Parasite Inhibition Research Articles

An evaluation of fusion partner proteins for paratransgenesis in Asaia bogorensis.

Mosquitoes transmit many pathogens responsible for human diseases, such as malaria which is caused by parasites in the genus Plasmodium. Current strategies to control vector-transmitted diseases are increasingly undermined by mosquito and pathogen resistance, so additional methods of control are required. Paratransgenesis is a method whereby symbiotic bacteria are genetically modified to affect the mosquito's phenotype by engineering them to deliver effector molecules into the midgut to kill parasites. One paratransgenesis candidate is Asaia bogorensis, a Gram-negative bacterium colonizing the midgut, ovaries, and salivary glands of Anopheles sp. mosquitoes. Previously, engineered Asaia strains using native signals to drive the release of the antimicrobial peptide, scorpine, fused to alkaline phosphatase were successful in significantly suppressing the number of oocysts formed after a blood meal containing P. berghei. However, these strains saw high fitness costs associated with the production of the recombinant protein. Here, we report evaluation of five different partner proteins fused to scorpine that were evaluated for effects on the growth and fitness of the transgenic bacteria. Three of the new partner proteins resulted in significant levels of protein released from the Asaia bacterium while also significantly reducing the prevalence of mosquitoes infected with P. berghei. Two partners performed as well as the previously tested Asaia strain that used alkaline phosphatase in the fitness analyses, but neither exceeded it. It may be that there is a maximum level of fitness and parasite inhibition that can be achieved with scorpine being driven constitutively, and that use of a Plasmodium specific effector molecule in place of scorpine would help to mitigate the stress on the symbionts.

Copper (I)-Chloroquine Complexes: Interactions with DNA and Ferriprotoporphyrin, Inhibition of β-Hematin Formation and Relation to Antimalarial Activity

A new Cu(I)-chloroquine (CQ) complex [Cu(CQ)(PPh3)2]NO3 (1) was synthesized and characterized, and its mechanism of action studied concomitant with the previously reported complex [Cu(CQ)2]Cl (2). These copper (I) coordination compounds can be considered as potential antimalarial agents because they show better inhibition of the CQ-resistant strain in in vitro studies than CQ alone. In comparison with other metal-CQ complexes, only the gold complex was similar to (1), i.e., more active than CQ against both CQ-susceptible (3D7) and CQ-resistant strains (W2). These two copper (I)-compounds also demonstrated higher antiplasmodial activity against W2 than other copper complexes reported to date. This suggests that the incorporation of the copper metal center enhanced the biological activity of CQ. To better understand their significant growth inhibition of the Plasmodium falciparum parasite, the interaction with two essential molecular targets for the survival and proliferation of the malarial parasite were studied. These were the ferriprotoporphyrin group and the DNA, both important targets for current antimalarial drugs at the asexual erythrocytic stages. Both compounds (1,2) exhibited significant interactions with these targets. In particular, interactions with the DNA were dominated by the intercalator properties of the CQ ligand but may have also been affected by the presence of copper. Overall, these compounds were better parasitic inhibitors than chloroquine diphosphate (CQDP) alone or other previously reported metal-CQ complexes such as platinum, ruthenium and gold.

Wild Egyptian medicinal plants show in vitro and in vivo cytotoxicity and antimalarial activities

BackgroundMedicinal plants have been successfully used as an alternative source of drugs for the treatment of microbial diseases. Finding a novel treatment for malaria is still challenging, and various extracts from different wild desert plants have been reported to have multiple medicinal uses for human public health, this study evaluated the antimalarial efficacy of several Egyptian plant extracts.MethodsWe assessed the cytotoxic potential of 13 plant extracts and their abilities to inhibit the in vitro growth of Plasmodium falciparum (3D7), and to treat infection with non-lethal Plasmodium yoelii 17XNL in an in vivo malaria model in BALB/c mice.ResultsIn vitro screening identified four promising candidates, Trichodesma africanum, Artemisia judaica, Cleome droserifolia, and Vachellia tortilis, with weak-to-moderate activity against P. falciparum erythrocytic blood stages with mean half-maximal inhibitory concentration 50 (IC50) of 11.7 μg/ml, 20.0 μg/ml, 32.1 μg/ml, and 40.0 μg/ml, respectively. Their selectivity index values were 35.2, 15.8, 11.5, and 13.8, respectively. Among these four candidates, T. africanum crude extract exhibited the highest parasite suppression in a murine malaria model against P. yoelii.ConclusionOur study identified novel natural antimalarial agents of plant origin that have potential for development into therapeutics for treating malaria.

