Development of a saprophytic fungal inoculum for the biodegradation of sub-bituminous coal

The rhizospheres of the weeds Ageratum conyzoides, Axonopus compressus, Emilia coccinea, Synedrella nodiflora, Urena lobata and Sida acuta from a sub-bituminous coal mining site and a control site, without coal discards, were screened for new fungi with ability to degrade sub-bituminous coal in the laboratory. The isolates were identified by cultural and molecular methods. Seventeen out of the sixty-one fungal isolates tested could utilize sub-bituminous coal as an energy source. Upon further evaluation, only seven of these were promising candidates for coal biodegradation, and they were assayed for their biosolubilization and depolymerization activities to determine their mechanisms of coal biodegradation. Based on the accumulation of humic acid (HA), which is the marker for biosolubilization, Mucor circinelloides and Aspergillus tubingensis were the most active. On the other hand, Cunninghamella bertholletiae, Simplicillium subtropicum, Penicillium daleae and Trichoderma koningiopsis were the highest producers of fulvic acid (FA), the indicator of depolymerization. Purpureocillium lilacinum produced the lowest yields of both HA and FA compared to the other six coal-degrading candidates. The presence of laccase in Trichoderma koningiopsis, Penicillium daleae and Simplicillium subtropicum suggests a role for this enzyme in the enhancement of the coal biodegradation process. However, the inability to amplify the laccase gene in Cunninghamella bertholletiae indicates that another enzyme probably aids its coal bioconversion. The current investigation highlights the potentials of these strains in harnessing biotechnological processes of sub-bituminous coal conversion into value-added products, which could be extended to the bioremediation of coal-polluted soils. The fungi with the highest coal bioconversion capabilities belonged to Ascomycota and Zygomycota and were found in the rhizospheres of the weeds Emilia coccinea, Ageratum conyzoides and Axonopus compressus.

In Vitro Assessment of the Cercaricidal Activity of Sida acuta Burm. F. and Sida rhombifolia Linn. (Malvaceae) Hydroethanolic Extracts, Cytotoxicity, and Phytochemical Studies

Despite the global efforts, schistosomiasis remains a public health problem in several tropical and subtropical countries. One of the major challenges in the fight against schistosomiasis is the interruption of the parasite life cycle. Here, we evaluated the anticercarial, cytotoxicity, and phytochemical profiles of Sida acuta (HESa) and Sida rhombifolia (HESr) hydroethanolic extracts (Malvaceae). Schistosoma mansoni cercaria was collected from fifteen Biomphalaria pfeifferi-infected snails. Twenty-five cercariae were incubated in duplicate with different concentrations (31.25–1,000 μg/mL) of HESa or HESr. The cercaria viability was monitored at 30 min time intervals for 150 min, and the concentration-response curve of each plant extract was used to determine their respective lethal concentration 50 (LC50). Additionally, the cytotoxicity profile of each plant extract was evaluated on the Hepa 1–6 cell line at a concentration range of 15.625–1,000 µg/mL using the WST-8 assay method and its inhibitory concentration 50 (IC50) was calculated. Moreover, phytochemical characterization of each plant extract was carried out by HPLC-MS. Both extracts exhibited cercaricidal activity in a time- and concentration-dependent manner. At 30 min time point, HESa (LC50 = 28.41 ± 3.5 µg/mL) was more effective than HESr (LC50 = 172.42 ± 26.16 µg/mL) in killing S. mansoni cercariae. Regarding the cytotoxicity effect of both extracts, the IC50 of HESa (IC50 = 109.67 µg/mL) was lower than that of HESr (IC50 = 888.79 µg/mL). The selectivity index was 3.86 and 5.15 for HESa and HESr, respectively. Fifteen compounds were identified from HESa and HESr after HPLC-MS analysis. N-Feruloyltyramine, a polyphenol, and thamnosmonin, a coumarin, were identified in both extracts. HESa and HESr displayed cercaricidal activity and were not toxic on Hepa 1–6 cell line. Based on the selectivity index of these extracts, S. rhombifolia extract could be more effective on S. mansoni cercariae than S. acuta extract. This study could provide baseline information for further investigations aiming to develop plant-based alternative drugs against S. mansoni.

