Adebukola Adisa Ekoh-Jolly1

1 Department of Medical Laboratory Science, University of Medical Sciences, Ondo city, Ondo State, Nigeria

Isaiah Nnanna Ibeh2 Erhabor Ikponmwosa2

2 Department of Medical Laboratory Science, University of Benin, Benin City, Nigeria

Samuel Ayobami Fasogbon3*

3Department of Laboratory Accreditation, Medical Laboratory Science Council of Nigeria, Abuja, Nigeria

Ahmed O. Adebayo4

4Department of Medical Laboratory Science, Lagos State College of Health Technology, Yaba-Lagos, Nigeria

Loveth O. Fasogbon5; Monica Osato Solade5

5Department of Medical Laboratory Science, College of Medicine, Ambrose Alli University, Ekpoma, Nigeria

Cyril A. Egbo6

6Department of Community Health, College of Medicine, University of Benin, Benin city, Nigeria

Samuel Ehiremen7

7Department of Anatomy, faculty of Basic medical Sciences, Olabisi Onabanjo University, Ago-iwoye, Nigeria.

Ifeoma G. Onukwue8

8National Quality External Assessment Laboratory, Medical Laboratory Science Council of Nigeria, Zaria, Nigeria.

Ayobami B. Olayera9

9Department of Haematology and Immunology, University of Nigeria Teaching Hospital,

Ituku-Ozalla, Enugu State, Nigeria4

All Corresponding to: Samuel Ayobami Fasogbon,, samfash4best@gmail.com

ABSTRACT

Background: The use of alcohol-based hand sanitizer that contains at least 60% alcohol prevents the spread of infections and thus prevents the spread of antimicrobial resistance. Aim: The aim of this study was to evaluate the antimicrobial efficacy of different brands of hand sanitizers sold in Nigeria. Materials and Methods: Antimicrobial efficacy of eleven popular brands of alcohol-based hand sanitizers sold in Nigeria was evaluated and comparatively analyzed using a combination of Agar well diffusion (Mueller-Hinton Agar for bacteria and Sabouraud Dextrose Agar for fungi), Broth dilution (Nutrient broth) and In vivo Viable Microbial count reduction techniques. A total of 330 clinical isolates comprising 55 each of Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris and Candida albicans obtained from culture collection were used as test organisms. Results and Discussions: This study revealed a very high significant difference (P<0.001) on the susceptibility pattern of the test organisms against the various hand sanitizers used. The study showed a very high significant difference between the activity of the various hand sanitizers and the positive control (P<0.001). All hand sanitizers used in this study apart from Purit are less active against Proteus vulgaris but are more active against Staphylococcus epidermidis. The findings in this study showed that isopropyl alcohol based-gel form hand sanitizer had 82.2% efficacy while ethanol-based gel form hand sanitizers had 76.9% efficacy. This study revealed Significant difference (P<0.001) on Minimum Inhibitory Concentration (MIC) of all hand sanitizers used against test organisms except for Evree hand sanitizer which showed no significant difference (P>0.05) on Minimum Inhibitory concentration against test organisms. Of all the hand sanitizers used in this study only Purit has bactericidal activity against Proteus vulgaris not lower than 95% concentration. Conclusion: The results of this study suggest high significant difference in the antimicrobial efficacy of various hand sanitizers sold in Nigeria.

Keywords: Anitimicrobial, Concentration, COVID-19, Efficacy, Sanitizers.

INTRODUCTION

Globally, there are 119,094,168 corona virus cases, 2,640,822 deaths, 94,632,462 recovered and 21,820,884 active cases as at 12th March, 2021 [1][2]. The genome sequence of virus SARS-CoV-2 is similar to SARS Coronavirus (SARS-CoV) [3][4]. The CoVs are members of the same genus Beta coronavirus, sharing similar morphology in the form of enveloped, positive single-stranded RNA viruses [5]. They can be deactivated by certain lipid solvents such as ethanol, ether (75%), chlorine-containing disinfectants, and chloroform, except chlorhexidine [5]. Ethyl alcohol, at concentrations of 60%–80%, is effective viricidal agent inactivating all the lipophilic viruses (e.g., influenza, herpes and vaccinia virus) and many hydrophilic viruses (e.g., adenovirus, enterovirus, rhinovirus, and rotaviruses excluding hepatitis A virus (HAV) or poliovirus) [6].

Regular hand washing with soap and water prevent the spread of infections and decrease the risk of getting sick. When there is non-availability of soap and water, CDC recommends the use an alcohol-based hand sanitizer that contains at least 60% alcohol [6]. Hand hygiene prevents the spread of antimicrobial resistance.

Alcohol-based hand sanitizers generally contain one or more alcohols (ethanol or 1-propanol or 2-propanol) in quantitative ratios that are deferred by brand. The alcohols present in the Alcohol-based hand sanitizers formulations triggers bactericidal action by denaturation of microbial proteins [8]. Some Alcohol-based hand sanitizers may as well contain other chemical constituents that work in synergy with the bactericidal action of the alcohols. Other chemical compounds may confer independent bactericidal activities through unknown mechanisms.

