agent.\u003c/p\u003e \u003cp\u003e \u003cem\u003eJusticia secunda\u003c/em\u003e is taxonomically classified in the family Acanthaceae and is notably utilized for treating anaemia in tropical regions of Africa [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. It is commonly known as 'Christ blood' and 'Blood herb' in Nigeria. The herbaceous plant has green leaves and pink inflorescence (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. According to Irinmwinuwa and Afonne [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], the ethanol leaf extract of \u003cem\u003eJ. secunda\u003c/em\u003e contains saponin (9.2%), tannins (9.0), flavonoids (7.0%) and alkaloid (2.4%). It has also been established that \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e has hematinic effect in rats with anaemia induced by phenylhydrazine [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Likewise, it exhibited anti-inflammatory and antioxidant activity in an \u003cem\u003ein vitro\u003c/em\u003e study using heat-induced bovine serum albumin (BSA) denaturation and erythrocyte membrane stabilization assay [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThis study evaluates the phytochemical constituent and lethal dose of the ethanol leaf extract of \u003cem\u003eJ. secunda\u003c/em\u003e, as well as the effect of the extract on the parasitaemia level, hematological profile, pro-inflammatory (TNF-α) and anti-inflammatory (IL-10) cytokines, liver biochemicals, and histopathology of the liver in \u003cem\u003ePlasmodium berghei\u003c/em\u003e NK65-infected mice.\u003c/p\u003e"},{"header":"2 Methods","content":"\u003cp\u003e\u003cstrong\u003e2.1 Collection of Plant Sample\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMatured leaves were harvested from stands of \u003cem\u003eJ.\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e plants from three different sites in Lusada, Ado Odo Ota Local Government Area (LGA) Ogun State, Nigeria.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2 Preparation of Plant Extract\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCold maceration technique was adopted for the extraction by simply soaking five hundred grams (500g) of the powdered leaves in five (5) litres of eighty percent (80%) ethanol for three days, and the bottle containing the mixtures was agitated at intervals. The liquid extract was filtered into a clean container using a piece of chiffon material after 72 hours. The filtrate was poured into an evaporating dish and placed on a water bath to obtain a solid and more concentrated form of the extract [15].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.3 Evaluation of Phytochemical Constituent of Plant Extract\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA standard method [16] was adopted in the phytochemical screening of the extract. The Gas Chromatography-Mass Spectrometry (GC-MS) analysis was conducted using a 7890B GC System (Agilent Technologies, USA) coupled with a 5977A Mass Selective Detector (Agilent Technologies, USA). The leaf extract of \u003cem\u003eJ.\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e was diluted in methanol, and 2\u0026mu;L of the mixture was injected into the GC-MS machine using a micro-syringe. The experimental parameters for the GC-MS system were: initial oven temperature: 70\u003csup\u003eo\u003c/sup\u003eC, Equilibration Time: 1 min, Max Temperature: 325 ̊C, Slow Fan: Disabled, Oven Program: 5\u0026deg;C/min On 250\u0026deg;C for 1 min, #1 then 30\u0026deg;C/min to 300\u0026deg;C for 0 min, Post Run: 50 \u0026deg;C. Cryo: Off, Front SS Inlet He \u0026ndash; Mode: Split. Heater: On 250\u0026deg;C, Pressure: On 11.089psi, Total Flow on 19.204mL/min, Septum Purge Flow: On 3 mL/min, Gas Saver: off, Purge flow to Split vent: on 15mL/min at 0.75min. Holdup time: 1.2386min, Flowrate: 1.0mL/minute\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.4 Maintenance of Experimental Animals\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSeventy-five (75) male Swiss albino mice with body weight between 20 and 30g were obtained for this study. Chicken grower\u0026rsquo;s marsh and water was provided at all time for the experimental mice [17]. The overall maintenance of the experimental animals was in accordance with the procedures approved by the ethical committee of the institution where the experiment was conducted.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5 Determination of Lethal Dose of Plant Extract\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe median lethal dose of the ethanol leaf extract of \u003cem\u003eJ.\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e was determined per the Up and Down method described by the Organization for Economic Co-orporation and Development [18].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6 Evaluation of Antiplasmodial Activity of Plant Extract\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe infected experimental mice were inoculated intraperitoneally with 0.2mL of blood from donor mouse infected with \u003cem\u003eP.\u003c/em\u003e \u003cem\u003eberghei\u003c/em\u003e NK65 (30% parasitaemia) and normal saline (1:9) containing 10\u003csup\u003e5\u003c/sup\u003e \u003cem\u003eP.\u003c/em\u003e \u003cem\u003eberghei\u003c/em\u003e parasitized erythrocytes. The seven-day curative test was evaluated seventy-two (72) hours after inoculation and confirmation of parasitemia in thirty (30) mice, and they were randomly assigned into six (6) groups. The suppressive test was conducted, and the inception was precisely three (3) hours after the infection of mice for a four-day duration using thirty (30) mice. These tests were performed using the method of Onyegeme-Okerentaa \u003cem\u003eet\u003c/em\u003e \u003cem\u003eal\u003c/em\u003e. [19], with a slight modification. Groups 1\u0026ndash;3 were the control groups treated with 1 mL of normal saline (the negative control), 25 mg/kg of chloroquine (the positive control), and the uninfected and untreated groups (the normal control), respectively. Groups 4-6 were given 500 mg/kg, 1000 mg/kg, and 1,500 mg/kg of the crude extract based on the LD\u003csub\u003e50\u003c/sub\u003e for the ethanol leaf extract of \u003cem\u003eJ.