Original Research Article

ANTIMICROBIAL SUSCEPTIBILITY EVALUATION OF MICROORGANISMS ISOLATED FROM POULTRY WATER IN AWKA METROPOLIS

UGOCHUKWU CHUKWUMA OKAFOR, PRISCA-MARIS UGWUEGBULEM

Journal of Biochemistry International, Page 1-9

The research was aimed at determining the microorganisms that are responsible for the contamination of poultry water in Awka metropolis and to evaluate the susceptibility of these organisms to some antimicrobial agents in the case of infections arising from water given to poultry. Three (3) poultry water samples were collected from three different poultry farms all in Awka metropolis and were microbiologically analyzed using standard methods. The bacterial isolates include Salmonella spp., Escherichia coli and Staphylococcus spp. Five fungal species were isolated which were identified as Aspergillus sp., Mucor spp., Fusarium spp., Cryptococcus spp., Candida spp. Using the standard disc diffusion method, the antimicrobial susceptibility pattern of isolates was determined against antimicrobial agents. The effectiveness of the antimicrobials or the susceptibility of the isolates were defined in percentage as follows: Rocephin (7.7%), Pefloxacin (10.9%), Gentamycin (9.3%), Ciprofloxacin (8.8%), Streptomycin (10.4%), Ampiclox (9.5%), Septrin (11.0%), Zinnacef (10.7%), Erythromycin (11.8%), Amoxacillin (10.0%), Nitrofurantoin(12.0%) (for antibacterial) and Nystatin (37.0%), Fluconazole (27.0%), Ketoconazole (36.0%) (for antifungal). For the antibiotics, Isolate-1 was more susceptible to Erythromycin and Rocephin but resistant to Streptomycin. Isolate-2 was more susceptible to Nitrofurantoin, but resistant to SXT(0.0%), GET(0.0%) and AMC(0.0%),. Isolate-3 was more susceptible to Nitrofurantoin and Erythromycin but least susceptible to GET(10.9%). For the antifungals, Isolate-1 was more susceptible to FLU(51.0%) but least susceptible to NYS(21.5%). Isolate-2 was more susceptible to KET(42.8%) but least susceptible to FLU(23.7%). Isolate 3 was more susceptible to FLU(40.8%) but least susceptible to KET(28.7%). Isolate-4 was more susceptible to NYS(55.6%), resistant to FLU(0.0%) but least susceptible to KET(44.4%). Isolate-5 was more susceptible to NYS(50.5%) but least susceptible to KET(21.0%). Fungal resistance was seen among Fluconazole. Thus, Nitrofurantoin, erythromycin and Nystatin should be the first line of prescription in cases of poultry infections and diseases caused by bacteria and fungi respectively in the poultry houses.

Original Research Article

MEMBRANE STABILIZING EFFECT AND ANTI-SICKLING POTENTIALS OF Trametes hirsuta (HAIRY BRACKET MUSHROOM) EXTRACT ON HUMAN HbS ERYTHROCYTES

REGINALD CHIBUEZE OHIRI, ROBINSON AMARACHUKWU OHANADOR

Journal of Biochemistry International, Page 10-17

The role of fungi based nutraceuticals in the treatment and management of Sickle cell has been under exploited and thus necessitates for more research. This research, studies the potential of decoction extract of Trametes hirsuta mushroom in lysis reduction and stabilization of erythrocyte membrane integrity. Freshly harvested, washed and air-dried T. Hirsuta was ground and 1.5 dm3 of distilled water was added to 1.5 kg of the ground mushroom. The filtrate obtained after boiling for 30 min. and cooling for 2 hrs was dried using an autodesicator to give decoction extract. Volumes of 5.0 mg/mL, 10.0 mg/mL and 15.0 mg/mL of the extract were obtained via serial dilutions, while HbS erythrocytes were obtained by centrifuging 5.0 mL of HbS-blood mixed in 5.0 mL of 0.9 % physiological saline. HbS erythrocytes treated with 15 mg/mL and 10 mg/mL had the highest and the lowest lysis at 50% mark. Lowest median corpuscular fragility was observed in erythrocytes suspended in 10.0 mg/mL of extract, while negative percentage stability were observed in erythrocytes suspended in 10.0 mg/mL and 5.0 mg/mL of extract. The control had the highest hemoglobin polymerization, while morphological stability was highest (89.40 + 4.61 %) after 5 min. of culturing in 10.0 mg/mL of extract. The ability of  this mushroom extract to stabilize erythrocyte membrane integrity and reduce the progression of lysis shows that this mushroom can be exploited as a possible nutraceutical means in both the management and treatment of sickle cell anemia and its associated crises.