Due to the continuing worldwide COVID-19 pandemic, the demand for hand sanitizers remains at a very high level. In Nigeria, many Pharmaceutical companies, cosmetics industries, even a soft drink bottling company and a University went to the production of hand sanitizers to meet market demand and for profit making. Majority of manufacturers claim 99.9% effectiveness of their hand sanitizers.

Different categories of hand sanitizers (alcohols, quaternary ammonium compounds and triclosan) possess different levels of sensitivities and therefore varying resistance by various infectious agents. For example, some species of Staphylococcus aureus carry a gene that allows resistance to some hand sanitizers [6]. Some hand sanitizers are fungistatic, others are bacteriostatic against gram-positive bacteria and bacteriostatic against some gram-negative bacteria [6]. Like ethanol hand sanitizers, those of the quaternary ammonium compounds are not active against non-enveloped viruses. More so, some are safer, more economical, less messy and more enjoyable to use for kids. This study intends to look into the usage and effectiveness of these different brands of hand sanitizers.

This study examined the antimicrobial efficacy of eleven brands of hand sanitizers sold in Nigeria against Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris and Candida albicans, and also to assess and compare their antimicrobial effectiveness.

MATERIALS AND METHODS

Type of Study

This is a cross sectional descriptive study and was conducted to comparatively analyze some hand sanitizers sold in Nigeria.

Study Location

The research was conducted in the Biology Laboratory of the Department of Biological Sciences, Faculty of Science, University of Medical Science, Ondo, Nigeria.

Ethical Considerations

Written/verbal informed consent was received from subjects who participated in the study at the study center. The Ethical committee of the Edo state Ministry of health, Benin City, reviewed and approved the study protocol, HA. 737/83.

Population and Subjects

The population of this study was centered on users of hand sanitizers among health workers, students and civil servants in Benin City and Ondo town of Nigeria.

Test Organisms

A total of Three hundred and thirty clinical isolates comprising fifty-five each of Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris, and Candida albicans obtained from Don Bosco Health Centre, Akure, Ondo State, Central Hospital, Benin City and Stella Obasanjo Hospital, Benin City, Edo State were used as test organisms in this study. These micro organisms were chosen because they are commonly implicated in community-acquired infections [8][9].

Test Subjects

A total of Two hundred Subjects comprising of Health Workers, Civil Servants and Students participated in this research after they consented to participate.

Hand Sanitizers

Eleven brands of alcohol-based hand sanitizers (10 waterless gel form – Lifebouy, Purit, Dettol, Carex, Tetmosol, Sanitol, Wind, Seed, Tribute, Evree, and one spray form – 2sure) commonly sold and used in Nigeria were chosen for this study based on their popularity and availability at retail outlets across Nigeria. Each hand sanitizer was stored as recommended by its manufacturer and was used well before the expiration date.

Test Organisms Standardization

Turbidity variability of Bacterial suspensions may affect result, thus the need for standardization of microbial testing; McFarland standards was used as a reference to adjust the turbidity of bacterial suspensions. The McFarland 0.5 turbidity standard was prepared by adding 0.5 ml of 1.175% w/v barium chloride dihydrate (BaCl2·2H2 O) solution to 99.5 ml of 15 w/v sulfuric acid (H2 SO4) [10]. This was mixed uniformly and aliquoted into test tubes identical to the ones used in preparing inoculum suspensions of the test organisms. The accuracy of the density of the standard was verified using a spectrophotometer. The absorbance of the 0.5 McFarland standards at wavelength of 625 nm was 0.08–0.10. The tubes were stored in a well-sealed container in the dark at room temperature until when required [11].

A sterile loop was used to pick a loopful of inoculum from a pure culture of the test organism. This was then transferred and suspended into a tube containing sterile normal saline (NaCl 8.5 g, distilled water 1 L). The tube was compared with the turbidity standard, and the density of the organism was adjusted by adding more bacteria or sterile saline until standardization was attained [12].

Susceptibility of Test Organism to Hand Sanitizers

Disk agar diffusion technique (well-variant) as described by Valgas et al. was used for determining antimicrobial efficacy of hand sanitizers [13]. Sterile Mueller-Hinton agar plates were inoculated with standardized test organisms. A sterile cotton swab was dipped into a test tube containing the inoculum and was rotated properly to allow maximum contact. Excess inoculum was removed by pressing and rotating the swab firmly against the inside wall of the tube above the liquid level. The swab was then streaked over the surface of the medium three times while rotating the plate at 60° angle after each application. The swab was also passed around the edge of the agar surface. The inoculum was left to dry for a few minutes at room temperature with the lid closed.

With the aid of a sterile 4mm cork borer, 3 equally spaced holes were bored in the agar plate. The agar plugs were discarded using a sterile needle. A 100 microliters volume of undiluted hand sanitizer was then introduced into the first well, the second well was filled with an equal volume of sterile water to serve as negative control, while the third well was filled with 90% ethanol as positive control. This was done for all the test organisms and hand sanitizers. The plates were incubated for 24hours at 37°C in an upright position. They were then examined for zones of inhibition which indicated the degree of susceptibility or resistance of the test organism to the antibacterial agent. The point of abrupt diminution growth which corresponds to the complete growth inhibition was taken as the zone edge. The test was carried out five times, and the average of all readings was taken as the zone of inhibition in each case.

