Effects of Processing on Proximate Composition of Hibiscus rosa-sinensis Leaf | Chapter 14 | Recent Advances in Biological Research Vol. 3

Leaves of Hibiscus rosa-sinensis are processed using different methods depending on the intended application. Using three different processing methods, we investigated the effects of processing on the proximate constitution of the leaf. Result demonstrated that the fresh raw leaf had moisture content of 82.30 ± 0.42%, which were significantly (p<0.05) reduced by drying but not extraction and blanching. The protein content of the raw leaf was low (1.80 ± 0.10%). Extraction and blanching reduced the protein content, whereas drying increased the protein content significantly (p < 0.05) for raw dried leaf powder and blanched leaf products. The raw leaf contained vitamins A, B2, C and E, which were significantly reduced by extraction and blanching, but were concentrated by drying. Anti-nutrient contents of the raw leaf were low and were reduced to negligible levels by the processing techniques employed. Comparing the nutrient and chemical constituents with recommended dietary allowance (RDA) values, we found that the leaves contain an appreciable amount of nutrients, minerals, vitamins, proteins and phytochemicals and low degree of toxicants. These findings suggested that the treatment method employed in processing this leaf affected the proximate composition, and this should be considered in utilization of this leaf (and other leaves) product in various food and pharmaceutical formulations. Various heat processing techniques applied during the preparation of the processed products from Hibiscus rosa-sinensis leaves, caused adverse effects on the chemical composition of the processed leaf products. This was evident especially for the vitamins and minerals constitution of the processed products. More so, blanching and drying caused a significant reduction in the nutrients and anti-nutrient composition of the formulated samples. While the best processed samples were the dried powdered products, especially the RDLP, whereas the worst processed samples were the extracts, notably B2LE. It is recommended that other processing techniques such as freezing, solar and spray drying and ethanol extraction can also be applied in order to determine their effects on nutrient retention and anti-nutrient reduction on the plant leaves and compare it with the results of this study.

Author  Details:

Ifeyinwa Mirabel Eze

Department of Food Science and Technology, University of Nigeria, Nsukka 410001, Nigeria.

Daniel Don Nwibo

Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan, Institute of Medical Mycology, Graduate School of Medicine, Teikyo University, 359 Otsuka, Hachioji, Tokyo 192-0395, Japan and Department of Chemistry, University of Nigeria, Nsukka 410001, Nigeria.

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In-vitro Anti-Bacterial Activity of Extracts of Euphorbia abyssinica (Desert Candle) Stem-Bark and Latex | Chapter 13 | Recent Advances in Biological Research Vol. 3

Introduction: Plants are a limitless gift of nature to humans and they possess very appreciative values and roles. They have stood the test of time in the life of man since creation. All over the world, they are hugely exploited for food, fuel, timber, medicine etc. The natural endowment of plants with numerous metabolites and bioactive compounds makes them good sources of therapeutic agents capable of replacing synthetic antibiotics; For example, Salversan and Penicillin are synthetic drugs formerly used for the treatment of Syphilis and Staphylococcus aureus infections, respectively, but which became less preferred because these pathogens developed resistance to the drugs.

Aim: This study was aimed at evaluating Euphorbia abyssinica (Desert Candle), a medicinal plant extensively used in folklore medicine among the Kendem people of South-west Cameroon for antibacterial activity and extracts analyzed for phytochemical composition.

Study Design: The completely randomized block design was used and data analyzed using of two way analysis of variance. Significant means were separated using Duncan’s New Multiple Range Test.

Place and Duration of Study: This study was carried out in the Department of Microbiology, University of Nigeria Nsukka, Enugu State, Nigeria, between April 2011 and August 2012.

Methodology: Extraction was done using absolute methanol, 50% methanol (in water) and water as solvents. Qualitative analysis methods were used to assay the phytochemical constituents. Agar-well diffusion, macro broth dilution and agar dilution and time-kill assay were the susceptibility test methods adapted.

