{'fraction_non_alphanumeric': 0.027301092043681748, 'fraction_numerical': 0.019110764430577222, 'mean_word_length': 4.988323213451658, 'pattern_counts': {'":': 0, '<': 0, '<?xml version=': 0, '>': 1, 'https://': 0, 'lorem ipsum': 0, 'www.': 0, 'xml': 0}, 'pii_count': 0, 'substrings_counts': 0, 'word_list_counts': {'cursed_substrings.json': 0, 'profanity_word_list.json': 0, 'sexual_word_list.json': 0, 'zh_pornsignals.json': 0}}
Antibacteria Assesement of Aframomum melegueta [roscoe] k. Schum [alligator pepper] Fruit on Bacteria Pathogen Isolated from the Urinary Tract

This study aims at assessing the Antibacterial activity of Aframomum melegueta against Urinary Tract bacteria. The study was carried out at the Microbiology department laboratory of the Federal University of technology, Akure, Ondo state, Nigeria, between February and June, 2018. Agar diffusion method was used in the susceptibility test, whie tube dilution method was used determination of Minimum Inhibitory concentration. The phytochemical analysis showed the presence of alkaloids, flavinoids, saponins and tannins in both the Methanol and Pet-ether fruit extract, the result also showed that the concentration of all these compounds are higher in the Methanol extract than in the Pet-ether extract. The in-vitro Susceptibility test showed that E. coli, P. mirabilis, S. aureus were sensitive to methanol extract of Aframomum. melegueta at 100 mg/ml with E. coli showing the highest zone of Inhibition of 13.67 ± 0.24 mm, while all the organisms were Original Research Article Oladumoye et al.; JAMB, 20(2): 1-8, 2020; Article no.JAMB.49337 2 resistant to Pet-ether extract at this concentration of 100 mg/ml except for E. coli with a zone of inhibition of 10.93 ± 0.07 mm at a highest concentration of 400 mg/ml. S. aureus was sensitive to Pet-ether showing a zone of inhibition of 16.97±0.09 mm while E. coli had 20.33 ± 0.23 mm, K. pneumoniae and P. aeruginosa were resistant to both extract at all the tested concentration. Methanol Extract had MIC values of 50 mg/ml, 100 mg/ml and 50 mg/ml for E. coli, P. mirabilis, S. aureus respectively while the Pet-ether extract had an MIC values of 100mg/ml and 200 mg/ml for E. coli, and S. aureus respectively. The MBC values for Methanol Extract were 100 mg/ml, 200 mg/ml and 50 mg/ml for E. coli, P. mirabilis, S. aureus respectively while that of Pet-ether extract was 100 mg/ml for Escherichia coli.