Determination of Minimum Inhibitory Concentration (MIC)

Minimum inhibitory concentration is the lowest concentration of an antimicrobial agent that completely inhibits the growth of a test organism as seen by the unaided eye [14]. To determine the MIC, increasing concentrations (5% – 100% with 5% intervals) of each Hand sanitizer were prepared in 9ml tubes of sterile nutrient broth. Exactly 100microlitre of each standardized test organism was then introduced into each tube of Hand Sanitizer. A tube containing only nutrient broth and bacteria without hand sanitizer served as negative control while another tube containing just the sanitizer and broth without bacteria served as positive control. Each tube was incubated for 18hours for bacteria and 48hours for fungi, then examined for visible growth or turbidity. The concentration of the hand sanitizer in the tube in which no visible growth was observed when compared with the controls was taken as the MIC.

Determination of Minimum Cidal Concentration

Minimum bactericidal concentration is the lowest concentration of an antimicrobial that can kill the test organism [11]. To determine the cidal for each Hand sanitizer, samples from the test tubes used in MIC test that showed no visible growth after the period of incubation were inoculated on sterile nutrient agar plates (which had no antimicrobial incorporated) for bacterial and Sabouraud Dextrose Agar for fungi in them using sterile swab sticks. The plates were incubated at 370C for 18 to 24hours and were then observed for growth. The concentration at which absence of growth was observed (bactericidal activity) was then taken as MBC.

In vivo Reduction of Viable Bacteria and Fungi counts on Hands of Subjects

The Hand sanitizers evaluated for their efficacy in reducing baseline bacteria and fungi counts on the hands of subjects. Ten individuals were selected for each products and verbal informed consent was obtained from each subject prior to the conduction of the experiment. Subjects did not apply antimicrobial substance to their hands prior to the experiment. Sterile nutrient agar and Sabouraud Dextrose agar plates were divided into two halves with one half labeled BF (before) and the other labeled AF (After). Subjects were asked to gently make an impression on the surface of the BF side of the agar with the three unwashed fingers. After this, 3ml of the hand sanitizer was then applied to the hands and rubbed thoroughly on the palms, fingers and the back of the hands until the hands became completely dried. Subjects were then asked to repeat the finger impression on the AF part of the plate. This was done by all subjects. The plates were incubated at 370C for 24hours and the numbers of colonies were counted. The percentage of CFU reduction was calculated as follows:

%CFU reduction = CFU/mL on Bf Section – CFU/mL on AF Section x 100 CFU/mL on BF Section [15].

Statistical Analysis

Analysis of Variance (ANOVA), Duncan Multiple Range Test, Unpaired T-Test and Chi-Square were used to Test for Significant Difference on Susceptibility Pattern of Test Organisms to the various Hand Sanitizers.

Paired T-test was used to test the significant difference of Percentage CFU Reduction of Viable Bacteria and Fungi from hands of Subjects by the hand Sanitizers.

Minimum Inhibitory Concentration (MIC), Minimum Cidal Concentration (Bacteriocidal) (MBC) and Fungicidal (MFC) of the Hand Sanitizers against Test Organisms are presented in Charts.

RESULTS

Data Analysis and Presentation

In this study, a total of Three hundred and thirty clinical isolates comprising fifty-five each of Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris, and Candida albicans were tested against eleven alcohol-based hand sanitizers commonly sold and used in Nigeria (Table 4.1) for their efficacy.

It was observed that all the hand sanitizers used in this study have varying inhibitory activities against all test organisms and that each of the hand sanitizers used had different level of inhibition against each of the test organisms. A sanitizer very active against a particular test organism is less active against another test organism (Table 4.2).

The study revealed that all the hand sanitizers used in this study have varying Minimum Inhibitory concentration (MIC) against the susceptible test organisms (Table 4.3).

It was observed that not all the hand sanitizers used in this study had cidal acitivity against test organisms and where it did have, the Minimum cidal concentrations were mostly 100% (Table 4.4)

The study revealed varying In-vivo reduction of Viable Bacteria and Fungi from hands of subjects by three hand sanitizers that have cidal activities on test organisms. It was observed that Purit hashigher (82.2%) viable bacterial count reduction than fungi (31.8%). The study also revealed that Evree has equal in-vivo reduction of viable bacteria and fungi (76.9%) while Lifebuoy has highe in-vivo reduction of viable fungi (64.5%) than bacteria (41.6%) (Tables 4.5 and 4.6)

Purit and Lifebouy have inhibitory activity against test organisms not below 60% concentration while Dettol has inhibitory activity not below 70% concentration. Sanitol, Seed and Tribute have inhibitory activity not below 75% concentration. Carex, wind and Evree have inhibitory concentration not below 80% concentration while Tetmosol and 2Sure hand sanitizers have inhibitory activity against test organisms not below 90% concentration

.

DISCUSSION

The use of alcohol-based hand sanitizers has become more popular with the emergence of Coronavirus pandemic. Many brands of alcohol-based hand sanitizers flooded the Nigerian markets with the manufacturers of various hand sanitizers claiming 99.9% effectiveness against infectious organisms. This study became necessary due to increasing reliance on alcohol-based hand sanitizers to prevent community spread of infectious diseases.