Results: The phytochemical constituents detected were alkaloids, flavonoids, tannins, cardiac glycosides, carbohydrates and steroids, and saponins. The 50% methanol extract of the stem-bark was highly active against Staphyloccocus aureus, Escherichia coli, Salmonella typhi and Pseudomonas aeruginosa and compared favorably with the Gentamycin control drug. The inhibition zone diameters (IZDs) obtained with 50% methanol extract measured 23 mm for S. aureus and 19 mm for P. aeruginosa compared to 18 mm achieved with the absolute methanol extract for both        S. aureus and P. aeniginosa. For the aqueous extract the overall IZD range of 10±1.60-13±2.16 mm. The susceptibility patterns obtained using both dilutions (agar and macro-broth) methods were similar to that obtained with the agar diffusion method above. S. aureus (with MIC, 10.93±1.00-; MBC, 25-mg/mL, agar dilution or MIC, 3.9±1.60 -, MBC, 12.5-mg/mL, macro broth dilution methods, respectively). It was considered to be the most significantly susceptible bacteria strain tested (significant mean value 3.933), while E. coli was the least susceptible (with MIC, 50±0.00-, MBC, 100-mg/mL, in the agar dilution; MIC, 25±0.00-, MBC, 50-mg/mL in the broth dilution and a significant mean value of 14.70). The stem-bark extracts was also significantly more active than the latex extracts P= .05 with significant mean values of 13.48 and 19.53 respectively. In the time-kill assay, all (100%) the organisms tested were killed by 50% methanol extract of E. abyssinica at concentrations equivalent to 1MIC- 4MIC.

Conclusion: E. abyssinica extracts showed considerable antibacterial activity against the bacterial species tested. These findings authenticate the folklore use of Euphorbia abyssinica for broad spectrum treatment of bacterial infections. The determination of the antimicrobial activity of Euphorbia abyssinica stem (bark and Latex extracts) extract included the 50% methanol, absolute methanol and aqueous extracts of these plant parts. The antimicrobial activity variously exhibited by the 50% methanol extracts of all the two plant parts tested, is significant. This is because it validates the popular traditional uses of dilute alcohol concoctions of medicinal plant preparations in ethno medicinal practice in south-West region of Cameroon. Secondly, the results indicated that these herbs used in traditional medicine have selective antimicrobial activities. Thus, the microorganisms which were susceptible to these extracts are those often associated with wound and ear infections, urinary and gastrointestinal tract infections as well as pyrexia of unknown origin. This explains the discriminate uses of these plants in the treatment of particular ailments. These findings provide evidence that E. abyssinica is a strong candidate in microgram concentrations while the plant extracts were effective in milligram concentrations. Therefore actual comparison between the control drugs and the extracts would await isolation, purification and determination of molar concentrations of the pure active ingredients of these plants extracts.

Author  Details:

Jacqueline Ebob Tarh

Department of Biological Sciences, Cross River University of Technology, Calabar, Nigeria.

Christian Ukwuoma Iroegbu

Department of Biological Sciences, Cross River University of Technology, Calabar, Nigeria.

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Phenomenon of Bacterial Pathogenicity as a Function of Bo-molecules | Chapter 12 | Recent Advances in Biological Research Vol. 3

The bacterial pathogenicity phenomenon is the poly-functional biological potency of germs that are realized by factors (determinants) of pathogenicity (PF) in multi-cellular organism.  

Biological functions are responsible for bacterial pathogenicity in a multi-cellular host organism: the adhesive function, the function of invasion and penetration into the cell, the function of evasion of host defense, and the damage function. Factors of pathogenicity are representative bio-molecules possessed different functional activity. The ligand – receptor interaction of bacterial PF and receptors on eukaryotic cells is the basis of specific lesions caused by the pathogen.

