The leaves and seeds of Piper guineense (Uziza) are widely used in homes to impact taste, aroma and flavor to

food. In folk medicine, the Piper guineense seeds and leaves are used as Post- partum recovery after delivery. It

extracts were used to estimate the phytochemical components, mineral and proximate properties. The bioactive

compounds were determined using high performance liquid chromatography (HPLC) and Gas chromatograph-

Gas spectrometry (GC- GM). The susceptibility patterns of the crude methanol extract were determined using

agar diffusion technique for bacterial pathogens while macro-broth techniques were used to estimate the fungal

pathogens. The toxic effect of crude methanol extract was investigated using 30-albino rats. The rats were group

into five containing six (6) rats per group . Group A and B were administered with 500mg/ml body weight and

200mg/ml body weight of crude methanol seed extract while Group C and D were administered 1500mg/ml and

1000mg/ml per body weight of crude methanol extract of piper guineense leaves extract and group E were

administered with normal saline and serve as net control. The experiment lasted for thirty (30) days. The

haematological indices, Liver function test, Electrolytes and Urea were estimated while the visceral organs were

Piper guineense seeds and leaves both showed activity against Staphylocous aureus , proteus mirabilis and

Escherichia coli while antifungal activity of these crude methanol extract inhibited the growth of Aspergillud

flavus, Mucor fragilis and Penicillum notatum. The crude methanol fractions of the seeds were more potent in

inhibiting both bacterial and fungal isolates. The crude methanol extracts had no effects on the hematological

indices , liver function test and Aspartate Transaminase (AST) value in both seeds and leaves while the

electrolyte (Na+, Ca

that crude methanol extract of Piper guineense seeds affected the colon, Jejenum, liver and kidney with mild

increase in inflammatory cells , liver necrosis while that of leaves, caused damage to colon , Jejunum inducing

disruption and erosion of lining of crypts of liberkuhn. The findings in this study showed that crude methanol

stem bark, leaves, flowers, seeds, fruits and those grouped as spices. Medicinal plants contain vital nutrients such

as proteins, vitamins, minerals, carbohydrates, fibers and chemical components. Some of these plants are inform

of spices and/or fruits, seeds and flowers that are useful to mankind. Piper guineense leaves and seeds are

commonly used as spices due to its aroma and flavor impacts on foods. In folk medicine, the plant are used in

treatment of diabetes, ulcer and enhances male fertility (Memudu, et al., 2015). Traditionally, both the leafy

vegetables and seeds of Piper guineense are used as spices for preparing soup for post parturient women.(Udeh,

et al., 1999). The plant is effectively used traditionally to terminate pregnancies under folk medicine practices

(Iwu, 2014).

Piper guineense is a native of the tropics of western and central African regions and is common in southern

Nigeria (Balofumi, et al., 2016). The plant belongs to Piperaceae family and of 20meters higher climbing vine

treating microbial infections is high rate of resistance of these antimicrobial agents against the microorganisms.

Multiple resistant organisms render therapy more precarious and costly and sometime unsuccessful. Individuals

may be victim of multiple drug resistant infections because all available drugs have failed especially in

developing world (Levy, 2002). This scenario presents a health challenge among the populace and thus the search

for alternative antimicrobial agents especially from medicinal plants. Basically the wide folk medicinal use of

leaves and seeds of Piper guineense within the populace is indication of its future application in the health system.

Previously, it has been reported that the leaf methanol extract has antioxidant and protective effects on lead

induced testicular damage in wistar rat (Nwosu, et al., 2022). It has also been shown that the leaves/seeds are

used in the treatment of asthma, rheumatism and weight control (Jabeen, et al., 2010). Different authors has

reported phytochemical, nutritional and antimicrobial properties of the plant based on different soil environments

suggest that Piper guineense seed may not be harmful at moderate dose; but high doses could be toxic in

experimental animals.

Fresh leaves/Seeds of Piper guineense (African Black pepper) were bought at New Market Enugu, Enugu North

Local Government area of Enugu State in June 2023. The leaves were carefully separated and sorted from their

stalks. Both Piper guineense Leaves and Seeds were thoroughly washed with distilled water to remove dirt and

microbial contaminant, sieved to removed excess water; Air-dried in the dark for three{3} weeks, grounded

into fine powder using mill and stored in an air–tight plastic container for later use at room temperature.

Methanol Extraction

continued until the samples become lighter. The crude extract is then concentrated using water bath and the

components characterized using standard methods {Okigbo, et al., (2010) and Nwankwo, et al., (2011).