Investigations of the anti-inflammatory, analgesic, and anti-malarial activities of extract from the leaf of Halea ciliata (Abura leaves)

Background: Halea ciliata (Abura leaves) is a medicinal plant and has long been used for the treatment of several diseases including malaria, pains and inflammation, However, no complete pharmacological study to confirm these effects have been reported. In this study, the anti-inflammatory, analgesic, and antimalarial effects of Halea ciliata were investigated. Methods: The leaf of Halea ciliata was extracted with ethanol. The anti-inflammatory activity was tested using the egg albumin induced paw oedema while the analgesic activities were investigated using the hot-plate and Tail flick tests in mice. The anti-malaria activities were evaluated in Plasmodium berghei infected mice. Results: The crude methanol extract of Halea ciliata had LD50 greater than 5000 mg/kg and contains alkaloid, tannin, flavonoid, glycoside, steroid, terpenoid and saponin. In vivo analysis revealed that the crude extract demonstrated 78.64±4.32 % and 78.09±2.34% inhibition of the Plasmodium berghei parasite, while chloroquine, exhibited 92.12±3.45 % activities. The extract also significantly (p<0.05) suppressed the egg albumin induce paw oedema with percentage inhibition of 13.55±0.34, and 11.88±1.34 at 200 and 400 mg/kg respectively, while the standard drug caused 64.40±2.34 inhibition of paw oedema when compared to the non-treated controls. Similarly, the crude extract demonstrated significant analgesic activities in both the hot plate and tail-flick models. Conclusion: These results suggest that crude methanol extract of Halea ciliata has potent anti-malaria, anti-inflammatory and anti-nociceptive activities.

Double drugging of prolyl-tRNA synthetase provides a new paradigm for anti-infective drug development.

Toxoplasmosis is caused by Toxoplasma gondii and in immunocompromised patients it may lead to seizures, encephalitis or death. The conserved enzyme prolyl-tRNA synthetase (PRS) is a validated druggable target in Toxoplasma gondii but the traditional ‘single target–single drug’ approach has its caveats. Here, we describe two potent inhibitors namely halofuginone (HFG) and a novel ATP mimetic (L95) that bind to Toxoplasma gondii PRS simultaneously at different neighbouring sites to cover all three of the enzyme substrate subsites. HFG and L95 act as one triple-site inhibitor in tandem and form an unusual ternary complex wherein HFG occupies the 3’-end of tRNA and the L-proline (L-pro) binding sites while L95 occupies the ATP pocket. These inhibitors exhibit nanomolar IC50 and EC50 values independently, and when given together reveal an additive mode of action in parasite inhibition assays. This work validates a novel approach and lays a structural framework for further drug development based on simultaneous targeting of multiple pockets to inhibit druggable proteins.

Investigation of Antiparasitic Activity of Two Marine Natural Products, Estradiol Benzoate, and Octyl Gallate, on Toxoplasma gondii In Vitro.

Toxoplasmosis, caused by Toxoplasma gondii, is a common disease worldwide and could be severe and even fatal in immunocompromised individuals and fetuses. Limitation in current available treatment options drives the need to develop novel therapeutics. This study assessed the anti-T. gondii potential of 103 marine natural products. A luminescence-based β-galactosidase activity assay was used to screen the marine natural products library. Afterward, those compounds that displayed over 70% parasite inhibition ratio were further chosen to assess their cytotoxicity. Compounds exhibiting low cytotoxicity (≥80% cell viability) were applied to evaluate the inhibition efficacy on discrete steps of the T. gondii lytic cycle, including invasion, intracellular growth, and egress abilities as well as the cell cycle. We found that both estradiol benzoate and octyl gallate caused >70% inhibition of tachyzoite growth with IC50 values of 4.41 ± 0.94 and 5.66 ± 0.35 μM, respectively, and displayed low cytotoxicity with TD50 values of 34.11 ± 2.86 and 26.4 ± 0.98 μM, respectively. Despite their defects in inhibition of invasion and egress of tachyzoite, the two compounds markedly inhibited the tachyzoite intracellular replication. Flow cytometric analyses further suggested that the anti-T. gondii activity of estradiol benzoate, rather than octyl gallate, may be linked to halting cell cycle progression of tachyzoite from G1 to S phase. Taken together, these findings suggest that both estradiol benzoate and octyl gallate are potential inhibitors for anti-T. gondii infection and support the further exploration of marine natural products as a thinkable source of alternative and active agents against T. gondii.