Phytochemistry and In vitro Antimicrobial Activity of Five Plant Species against Nine Common Human Pathogens

Momordica charantia, Senna podocarpa, Senna alata, Ocimum gratissimum, and Sida acuta which have reportedly been used in folklore for the treatment of various diseases were studied for their antimicrobial activity. Crude methanolic extracts of the leaves were screened for phytochemicals after which they were tested in vitro for activity against clinical isolates of Staphylococcus aureus, Klebsiella oxytoca, Pseudomonas aeruginosa, Proteus vulgaris, Bacillus subtilis, Enterobacter aerogenes, Aspergillus niger, A, flavus and Candida albicans. Phytochemicals present in the extracts included saponins, alkaloids, anthraquinones, tannins, flavonoids and cardiac glycosides. These extracts also showed varying degrees of activity against tested organisms at a concentration range of 15.6 to 500 mg/ml. However, S. acuta showed activity against P. vulgaris, E. aerogenes and As. flavus only. The MIC of the extracts against test organisms ranged from 31.25 to 250 mg/ml. The significant antimicrobial susceptibility of the plant extracts against gram positive bacterial pathogens and some pathogenic yeasts, may not be due to the presence and synergistic interactions of secondary metabolites found in these plant extracts. This would have implications in health, particularly in developing countries where a singnificant percentage of the population are still using traditional plant extracts for health care. Dhaka Univ. J. Pharm. Sci. 20(2): 139-148, 2021 (December)

Antimicrobial activity of Sida acuta, Phyllanthus amarus and Phyllanthus muellerianus against microorganisms implicated in urinary tract infections

Increasing level of antimicrobial resistance among bacterial pathogens causing Urinary Tract Infection (UTI) is one of the most significant public health challenges globally. Hence, the search for alternatives from medicinal plants. This study investigated the efficacy of Phyllanthus amarus (PA), Phyllanthus muellerianus (PM) and Sida acuta (SA) leaf extracts on microorganisms implicated in UTI. Mid-stream urine samples collected from 100 patients clinically diagnosed with UTI were cultured. The microorganisms isolated were identified using their morphological and biochemical characteristics. Methanol leaf extracts of the three plants were obtained by cold maceration in 60% methanol. Crude extract of PM was thereafter purified by solvent partitioning. Antibiotic susceptibility test was determined using the Kirby Bauer disc diffusion. Antimicrobial effects of the extracts and oil was ascertained using agar well diffusion. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentrations (MBC) were also determined. Rate of kill and mechanism of action of the purified extract of PM on isolates were investigated. Cytotoxicity of plant extracts were assayed on brine shrimps while synergism of the purified extract with ciprofloxacin was ascertained using overlay inoculum susceptibility disc method. Antioxidant and phytochemical analyses of the extracts were conducted using standard methods. Phytochemical analysis of the leaf extracts showed the presence of alkaloids, tannins, saponins and steroids. Antioxidant assay also indicated SA had the highest total flavonoids and phenol content of 339.86 mgQUE/g and 27.63 mgGAE/g. Microorganisms isolated include: Escherichia coli (24%), Proteus mirabilis (24%), Staphylococcus aureus (19%), Klebsiella pneumoniae (13%), Candida albicans (11%), Enterobacter sp. (5%) and Citrobacter sp. (4%). The crude extract of PA had zone of inhibition ranging from 16.7 ± 1.53 mm to 24 ± 1.00 mm while SA crude extract had 14.7 ± 1.53 mm to 27 ± 2.00 mm. PM crude extract had inhibition zones of 17 ± 1.00 mm to 22.3 ± 2.12 mm. The MIC and MBC ranged from 6.25 mg/ml to 50 mg/ml and 12.5 mg/ml to 50 mg/ml respectively. Ethyl acetate fraction of PM showed the highest percentage yield and had a zone diameter range from 13.5 ± 1.00 mm to 28 ± 1.53 mm with MIC and MBC ranges of 6.25 mg/ml – 12.5 mg/ml and 25 mg/ml to 50 mg/ml respectively. Synergism with ciprofloxacin was observed at 25% of the microorganisms, 50% antagonism and 25% additively. Toxicity analysis showed lethal dose concentrations of 19.05 mg/ml, 25.12 mg/ml and 130.11 mg/ml for PM, PA and SA respectively. The findings of this study suggest that the methanol extracts of the medicinal plants used in this study does possess a potent lead molecule in combating microorganisms causing UTI. Key words: Antimicrobial activity, Phyllanthus muellerianus, Phytochemicals, Toxicity, UTI,