Author  Details:

Yurii V. Ezepchuk

The Gamaleya Institute for Epidemiology and Microbiology, Moscow, Russia and The University of Colorado, Denver, USA.

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A Review on Gongronema latifolium, an Extremely Useful Plant with Great Prospects | Chapter 11 | Recent Advances in Biological Research Vol. 3

Gongronema latifolium is a plant that has a wide range of nutritional and ethnomedical uses in different tropical African communities. Scientific reports on the chemical composition and bioactivity (anti-inflammatory, antimicrobial, antidiabetic, antioxidant, anticancer and allelopathic properties) of the plant material by different authors are discussed in this review. Future prospects of the plant extracts in the areas of herbal formulations, food preservation, alcoholic fermentation and beer production, drug discovery and allelopathy are also highlighted.

Author  Details:

Olugbenga Morebise

Faculty of Basic Medical Sciences, All Saints University School of Medicine, Roseau, Commonwealth of Dominica.

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Partial Purification and Antimycobacterial Screening of the Ethyl Acetate Extract of Alcaligenes faecalis BW1 | Chapter 10 | Recent Advances in Biological Research Vol. 3

Aims: The focus of this study was to evaluate the antimycobacterial activity of Alcaligenes faecalis BW1 extract and to purify it partially.

Study Design: Partial purification of A. faecalis BW1 extract was performed by using thin layer chromatography and active substances responsible for the biological activity were localized.

Place and Duration of Study: The study was carried out at laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco, during the period from January 2011 to July 2011.

Methodology: Crude extract of A. faecalis BW1 was obtained by using ethyl acetate as an organic solvent and its antimycobacterial effect was investigated by agar discs diffusion method. The extract was then fractionated by thin layer chromatography and the bioactivity was assessed with a bioautography technique followed by spots elution tests.

Results: The results showed that A. faecalis BW1 produced compounds with antimycobacterial activity. All the detected spots by thin layer chromatography inhibited the growth of M. smegamtis.

Conclusion: Various metabolites of A. faecalis BW1 are responsible for the sought effect or they could act synergistically to inhibit mycobacterial growth. These compounds could be used after their total purification in further work against mycobacterial infections.

Author  Details:

Ilham Zahir

Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco and Polyvalent Laboratory in Research and Development, Department of Biology, Polydisciplinary Faculty, Sultan Moulay Slimane University, Beni Mellal, Morocco.

Abdellah Houari

Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco.

Mohammed Iraqui

Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco.

Saad Ibnsouda

Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco.

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Nosocomial Infections in Sub-Saharan Africa | Chapter 09 | Recent Advances in Biological Research Vol. 3

The history of nosocomial infections can be traced to the origin of hospitals themselves and have been defined by the WHO as infections that develop in a patient during his/her stay in a hospital or other types of clinical facilities which were not present at the time of admission. Nosocomial infections are a major public health problem globally and are on the increase despite efforts in hospital infection control measures and contribute significantly to morbidity and mortality. Naturally, any micro-organism has the potential to cause infection in hospitalized patients however, only a few including Staphylococci, Escherichia coli, Pseudomonas aeruginosa, Enterococci, fungi and to a lesser extent, viruses and parasites are responsible for the majority of nosocomial infections. In sub-Saharan Africa, data available show that the incidence of nosocomial infections ranges from 2-49% with patients in intensive care units having the   highest rate ranging from 21.2-35.6%. The prevalence of nosocomial infections have  been reported  to vary  between 1.6%-28.7% in Burkina Faso, United Republic of Tanzania, Ghana, Mali, Cameroon, Gabon, Uganda, Burundi, Democratic republic of Congo and Senegal. In Nigeria and Ethiopia, the total accruing occurrence in surgical wards has been reported to vary from 5.7%-45.8% with the later having an incidence as high as 45.8% and an incidence density equal 26.7 infections per 1000 patient days in paediatric surgical patients. In addition, 3.4 -10.9% of hospital-associated infections often  result to mortality in most developed countries though these figures are suspected to be higher in developing countries of sub-Saharan Africa including Nigeria. However, simple and effective control programmes together with effective training of healthcare workers will go a long way in reducing the endemic nature of nosocomial infections in sub Saharan Africa. This paper highlights the natural history, distribution, risk factors of nosocomial infections especially in sub Saharan Africa as well as its contributory factors. Nosocomial infections are endemic in sub Saharan Africa and are further enhanced by emerging and re- emerging resistant agents. Simple and effective control programme together with computer-based epidemiological surveillance carried out as a global project with considerable inputs from developing countries for monitoring will enable the development of nosocomial infections to be halted if not eliminated. In addition, it is necessary to review the current infection control practices in all hospitals particularly in developing countries including Nigeria so as to incorporate molecular techniques which have been proven to be effective in keeping the spread of nosocomial infections under check. The training and re-training of health care givers on principles of infection control is strongly recommended. Also, the principles of infection control should be incorporated into student nurses, medical students, and other paramedical curriculum as well as employment of adequately competent health workers to avoid over labour which sometimes cause workers to be inefficient resulting in disease outbreaks. Finally, hand washing and other standard infection control practices should be adhered to so that nosocomial infections can be controlled effectively.