Original Research Article

EVALUATION OF THE PEPSIN INHIBITORY POTENTIAL VIA RED KIDNEY BEANS EXTRACT

NANCY A. A. TAHA

Journal of Biochemistry International, Page 35-40

The physiological function of pepsin inhibitors could be the prevention of unwanted proteolysis and consequently control of protein turnover and metabolism. Where pepsin enzyme inhibitors reduce dietary proteins digestion and absorption. So, the aim of this research work was to inactivate pepsin presented in various crops. Pepsin inhibitors were extracted from red kidney beans seeds. The substrate was BAPA (Nα-benzoyl-L-arginine 4-nitroanilideHydrochloride) a chromogenic substrate. Phytochemical screening of red kidney beans seeds extract confirmed the presence of some phytochemical constituents e.g., saponins, glycosides, ascorbic acid, reducing compounds, carotenoids, carbohydrates, proteins, alkaloids and tannins. On the other hand, the protein concentration in the crude extract of inhibitor extracted from red kidney beans seeds was 14.05 mg/ml (17.90 mg/g, DW).

Also, the obtained results illustrated that the crude extract of red kidney beans showed inhibitory activity against pepsin enzyme. Pepsin inhibitor activity was 151.25 TUI/mg dray sample.  Generally, it can be concluded that the pepsin inhibitor extracted from red kidney bean has the potential for usage as a protease inhibitor.

Original Research Article

NUTRITIONAL AND TOXICOLOGICAL STUDIES ON Periophthalmus koelreuteri FROM OKPOAMA-BRASS, BAYELSA STATE

FELAGHA, I., AMADI, B. A., MONANU, M. O., GODWIN, S. O.

Journal of Biochemistry International, Page 41-49

Mudskippers have been used extensively in ecotoxicological studies due to their tendency to bioaccumulate heavy metals and other environmental toxicants. The present study investigated the proximate, fatty acids, minerals and heavy metals composition of Periophthalmus koelreuteri (mudskipper) from Okpoama-Brass, Bayelsa State. Proximate composition was determined by AOAC methods, fatty acids composition was determined by gas chromatography and mineral/heavy metals were determined by atomic absorption spectrophotometry. Health risks associated with consumption of mudskippers (estimated daily intake-EDI, target hazard quotient-THQ and carcinogenic risk-CR) were determined by calculation. Proximate composition of samples were as follows: 52.50±0.20% crude protein, 7.50±0.02% crude fat, 0.29±0.01% carbohydrates, 15.71±0.015% crude fibre, 18.00±1.00% ash and 6.00±0.04% moisture. The following fatty acids were detected: lauric acid, palmitic acid, margaric acid, stearic acid, oleic acid, linoleic acid and arachidonic acid. Minerals detected includes Na, K, Ca, Mg, Fe, Zn, Cu, Co and Mo and heavy metals (Cd>Cr>Pb). Health risk assessment showed the following: EDI for Cd was 0.001 mg/100g-1bw/day; EDI for Cr in the samples was 0.000628mg/100g-1bw/day. THQ value for Cd was >1 hence consumption of mudskipper (with respect to Cd) is not advised as it may pose health threats to the consumers. THQ for Cr and Pb were <1 hence there are no health risks associated with respect to these metals. CR values for all heavy metals (Cd, Cr and Pb) in the sample were within the permissible limits. Mudskipper is a good source of protein, fatty acids and minerals however it accumulates toxic heavy metals from the environment.

Original Research Article

EVALUATION OF ANTI QUOROM SENSING ACTIVITIES USING Emblica officinalis ETHANOLIC LEAF EXTRACTS AGAINST MARINE MICROBES

BRISHTI NANDI, K. VIJAYALAKSHMI, ATHMAJA V. CHRISTOPHY, G. HARSHAVARDHINI, P. B. RAMESH BABU

Journal of Biochemistry International, Page 59-66

Biofilm formation in pathogenic bacteria displays a major role in various human infectious diseases posing a major health problem in several countries. Biofilms contain colony of bacteria which are 1,000 folds more resistant to antibiotics and also they develop more insensitivity to innate and acquired immune responses. Quorum sensing (QS) is characterized well due to the interaction of bacterial cells, which is a dependent mechanism and acts its stimulus response related to population density. In the present study we tested the antiquorum sensing activity with leaf extracts of commonly used dietary supplement Emblica Officinalis      (Amla) against marine microbes. In our method, we cultured Klebsilla, Proteus, E.coli, Shigella, Salmonella typhi from marine water source for quorum sensing activities and inhibitory effects through antimicrobial activities. Marine bacteria were isolated, identified based on physical and biochemical characterization. In our results we investigated the inhibitory effects of amla against pathogenic bacteria using swarming method. Further highest inhibitory bacterial isolates were examined under scanning electron microscopy (SEM). We conclude that amla exhibits potential inhibitory effects in biofilms formation from samples obtained from marine waters and further studies are needed in QS gene expression in more number of marine microbial samples.