ETHNO BOTANICAL APPROACHES, PHARMACOLOGICAL AND PHYTOCHEMICAL BENEFITS OF GENUS SIDA USED IN TRADITIONAL MEDICINES

The four species of Sida genus viz: Sida cordifolia L., Sida acuta Burm. f., Sida cordata Burm. f. and Sida rhombifolia L. are medicinal herbs plants reported for its used in Indian system of traditional medicines of Ayurveda and Siddha. The main objective of the review is to deliver detailed information on botanical descriptions, traditional uses, ethno medicinal benefits, pharmacological activities and meaningful knowledge on these plants for future research. Plants were collected from Agra region and identified by RARI, Jhansi, Ministry of AYUSH, Govt. of India for exact information. Scientific literature collected from journals, books and libraries, electronic sources like PubMed, Web of Science, Science Direct, Springer, Scopus and major search engines to obtain get relevant information. Several parts of Sida plants used by the traditional system of herbal medicine which are traditionally used in diarrhea, leucorrhoea, gonorrhea, asthma, wheezing, fever, cold, flu, headache, weight loss, sexual strength, hair strength, hypertension, diuretic, piles, ulcer, cancer, aphrodisiac, rheumatism, urinary, venereal, skin, respiratory and heart diseases. Pharmacological activity viz: antioxidant, antibacterial, antimicrobial, anti-inflammatory, anticancer and anti-diabetic property are present, and rich in saponins, alkaloids, flavonoids, coronaric acid, coumarins, ephedrine, pseudoephedrine and ethno medicinal property. This review will serve as a database to provide knowledgeful information about the medicinal significance, traditional uses, ethno medicinal and ethno pharmacological benefits and contribution of Sida plant

Sida acuta Burm.f. leaves ethanol extract ameliorates haematological and biochemical alterations induced by Plasmodium berghei ANKA-65 in mice

Background Malaria has continued to be a threat to man and his wellbeing, especially Africans and Asians. New antimalarial drugs are urgently needed to mitigate malaria treatment failure due to resistant Plasmodium species. Medicinal plants used by indigenous Nigerians for treating fever and malaria such as Sida acuta Burm.f. (Malvaceae) could be a promising source of lead compounds for developing new generations of antimalarial drugs. The effects of ethanol extract of S. acuta leaves (EESAL) on malaria parasitemia, haematological and biochemical status of P. berghei-infected mice were investigated, using the 4-day curative test. Methodology EESAL was prepared by maceration method. The phyto-constituents and acute toxicity profile of the extract were evaluated using standard protocols. In addition, malaria parasitemia and chemo-suppression, and indicators of haematological and biochemical status of P. berghei-infected mice treated with EESAL were assessed. Results At 200, 400 and 600 mg/kg/d b.w., p.o doses for 4 consecutive days, EESAL significantly (p < 0.05) decreased parasitaemia and suppressed malaria parasite by 89.64%, 95.95% and 97.38%, respectively comparable to negative control. The reduction in percentage malaria parasitemia by EESAL is comparable to Artemether (140 mg/kg/d b.w., p.o) used as standard antimalarial drug in this study. The packed cell volume (PCV), haemoglobin (Hb) concentration, and red blood cell (RBC) and white blood cell (WBC) counts of negative control are significantly (p < 0.05) higher than normal control. However, parasitized-EESAL-treated mice have significantly (p < 0.05) higher PCV value, Hb concentration and RBC and WBC counts than negative control. Similarly, treatment of parasitized mice with EESAL restored some indicators of the antioxidant, lipid peroxidation, lipid profile and liver status altered by malaria. In addition, EESAL was tolerable up to 5000 mg/kg b.w., p.o. Conclusion These results indicate that the EESAL possesses antimalarial activity and normalizes alterations in haematological and biochemical status of malaria-infected mice.

Evaluation of Immunomodulatory Activity of Extract and Fractions of Sida acuta Burm. f. Leaves in Mice and GC-MS Analysis of n- Hexane Fraction

The leaves of Sida acuta Burm. f. (Malvaceae) has been reported to possess potent anti- inflammatory, anti- plasmodial and anti-microbial activities. The relationship of these bioactivities and immune responses lead to the evaluation of the immunomodulatory activity of Sida acuta Burm. f. leave extract and fractions. This our study was done to determine the immunomodulatory activity and chemical study of methanol leave extract and fractions of Sida acuta Burm. f. The immunomodulatory evaluation was done by invivo Delay Type Hypersensitivity reaction (DTHR) in the body and in vitro measurement of phagocytosis of killed Candida albicans by the phagocyte polymorphonuclear leucocytes using slide method. Acute toxicity, phytochemical and GC-MS analysis were also performed. The DTHR tested in the blood with T-cells in mice showed that the extract and its fractions caused a delayed hypersensitivity response in 24hrs which was very significant (P ? 0.05) in the n- hexane fraction of the extract when compared to the control group at the dose of 100mg/kg. The in vitro studies showed a very significant difference (P ? 0.05) in the positive control group (LEVA) at concentration of 50, 100 and 200µg/ml, in crude extract (SrE) at concentrations of 50, 100 and 200µg/ml, n- hexane fraction 50, 100 and 200µg/ml, Ethyl acetate fraction at 200µg/ml and Absolute methanol fraction at 100µg/ml and also have high percentage phagocytic stimulation (PPS). The acute toxicity test did not cause clinical signs or death within 24hours post treatment in all doses tested and highest dose of 5000mg/kg. Phytochemical analysis revealed the presence of flavonoids, saponins, alkaloids, triterpenoids, tannins, steroids and cardiac glycosides. GC-MS analysis of fraction with highest activity was carried out on n-hexane fraction which showed the presence of some compounds like hexadecanoic acid, 2-hydroxy-1 (hydroxymethyl) ethyl ester, 3,4-seco-5alpha-cholestan-3-oic acid,4-hydroxy-4-methyl epsilon-lacto