This study revealed a very high significant difference (P<0.001) on the susceptibility pattern of the test 0rganisms (Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris, and Candida albicans) against the various hand sanitizers used in this study, this is in line with the finding of [16], who also reported significant difference (P<0.001).

The study also show a very high significant difference between the activity of the various hand sanitizers and the positive control (P<0.001). This finding is different from the finding of [17] who reported no significant difference between the activity of the various hand sanitizers and the positive control. Though he did not revealed the brand name of the hand sanitizers used in his study, they were also alcohol-based hand sanitizers, the difference may be due to other ingredients in the hand sanitizers, mores so [17] used a commercial hand sanitizer as his positive control while 90% ethanol was used as positive control in this study.

All hand Sanitizers used in this study apart from Purit are less active against Proteus spp (a Gram-negative bacilli) but are more active against Staphylococcus epidermidis (a Gram-positive cocci), this may be due to differences in their cell wall components.

The findings in this study that isopropyl alcohol based-gel form Hand sanitizer (Purit) has higher efficacy than ethanol-based gel form hand sanitizers is the same with what was observed by [15] that isopropyl alcohol solution has higher efficacy than ethanol solution, but contrary to findings of [18] which was in reverse.

This study also showed Significant Difference (P<0.001) on Minimum Inhibitory Concentration (MIC) of all Hand Sanitizers used against Test Organisms except for Evree Hand Sanitizer which show no significance difference (P>0.05) on Minimum Inhibitory concentration against test organisms.

This study revealed that none of the hand sanitizers sold in Nigeria, used in this study have inhibitory activity (MIC) below 60% concentration which is lower than below 75% concentration that was reported by [17].

2Sure the only Liquid form hand sanitizer used in this study has lower antimicrobial efficacy than other gel form hand sanitizers used, this is the same with the report of [18].

Of all the hand sanitizers used in this study only Purit has bactericidal activity against Proteus spp not lower than 95% concentration. Lifebuoy, Dettol, Sanitol, Seed, Tribute and Evree have bactericidal effects not lower than 100% concentration on other test organisms. Carex, Temosol, 2Sure and Wind hand Sanitizers have no bactericidal effect on test organisms.

This study showed that Purit, Lifebuoy, Tetmosol and Evree have fungicidal effects on Candida albicans at concentration not lower than 100%. Dettol, Carex, Sanitol, 2Sure, Wind, Seed and Tribute hand Sanitizers sold in Nigeria, used in this study did not have fungicidal effect on test organism.

The mean percentage in vivo reduction of viable bacteria and fungi counts observed in this study is higher than what was observed by [17] but lower than 99.9% claim by manufacturers of hand sanitizers.

Conclusion

Based on the findings of this study, the Manufacturers’ claim of 99.9% of antimicrobial efficacy of various hand sanitizers sold in Nigeria, used in this study is in doubt. There is significant difference on the antimicrobial efficacy of various hand sanitizers sold in Nigeria, used in this study and the various hand sanitizers sold in Nigeria are more active against Gram positive bacteria than Gram negative bacteria with more potency against bacteria than fungi. Isopropyl alcohol based-gel form Hand sanitizer (Purit) has higher efficacy than ethanol-based gel form hand sanitizers

Recommendation

The revelation of this study suggests that no hand sanitizer sold in Nigeria, used in this study have 99.9% antimicrobial efficacy, therefore hand washing with water and soap should be practiced more often to physically remove the infectious microbes from the hands then combine with rubbing hand with hand sanitizers.

Conflict of Interest

Authors declare no conflict of interest

REFERENCES

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2. NCDC, (2021). COVID-19 Coronavirus 2019-n Cov Statistics Update (Live): 4,122,912 Cases and 280,337 Deaths. Available online: https:// virusncov. com/ (accessed on 8 May 2020).

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5. Cascella, M., Rajnik, M., Cuomo, A., Dulebohn, S.C. and Di Napoli, R. (2020). Features Evaluation and Treatment of Coronavirus (COVID-19); StatPearls Publishing: St. Petersburg, FL, USA. pp. 4 – 10.

6. CDC, (2020). COVID-19 Coronavirus 2019-n Cov Statistics Update (Live): 4,122,912 Cases and 280,337 Deaths. Available online: https:// virusncov. com/ (accessed on 8 May 2020).

7. Paulson, D.S., Fendler, E.J., Dolan, M.J. and Williams, R.A. (1999). A close look at alcohol gel as an antimicrobial sanitizing agent. American Journal of Infection Control. 27: 332 – 338.

8. Montravers, P., Alain, L., Dubreuil, L. and Benchimol, D. (2009). Clinical and microbiological profiles of community-acquired and nosocomial intra-abdominal infections: Results of the French prospective, observational EBIIA study. Journal of Antimicrobial Chemotherapy. 63(4): 785 – 794.

9. Rosenthal, K.L., Hendrick, M.J. and Rankin, S.C. (2008). Methicillin-resistant Staphylococcus aureus associated dermatitis in a Congo African grey parrot (Psittacus erithacus erithacus). Medical Journal of Avian Mycology/Bacteriology.12(9): 4 – 10.