INTRODUCTION
Infectious diseases are the number one cause of death due to illnesses across the world and account for approximately one-half of all deaths countries in tropical. According to World Health Organization [WHO] report, about 15 million [>25%] of 57 million annual deaths worldwide are the direct result of infectious diseases [1]. Of these infectious diseases, microorganisms are the commonest organisms responsible for morbidity and mortality [2,3]. As such, bacterial and fungal diseases continue to remain a major public health problem [4]. Urinary tract infections [UTI] are one of the most common infectious diseases diagnosed in outpatients as well as in hospitalized patients, and can lead to significant mortality UTIs account for more than 8 million visits to physicians' offices, 1.5 million emergency room visits, and 300,000 hospital admissions in the United States annually [5]. Typical symptoms associated with UTI include the triad of dysuria [painful urination], urgency [the enhanced desire to void the bladder] and frequency [increased rate of urination] [5].
Due to indiscriminate use of synthetic antimicrobial drugs, microorganisms resistant and or multi resistant to major class of antibiotics have emerged in recent years and the situation is exacerbated too [6,7]. Moreover, high cost and adverse side effects of popular synthetic antibiotics are major burning global issues [8]. To this regards, antibiotics resistance has resulted in morbidity and mortality while the high cost and adverse side effects have increased health care costs. Hence, recent attention has been paid to biologically active extracts and compounds from plant species used in herbal medicines [9,10]. In another words, increasing capability of microbes to develop multidrug resistance has no doubt encouraged search for new, safe and effective bioactive agents of plant origin considering the fact by previous authors [11], that traditional medicine is an important source of potentially useful new compounds for the development of chemotherapeutic agents.
Medicinal plants have received huge attention both in the developed and developing nations. Their economic importance has drawn attention of various world bodies mostly; the World Health Organization [WHO] which released a special document concerning collection practices for medicinal plants [12,13]. The use of medicinal plants has always been part of human culture and is wide spread in Africa. In some countries, like Ghana, government encourages the use of indigenous forms of medicine rather than expensive imported drugs. Also in Nigeria, a large percentage of the populace depends on herbal medicines because the commercially available orthodox medicines are becoming increasingly expensive and out of reach [14]. Aframomum melegueta is a tropical herbaceous perennial plant of the genus Aframomum belonging to the family Zingiberaceae [ginger family] of the angiosperms in the Kingdom plantae. The seeds have pungent peppery taste due to aromatic ketones [15,16]. It is a plant with both medicinal and nutritive values,found commonly in rain forest. It is widely spread across tropical Africa.

Collection and Identification of Plants
Fresh whole seed pod of Aframomum melegueta were collected from Oja Oba in Akure south local Government, Akure, Ondo state. The plants were identified by the department of Agricultural science, Federal University of Technology, Akure, Ondo state. The fruit of Aframomum melegueta were then washed with distilled water to remove dirts, and then air-dried at room temperature for 21 days until it is well dried to be milled using an electric blender.

Standardization of Test Orgnainsms
A loopful of test organisms were inoculated into 5.0 ml of nutrient broth and incubated at 37C for 24 hours. 0.2 ml from the 24 hours culture of the organisms was then dispensed into 20 ml sterile nutrient broth and incubated for 3-5 hours to standardize the culture to 10 -6 cfu/ml the turbidity of the broth was then compared with MacFarand Standard [17].

Preparation of Plant Extract
A 50 g of the blended materials [Aframomum melegueta] was soaked in two hundred and fifty millilitres [250 ml] of 90% ethanol in a sterile conical flasks and allowed to soak at ambient temperature for 72 hrs. With frequent agitation, this process was repeated in distilled water. It was then be filtered and the filtrate evaporated to dryness in a water bath at a controlled temperature of 40C, the crude extract was stored in the refrigerator at a temperature of 4C for further analysis [18].

Qualitative Assay for Phytochemicals
The methods described in [19,20] was adopted for the qualitative phytochemical assay conducted on the extracts.

Quantitative Assay for Phytochemicals
The methods described in [21,22] were adopted for the quantitative phytochemical assay conducted on the extracts.

In-vitro Antibacterial Screening of Crude Extracts
The Agar diffusion method was used, sterile nutrient agar plates was prepared and 1 ml of the test organism was mixed properly with 19 ml of nutrient agar and each plate was then labelled appropriately and allowed to solidify. A sterile cork borer [7 mm] was used to bore three holes of equidistance from each other on each plate. Molten agar was used to seal the base of each hole after which different holes was filled with different concentrations of the various extracts. The plates were left on the bench for 30 minute to allow diffusion of the extracts before incubation at 37C for 24 hours. The zones of clearance [Inhibition] produced around the holes after incubation were observed, measured and recorded appropriately [23].

Minimum Inhibitory Concentration (MIC)
The tube dilution method described by Ajaiyeoba et al. 2003 was adopted. The MIC was determined by serially diluting extracts. 5 ml of each of the dilution representing a known concentration of the extract was introduced into 5 ml of sterile nutrient broth in test tubes. The mixture was then inoculated with 0.1ml of the test of organism previously standardized to 10 6 and then incubated at 37C for 24 hours. The least concentration of the extract with no turbidity was taken as the minimum inhibitory concentration (MIC) [23].