Author  Details:

Elizabeth N. Mbim

Department of Microbiology, Faculty of Biological Sciences, University of Calabar, P.M.B. 1115, Calabar, Nigeria.

Clement I. Mboto

Department of Microbiology, Faculty of Biological Sciences, University of Calabar, P.M.B. 1115, Calabar, Nigeria.

Bassey E. Agbo

Department of Microbiology, Faculty of Biological Sciences, University of Calabar, P.M.B. 1115, Calabar, Nigeria.

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Antibacterial Activity of Flavonoids Extracted from Seeds of Pongamia pinnata Linn against Methicillin Resistant Staphylococcus aureus | Chapter 08 | Recent Advances in Biological Research Vol. 3

Introduction: Methicillin resistant Staphylococcus aureus (MRSA) is defined as the resistant to penicillinase-stable penicillin’s, thus the acronym MRSA is still under use even though methicillin is no longer the agent of choice for treatment. The use of vancomycin for MRSA remains as the treatment of choice but concerns with rising resistance to glycol peptides call for the restrictive use of these drugs. The resistance mechanism and the genes that mediate resistance have presumably evolved in organism that produce antibiotics such that the antibiotic produced is not effective against the producing organism.

Aims: To assess the antibacterial property of seed crude extracts of Pongamia pinnata Linn and isolated flavonoids component from crude extract against Methicillin resistant Staphylococcus aureus obtained from clinical isolates.

Study Design: Observational study.

Place and Duration of the Study: Department of Allied health sciences, Department of Biochemistry and Department of Microbiology in Sri Devaraj Urs Academy Of Higher Education and  Research, Tamaka, Kolar, between February 2014 and march 2015.

Methodology: Confirmed clinical isolates for MRSA were collected from Microbiology department to test the efficacy of crude extracts of seeds from Pongamia pinnata L. Methanolic crude extract has been preferably used for isolation of flavonoid content using Dimethyl Sulfoxide [DMSO] and methanol as ideal solvents during extraction process by column chromatography technique. Agar well diffusion method was performed to determine the antibacterial activity of crude seed extracts of Pongamia pinnata and isolated flavonoids by using quercitin as positive control for flavonoids. Vancomycin a glycopeptide powder used as gold standard for comparing bactericidal activity of quercitin, flavonoids and crude extracts of P. pinnata on MRSA.

Results: The highest antibacterial activity (75-89%) was observed in crude extract of Pongamia pinnata in comparison to vancomycin considered as cent percent. Extracted flavonoids showed activity (66-92%) with respect to crude extract and (50-84%) with vancomycin and the activity (71-92%) with respect to quercitin when tested with concentration ranging from 25-400 µg/ml.

