ABSTRACT
Medicinal plants contain active ingredients with therapeutic values. Helianthus annuus L (Asteraceae) commonly referred to as “sunflower” has been reportedly used in managing and treating some ailments over the years. This study investigated the in vitro and in vivo antibacterial effects of the crude extracts of Helianthus annuus seeds on Salmonella spp and Eschericha coli. The phytochemical screening of the methanolic and ethanolic crude extracts revealed the presence of alkaloids, saponins, tannins, flavonoids and phenols. The effect of the crude extracts against the test organisms ranges from 7.33±0.67 mm to 25.67±0.33 mm at concentrations ranging from 60 mg/ml to 480 mg/ml. The minimum inhibitory concentration of the extracts ranges from 0.07±0.03 mg/ml to 2.40±0.06 mg/ml. The minimum bactericidal concentration of the extracts ranges from 0.48±0.01 mg/ml to 2.40±0.12 mg/ml. The acute toxicity study revealed LD50>2000 mg/kg. The sub acute toxicity study showed that there was a significant decrease in Alanine transaminase, Aspartate transaminase and Alanine phosphatase levels in most of the treated groups compared to the control group. Total cholesterol, triglyceride and low density lipoprotein cholesterol were significantly reduced compared to the control while a significant increase in high density lipoprotein was observed in the treatment groups. The serum biochemical protein components were comparable to the control. The result of the haematological study revealed that mean cell volume, haemoglobin and packed cell volume levels were significantly increased in most of the treated groups compared to the control group. There were no features of acute or chronic damage by the extracts on the hepatic and renal tissues. The in vivo antibacterial effect of the extracts revealed decrease in total bacteria load in treated rats. Therefore, H. annus seeds extracts possess antibacterial effects against Salmonella spp and E.coli and is considered to be relatively safe. Further studies should be carried out to identify the exact active compound or compounds that possess bactericidal activity. There is need to establish the validity and mechanism of action of the hepato-protective effect of H.annuus seed extracts inferred in this study.
CHAPTER ONE
1.0 INTRODUCTION
1.1 Background to the Study
Medicinal plants contain active ingredients in one or more of its parts with therapeutic value or play the role of precursors for chemo-pharmaceutical synthesis (Bassam, 2012). Over the years, several medicinal plant parts have been utilized in managing and treating certain disease conditions, however only a few have been successfully evaluated and scientifically validated for the claimed effects (Saravanakumar et al., 2010). Among the plants used in traditional medicine is Helianthus annuus.
Sunflower (Helianthus annuus L) is majorly cultivated in certain areas in the world for its seed, which is an important source of edible oil (Eze et al., 2015). It is a carrier oil and also used in the production of biodiesel and margarine, as it is less expensive compared to olive oil. Sunflowers have the potential to play vital role in phytoremediation as it can be used to extract toxic chemicals like heavy metals, such as lead, arsenic and uranium from soil, as well as engaged in rhizofiltration to neutralize radionuclides and other toxic compounds and materials as well as harmful bacteria from water (Adler, 1996). Extracts from the leave as well as other preparations made from different parts of plant, are used in the treatment of high fevers, as a poultice on sores, swellings, snakebites and spider bites, in the treatment of malaria, lung ailments, diabetes (Saini and Sharma, 2011).
Helianthus annuus seed oil, and herbal tincture have been reported to have anti- inflammatory, antioxidant, antitumor, antipyretic, antihypoglycemic, cathartic, diuretic and antimicrobial activity (Aboki et al., 2012). It has shown antimicrobial properties against diverse microorganism such as Staphylococcus aureus, Escherichia coli, Bacillus subtilis among others. The effect of polar oil extract from the seeds of sunflower (Helianthus annuus) in Napkin dermatitis and its antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli and Proteus vulgaris were studied (Ahmad et al., 2015). Sunflower oil is highly absorbable through the dermal tissues, providing significant nourishment and moisturizing effects. As a result of this, it is therefore, famous as an over-the-counter ingredient as well as ingredients used in the manufacturing of homemade beauty products such as lotions, creams and massage oils. Oil extracted from sunflower plants may also possess a protective barrier property which resists infections in developing infants. Infants administered this oil subcutaneously on a daily basis were 69 % more likely not to develop infections in the hospital (Ahmad et al., 2015).
1.2 Statement of the Research Problem
Antimicrobial resistance (AMR) is a complex and growing international public health problem (WHO, 2015). Antimicrobial resistance is estimated to account for more than 700,000 deaths annually. Antimicrobial resistance is projected to increase to 10 million deaths with estimated cost of about US$100 trillion by the year 2050 (O’Neill, 2016). Antimicrobial resistance affects every country irrespective of income and development (O’Neill, 2016). The rapid spread of multidrug resistant pathogens is of global threat. Drug-resistant pathogens accounts for 214,000 neonatal sepsis leading to deaths annually (Laxminarayan, 2016). Antimicrobial resistance is not restricted to communicable diseases, it also has potential consequences on non-communicable diseases (NCDs). Antimicrobial resistance poses threat to surgical interventions, cancer treatment and organ transplant (Laxminarayan, 2013). About 50 % of pathogens implicated in surgical site infections are resistant to standard prophylactic antibiotics (Teillant et al., 2015). Cancer patients who have undergone chemotherapy also suffer infections caused by pathogens which are resistant to commonly used antibiotics (Teillant et al., 2015).