Evaluation of anti-malaria potency of wild and genetically modified Enterobacter cloacae expressing effector proteins in Anopheles stephensi

BackgroundMalaria is one of the most lethal infectious diseases in tropical and subtropical areas of the world. Paratransgenesis using symbiotic bacteria offers a sustainable and environmentally friendly strategy to combat this disease. In the study reported here, we evaluated the disruption of malaria transmission in the Anopheles stephensi-Plasmodium berghei assemblage using the wild-type (WT) and three modified strains of the insect gut bacterium, Enterobacter cloacae.MethodsThe assay was carried out using the E. cloacae dissolvens WT and three engineered strains (expressing green fluorescent protein-defensin (GFP-D), scorpine-HasA (S-HasA) and HasA only, respectively). Cotton wool soaked in a solution of 5% (wt/vol) fructose + red dye (1/50 ml) laced with one of the bacterial strains (1 × 109cells/ml) was placed overnight in cages containing female An. stephensi mosquitoes (age: 3–5 days). Each group of sugar-fed mosquitoes was then starved for 4–6 h, following which time they were allowed to blood-feed on P. berghei–infected mice for 20 min in the dark at 17–20 °C. The blood-fed mosquitoes were kept at 19 ± 1 °C and 80 ± 5% relative humidity, and parasite infection was measured by midgut dissection and oocyst counting 10 days post-infection (dpi).ResultsExposure to both WT and genetically modified E. cloacae dissolvens strains significantly (P < 0.0001) disrupted P. berghei development in the midgut of An. stephensi, in comparison with the control group. The mean parasite inhibition of E. cloacaeWT, E. cloacaeHasA, E. cloacaeS−HasA and E. cloacaeGFP−D was measured as 72, 86, 92.5 and 92.8 respectively.ConclusionsThe WT and modified strains of E. cloacae have the potential to abolish oocyst development by providing a physical barrier or through the excretion of intrinsic effector molecules. These findings reinforce the case for the use of either WT or genetically modified strains of E. cloacae bacteria as a powerful tool to combat malaria.Graphical

Antimalarial Activity of Eco-friendly Green Synthesis of Gold Nanoparticles Using Coccinia grandis (L.) Voigt. against Plasmodium berghei

Malaria is one of the life-threatening infectious diseases caused by the Plasmodium parasites infect red blood cells of humans. The major problem associated with malaria treatment and control is the spread and development of resistance against chemical insecticides. Hence, the plant-based formulations may serve as an alternative source for the development of new drugs for the treatment of malaria. The present study is an attempt to evaluate the antiplasmodial activity of synthesized gold nanoparticles (Au NPs) using the aqueous leaves extract of Coccinia grandis against Plasmodium berghei and characterized by UV-Visible (UV-Vis) spectroscopy, Fourier transform infrared, X-ray diffraction, Scanning electron microscopy with Energy dispersive X-ray spectroscopy, and Transmission electron microscopy (TEM) with selected area (electron) diffraction analyzes. The UV-Vis spectra of Au NPs showed a surface plasmon resonance peak range of 500–600 nm. The TEM image revealed the spherical shape with a mean particle size of 2.82 ± 2.5 nm. The in vivo antimalarial activity of synthesized Au NPs showed a significant (P &lt; 0.001) chemosuppressive effect, with parasite suppression of 88.75%, compared to aqueous extracts of C. grandis showed 69.45% against P. berghei at 100 mg/kg body weight. To sum up, further phytochemical research is required in the future to standardize C. grandis compounds with strong antiplasmodial activity and devise the best alternative nano herbal formulation to replace the synthetic drugs currently in use.