\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e determined in this study. The oral treatment was done once a day.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.7 Collection of Blood and Liver Samples\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe animals were sacrificed after inhaling vapour from ketamine-damped cotton wool at the end of the experiment. The blood of experimental animals was collected in EDTA bottles for haematological and biochemical analysis through cardiac aspiration. A portion of the liver was placed in cold phosphate buffer for antioxidant and MDA analysis. A small portion of liver of mice was preserved in phosphate-buffered formalin.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.8 Determination of Parasitaemia and Percentage Chemosuppression\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe number of parasitized erythrocytes in ten (10) slides prepared with thin blood smears was counted, and the average was computed to give the parasitemia of each mouse [20].\u003c/p\u003e\n\u003cp\u003ePercentage parasitaemia and chemosuppression was calculated as:\u003c/p\u003e\n\u003cp\u003eParasitaemia (%) = (Number of parasitized RBC/Total number of RBC) x 100\u003c/p\u003e\n\u003cp\u003e\u003cimg 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mean % parasitaemia in negative control\u003c/p\u003e\n\u003cp\u003eB= mean % parasitaemia in treated group\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.9 Assessment of Haematological Parameters\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Red Blood Cell (RBC), White Blood Cell (WBC), Packed Cell Volume (PCV) and Haemoglobin (HGB) concentration were analyzed using a Biobase Bk6100 haematology analyzer [21] in blood samples collected in Ethylene Diamine Tetracetic Acid (EDTA) bottles.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.10 Assessment of Plasma and Liver Biochemicals\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe liver homogenate was used to evaluate the levels of Malondialdehyde (MDA), Superoxide dismutase (SOD), Catalase (CAT) and reduced Glutathione (GSH) using a Sigma Aldrich Assay kit. Blood in EDTA bottles was used to check the concentrations of Alanine aminotransferase (Randox kit), Interleukin-10 and Tumour Necrosis Factor-\u0026alpha; using ELISA kits by FineTest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.11 Liver Histopathological Studies\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe liver of mice preserved in phosphate-buffered formalin was prepared for histological study using standard histotechnique and viewed with a microscope at 400 \u0026times; magnification. The photomicrographs were taken using a digital camera [22].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.12 Data Analyses\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOne-way Analysis of Variance (ANOVA) was used to determine statistical differences in the antiplasmodial activity, haematological and biochemical parameters in the \u003cem\u003ein\u003c/em\u003e \u003cem\u003evivo\u003c/em\u003e suppressive and curative test. Tukey HSD test was used to rank the difference between significant parameters at \u003cem\u003ep\u003c/em\u003e\u0026lt;0.05. Data were represented in tables and summarized in form of mean \u0026plusmn; standard error. Statistical analyses were done using \u0026apos;R\u0026apos; statistical package (version 4.1.2) for windows.\u003c/p\u003e"},{"header":"3 Results","content":"\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1 Phytochemical Characterization and Acute Toxicity\u003c/h2\u003e\n \u003cp\u003eThe phytochemical screening detected eight different classes of phytochemicals (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e), and the GC-MS technique confirmed the presence of thirty different compounds, as shown in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. Treatment of mice with limit dose of ethanol leaf extract of \u003cem\u003eJ. secunda\u003c/em\u003e (5000mg/kg) caused no mortality and signs of overtoxication. This suggests that the LD\u003csub\u003e50\u003c/sub\u003e of the extract is greater than 5000mg/kg.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003ePhytochemical screening of ethanol leaf extract of \u003cem\u003eJusticia secunda\u003c/em\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"3\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eS/No\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePhytoconstituent\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eInference\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAlkaloids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCardiac glycosides\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSaponin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePhenolic compounds\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTannins\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSteroids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCarbohydrates\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eFlavonoids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTerpenoids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAnthraquinones\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\"\u003eKeys: (+)\u0026thinsp;=\u0026thinsp;Present, (-)\u0026thinsp;=\u0026thinsp;Absent\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"char\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eGC-MS profile of ethanol leaf extract of \u003cem\u003eJusticia