Fractionation

The crude extract was adsorbed in a silica gel (70-230 mesh size] and removed fractionated using methanol

fractionate (MF) which was used to increase the solvent polarities. The percentage yield was 16.75%. The crude

extract was further fractionated using column chromatograph. Fifty (50) ml of the methanol extract was subjected

to column chromatograph on silica gel (100-200 mesh merck) parked and eluted, therefore pure methanol crude

extract was obtained.

Bacteria Isolates from different clinical samples were used for the susceptibility test. The bacteria included

used because the extract could not dissolve in sterile water. The selected bacterial Isolates were prepared by

inoculating the organisms into peptone water, incubated at 37

bacteria (Escherichia coli, Klebsiella pneumonie, Proteus mirabilis) to three (3) hours in case of slowing

bacteria; Staphylococcus aereus and Streptococcus pyogenes.

Reagents include urea acid reagent (Reagent No 68) and Diacetylmonoxime, 4g/l (reagent No 21) which can be

stored at room temperature. Equal volumes of urea acid regent and diacetyl monoxime reagent were mixed.

Tubes were rebelled accordingly.

The contents of each tube were mixed well and incubated at 100

C

each tube was stoppered using loose flitting cap. The tubes were removed and cooled the contents in a container

of coloured water for about 5miuntes without allowing water to enter the tubes nor allowed exposure to light for

I hour he absorbance’s of the solutions in a colorimeter were read using a green filter 520nm (eg 11 ford No

604) (or a spectrophotometer set at wavelength530nm. The instrument was zeroed with a blank solution (Tube

B). The concentration of urea in the control and rat’s samples can be calculation. By reading the values from

the calibration graph providing the reading of the 10mmol/L stand and agree with the calibration or using the

formular

Urea mmol/L =

zeroed with distilled water .Calculate the concentration of the total bilurubin in the control and rats samples.

The leaves crude methanol extract of Piper guineense had inhibitory activity against Aspergillus niger, Mucor

fragilis , Candida albicans at a concentration of 3.12mg/ml while showing inhibition effect at higher

The methanol fractions of the Piper guineense seeds was effective against Staphylococcus aureus, Klebsiella

oxytica, Pseudomonas aeuroginosa at a concentration of 3.125mg/ml with zone of inhibition of 10mm

respectively Proteus mirabilis, Klebsiella pneumonia, Streptococcus pyogenes and Escherichia coli were

controlled by the fraction at a concentration 6.25mg/ml..(Table 1.5)

Table 1.3: Anti-Fungal activity of crude seed methanol extract of Piper guineense.

Table 1.4: Anti-fungal activity of crude methanol extract of Piper Guineense Leaves

Table 1.4: Anti-bacterial activity of fractionation extract of Piper guineense (Uziza)seeds

The antimicrobial activity of the Piper guineense leaves of crude methanol fraction inhibited the growh of

Staphylococcus aureus and Klebsiella pneumonia; Klebsiella oxytiaa and Eschicheria coli at a concentration of

3.125mg/ml with at 10mm and 15mm in diameter respectively. (Table 1.5)

Activity of Piper guineense seeds and leaves fractions on fungal isolates.

Both the Piper guineense leaves and seeds fractions inhibited the growth of Mucor fragilis, Aspergillus niger,

Table 1.7: Anti-fungal activity of fraction extract of Peper guineense(Uziza) Seeds and leaves

The damaging effect of crude methanol extract of Piper guineense indicated that packed cell volume (PCV),

hemoglobin (Hb) and Red blood cells (RBC) were not affected by the seeds and leaves of methanol extract

though there were minor increases of the PCV and Hemoglobin compared with the control.

The methanol leaves extract administered at 500mg/kg weight indicated that PG was 53+ 2.86% (percent) when

compared with the control which 47.8+16% in the same way the leaves administered at 1500 mg/kg increased

observed that seeds receives 200mg/ml of the extract has a mass reduction of TWBC 3.69+0.92. All the TWBC

were statistically significant when compared with the control. The lymphocytes showed that marked reduction

in Group A and Group B that were administered with seeds while compared with the methanol leaves extract:

aureus. Thus the zones of inhibition of the crude methanol extract of Piper guineense ranges from 28mm to

10mm (Klebsiella oxytica) in diameter while the least efficacy was observed in Proteus mirabilis with a range

showed Proteus mirabilis zone of inhibition at 16mm in diameter but disagreed with Okeke, et al., [2001] who

reported that Piper guineense seeds was effective to Proteus vulgaris but not to Proteus mirabilis.