A Study of the Antimalarial Activity of Combined Plant Extracts Traditionally Used in Kenya

Introduction: Due to resistance and cost of current antimalarial drugs in use, there is need to continue searching for effective and affordable treatment for this disease. Traditional medicinal plants have been focused on as source of such drugs.&#x0D; Methodology: For this study, combined crude extracts from six plants used in Kenya were tested in vitro against chloroquine resistant Plasmodium falciparum W2 strain using the [3H] hypoxanthine uptake assay and in vivo using Plasmodium berghei on mice. Cytotoxicity tests were done using the MTT based colorimetric assay on Vero 199 and selectivity index used to assess safety of the extracts with the best activity.&#x0D; Results: In vitro results with combinations involving Zanthoxylum chalybeum with four of the extracts indicated activity below IC50 10µg/ml. A combination of S. pinnata and Toddalia asiatica extracts exhibited the best activity in vivo having a parasite growth suppression of 81.3% at a dose of 500mg/kg. S. pinnata and T.asiatica combination had CC50 values of 102.93±5.96µg/ml and &gt;1000µg/ml with selectivity indices of 9.52 and 15.24 respectively, indicating good therapeutic potential due to their safety margins.&#x0D; Conclusion: The results obtained from both in vitro and in vivo tests support the use of the plants selected in traditional treatment of malaria. A combination of methanol fruit extract of T. asiatica and methanol whole plant extract of S. pinnata showed promising parasite growth suppression against P. berghei that was comparable with Artemether and was synergistic on in vitro testing against chloroquine- resistant W2 strain of P. falciparum.

Antimalarial, Antioxidant Activities and Chemoprofile of Sansevieria liberica Gerome and Labroy (Agavaceae) Leaf Extract.

Background Sansevieria liberica Gerome and Labroy (Agavaceae) is a religious and ornamental perennial plant with highly valued medicinal usage in Nigeria. Sansevieria liberica is used in the management of malarial fever. The ease of development of resistance to available antimalarial drugs has resulted in increased clinical failure and mortality. The study investigated the antimalarial effects of Sansevieria liberica (SL) leaf extract in mice infected with Plasmodium berghei. Materials and Methods The ability of SL leaf extract to suppress the growth of malaria parasites in early (suppressive) and established (curative) infections was established using animal models. The mean survival time (MST) was determined. The antioxidant potential was established using two standard in vitro models. High-performance liquid chromatography (HPLC) and phytochemical analysis methods were used to create a chemoprofile fingerprint of SL hydroethanolic leaf extract. Results At 200, 400, and 400·mg kg−1, SL produced 68.71, 70.74, and 75.09% parasite suppression in the suppressive model while the curative model gave a percentage of cure of 71.09, 72.60, and 62.09, respectively. The animals lived longer compared to both negative and positive controls but were not fully protected. The IC50 values of SL and vitamin C were calculated to be 3.599 µg mL−1 and 3.08 µg mL−1, respectively. The reducing power of vitamin C was significantly (P < 0.05) higher than that of SL extract. Some flavonoids were established as possible marker compounds for SL leaf extract. Discussion and Conclusions. The antimalarial assay results demonstrated that the use of SL in folk medicine may have scientific support.

Antiplasmodial Activities of Some Locally Prepared Herbs Against Plasmodium falciparum Obtained from Asymptomatic School Children in Zaria, Kaduna State, Nigeria