10. NCCLS, (1999). Performance standards for antimicrobial susceptibility testing. www.scrip.org.

11. Cheesbrough, M. (2006). District Laboratory Practice in Tropical Countries. 2nd edition. New York: Cambridge University Press. pp. 137 – 141.

12. Vandepitte, J., Verhaegen, J., Engbaek, K., Rohner, P., Piot, P. and Heuck, C.C. (2003). Basic Laboratory Procedures in Clinical Bacteriology. 2nd edition. Geneva: WHO; 2003.

13. Valgas, C., De Souza, S.M., Smania, E.F. and Smania, A. (2007). Screening methods to determine antibacterial activity of natural products. Brazil Journal of Microbiology. 38: 369 – 380.

14. CLSI. (2006). Performance standards for antimicrobial disk susceptibility tests. Approved standard. 9th Edition Document M2-A9. Clinical and Laboratory Standards Institute, Wayne, PA.

15. Oke, M.A., Bello, A.B., Odebisi, M.B., El-Imam, A.A. and Kazeem, M.O. (2013). Evaluation of antibacterial efficacy of some alcohol-based hand sanitizers sold in Ilorin (North-Central Nigeria). Ife Journal of Science. 15(1): 111 – 117.

16. Vardhaman, M., Gundabaktha, N., Arun, S. and Vishwakarma, K.G. (2016). Comparative assessment of antimicrobial efficacy of different hand sanitizers: An in vitro study. Dental Research Journal. 13(5): 424 – 431.

17. Imuetinyan, O. (2017). Comparative analysis of the antimicrobial efficacy of some hand sanitizers in use in Benin Metropolis. Journal of Microbiology.23:3-6.

18. Ochwoto, M, Muita, L., Talaam, K., Wanjala, C., Ogeto, F., Wachira, F., Osman, S., Kimotho, J. and Ndegwa, L. (2017). Anti-bacterial efficacy of alcoholic hand rubs in the Kenyan market. Antimicrobial Resistance & Infection Control. 6(1): 17 – 20.

The post Antimicrobial Efficacy of Various Hand Sanitizers Sold in Nigeria appeared first on Nigerian Biomedical Science Journal.

Joseph. Emberga. Toryila, Ayuk Hughes Bisong, Sheba John Ishaku

Department of Physiology, Faculty of Basic Medical Sciences, Bingham University, Karu,

All Corresponding to: Joseph, joetoryila@gmail.com

ABSTRACT

Background: Infertility is a worldwide health concern in the developing world, affecting 8-15% of couples in their reproductive age. Musa paradisiaca (BANANA) is a crop in the genus Musa and all members of the genus are indigenous to tropical and subtropical countries. Banana is wildly consumed by Africans especially by Nigerians. The study was aimed at determining the effect of Musa paradisiaca (BANANA) flour supplement on the reproductive system of male wistar rats. Methodology: Twenty (20) male wistar rats weighing between 85-170g were used for this study and were randomly divided into four (4) groups. Group one served as the control, group 2 and 3 were treated with commercial feed supplemented with banana flour in the ratio 20:80 ratio and 5ml/kg of body weight of lime juice orally for four weeks .Group 4(Positive control) was given 5ml/kg body weight of lime juice orally for four weeks. The animals were sacrificed by cervical dislocation and semen samples were collected. Sperm motility, sperm morphology, sperm count and testosterone levels were investigated. Resuls: The result shows that there was increase in sperm motility in treated groups 2 (65.00±13.23) and 3 (68.33±7.64) compared to group 4 and the control group (60.0±20.0), significant increase (P≤ 0.05) in sperm count in treated groups 2 (2100.00±208.44) and 3 (2700.00±754.44) compared to group 4 and the control group (1933.33±120.43), There was a significant increase (P≤ 0.05) in sperm mortality in treated groups 2 and 3) compared to group 4 and the control group. There was a significant increase (P≤ 0.05) in testosterone level in treated groups 2 (5.73 ± 4.38) and 3 (7.56 ± 5.07) compared to control the group (1.23 ± 0.32). There was a significant increase in sperm abnormalities in group 4 (11.67±7.63) when compared to group 2, 3 and the control (P≤ 0.05). Conclusion: The effect of banana flour supplement on the reproductive system of male wistar rats was investigated and it was shown to increase sperm motility, sperm count, testosterone levels in all treated groups, while decreasing sperm mortality and sperm abnormalities.

INTRODUCTION

Infertility is a couple`s inability to have a child after one year of unprotected sexual intercourse (WHO, 2016). Infertility is a worldwide health concern in the developing world, affecting 8-15% of couples in their reproductive age (Otubu, 2006). The prevalence of infertility in a rural Nigerian community is determined by a systematic random sampling of the population. The overall prevalent rate was 30.3%, giving indices of 9.2% for primary infertility and 21.1% for secondary infertility (Adebayo et al,2021). Infertility leads to stigmatization, marital instability, and enormous psychological stress (Adebayo et al,2021). Infertility is a public health concern in many developing nations of the world due to its high prevalence and especially because of its serious social implications. Infertility has social, economic and personal effects, which go beyond childlessness, and women bear the major brunt of the burden. It is a major cause of marital disharmony and separation, and personal misery in some other population groups (Ola. 2009). The highest prevalence is in low resource countries, particularly in sub-Saharan Africa (Sharma et al 2008). Male infertility can be caused by poor penile erection, abnormal sperm quality and volume, abnormal ejaculation, other causes such as plants like lime, which has been described as a natural spermicide; a contraceptive substance that reduces sperm concentration to prevent pregnancy.