secunda\u003c/em\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"5\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePeak\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eRetention Time (Minute)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLibrary I.D\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePeak area (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eChemical Abstract Service Number\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.4652\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17-Pentatriacontene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.1691\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e006971-40-0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOxalic acid, cyclobutyl heptadecyl ester\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.2012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1000309-70-7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20.0166\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHexadecanoic acid, methyl ester\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.6316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e000112-39-0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e21.4568\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3-Chloropropionic acid, heptadecyl ester\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.2485\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1000283-05-1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e23.0506\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9,12-Octadecadienoic acid, (linoleic) methyl ester, (E,E)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.1048\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e002566-97-4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e23.2025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecis-13-Octadecenoic acid, methyl ester\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6.3996\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1000333-58-3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e23.7984\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMethyl stearate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.8041\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e000112-61-8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e25.1498\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCarbonic acid, but-3-en-1-yl hexadecyl ester\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.1945\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1000383-23-7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e25.279\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAspidospermidin-17-ol, 1-acetyl-19,21-epoxy-15,16-dimethoxy-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.2787\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e002122-26-1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.5241\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8-Hexadecenal, 14-methyl-, (Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.1543\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e060609-53-2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e26.5546\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCyclododecane\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.5526\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e000294-62-2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e26.7618\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9,12-Octadecadienal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.6438\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e026537-70-2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e26.8969\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ep-Menth-8(10)-en-9-ol, cis-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.1951\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e015714-13-3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e26.9881\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOxirane, [(dodecyloxy)methyl]-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.7356\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e002461-18-9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.3173\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOleic Acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.1944\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e000112-80-1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.7978\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22-Stigmasten-3-one\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.3243\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e004736-95-2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.2861\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(S)(+)-Z-13-Methyl-11-pentadecen-1-ol acetate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.9133\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1000130-84-8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.5241\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8-Hexadecenal, 14-methyl-, (Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.2379\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e060609-53-2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9265\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3-Cyclohexylthiolane,S,S-dioxide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.317\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e071053-08-2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9874\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecis-7, cis-11-Hexadecadien-1-yl acetate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.3175\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e052207-99-5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e29.3867\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecis-9-Hexadecenal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.7726\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e056219-04-6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e29.8609\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9-Octadecenoic