The efficacy of current or any intended antimalarial can only be resolute by cultivation and susceptibility studies. The aim of this research was to cultivate Plasmodium falciparum in vitro and comparing the antiplasmodial effects of standard antimalarial medications including herbaceous preparation. Asymptomatic pupils attending some schools in Zaria, Kaduna state, Nigeria were recruited into this research and blood samples were collected from them. Microscopy was done after thin and thick blood films were prepared and stained. The antiplasmodial activities of antimalarial drugs as well as herbal preparation were determined after the successful culturing of red blood cells in the Jatropha curcas medium.The phytochemical constituents of the herbs that made up the concoction were determined. The incidence of asymptomatic Plasmodium infection amongst school kids was 17.5%. There was no statistical association of location, gender and age with the obtained prevalence. Presence of alkaloids was observed in every plant screened while the same was also observed for tannins except in Enantia chloranta.Steroids and phlobatanins were observed in Citrus aurantifolia while there was presence of saponins in all the plant extracts except Cymbopogbon citratus. All the plants except Enantia chloranta contained flavonoids. There was presence of terpenoids in all screened plants except Azadirachta indica and Cymbopogbon citratus while cardiac glycosides were found in every plant except Magnifera indica and Enantia chloranta. Results obtained from the in vitro cultivation of the Plasmodium falciparum with Athemether/lumefanthrine, amodiaquine and herbal concoction showed average percentage parasite inhibition of 80%, 37.8% and 38.6% respectively. This implies that Arthemether/lumefanthrine was capable of inhibiting the growth of the parasite best. The herbal concoction also inhibited growth (38.6% inhibition). There is need for additional investigation on a wider variety of plants to explore their antiplasmodial activities since there is evidence that it works, and it is quite available and affordable.

In vitro and In vivo antimalarial activities of Avicennia africana P. Beauv. (Avicenniaceae) ethanolic leaf extract

BackgroundThe emergence of widespread drug-resistant strains of the malaria parasites militates against strives for more potent antimalarial drugs. AimThe present study evaluated the antimalarial activity of A. africana ethanolic crude extract in vitro and in vivo against Plasmodiumberghei -infected mice in anticipation of acquiring scientific evidence for it used by mangrove dwellers to treat malaria in Ghana. MethodologyThe pulverized dried leaves were extracted with 70% ethanol (v/v) and screened for phytochemicals using standard protocols. The in vitro antimalarial activity was investigated against chloroquine-sensitive Plasmodium falciparum (Pf3D7 clones), MRA-102, Lot:70032033, via SYBR® Green I fluorescent assay method using positive control Artesunate (50–1.56 × 10−3 μg/mL). In the in vivo studies, doses (200–1500 mg/kg) of AAE were used in the 4-day suppressive and curative tests, using P. berghei-infected mice. Artemether/lumefantrine (1.14 mg/kg) and normal saline were used as positive and negative control respectively. ResultsThe phytochemical analysis revealed the presence of alkaloids, saponins, flavonoids, glycosides, tannins, terpenoids and phytosterols. The extract showed an IC50 of 49.30 ± 4.40 μg/mL in vitro and demonstrated complete parasite clearance at dose 1500 mg/kg in vivo with a suppressive activity of 100% (p < 0.0001) in the 4-day suppressive test. The extract demonstrated high curative activity (p < 0.0001) at 1500 mg/kg with 100% parasite inhibition and the oral LD50 > 5000 mg/kg in mice. ConclusionThe results demonstrated that A. africana crude extract has antimalarial activity both in vitro and in vivo supporting the traditional use of the plant to treat malaria.

In vitro screening of novel anti-Babesia gibsoni drugs from natural products.

Babesia gibsoni is a tick-transmitted intraerythrocytic apicomplexan parasite that causes babesiosis in dogs. Due to the strong side effects and lack of efficacy of current drugs, novel drugs against B. gibsoni are urgently needed. Natural products as a source for new drugs is a good choice for screening drugs against B. gibsoni. The current study focuses on identifying novel potential drugs from natural products against B. gibsoniin vitro. Parasite inhibition was verified using a SYBR green I-based fluorescence assay. A total of 502 natural product compounds were screened for anti-B. gibsoni activity in vitro. Twenty-four compounds showed high growth inhibition (>80%) on B. gibsoni and 5 plant-derived compounds were selected for further study. The half-maximal inhibitory concentration (IC50) values of lycorine (LY), vincristine sulfate (VS), emetine·2HCl (EME), harringtonine (HT) and cephaeline·HBr (CEP) were 784.4±3.3, 643.0±2.8, 253.1±1.4, 23.4±1.2, and 108.1±4.3nM, respectively. The Madin-Darby canine kidney (MDCK) cell line was used to assess cytotoxicity of hit compounds. All compounds showed minimal toxicity to the MDCK cells. The effects of hit compounds combined with diminazene aceturate (DA) on B. gibsoni were further evaluated in vitro. VS, EME, HT or CEP combined with DA showed synergistic effects against B. gibsoni, whereas LY combined with DA showed an antagonistic effect against B. gibsoni. The results obtained in this study indicate that LY, VS, EME, HT and CEP are promising compounds for B. gibsoni treatment.