Banana is an elongated, edible fruit botanically a berry (Armstrong et al., 2013) produced by several kinds of large herbaceous flowering plants in the genus Musa (Armstrong et al., 2013). Researches into natural diets like banana (plantain) showed that its consumption by men could enhance some reproductive functions, and also alleviate certain reproductive dysfunctions (Yakubu et al., 2007) (Ojewole et al., 2003). Bananas are considered nutritive with high content of vitamins A, B1 and C (Margard and Briav, 1979). Generally, bananas contain a considerable amount of mineral elements and could therefore serve as a good source of mineral supplement in human/animal diets. It had a medicinal use in people with high blood pressure and stroke and also reported to help to stimulate the production of hemoglobin and also have hypolipidemic effects

Musa paradisiaca promotes healthy digestion, improves affective state, helps in the retention of and serves as good sources of calcium, phosphorus and nitrogen, which build and regenerate tissues in the body, and is also a rich source of iron and vitamins, especially Vitamins C and E. The fruit is also rich in potassium which are essential for production of sex hormones and enhancing the male libido and improve fertility by enhancing semen quality (Hossain et al., 2015). Enzyme bromelain in banana increases sex drive and reverses impotence in men (also found in pineapple). These phytochemicals also have stimulatory effects on sex hormone and semen production (Hossain et al., 2015), as well as sperm motility. Hence the aim of this research was to ascertain the effect of banana flour supplements on the reproductive system of male wistar rats using lime juice to induce sperm mortality.

MATERIALS AND METHODS

Preparation of the banana flour

Green plantain fruits (ripe); were obtained from a farm. The fruits were cut longitudinally into chips of about 5 mm thickness and air dried for 4 days after which they will be grinded and made into flour..Commercial feed supplemented with banana flour in the ratio 20:80 (Manuela et al., 2018).

ANIMAL GROUPING

Twenty (20) male wistar rats weighing between 85-170g were used for this study were divided randomly into four groups:

Group 1: Served as Control group.

Group 2: Commercial feed supplemented with banana flour in the ratio 20:80 ratio and 5ml/kg lime juice orally.

Group 3: Commercial feed supplemented with banana flour in the ratio 20:80

Group 4: Given 5ml/kg body weight of lime juice orally (Ojewole et al., 2009)

The treatment lasted for a period of 14 days.

Twenty four (24) hours after the 14th day of treatment, the rats were euthanized by cervical dislocation and the testes and epididymis excised using a midline abdominal incision. The left caudal epididymis was immediately transferred into sterile bottles containing 2 ml of normal saline for semen analysis. Semen analysis was carried out on sperm motility, sperm morphology and sperm count according to the method of Dolt and Foster, 1972. Blood sample was collected by cardiac puncture and Serum Testosterone level was determined using ELISA technique.

RESULTS

The results indicate significant increase in testosterone level in treated groups 2 and 3 compared to control the group (P≤ 0.05). There was a decrease in testosterone level in lime juice treated group 4 compared to the control group.

Semen Analysis

There was significant increase in sperm motility and sperm count in treated groups 2 and 3 compared to the control group (P≤ 0.05). There was significant decrease in sperm motility and sperm count in lime juice treated group 4 compared to the control group. The result indicates no difference in abnormalities in treated groups 2 and 3 when compared to the control group. However there was an increase sperm abnormality in lime juice treated group 4 when compared to the control group (P≤ 0.05).

Semen Analysis

The results indicate significant increase (P≤ 0.05) in sperm motility in treated groups 2 (65.00±13.23) and 3 (68.33±7.64) compared to the control group (60.0±20.0) due to the banana flour supplement treatment. There was significant decrease in sperm motility in lime juice treated group 4 (60.00±10.00) compared to the control group (60.0±20.0). The results indicate no difference in sperm abnormalities in treated groups 2 (5.00±5.00) and 3 (5.00±5.00) compared to the control group (5.00±5.00). However there was a significant increase (P≤ 0.05) in sperm abnormalities in lime juice treated group 4 (11.67±7.63) when compared to control group (5.00±5.00). There was significant increase (P≤ 0.05) in sperm count in treated groups 2 (2100.00±208.44) and 3 (2700.00±754.44) compared to the control group (1933.33±120.43) due to the banana flour supplement treatment. However was there was a significant decrease (P≤ 0.05) in sperm count in lime juice treated group 4 (316.7±497.17) compared to the control group (1933.33±120.43).

The results indicate significant increase (P≤ 0.05) in testosterone level in treated groups 2 (5.73 ± 4.38) and 3 (7.56 ± 5.07) compared to control the group (1.23 ± 0.32) due to the banana flour supplement treatment. There was a decrease in testosterone level in lime juice treated group 4 (0.77 ± 0.90) compared to the control group (1.23 ± 0.32).