acid (Z)-, 2,3-dihydroxypropyl ester\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.1331\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e000111-03-5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e30.1094\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eZ-8-Methyl-9-tetradecenoic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.1701\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1000130-84-5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e30.2394\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1-Nonadecene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.2528\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e018435-45-5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e30.394\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCyclododecanol, 1-ethenyl-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.5624\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e006244-49-1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e30.9115\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCyclododecane, ethyl-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.3424\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e028981-49-9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e31.2385\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2-Methyl-Z,Z-3,13-octadecadienol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.9236\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1000130-90-5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e31.5878\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eE-9-Tetradecenal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.9144\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1000131-35-7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e32.575\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026beta;-Sitosterol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e19.7234\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e000083-46-5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e34.706\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSqualene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.1521\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e000111-02-4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2 Antiplasmodial Activity of the Crude Extract\u003c/h2\u003e\n \u003cp\u003eThe crude extract was tested for curative and suppressive antiplasmodial activity as presented in Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e and the result was expressed in the form of percentage parasitaemia and percentage chemosuppression for the infected controls and the treatment groups. The extract significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) reduced parasitaemia level in infected mice for both curative and suppressive tests compared with the negative control. A dose-dependent chemosuppression was observed for the experimental groups, which was lower than the standard drug with 100% chemosuppression.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eAntiplasmodial activity of ethanol leaf extract of \u003cem\u003eJusticia secunda\u003c/em\u003e against \u003cem\u003eP\u003c/em\u003e. \u003cem\u003eberghei\u003c/em\u003e in mice\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"5\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCurative test\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSuppressive test\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eExperimental groups\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e% Parasitaemia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e% Chemo-suppression\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e% Parasitaemia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e% Chemo-suppression\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e41.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.40\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.29\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e53.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.70\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e52.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.959\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e60.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.46\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e62.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.81\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e71.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.67\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e75.18\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003ep\u003c/em\u003e-value\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cstrong\u003eNOTE:\u0026nbsp;\u003c/strong\u003eData are expressed as mean \u0026plusmn; SEM, n=5\u003c/p\u003e\n \u003cp\u003eStatistical analysis performed amongst Negative control (C-), Positive control (C+), Normal control (NC),\u0026nbsp;500mg/kg(T1), 1000mg/kg(T2) and 1,500mg/kg(T3) dose of ethanol leaf extract of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e, means with different superscript within a column are statistically significant at \u003cem\u003ep\u003c/em\u003e\u0026lt;0.05\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003e3.3 Haematological Profile of Experimental Mice\u003c/h2\u003e\n \u003cp\u003eThe haematological profile of mice measured at the end of the experiment shows that the extract significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) prevented the reduction in Packed Cell Volume (PCV), Red Blood Cell (RBC) and Haemoglobin (HGB) for both curative and suppressive test as shown in Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The group of mice that received chloroquine (4.