The crude methanol extract of Piper guineense seeds and leaves had effects on common bacterial isolates which

can be explored for treatment of human illness. The mechanism of this killing effects were based on disrupting

their cell membrane and other vital processes for instance Tannis have been found to form irreversible complexes

with proline rich protein (Shamada 2006) resulting in inhibition of cell protein synthesis. Parekh and Chanda,

[2007] reported that taninis are known to react with protein to provide typical tanning effect. .Piperine (alkaloids)

had shown to have properties of antibacterial activity Heinrich et al.,2021).

The crude methanol seeds extract showed inhibitory effects against all fungi at a concentration of 100mg/ml.

Mucor fragils, Aspergillus flavus, Candida albicans and Aspergillus fumigatus had the highest inhibitory effect

maximum effect. The crude methanol extract fractions of the seeds showed killing effects against bacterial

pathogens at concentration of 100mg/ml. Staphylococcus aureus was inhibited at a range 32mm-100mm in

diameter while the least inhibited was Proteus mirabilis. Escherichia coli (E coli) growth was inhibited at a range

of 25mm-8mm in diameter Pseudomonas aeroginosa was inhibited at 32mm. Pseudomonas aeroginosa,

methanol extract of Piper guineense seeds fraction was effective against bacterial pathogens that do cause human

disease (Irshad, et al.,.2017; Subramani, et al., 2017).

Similarly, the fractions had inhibitory effects on fungal agents at the least concentration of 3.125mg/ml. The

fungi inhibited include Mucor, fragilis, Aspergillus niger, Aspergillius flavus, Candida albicans, Aspergillus

fumigatus and Penicillum notatum. This indicated that purifying the seed and extracting the essential oil or

ingredient will be helpful as antifungal agent.

was potent inhibitors of many hydrolytic enzymes such as pecteolytic macerating enzymes used by plant

pathogens. Tannins have been found to form irreversible complexes with proline rich protein (Shumuda, 2006

and Echo et al 2012] will result in the inhibition of cell protein synthesis herbs that have tannis as their main

component are as astringent in nature and are used for treating intestinal disorder such as diarrhea and dysentery.

Thus the presence of tannis is a valuable component in killing of bacteria and fungi. Other components such as

flavonoids, saponins and alkaloids are also important in folk medicine. Therapeutically, alkaloids are particulary

well known as asthetic cardio proletic and anti-inflamatory operation. Well known alkaloids use in clinical

setting include morphine, stychine, quinine, ephedrine, nicotine (Hajar, 2015). In this study, Piper guineense had

effect against bacterial and fungal pathogens. This may be due to combined effect of alkaloids with other

components. It has been reported that alkaloids induces bacterial and fungal synthesis by disrupting bacterial

is because saponins act as permeabilising plasma membrane their amphipathic properties enable them to

The effect of saponins can be reduced due to glycosylation of saponin (Sandrock and Vanetter 1998).Miorrissey

and Osbourn 1999). The loss of a single sugar formed the oligosaccharide chain can pair the ability to complex

with steroid. Arneson and Durbinm 1967). May fungi can hydrolysed sugar to saponin thereby reducing anti-

fungal activity. This effect was seen with the fungal agent Penicillum notatum and Trichophyton ssoudanese

where the leaves methanol extract show poor inhibitory effect. The high content of carbohydrate and protein in

the methanol extract of Piper guineense seeds and leaves induces balancing effect on the antimicrobial activity

individual from any toxic effects. It is normal to test the efficacy of the medicinal plant but in human usage and

its domesticated animal. Toxic effect in vivo may impair the function of the cells and tissues of the body system.

In this study, the toxic effects of the seeds and leaves of Piper guineense were experimented in an experimental

albino rats and its effect on histological kidney and liver biomarkers and tissue damages were investigated. The

experimental rats were grouped into five; Group A and B received 500mg/ml and 2000mgt/ml concentration of

methanol seeds extracts while Group C and D received 1500mg/ml and 1000mg/ml crude methanol leaves

extract respectively while Group E remain normal control that receive normal saline. It was observed that there

was increase in weight. The increase in weight of the experimental rats after the experiment may be as a result

of the added advantage of the high carbohydrate and protein content of Piper guineense seeds and leaves. This

increase in weight was statistically significant which indicates the overall effect of the Piper guineense seeds and