Original Research Article

EFFECT OF MIXTURE OF LEAF EXTRACT OF Ocimum gratissimum AND Vernonia amygdalina ON CADMIUM INDUCED HEPATOTOXICITY IN WISTAR RATS

OGHENETEKEVWE, EFEKEMO, ORORORO O. CLEMENT

Journal of Biochemistry International, Page 73-81

The search for potential ameliorative substances for cadmium toxicity is still in progress, thus, this study investigated the invivo effect of oral administration of the leaf extract of Vernonia amygdalina (Bitter leaf) and Ocimum gratissimum (Scent leaf) on biochemical parameters; Alanine transaminase (ALT), Aspartate transaminase (AST), Alkaline phosphatase (ALP), Albumin, Total protein and oxidative stress biomarkers Malondialdehyde (MDA), Reduced glutathione (GSH), Superoxide dismutase (SOD) and catalase (CAT) in the liver of cadmium exposed Wistar rats. A total of sixteen female rats weighing 120-150g were used and they were divided into four groups with four rats each. The experimental period was for four weeks. Group 1 served as control, Group 2 (Cadmium 10mg/kg body weight), Group 3 (Extract 200mg/kg per body weight). Group 4 (Cd 10mg/kg + Extract 200mg/kg body weight). The samples were administered orally by gastric intubation through the use of a cannula attached to a 1ml syringe. Blood and organ samples were collected and analyzed. Results obtained from administered groups were all compared to the control. Serum biochemical markers showed significant (P≤0.05) increase in the activities of AST, ALT, ALP, but there was no significant change in Total protein and Albumin when values were compared to the control (Group 1). Comparing Group 2 to Group 4 showed significant decrease in enzyme activities, but no significant change in the concentration of total protein and albumin. Oxidative stress markers showed significant increase (P<0.05) in concentration of MDA in the liver and significantly (P<0.05) decreased the activity of SOD and CAT and concentration of GSH in cadmium control (Group 2) when compared with the control (Group 1). Comparing Group 2 to the Group 4 (cadmium and extract) showed significant (P<0.05) increase in activity of SOD and CAT and concentration of GSH, while MDA had significant (P<0.05) decrease. The results showed that the leaf extract had some antioxidant and hepatoprotective potentials, which can help in ameliorating cadmium induced stress to the liver.

Short Research Article

PRODUCTION AND CHARACTERIZATION OF THERMOSTABLE LIGNOLYTIC ENZYMES PRODUCED FROM Staphylococcus saprophyticus EXPOSED TO LOW-DENSITY POLYETHYLENE (LDPE)

E. OGUNJEMITE OLUWADAMILOLA, ZIGE TIMINIBEFI, A. O. MOMOH

Journal of Biochemistry International, Page 18-27

Enzymatic degradation has been revealed as an effective method of eradicating polyethylene pollution and different studies have opined lignolytic enzymes to be involved in the process of low-density polyethylene biodegradation. This study focused on screening and characterization of manganese peroxidase and laccase produced by Staphylococcus saprophyticus exposed to low- density polyethylene. The result showed that Staphylococcus saprophyticus produced optimum manganese peroxidase and laccase activities on the third and second days. A study of the physicochemical properties of manganese peroxidase and laccase revealed that their activities were optimum and stable at acidic pH (pH 3 and pH 3 respectively). Manganese peroxidase had optimum temperature activity at 50 oC and was stable at 60°C while laccase was active at 70°C and most stable at 80°C. Staphylococcus saprophyticus produced thermostable acidophile lignolytic enzymes, therefore it can be used for polyethylene biodegradation.