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.189) had a statistically (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) reduced White Blood Cell (WBC) than and the negative control (5.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.356) and experimental groups treated for seven days (4.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.362, 4.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.252, 4.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.218).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\n \u003ch2\u003e3.4 Blood Plasma Biochemicals of Experimental Mice\u003c/h2\u003e\n \u003cp\u003eThe immunomodulatory and hepatoprotective potentials of the plant extract was assessed by analysing the biochemical components of plasma such as TNF-\u0026alpha;, IL-10 and ALT. The outcome of this biochemical assay for the therapeutic and inhibitory test is presented in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. The infected mice treated with ethanol leaf extract of \u003cem\u003eJ. secunda\u003c/em\u003e had significantly higher (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) concentrations of TNF-\u0026alpha; and IL-10 compared to negative control for both curative and suppressive test. Furthermore, the plasma concentration of ALT in negative control was significantly higher (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) than groups that received plant extract treatment.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003e\u003cstrong\u003e3.5 Antioxidant and Lipid Peroxidation Status of the Liver\u003c/strong\u003e\u003c/h2\u003e\n \u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e shows the liver\u0026rsquo;s antioxidant and lipid peroxidation profile of experimental mice for the curative and suppressive test. The groups treated with plant extract exhibited a significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) increase in the bioactivity of antioxidants, including SOD, CAT, and GSH, compared to the negative control. The SOD concentration was, however, significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lower in the extract-treated group (3.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.447u/ml, 3.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.526u/ml, 3.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.579u/ml) and negative control (3.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.481u/ml) than in the normal control (7.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.462u/ml) and positive control (6.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.331u/ml). In addition, the hepatic malondialdehyde (MDA) concentrations were significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) reduced in treatment groups compared to the negative control.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\n \u003ch2\u003e3.6 Liver Histology of Experimental Mice\u003c/h2\u003e\n \u003cp\u003eThe liver of negative control shows mass vascular congestion when compared to liver sections of other groups for the curative test (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). The positive control and normal control have normal architecture of a liver photomicrograph, while the 500mg/kg, 1000mg/kg and 1,500mg/kg have a moderate hepatic necrosis, moderate vacuolation and slight Kupffer cell hyperplasia respectively.\u003c/p\u003e\n \u003cp\u003eComparing the histological sections of the treatment groups to those of the negative control group for the suppressive test (Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e), the treatment groups showed slight vacuolation and necrosis (500 mg/kg), slight Kupffer cell hyperplasia (1,000mg/kg and 1,500 mg/kg) and the negative control had slight vascular congestion. Also, the liver photomicrographs of the positive and normal control exhibit normal features in their hepatic histology.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4 Discussion","content":"\u003cp\u003eEight (8) different secondary metabolites were present in the ethanol leaf extract of \u003cem\u003eJ. secunda\u003c/em\u003e namely: alkaloids, cardiac glycosides, phenolic compounds, tannins, steroids, carbohydrates, flavonoids and terpenoids. In addition, several phytochemical compounds with antioxidant, anti-inflammatory and anti-plasmodial activity were detected in the extract. For instance, β-sitosterol has anti-inflammatory properties [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], Squalene has antiparasitic and anti-inflammatory properties [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], 2-Methyl-Z,Z-3,13-octadecadienol has antimicrobial properties [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], 17-Pentatriacontene has anti-inflammatory properties [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e], hexadacenoic acid methyl ester has anti-inflammatory properties [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], 9,12-Octadecanoic (linoleic) acid methyl ester has anti-plasmodial and anti-inflammatory [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e], Methyl stearate has anti-inflammatory properties [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], cis-13-Octadecenoic acid, methyl ester has anti-inflammatory activity [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e], 16-dimethoxy-aspidospermidin-17-ol has anti-plasmodial activity [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e], 8-Hexadecenal 14-methyl-, (Z)- has antioxidant activity [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e], Oleic acid has antioxidant activity [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e], cis-9-Hexadecenal has anti-inflammatory properties [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e] and cyclododecane, ethyl has