Treating drug-resistant infections is expensive and takes longer period and has lower success rate than treating drug-susceptible infections. Therefore, AMR negatively impacts on national economic performance (Smith et al., 2015). The challenges of AMR and its magnitude within the African Region including Nigeria have been monitored by surveillance of drug resistance. In the past decades, the widespread of antibiotic used against microbial pathogens of human and animal origin has led to resistance to many antimicrobial agents which constitute a major health threat around the globe (WHO, 2013).
One of the factors contributing to AMR is misuse of antibiotics. However, lack of tests for other infections in patients who test negative for malaria has led to indiscriminate use of antibiotics (Do et al., 2016). According to Leopold et al. (2014), there is high level of resistance to commonly used antibiotics in the sub-Saharan African region. For example, 90% of Gram negatives were resistant to chloramphenicol, a commonly used antibiotic while resistance to third-generation cephalosporins (ceftriaxone) was less common (Leopold et al., 2014).
In Nigeria, resistance to Ampicillin increased from 70 to 90 %, Co-trimoxazole from 77 to 85 %, Chloramphenicol from 71 to 77 % and Streptomycin from 71 to 79 % between 1990 and 2000 in finding the root cause of antimicrobial resistance (Ifeanyi, 2012). Methicillin resistant Staphylococcus aureus (MRSA), Vancomycin Resistant Staphylococcus aureus (VRSA), carbapenem resistant Enterobacteriaceae, Multi drug and extensive-drug resistant Mycobacterium tuberculosis (MDR-TB and XDR-TB) have also been reported (Nannini, 2013).
The transfer of genetic elements such as plasmids, transposons, and integrons has contributed to the rapid spread of AMR among bacterial species. According to Harbottle (2011), AMR genes have been found on mobile elements, leading to multidrug resistance which can be transferred to a susceptible recipient via a single genetic event. The emergence of multidrug-resistant bacteria can only be detected through systematic screening in quality assured microbiology laboratories ( Xavier et al., 2010; Liu et al., 2016). The health and economic consequences of AMR has led to collaborative global action to improve access to antimicrobial medicines and to promote availability of new product (Mendelson, 2015). Therefore, the use of herbal traditional medicines has emerged as effective alternatives.
1.3 Justification for the Study
Helianthus annuus has been found to possess broad spectrum of in vitro antimicrobial activity against pathogenic microorganisms such as Staphylococcus aureus, Salmonella typhi, Escherichia coli, Candida albican, Aspergillus fumigatus and Rhizopus stolonifera (Subashini and Rakshitha, 2012; Eze et al., 2015). As evident from the antibacterial study of H. annuus seeds by Rubab et al. (2016), there are claims that it may contain important chemical substances that confer this plant as medicinal agent possessing antibacterial activity. According to Sharma (2014), H.annuus contains various alkaloids, flavonoids, volatile oils and terpenoids which are essential for various antimicrobial activities, antitumor activity and antioxidant activity.
H. annuus seeds are cheap, readily available, accessible and nutritious which possess promising role in variety of infections, inflammations, cancers and cardiac diseases (Ruchika, 2014). Previous studies by Aboki et al. (2012) used Soxhlet extraction method and n-hexane (as solvent of extraction) at 60-65˚C, to determine antimicrobial activity of H. annuus extracts. In this study, cold maceration method was employed. Several studies have proved that the methanolic crude extracts of H. annuus have antimicrobial activity against wide range of pathogenic microorganisms.
1.4 Aim and Objectives of the Study
The study investigated the antibacterial effect of crude extracts of Helianthus annuus seeds on Salmonella spp and Escherichia coli.
The objectives of the study were to determine the:
i. phytochemical constituents of the crude extracts of Helianthus annuus seeds.
ii. in vitro antibacterial effect of the crude extracts of Helianthus annuus seeds onSalmonella spp. and E.coli
iii. minimum inhibitory concentration (MIC) and minumum bactericidal concentration (MBC) of the crude extracts of Helianthus annuus seeds.
iv. toxicity of the crude extracts of Helianthus annuus seeds.
This material content is developed to serve as a GUIDE for students to conduct academic research
ANTIBACTERIAL ACTIVITY OF HELIANTHUS ANNUUS SEEDS EXTRACT ON SALMONELLA SPECIES AND ESCHERICHIA COLI
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