Conclusion: This study showed that seed extracts of Pongamia pinnata L and its phytochemical compound flavonoids showed potential antibacterial activity against MRSA using quercitin and vancomycin. Flavonoids occupy the first grade antimicrobials in combating methicillin resistant staphylococcus aureus infections. These infections which are prominent in ICU units and HICU units can be drastically controlled without any side effects.

Author  Details:

Mary Shobha Rani Inala

Department of Cell Biology and Molecular Genetics, Sri Devaraj Urs Academy of Higher Education and Research, Kolar, Karnataka, India.

C. D. Dayanand

Department of Biochemistry/Allied Health Sciences, Sri Devaraj Urs Academy of Higher Education and Research, Kolar, Karnataka, India.

P. M. Beena

Department of Microbiology, Sri Devaraj Urs Academy of Higher Education and Research, Kolar, Karnataka, India.

A. V. M. Kutty

Department of Biochemistry/Allied Health Sciences, Sri Devaraj Urs Academy of Higher Education and Research, Kolar, Karnataka, India.

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Tolerance Tests of Alcaligenes faecalis BW1 Extract | Chapter 07 | Recent Advances in Biological Research Vol. 3

Aims: To highlight whether metabolites of Alcaligenes faecalis BW1 extract can be administered orally for their possible antimycobacterial effects.

Study Design: Study of the influence of certain parameters on the extract of Alcaligenes faecalis by using either discs or well diffusion methods against M. smegmatis.

Place and Duration of Study: Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco From April to August, 2012.

Methodology: The impact of acidic pH of gastric juice, bile, hydrogen peroxide, pancreatic enzymes and lysozyme on the antimycobacterial activity of Alcaligenes faecalis BW1 extract was evaluated by agar diffusion method. Detection whether or not antibacterial metabolites having a synergistic effect with rifampicin against M. smegmatis was also explored.

Results: Antibacterial metabolites of Alcaligenes faecalis BW1 extract resist to the action of gastric pH, gallbladder bile and hydrogen peroxide. In addition, they are not affected by pancreatic enzymes and lysozyme. Moreover, they have a synergistic effect with rifampicin against M. smegmatis.

Conclusion: Anti-mycobacterial metabolites of Alcaligenes faecalis BW1 extract are compatible with rifampicin and could be administered orally as antitubercular agents after their purification, identification in further work.

Author  Details:

Ilham Zahir

Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco and Polyvalent Laboratory in Research and Development, Department of Biology, Polydisciplinary Faculty, Sultan Moulay Slimane University, Beni Mellal, Morocco.

Abdellah Houari

Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco.

Mohammed Iraqui

Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco.

Saad Ibnsouda

Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco.

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Isolation, Identification and Pathogenicity of Fungal Organisms Causing Postharvest Spoilage of Tomato Fruits during Storage | Chapter 06 | Recent Advances in Biological Research Vol. 3

Tomato is one of the most widely grown and extensively consumed horticultural crops in the world. Isolation, identification and pathogenicity of fungal organisms causing postharvest spoilage of tomato fruits during storage was carried out. Tomato fruits showing symptoms of rot were collected from the store house. Small sizes were cut and surface sterilized in 1% of Sodium hydrochloride and rinsed in several changes of sterile distilled water. They were plated on Potato Dextrose Agar (PDA) and observed for fungal growth. Identification was done macroscopically and microscopically. For pathogenicity, healthy tomato fruits were plugged with pure cultures of the fungal isolates and disease incidence and severity were evaluated. Five fungi namely Aspergillus flavus, Penicillium waksmanii, Botryodiplodia theobromae, Fusarium oxysporum and Colletotrichum asianum were isolated. Incidence of decay on healthy tomato fruits was 100% for all fungal isolates while the control was 0%. T-test revealed significant differences between the inoculated and the controls at 1% and 5% levels of probability. Severity of decay ranged from 51–53% for all fungal isolates, while the controls showed 0%. T-test revealed significant differences between the inoculated and the control at 1% and 5% levels of probability. Pathogenic microorganisms on tomato are a potential health hazard to man and animals following ingestion.