antioxidant activity [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThese myriads of phytochemicals may have conferred on this plant its popularity and wide use in African traditional medicine [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. The phytochemical screening contrasts with the findings of a study conducted on a similar extract of \u003cem\u003eJ. secunda\u003c/em\u003e leaves collected from a farm in south-east Nigeria that is devoid of steroids and glycosides [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]; however, hexadecenoic acid methyl ester and octadecanoic acid methyl ester was also detected in extract of \u003cem\u003eJ. secunda\u003c/em\u003e [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. The difference in phytochemical constituents in both studies could be attributed to ecological variation [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. According to Hodge and Sterner's [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e] toxicity scale, this plant extract is practically non-toxic. A study [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e] that processed the leaves of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e for three weeks and macerated them in ethanol for 24 hours reported an LD\u003csub\u003e50\u003c/sub\u003e of 3,800 mg/kg body weight in rats due to the cyanide content of the extract. It is very likely that the selection of mature leaves and the longer processing duration of the extract for this present study may be reasons for the reduced toxicity observed [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOne of the major foci of this study is to include \u003cem\u003eJusticia secunda\u003c/em\u003e in the record of antimalarial study, and the crude extract has a percentage chemosuppression of 71.21% (1,500 mg/kg) and 75.18% (1,500 mg/kg) in the suppressive and curative tests, respectively. The result is less than the antimalarial activity of the ethanol leaf extract of \u003cem\u003eJusticia carnea\u003c/em\u003e (800 mg/kg dose) with a percentage chemosuppression of 82% [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. The efficacy of plant extracts in treating malaria disease depends on several factors, such as the \u003cem\u003ePlasmodium\u003c/em\u003e species, the dosage, the duration of treatment, and the host immune response [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Experimental research on 16-dimethoxy-aspidospermidin-17-ol [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] and linoleic acid-methyl esters [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] suggest that these phytochemicals inhibit the growth of \u003cem\u003ePlasmodium\u003c/em\u003e parasites by binding with the Fab-I enzyme responsible for fatty acid biosynthesis. This is to create awareness that \u003cem\u003eJ. secunda\u003c/em\u003e could be an alternative source of antimalarial compounds.\u003c/p\u003e \u003cp\u003eThe ethanol leaf extract of \u003cem\u003eJ. secunda\u003c/em\u003e prevented PCV, HGB, and RBC reduction in mice, but there was an increase in WBC in the curative test. A different study observed similar trend for these haematological parameters with a normal WBC in curative and suppressive tests on the antioxidant and antimalarial activity of the ethanol stem bark extract of \u003cem\u003eTerminalia macroptera\u003c/em\u003e in \u003cem\u003eP. berghei-\u003c/em\u003einfected mice [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. The haematinic properties of the extract in this study may be due to the presence of flavonoid compounds that have been reported to increase iron absorption and deposition in tissues while reducing iron excretion [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. In addition, many antimalarial herbal preparations may exert their anti-infective activity not only by affecting the parasite directly but may also stimulate the defensive system of the host through many other mechanisms [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe assessment of modulatory and hepatoprotective potentials reveals that TNF-α and IL-10 increased and the concentrations of ALT reduced in treatment groups. This outcome corroborates the TNF and IL-10-increasing potentials observed in \u003cem\u003eP. berghei-\u003c/em\u003einfected mice treated with the ethyl acetate leaf extract of \u003cem\u003eSonchus arvensis\u003c/em\u003e [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e] and the hepatoprotective nature of \u003cem\u003eJ. secunda\u003c/em\u003e in reducing plasma ALT in rats administered carbon tetrachloride [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Infection and immunological reactions to the extract may be reasons for the increased level of TNF-α in treated mice. The anti-inflammatory activity of β-sitosterol was harnessed in reducing inflammation in Zebra fish [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e] and the presence of this phytochemical in the extract utilized for this research may be the reason for the observed pharmacological effect. This is an indication that \u003cem\u003eJ. secunda\u003c/em\u003e may be utilized for treatment associated with pathogenesis of malaria infection.