Minimum Bactericidal Concentration (MBC)
This was an offshoot of the previously determined MIC. The MBC of the plants extracts were determined by sub-culturing from all the tubes that showed no turbidity from the MIC test into a sterile nutrient agar plate, the least concentration in which no growth is observed was taken as the MBC [23].

Phytochemical analysis
The results of this work showed that the extracts obtained from Aframomum melegueta using both polar     were sensitive to methanol extract at all the concentration tested similarly Escherichia coli was sensitive to pet-ether at all the concentration tested while Staphylococcus aureus was resistant at low concentration of 100mg/ml but sensitive to higher concentration. The Negative control DMSO showed no antimicrobial activity. The resistance of K. pneumonia and P. aeruginosa to the plant cannot be clearly ascertained by the scope of this work but There are three basic mechanisms by which organisms resist the action of antimicrobial agents: restricted uptake and efflux; drug inactivation and changes in targets. The outer membrane of P. aeruginosa is known to present a significant barrier to the penetration of antimicrobial agents [29], restricting the rate this could also be responsible for the resistance of the organism of penetration of small hydrophilic molecules and excluding larger molecules [29].
The MIC value of methanol extract for Escherichia coli, Proteus mirabilis and Staphylococcus aureus were 50 mg/ml, 100 mg/ml and 50 mg/ml while the value recorded for pet-ether were 100 mg/ml and 200 mg/ml for Escherichia coli and Staphylococcus aureus, the MBC value of methanol for Escherichia coli, Proteus mirabilis and Staphylococcus aureus 100 mg/ml, 200 mg/ml and 50 mg/ml respectively, this indicates that methanol extract is bactericidal at lower concentration on Staphylococcus aureus while it is bacteriostatic at lower concentration on Escherichia coli, Proteus mirabilis, pet-ether extract has an MBC value of 100 mg/ml on Escherichia coli but does   The biological functions of many of these phytochemicals detected and quantified have been documented as findings from [28,27,25] revealed the pharmacologic effects exerted by flavonoids which include protection against allergies, inflammations, free radicals and tumors. Anosike et al. [26] reported that Flavonoids are known to be synthesized by plants in response to microbial infection. They have effective antimicrobial activities in vitro against a wide array of microorganisms. Their activity is probably due to their ability to complex with extracellular and soluble proteins and also with bacterial cell, Schito [30] aslo reported the pharmaceutical importance of tanins, it was reported that Many human physiological activities, such as stimulation of phagocytic cells, host-mediated tumor activity, and a wide range of anti-infective actions, have been attributed to tannins. Their mode of action is to complex with proteins through nonspecific forces, such as hydrogen bonding as well as by covalent bond formation. They also complex with polysaccharides which are components of bacterial cell wall. Studies show that tannins can be toxic to filamentous fungi, yeasts, and bacteria. Thus, the mode of antimicrobial action of this plant may be related to the ability of these bioactive constituents to inactivate microbial adhesins, enzymes, and envelope transport proteins. Alkaloids also are of therapeutic significance. Pure isolated alkaloids and the synthetic derivative are used as the basic medicinal agents due to their analgesic, antispasmodic and antibacterial potentials [31], Likewise, the presence of phenolic compounds as well as tannins in the extracts indicates their potential as antimicrobial agents [27,22,25]. The study have shown that the methanol extract possess more antimicrobial activity in comparison to the pet-ether extract this is most likely due to the presence of higher concentration of phytochemical constituent in the methanol extract as shown in table 1 this is also consistent with the work of Anosike et al. [26].

CONCLUSION
This research have further established the the use of the Aframomum melegueta fruit in traditional medicine. The work have shown that the fruit has potential of treating Urinary Tract infection, it has a broad spectrum of activity against both gram Positive and gram Negative Bacteria. Although further research on the plant is recommended to further purify the active compound and then carry out toxicity studies in a bid to further improve its activity.