Evaluation on In Vitro Blood Clot Dissolving Potential of Aqueous Extract of Sida acuta Burm. F. Leaves

Clotting of blood is the vital processes and a perplexing interaction of various mechanisms of circulatory system due of failure of which is sometimes considered as a concern within the circulatory system causing acute myocardial or cerebral infarction which might cause demise. Sida acuta burm. f (Malvaceae) is abundantly growing small perennial shrub utilized by natives for diuretic, anthelmintic, calmative and wound healing properties, and are utilized in treating disorders like blood, bile, liver, nervous, urinary diseases and rheumatism. The present study was intended to evaluate the blood clot dissolving potential of Sida acuta leaf aqueous extract in vitro. The plant material as leaves were locally collected and subjected to phytochemical extraction with distilled water. The preliminary phytochemical tests total phenolic content was estimated by Folin-Ciocalteu’s method. In vitro thrombolytic activity of 3 different concentrations of aqueous extract was estimated on goat blood clot compared to the activity of streptokinase. The aqueous extract of S. acuta leaves are reported to be rich in alkaloids, flavonoids, tannins, terpenoids and glycosides while the total phenolic content was estimated to be 17.48 % in extract which are mostly responsible for any pharmacological activity. Compared to the thrombolytic activity of standard streptokinase which was 73 %, the aqueous leaf extract of S. acuta displayed considerable blood clot dissolving activity at concentration 10 mg/100µl, 5.0 mg/100µl, and 2.5 mg/100µl as 41 %, 34 % and 12 % respectively. This property of plant extract is promising which could be could be exploited in development of new biopharmaceutical and therapeutic agents after stringent further physiological compatibility and in vivo pharmacological studies. Keywords: Sida acuta, phytochemical extract, thrombolytic activity, streptokinase

Weed Flora Dynamics in Maize (Zea mays L.) Field as Influenced by Cover Crop Types and Planting Densities at Jalingo, Northeast, Nigeria

Aim: Competition from weeds is an obstacle to profitable maize production. Knowledge of weed species limiting productivity is essential for sustainable management. A two years field experiment was conducted to evaluate the effect of a leguminous cover crop, akidi (Vigna unquiculata sub-sp sequipedalis) and two non-leguminous cover crops (melon and sweet potato) planted at three densities on weed flora composition and shift in maize at Jalingo, North-eastern Nigeria. Study Design: There were 11 treatments replicated three times in a randomized complete block design. Methodology: Low, moderate and high densities (20,000, 30,000 and 40,000 stands/ha respectively) of Akidi, Melon and Sweet potato (A1, A2, A3, M1, M2, M3, S1, S2, S3) with weeded (C1) and unweeded (C2) checks constituted the treatments laid out in a randomized complete block design replicated three times. Weed flora abundance, percentage density, percentage frequency, frequency index and percentage contributions were determined from a 50cm x 50cm quadrat thrown twice per plot at various phases of maize growth. Results: Thirteen weed families made up of 11 broad leaves, one grass and one sedge were identified. The abundance was in the order Broad leaf > Grass > Sedges. The most abundant weed species were: Imperata cylindrica L (3.8), Mucuna utilis (3.0), Oldenlandia herbacea L. (2.6) Sida acuta Burm F, Tridax procumbens L. (2.5 each), Leucas martinicensis, Pennisetum pedicellatum (2.3 each). Commelina benghalensis, Cyperus difformis (3.5), Digitaria horizontalis, Fimbristylis littoralis (2.8 each). The influence of selected cover crops on individual weed percentage the contribution shows: Commelina benghalensis ( C2<C1 < M <S< A), Euphorbia hyssopifolia (C2<M/S < A<C1), Euphorbia heterophylla (M<S<A), Leucasmartinicensis (C1<C2<M<A<S), Mucunautilis (S<M<C1<C2<A) and Sida acuta (M<S<A<C2<C1) while grassy weeds, Imperata cylindrical (A<C2<S<M), Pennisetum pedicellatum (A<C2), Digitaria horizontalis (C1/ C2< A < M <S), Kyllinga squamulata (M/S <C2<A <C1). Commelina spp. increased in status from an accessory (21.2%) to abundance (66.7%) while others increased from rare to accessory (from 0.0 to 20-40%). Conclusion: Akidi was more effective on broad-leaved weeds while melon and sweet potato were more effective on grasses and Commelina benghalensis need to be monitored.