Musa paradisiaca (Banana) contains the enzyme bromelain and other chemicals such as calcium, potassium, alkaloids, flavonoids, vitamins A ,B1 andC . Bromelain, potassium and vitamins B are essential for the production of sex hormone (testosterone), enhancing the male libido and improving fertility by enhancing semen quality (Hossain et al., 2015). They also have stimulatory effects on sex hormone (testosterone) and semen production (Aiddiq et al., 2012), as well as sperm motility.

These results are in line with work done by Yakubu (2007) on the effects of banana fruits in the management of sexual dysfunction in male wistar rats. Although the exact mechanism of action of antioxidants was not stated, the banana fruit was reported to be efficient in the management of sperm concentration (Yakubu et al., 2007). Vitamin A has been studied in combination with other antioxidants including zinc, N-acetylcysteine (NAC), selenium, and vitamins C and E. Scott et al. 1998, reported a 30% increase in motility after a 3-month regimen of vitamin A supplementation combined with vitamins C and E and selenium

For men hoping to increase fertility, the goal is to make sure sperm are healthy. Antioxidant supplements, vitamin A,B1 and C contain in banana can help.

RECOMMENDATION

Infertility is on the increase, eating banana flour as a supplement has proven to affect the male reproductive system positively. Thus medical practitioners should look outside traditional methods for treating infertility in men. Further study should be done on the possible mechanism of action of banana flour supplement on the hypothalamic-pituitary-gonadal axis.

CONCLUSION

The effect of banana flour supplement on the reproductive system of male wistar rats was investigated and it was shown to increase sperm motility, sperm count, testosterone levels in all treated groups. Banana flour supplement should be considered for treatment of infertility in men.

REFERENCES

1. Ketiku AO (1973). Chemical composition of unripe (green) and ripe plantain (Musa paradisiaca). Journal of Science and Food Agriculture 24: 703-707.

2. Brugh V., and Lipshultz L., (2004). Male factor infertility.Journal of Medical Clinics of North America.

3. Caballero B., Finglas P., and Toldra F., (2015). Encyclopedia of Food and Health. Elsevier Science.

4. Corbier, P. Edwards, D.A. Roffi, J. (1992). “The neonatal testosterone surge: a comparative study”. Arch Int Physiol Biochim Biophys. 100 (2): 127–131

5. Hirsh, A. (2003). “Male subfertility”. British Medical Journal. 327 (7416): 669–672.

6. Holstein AF, Schulze W, Davidoff M. Understanding spermato-genesis is a prerequisite for treatment.

7. Reproductive Biology and Endocrinology. 2003; 1:107

Fellers, P.J. Nikdel, S. and Lee, H.S. (August 1990).

8. “Nutrient content and nutrition labeling of several processed Florida citrus juice products”. Journal of America Diet Association.

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10. Chimbatata N., and Malimba C., (2016). Infertility in sub-Saharan Africa. A qualitative review of literature. Journal of Social Science. .

11. Araoye O., (2003). Epidemiology of infertility: social problems of infertile couples. Journal of West African Medicine.

12. Van Balen F., and Bos H., (2009). The social and cultural consequences of being childless in poor-resource countries. Facts, Views and Vision in Obstetrics and Gynaecology. Journal of Medical Gynaecology.

13. Ola T., Aladekomo F., and Oludare B., (2008). Determinants of the choice of treatment outlets for infertility in Southwest Nigeria. Journal of Shawal Medicine.

14. Pantii A., and Sununu Y., (2014). The profile of infertility in a teaching hospital in north West Nigeria. Journal of Shawal Medicine.

15. Scott R et al. The effect of oral selenium supplemen- tation on human sperm motility. British Journal Urology. 1998;82(1):76–80

16. World Health Organization. (2010). “WHO Laboratory manual for the examination and processing of human semen, 5th edition”. Cambridge: Cambridge University Press.

17. Walker, W. H. (2009) “Molecular mechanisms of testosterone action in spermatogenesis”. Steroids 74: 602-607

18. Moench GL, Holt H. (1931) Sperm morphology in relation to fertility. American Journal of Obstetrics and Gynaecology ; 22: 199–210

19. Katz DF, Overstreet JW, Samuals SJ, Niswander PW, Bloom TD et al (1986). Morphometric analysis of spermatozoa in the assessment of human male infertility. Journal of Andrology; 7:203–10

The post Effect of Musa Paradisiaca (BANANA) Flour Supplement on the Reproductive System of Male Wistar Rats appeared first on Nigerian Biomedical Science Journal.