Author  Details:

Dr. Liamngee Kator

Department of Biological Sciences, Benue State University, Makurdi, Benue State, Nigeria

A. C. Iheanacho

Department of Agribusiness, University of Agriculture, Makurdi, Benue State, Nigeria.

Kortse P. Aloho

Department of Plant Breeding and Seed Science, University of Agriculture, Makurdi, Benue State, Nigeria.

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The Population Dynamics of the Mudskipper, Periophthalmus barbarus (LINNEAUS 1766) (TELEOSTEI, GOBIIDAE) and the Implication for Conservation and Management in the Mangrove Swamp of Iko River Estuary, Southeastern Nigeria | Chapter 05 | Recent Advances in Biological Research Vol. 3

The population dynamics of Periophthalmus barbarus in the mangrove swamp of Iko River estuary, southeast of Nigeria were obtained from a twenty four month length composition data ranging 4.6 – 14.5 cm total length (TL) (mean 9.1841± 1.6346: n = 2,876) corresponding to 1.16 – 50.6 g total weight (TW) (mean = 9.9626 ± 5.4796) the growth was exponential. The asymptotic length (L∞) of the Powell-Wetheral plot (L∞ = 15.03 cm) was seeded into FSAT II (FAO-ICLARM Stock Assessment Tools II) software to obtain best estimates of von Bertalanfy growth parameters as L∞ = 16.22 cm TL, growth coefficient (K) = 1.2 year-1, age of fish at zero length, to = 0.071, longevity, tmax = 2.5 years. The estimated growth performance index, φ’ = 2.449. Other FISAT II growth parameters were the amplitude of growth oscillation, C = 0.6 and the winter point, WP = 0.6, Rn = 0.3127. Mortality parameters were total mortality, Z = 479 year-1, natural mortality, M = 2.39 year-1 and fishing mortality, F was 2.40 year-1. Result indicate the fishery is optimally exploited with current exploitation rate, Ecur = 0.50 < Emax = 0.668 > Eopt = 0.5 which suggests stock optimal exploitation, corroborated by Z/K ration (3.184).  Ecur (0.5) means that 50% of the available stock is being fished annually. The length-at-first capture Lc = 7.33 cm TL and Lc/L∞ was 0.45, indicating the fish was yet to complete 55% of growth as at the time of capture at Lc, hence P. barbarus in the ecosystem is at the optimal level of exploitation as well as the presence of growth overexploitation. Thus to circumvent the consequences of growth overfishing, sustainable fisheries measures such as monitoring of fishing effort, use of selective gears and increase in mesh size should be encouraged, implemented and enforced. The study has revealed that P. barbarus population residing in the mangrove swamp of Iko River Estuary is experiencing exploitation rate close to the maximum sustainable yield amidst the presence of heavy fishing pressure. Moreover, the mudskipper fishery in this ecosystem is currently exhibiting growth overexploitation signs which could lead to severe implications on the population size and food security within vulnerable fishing households in the future. Therefore, urgent management interventions in the form of monitoring fishing efforts, return of captured juveniles back to the water body from the non-selective fishing gear and use of selective gear with large mesh size (to increase length at first capture) are needed to safeguard this important fish species from possible collapse in the future.

Author  Details:

Dr. Nsikak Abiaobo

Department of Zoology, Akwa Ibom State University, Ikot Akpaden, Mkpat Enin, P.M.B. 1167, Akwa Ibom State, Nigeria.

Dr. Mfom Tmothy Udo

Department of Fisheries and Aquatic Environment Management, University of Uyo, P.M.B. 1017, Akwa Ibom State, Nigeria.

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