Repurposing the Trypanosomatidic GSK Kinetobox for the Inhibition of Parasitic Pteridine and Dihydrofolate Reductases.

Three open-source anti-kinetoplastid chemical boxes derived from a whole-cell phenotypic screening by GlaxoSmithKline (Tres Cantos Anti-Kinetoplastid Screening, TCAKS) were exploited for the discovery of a novel core structure inspiring new treatments of parasitic diseases targeting the trypansosmatidic pteridine reductase 1 (PTR1) and dihydrofolate reductase (DHFR) enzymes. In total, 592 compounds were tested through medium-throughput screening assays. A subset of 14 compounds successfully inhibited the enzyme activity in the low micromolar range of at least one of the enzymes from both Trypanosoma brucei and Lesihmania major parasites (pan-inhibitors), or from both PTR1 and DHFR-TS of the same parasite (dual inhibitors). Molecular docking studies of the protein–ligand interaction focused on new scaffolds not reproducing the well-known antifolate core clearly explaining the experimental data. TCMDC-143249, classified as a benzenesulfonamide derivative by the QikProp descriptor tool, showed selective inhibition of PTR1 and growth inhibition of the kinetoplastid parasites in the 5 μM range. In our work, we enlarged the biological profile of the GSK Kinetobox and identified new core structures inhibiting selectively PTR1, effective against the kinetoplastid infectious protozoans. In perspective, we foresee the development of selective PTR1 and DHFR inhibitors for studies of drug combinations.

Comparative efficacy of amphotericin B-loaded chitosan nanoparticles and free amphotericin B drug against Leishmania tropica

BackgroundThe preparation of an effective drug delivery formulation is an urgent need to treat cutaneous leishmaniasis (CL). Pentavalent antimonials and Amphotericin B (AmB) are considered to treat leishmaniasis; however, their clinical usage is hampered by poor solubility, high cost, toxicity, and the emergence of drug-resistant Leishmania spp. The drug delivery systems (DDS) could be used as an alternative treatment option for the treatment of CL to circumvent these problems. We tested the antileishmanial efficacies of free AmB and amphotericin B-loaded chitosan nanoparticles (AmB-CNPs) under in vitro conditions.ResultsChitosan nanoparticles (CNPs) were synthesized using the ionic gelation method with negatively charged tripolyphosphate (TPP). During the synthesis of CNPs, AmB was incorporated into the nanoparticles (NPs). The NPs were characterized for their size, surface morphology, encapsulation efficacy (EE), drug loading content (DLC), and surface charge using different techniques. Their efficacy was evaluated against promastigotes and axenic amastigotes forms of Leishmania tropica using MTT assay. The synthesized AmB-CNPs displayed a spherical shape with a mean particle size of 118 nm, a positive zeta potential of (+ 6.21 ± 2.02 mV), and an encapsulation efficacy of 88%. Dynamic light scattering technique (DLS) shows that the average size of prepared AmB-CNPs was 95.5 nm. Free AmB presented very low efficacy (only 65% and 67% inhibition of the promastigotes and axenic amastigotes parasite load), whereas AmB-CNPs exhibited 90% and 84% parasite inhibition after 72 h incubation. The AmB-CNPs exhibited significantly higher efficacy than free AmB in terms of reduction in parasite viability. Half-maximal inhibitory concentration (IC50) measured values of the AmB-CNPs were significant lowers than free AmB.ConclusionsThe present data indicated that AmB-CNPs exhibited vigorous anti-leishmanial activity than free AmB by dose and time-dependent manner. This formulation can be used for local therapy of CL after in vivo efficacy conformational studies.

Predicting Parasite Dynamics in Mixed-Use Trans-Himalayan Pastures to Underpin Management of Cross-Transmission Between Livestock and Bharal.