\u003c/p\u003e \u003cp\u003eThe liver homogenate of the experimental groups sustained an increased bioactivity of antioxidants (SOD, GSH, and CAT). Yet, a reduced level of SOD was observed in mice treated days after infection. The lipid peroxidation biomarker (MDA) is equally reduced in mice treated with crude extract. Hepatic antioxidant and MDA concentrations follow the same pattern as those of experimental mice treated with an extract of \u003cem\u003eCroton membranaceus\u003c/em\u003e but differ with an increased SOD level [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. The infection also caused a reduction in hepatic SOD in mice infected with \u003cem\u003eP. berghei\u003c/em\u003e and treated with a stem-bark extract of \u003cem\u003eTerminalia macroptera\u003c/em\u003e [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. \u003cem\u003eJusticia secunda\u003c/em\u003e exhibited antioxidant properties in this study, which may be attributed to components of the extract with the relevant pharmacological activity [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe ethanol leaf extract of \u003cem\u003eJ. secunda\u003c/em\u003e reduced vascular congestion in the liver of mice, as the photomicrograph of the treatment groups only shows sections with Kupffer cell hyperplasia, vacuolation, and hepatic necrosis. Ibukunoluwa [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e] also reported similar histopathology in the antiplasmodial activity of polyherbal mixtures. The slight changes seen in the liver when used to test the efficacy of the plant extract may be due to the induced infection and the activation of immune cells in the organs. Ayawa \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e] also advanced cellular damage by immunological reactions as a reason for histopathological changes in the liver of mice.\u003c/p\u003e \u003cp\u003ePure compounds should be isolated from \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e extract to conduct trials on their pharmacological activity in relation to malaria infection and this could potentially uncover another effective antimalarial compound.\u003c/p\u003e"},{"header":"5 Conclusion","content":"\u003cp\u003eIn conclusion, the ethanol leaf extract of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e contains phytochemicals with numerous pharmacological activities, and is non-toxic at 5,000mg/kg. It exhibits a dose-dependent percentage of chemosuppression of \u003cem\u003eP\u003c/em\u003e. \u003cem\u003eberghei\u003c/em\u003e at 500mg/kg, 1,000mg/kg and 1,500mg/kg. The extract prevented the reduction of the studied haematological parameters such as RBC, HGB and PCV. The concentrations of TNF-α and IL-10 increased and ALT concentrations reduced in the plasma of mice treated with extract. Evaluation of liver biochemicals revealed that MDA concentration was reduced with increased bioactivity of the studied antioxidants for the infected mice treated with the plant extract. Vascular congestion was also reduced in the liver histology of mice infected and treated with the plant extract but shows increased infiltration of immune cells and necrosis. This present study infers that ethanol leaf extract of \u003cem\u003eJ. secunda\u003c/em\u003e contains phytochemicals that have been tested individually to treat malaria and inflammation, which can be isolated for treating malarial pathogenesis.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eALT \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Alanine aminotransferase\u003c/p\u003e\n\u003cp\u003eMDA\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Malondialdehyde\u003c/p\u003e\n\u003cp\u003eSOD\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Superoxide dismutase\u003c/p\u003e\n\u003cp\u003eCAT\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Catalase\u003c/p\u003e\n\u003cp\u003eGSH\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Reduced Glutathione\u003c/p\u003e\n\u003cp\u003eIL-10\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Interleukin-10\u003c/p\u003e\n\u003cp\u003eTNF-α \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Tumour Necrosis Factor-alpha\u003c/p\u003e\n\u003cp\u003eHGB\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Haemoglobin\u003c/p\u003e\n\u003cp\u003ePCV\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Packed Cell Volume\u003c/p\u003e\n\u003cp\u003eRBC\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Red Blood Cell\u003c/p\u003e\n\u003cp\u003eWBC \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;White Blood Cell\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMaintenance and procedures performed involving animals in this study follow the ethical standards of Ahmadu Bello University, Zaria Committee on Animal Use and Care (ABUCAUC) with the approval number ABUCAUC/2023/055.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed in this study are included in this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNone\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNone\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors’ contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eN.A.A, S.J.O, P.I.R, A.Y., and Y.A.W conceptualized the study, conducted the investigation and formal analysis.\u003c/p\u003e\n\u003cp\u003eN.A.A and M.H. prepared figures and tables.\u003c/p\u003e\n\u003cp\u003eAll authors reviewed the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA sincere gratitude to the Department of Pharmacology, Ahmadu Bello University (ABU), Zaria for providing the animal house and Swiss albino mice used for this research, Department of Pharmacognosy and Drug Development ABU, Zaria for providing the facility to achieve the extraction of the plant and the phytochemical screening, the Multi-User Science Research Laboratory ABU, Zaria for the Gas Chromatography-Mass Spectrometric analysis of the plant extract and the Teaching laboratory, Department of Human Anatomy ABU, Zaria for the successful investigation of histological, haematological and biochemical aspect of this work. We also appreciate Mr. S. Namadi from the Department of Botany, ABU, Zaria for the authentication of the plant (voucher number: \u0026nbsp;ABU06885). A special thank you to the laboratory staff of these departments for their assistance in successfully conducting this experiment.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eWorld Health Organization. (2022). World malaria report 2022. https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2022\u003c/li\u003e\n\u003cli\u003eOkoronkwo IL (2014) Social, cultural, political and economic issues connected to the causes and consequences of malaria. \u003cem\u003eJ. 