Short Research Article

EVALUATION OF THE PROPHYLACTIC POTENTIALS OF GOKO CLEANSER HERBAL MIXTURE (GCHM) IN ACETAMINOPHEN-INDUCED NEPHROTOXICITY

L. I. ANYANWU, G. O. UDEBUNU, P. K. IZUCHUKWU, P. C. EZEUGWU

Journal of Biochemistry International, Page 67-72

This study evaluated the potency of an herbal mixture – Goko Cleanser Herbal Mixture (GCHM) in the prevention of Acetaminophen induced Nephrotoxicity. A total of 30 adult wistar rats weighing 150g-200g were divided into 6 groups of 5 animals each. Animals were acclimatized for 7 days to laboratory conditions, fed with rat chow and distilled water ad libitum. All groups were pretreated for 11days (morning and night) as follows: Group1 (Normal-control) received 3ml/kg B.W distilled water, Group2 (Positive-control) not pretreated, Group3 (Standard-control) received Silymarin (100mg/kg b.w), Group4,5 and 6 received 1ml/kg b.w 2ml/kg b.w and 3ml/kg b.w GCHM respectively. On the 11th day, all Groups apart from Group1 were induced with 2mg/kg b.w Acetaminophen 30minutes after the normal drug administration. The normal drug administration continued to 14th day and on the 15th day, blood samples were collected after an overnight fast and the animals were sacrificed via Jugular-strangulation. The phytochemical constituents of the herbal mixture and the renal function makers were evaluated by standard methods. According to our data, Alkaloids, Terpenoids, Steroids, Glycosides, Anthocyanins, Anthraquinones, Phenols, Carotenoids, Flavonoids, Tannins, and Saponins were present. Rats in group2 had significant (p<0.05) increase in serum creatinine and urea levels but a significant (p<0.05) decrease in potassium and sodium levels indicating Nephrotoxicity. Groups 3,4,5 and 6 showed a significant (p<0.05) increase in serum sodium and potassium ion levels compared to Group1 and 2. We conclude that GCHM is rich in phytochemicals and could serve as a potential raw material in the development of therapeutics for kidney and related conditions.

Review Article

REVIEW ON APPLICABILITY OF mTOR INHIBITOR DOCKING SCORE ANALYSIS METHODS IN DRUG DEVELOPMENT OF LUNG CANCER

P. B. RAMESH BABU, S. NIVEDHITA, A. MANIKANDAN, N. S. SHARON, P. BALASHANMUGAM

Journal of Biochemistry International, Page 28-34

Protein kinases have been broadly considered as significant class of target in cancer therapeutics. The mammalian target of rapamycin (mTOR) belongs to a group of protein kinases regulating cell growth and protein synthesis. Rapamycin, a natural antifungal antibiotic, can that inhibit protein kinases by partner with its intracellular receptor FKBP12. The FKBP12-rapamycin complex ties straight forwardly to the FKBP12-Rapamycin Binding (FRB) space of mTOR. Side effects in patients consuming rapamycin include: severe mood swings, small purple spots over the body retention, face allergy and anxiety. Based on recent literatures, it is understood that drug like molecules from drug bank were used to screen for potential inhibitors by docking into the active site of mTOR. A few studies used in silico tool like GLIDE, which yielded useful information about several docked structures. The drug like molecules which were complexed with mTOR could be subjected to molecular simulation studies with molecules like DB00094, which was previously reported as fertility agent targeting follicle stimulating hormone receptor. Recent literature studies have demonstrated progress in identifying mTOR drug targets in treating lung cancers and in this paper we have updated the docking score methods using in silico tools.

Review Article

AN UPDATED REVIEW IN CORRELATING FAT MASS OBESITY ASSOCIATED GENETIC POLYMORPHISM IN WOMEN WITH POLYCYSTIC OVARIAN CONDITIONS

R. REHANA BABU, FLORIDA TILTON, ANEESH NAIR, K. VIJAYALAKSHMI, P. B. RAMESH BABU

Journal of Biochemistry International, Page 50-58

Women with Polycystic ovarian condition (PCOS) are reported to have obesity, which essentially influences the disorder's associated with insulin resistance, type 2 diabetes and cardiovascular disorders.  PCOS is a problem of endocrine abnormality in women between the ages of 14-44. In this paper we review on recent methods in genotyping of fat mass obesity (FTO) associated gene single nucleotide polymorphism and its relationship with PCOS. Literature survey indicated that in diagnosis of PCOS, blood tests were normally used in gene polymorphism investigations and DNA tests were used for genotyping examination. The quick detection by polymerase chain reaction (PCR) amplification is one of the commonest tests for identifying single nucleotide polymorphism (SNP) for FTO traits which is being routinely used to understand the frequency of SNP polymorphism in a given set of population in various countries and in ethnic groups. Worldwide, extensive investigations were carried out to analyze the genotypes of FTO traits and its association in understanding the etiology of PCOS. Such literature surveys are beneficial in developing simple PCOS diagnostic                         methods and finding new approaches in therapeutic interventions.  In this review paper, we highlight the genetic basis and importance of testing the FTO parameter in understanding the pathogenesis and the management of PCOS.