The complexities of multi-use landscapes require sophisticated approaches to addressing disease transmission risks. We explored gastro-intestinal nematode (GINs) infections in the North India Trans-Himalayas through a socio-ecological lens, integrating parasite transmission modelling with field surveys and local knowledge, and evaluated the likely effectiveness of potential interventions. Bharal (blue sheep; Pseudois nayaur), a native wild herbivore, and livestock share pasture year-round and livestock commonly show signs of GINs infection. While both wild and domestic ungulates had GINs infections, egg counts indicated significantly higher parasite burdens in bharal than livestock. However, due to higher livestock densities, they contributed more to the total count of eggs and infective larvae on pasture. Herders also reported health issues in their sheep and goats consistent with parasite infections. Model simulations suggested that pasture infectivity in this system is governed by historical pasture use and gradually accumulated larval development during the summer, with no distinct short-term flashpoints for transmission. The most effective intervention was consequently predicted to be early-season parasite suppression in livestock using temperature in spring as a cue. A 1-month pause in egg output from livestock could lead to a reduction in total annual availability of infective larvae on pasture of 76%, potentially benefitting the health of both livestock and bharal. Modelling suggested that climate change over the past 33 years has led to no overall change in GINs transmission potential, but an increase in the relative influence of temperature over precipitation in driving pasture infectivity. Our study provides a transferable multi-pronged approach to investigating disease transmission, in order to support herders' livelihoods and conserve wild ungulates.

Antimicrobial properties of tomato leaves, stems, and fruit and their relationship to chemical composition

BackgroundWe previously reported that the tomato glycoalkaloid tomatine inhibited the growth of Trichomonas vaginalis strain G3, Tritrichomonas foetus strain D1, and Tritrichomonas foetus-like strain C1 that cause disease in humans and farm and domesticated animals. The increasing prevalence of antibiotic resistance requires development of new tools to enhance or replace medicinal antibiotics.MethodsWild tomato plants were harvested and divided into leaves, stems, and fruit of different colors: green, yellow, and red. Samples were freeze dried and ground with a handheld mill. The resulting powders were evaluated for their potential anti-microbial effects on protozoan parasites, bacteria, and fungi. A concentration of 0.02% (w/v) was used for the inhibition of protozoan parasites. A high concentration of 10% (w/v) solution was tested for bacteria and fungi as an initial screen to evaluate potential anti-microbial activity and results using this high concentration limits its clinical relevance.ResultsNatural powders derived from various parts of tomato plants were all effective in inhibiting the growth of the three trichomonads to varying degrees. Test samples from leaves, stems, and immature ‘green’ tomato peels and fruit, all containing tomatine, were more effective as an inhibitor of the D1 strain than those prepared from yellow and red tomato peels which lack tomatine. Chlorogenic acid and quercetin glycosides were present in all parts of the plant and fruit, while caffeic acid was only found in the fruit peels. Any correlation between plant components and inhibition of the G3 and C1 strains was not apparent, although all the powders were variably effective. Tomato leaf was the most effective powder in all strains, and was also the highest in tomatine. S. enterica showed a minor susceptibility while B. cereus and C. albicans fungi both showed a significant growth inhibition with some of the test powders. The powders inhibited growth of the pathogens without affecting beneficial lactobacilli found in the normal flora of the vagina.ConclusionsThe results suggest that powders prepared from tomato leaves, stems, and green tomato peels and to a lesser extent from peels from yellow and red tomatoes offer potential multiple health benefits against infections caused by pathogenic protozoa, bacteria, and fungi, without affecting beneficial lactobacilli that also reside in the normal flora of the vagina.

In vitro antiplasmodial evaluation of ethanolic and n-hexane extracts of Parinari curatellifolia stem bark

Parinari curatellifolia and other Parinari species are used traditionally in many parts of Africa as a remedy for malaria among other diseases. To ascertain this folkloric claim, the antiplasmodial potential of ethanol extract of Parinari curatellifolia stem bark (EEPCSB) and n-hexane extract of Parinari curatellifolia stem bark (HEPCSB) on Plasmodium falciparum was studied. Parasites were grown in a 96-well plate containing Roswell Park Memorial Institute-1640. The wells were grouped into: control (untreated), artemether-treated, EEPCSB-treated and HEPCSBtreated groups. Treatments were administered to the tune of 10, 20, 40 and 80 μg/ml. Parasitemia was observed by microscopy after 24, 48 and 72h of incubation. EEPCSB and HEPCSB elicited dose and duration-dependent reduction (p&lt;0.05) in parasitemia when compared with the untreated group. The recorded percentage parasite inhibition by the extracts was lower (p&lt;0.05) when compared with artemether. There was no difference (p&gt;0.05) in plasmodium lactate dehydrogenase activity of EEPCSB-treated and artemether-treated groups. Findings from this study show that extracts of P. curatellifolia stem bark, especially EEPCSB, demonstrated excellent inhibitory activities against P. falciparum and can be a good source of compounds for the development of novel antimalarial drugs.&#x0D; Keywords: Parinari curatellifolia; Extracts; Plasmodium falciparum; Parasitemia; Antiplasmodial