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J\u003cem\u003e Med Plant Stud, 9\u003c/em\u003e(4), 1-8.\u003c/li\u003e\n\u003cli\u003eSidiki NN, Nadia NA, Cedric Y, Guy-Armand GN, Sandra TN, Kevin TD, Azizi MA, Payne VK (2023) Antimalarial and antioxidant activities of ethanolic stem bark extract of \u003cem\u003eTerminalia\u003c/em\u003e \u003cem\u003emacroptera\u003c/em\u003e in swiss albino mice infected with \u003cem\u003ePlasmodium\u003c/em\u003e \u003cem\u003eberghei\u003c/em\u003e. \u003cem\u003eJ Parasit Res, \u003c/em\u003e2023\u003c/li\u003e\n\u003cli\u003eZhang P, Liu N, Xue M, Zhang M, Xiao Z, Xu C, Fan Y, Liu W, Qiu J, Zhang Q (2023) Anti-Inflammatory and antioxidant properties of squalene in copper sulfate-induced inflammation in zebrafish (\u003cem\u003eDanio\u003c/em\u003e \u003cem\u003ererio\u003c/em\u003e). \u003cem\u003eInt J Mol Sci, \u003c/em\u003e24(10), 8518.\u003c/li\u003e\n\u003cli\u003eWei CC, Yen PL, Chang ST, Cheng PL, Lo YC, Liao VHC (2016) Antioxidative activities of both oleic acid and \u003cem\u003eCamellia\u003c/em\u003e \u003cem\u003etenuifolia\u003c/em\u003e seed oil are regulated by the transcription factor DAF-16/FOXO in \u003cem\u003eCaenorhabditis\u003c/em\u003e \u003cem\u003eelegans\u003c/em\u003e. \u003cem\u003ePloS one, \u003c/em\u003e11(6), e0157195.\u003c/li\u003e\n\u003cli\u003eIbukunoluwa MR (2017) \u003cem\u003eIn\u003c/em\u003e \u003cem\u003evivo\u003c/em\u003e anti-plasmodial activity and histopathological analysis of water and ethanol extracts of a polyherbal antimalarial recipe. \u003cem\u003eJ Pharmacogn Phytother, \u003c/em\u003e9(6), 87-100.\u003c/li\u003e\n\u003cli\u003eAyawa NG, Ramon-Yusuf SB, Wada YA, Oniye SJ, Shehu DM (2021) Toxicity study and anti-trypanosomal activities of aqueous and methanol whole plant extracts of \u003cem\u003eBrillantaisia\u003c/em\u003e \u003cem\u003eowariensis\u003c/em\u003e on \u003cem\u003eTrypanosoma\u003c/em\u003e \u003cem\u003ebrucei\u003c/em\u003e-induced infection in BALB/c mice. \u003cem\u003eClin Phytoscience\u003c/em\u003e, 7(1), 39.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Antiplasmodial, Phytochemicals, Immunomodulatory, Histopathology ","lastPublishedDoi":"10.21203/rs.3.rs-3879871/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3879871/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003ePlasmodium\u003c/em\u003e parasite that causes malaria has developed resistance to most antimalarial medications and this has prompted research on natural products. The plant, \u003cem\u003eJusticia\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e, is domesticated in the tropical region of Africa, and is used for the treatment of anaemia and other debilities.\u0026nbsp; We evaluated the antiplasmodial and immunomodulatory activity of ethanol leaf extract of \u003cem\u003eJ.\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e in mice experimentally infected with \u003cem\u003ePlasmodium berghei\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResult\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEthanol leaf extract of \u003cem\u003eJusticia\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e contains steroids, tannins, flavonoids, terpenoids, phenols, glycosides and carbohydrate as well as thirty (30) compounds with numerous pharmacological activities. The LD\u003csub\u003e50\u003c/sub\u003e was above 5,000mg/kg with no mortality. A dose-dependent percentage suppression of \u003cem\u003eP\u003c/em\u003e. \u003cem\u003eberghei\u003c/em\u003e was observed in the curative test for the three doses respectively (53.13%, 60.71%, 71.21%). The suppressive test for ethanol leaf extract of \u003cem\u003eJ.\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e also showed a similar trend (52.2%, 62.16%, 75.18%). The ethanol leaf extract of \u003cem\u003eJ\u003c/em\u003e. \u003cem\u003esecunda\u003c/em\u003e effectively prevented anaemia, reduced the level of ALT enzyme and also increased the level of TNF-α and IL-10 in the plasma of \u003cem\u003eP. berghei\u003c/em\u003e-infected mice. It also significantly reduced the oxidative stress and vascular congestion in the liver of mice infected with \u003cem\u003eP\u003c/em\u003e. \u003cem\u003eberghei\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study revealed that ethanol leaf extract of \u003cem\u003eJ.\u003c/em\u003e \u003cem\u003esecunda\u003c/em\u003e could be utilized to treat infection caused by \u003cem\u003ePlasmodium\u003c/em\u003e parasite and also ameliorate the pathogenesis of the disease.\u003c/p\u003e","manuscriptTitle":"Pharmacological Efficacy of Ethanol Leaf Extract of Justicia secunda in Swiss Albino Mice Experimentally Infected with Plasmodium berghei","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-24 12:00:45","doi":"10.21203/rs.3.rs-3879871/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f30489dc-539a-46f2-9051-a24471f74b06","owner":[],"postedDate":"January 24th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-06-10T11:43:06+00:00","versionOfRecord":{"articleIdentity":"rs-3879871","link":"https://doi.org/10.26502/ijabpt.202130","journal":{"identity":"international-journal-of-applied-biology-and-pharmaceutical-technology","isVorOnly":true,"title":"International Journal of Applied Biology and Pharmaceutical Technology"},"publishedOn":"2024-08-27 00:00:00","publishedOnDateReadable":"August 27th, 2024"},"versionCreatedAt":"2024-01-24 12:00:45","video":"","vorDoi":"10.26502/ijabpt.202130","vorDoiUrl":"https://doi.org/10.26502/ijabpt.202130","workflowStages":[]},"version":"v1","identity":"rs-3879871","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3879871","identity":"rs-3879871","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}