Curzerene antileishmania activity: Effects on Leishmania amazonensis and possible action mechanisms

Leishmaniasis is a set of infectious diseases with high rates of morbidity and mortality, it affects millions of people around the world. Treatment, mainly with pentavalent antimonials, presents significant toxicity and many cases of resistance. In previous works we have demonstrated the effective and selective antileishmanial activity of Eugenia uniflora L. essential oil, being constituted (47.3%) by the sesquiterpene curzerene. Considering the high rate of parasite inhibition demonstrated for E. uniflora essential oil, and the significant presence of curzerene in the oil, this study aimed to evaluate its antileishmania activity and possible mechanisms of action. Curzerene was effective in inhibiting the growth of promastigotes (IC50 3.09 ± 0.14 µM) and axenic amastigotes (EC50 2.56 ± 0.12 µM), with low cytotoxicity to RAW 264.7 macrophages (CC50 83.87 ± 4.63 µM). It was observed that curzerene has direct effects on the parasite, inducing cell death by apoptosis with secondary necrotic effects (producing pores in the plasma membrane). Curzerene proved to be even more effective against intra-macrophage amastigote forms, with an EC50 of 0.46 ± 0.02 µM. The selectivity index demonstrated by curzerene on these parasite forms was 182.32, being respectively 44.15 and 8.47 times more selective than meglumine antimoniate and amphotericin B. The antiamastigote activity of curzerene was associated with immunomodulatory activity, as it increased TNF-α, IL-12, and NO levels, and lysosomal activity, and decreased IL-10 and IL-6 cytokine levels detected in macrophages infected and treated. In conclusion, our results demonstrate that curzerene is an effective and selective antileishmanial agent, a candidate for in vivo investigation in models of antileishmanial activity.

In Vivo Antiplasmodial Activity and Toxicological Analyses of the Ethanolic Leaf and Twig Extract of Faurea speciosa Welw. (Proteaceae).

In Africa, medicinal plants are commonly used to treat malaria and other diseased conditions. The ethanolic leaf and twig extract of Faurea speciosa has been shown to possess promising antiplasmodial properties. This present study was aimed at investigating its antiplasmodial effect in vivo. Qualitative phytochemical screening was carried out on the plant samples using standard methods. The antiplasmodial effect against early infection, curative effect against established infection, and prophylactic effect against residual infection were studied in vivo in Plasmodium berghei-infected mice while the carrageenan-induced edema model in chicks was used for anti-inflammatory tests. The phosphomolybdenum and DPPH radical scavenging assays were used in the evaluation of antioxidant potential. Acute toxicity of the extract was evaluated using the Organization for Economic Cooperation and Development (OECD) guidelines. Phytochemical screening of plant samples revealed the presence of flavonoids, coumarins, tannins, saponins, and glycosides. Faurea speciosa leaf and twig extract exhibited significant antiplasmodial activities in the mouse model with parasite suppression rates of 66.63%, 71.70%, and 56.93% in the suppressive, curative, and prophylactic tests, respectively. A 55.50% reduction of edema in the anti-inflammatory test indicated moderate success in reducing inflammation. The total antioxidant capacity of the extract was determined to be 65.4 mg AAE/g of extract, while in the DPPH radical scavenging assay, the IC50 value was found to be 499.4 μg/mL. With the exception of an inconsistent rise in urea level, there was no significant difference in the other biochemistry parameters in the acute toxicity studied. The median lethal dose (LD50) of the extract was over 2000 mg/kg. The results of this study show that Faurea speciosa leaf and twig extract has promising antimalarial capabilities and is fairly safe at low concentrations.