Animoku Abdulrazaq A.1,5, Mesole Samuel Bolaji2, Okpanachi Omachonu Alfred2,

Yusuf Uthman Ademola3 Shember Pascal Bemseer4

1Department of Anatomy, College of Health Sciences, Kogi State University, Anyigba-Nigeria

2Department of Clinical Medicine, School of Medicine, Eden University-Zambia

3Department of Human Anatomy, School of Medicine, Mulungushi University-Zambia

4Department of Pharmacology and Therapeutics, College of Health Sciences, Benue State University, Makurdi-Nigeria

5Department of Anatomy, Ahmadu Bello University Zaria-Nigeria

All Corresponding to: Animoku Abdulrazaq, animokuaa@gmail.com

ABSTRACT

Background Mercury is a heavy metal contaminant of known toxicity with potential for global mobilization following its give off through air, soil, water and food Aim: The objective of this work was to examine the molecular changes in the DNA caused by a heavy metal pollutant mercuric chloride (HgCl2) in Wistar rats and the role of ascorbic acid administration. Methods: Twenty five adult Wistar rats (average weight 185 g) were randomly divided into five groups of five rats per group; a control group administered normal saline, mercuric chloride (HgCl2; 49.8 mg/kg), HgCl2 with distilled water, HgCl2 with low dose ascorbic acid (595mg/kg) and HgCl2 with high dose ascorbic acid (1,190 mg/kg) groups. All administration was carried out orally for a period of three to six weeks and the animals were humanely sacrificed. Results: The results from molecular studies showed that the level of migration of the genomic DNA were fastest in the mercury treated groups when compared to ascorbic acid and the control groups as characterized on agarose gel electrophoresis. While, significant decrease (p<0.05) in the DNA area and height were observed in mercury treated groups as compared to ascorbic acid and the control groups. Conclusion: The findings from this study showed that ingestion of mercuric chloride has potential deleterious effects on genomic DNA of Wistar rats. However, ascorbic acid administration demonstrate possible ameliorative potential against mercury induced molecular changes

Key words: Mercuric Chloride, Ascorbic acid, DNA, Agarose gel electrophoresis, Wistar rats

INTRODUCTION

Mercury intoxication has been a public health problem for many decades (Burger et al., 2011). Consideration of the role of environmental factors in determining the susceptibility to mercury has recently been renewed by evidence from epidemiological studies (Wang, et al., 2007). Populations are exposed to mercury through the consumption of fishes and sea foods (European Commision, 2005), dental amalgam, mining of silver, gold in industries (WHO, 2007). Mercury readily crosses the blood-brain barrier due to limited lipid solubility and can result in neurological symptoms; seizures, mental retardation, language disorders, vision, hearing and memory loss (WHO, 2003, 2007; Animoku et al., 2018). It is a potential factor in brain damage (Ibegbu et al., 2014), mental impairment, behavioral anomalies (Farina et al., 2011), but little information concerning the potential genotoxicity of mercury compounds (Barregard et al., 1991). However, heavy metals like mercury and lead had been associated with chromosomal aberrations, elongation of X-chromosome, pulverization and birth defects (Valverde et al., 2002), cell death by apoptosis and DNA fragmentation (Hassan et al., 2010). Mercuric salts can be absorbed through the skin of animals (Altmann et al., 2008), Mercury compounds rapidly enter the bloodstream (Berlin et al., 2007), and the level of mercury in the blood is higher after exposure to low dose than a high dose (Berlin et al., 2007). Elimination of mercury is via urine, feaces (WHO 2005), expired air, sweat, and saliva (Booth and Zeller, 2005). In Nigeria, fishes from Lagos Lagoon and the use of “Kohl” a traditional cosmetic had been reported as an agent of mercury toxicity (Onyeike et al., 2002). Presenting symptoms include headache, itching, dizziness, burning, excitability, irritability, restlessness, increased salivation, profuse sweating and frequent urination (Grant and Lipman, 2009; ATDRS, 2011).

Ascorbic acid is an essential nutrient for humans and some other animal species. Vitamin C is an antioxidant that protects our cells against free radicals (Padayatty et al., 2003), pneumonia (Hemila and Louhiala, 2007), prevents scurvy (WHO, 2001) and lowering the incidence of gout (Choi et al., 2009). Ascorbic acid is a well known non-enzymatic antioxidant that can break the chain of lipid peroxidation in cell membranes and scavenges free-radicals such as superoxide ion, hydrogen peroxide, nitric oxide, hydroxyl, alkoxyl and alkyl-peroxyl, collectively called reactive oxygen species (ROS) (Kacuk et al., 2003). Vitamin C is found in high concentration in immune cells and is consumed quickly during infections (Preedy et al., 2010). Ascorbic acid is absorbed in the body by both active transport and simple diffusion (Savini, et al., 2007). Examples of antioxidants are Ascorbic acid (Vitamin C), Vitamin E and Vitamin A (Vasudevan and Sreekumari, 2007). Sources of vitamin C are fruits, vegetables, liver, nutritional supplement, tablets, drinks (Wilson, 2005) and animal products (United Kingdom Food Standard Agency, 2007). Scurvy results from lack of vitamin C, it leads to the formation of brown spot on the skin, spongy gums, and bleeding from all mucous membranes (Stanner et al., 2004).

The aim of the study was to determine the role of ascorbic acid on mercury induced molecular changes and DNA alteration in Wistar rats.

MATERIALS AND METHODS

Twenty five (25) Adult Male Wistar rats of average weight 185g were used for this study. After acclimatization in the Animal House of the Department of Human Anatomy, Ahmadu Bello University, Zaria, the animals were grouped into five groups of five animals each (n = 5). Mercuric chloride (X-N202, May and Bakers, England) was utilized at LD50 of 166 mg/kg body weight as adopted from ATSDR (2011).While; the LD50 of ascorbic acid (S42238, Sam Pharmaceuticals, Nigeria) was adopted from MSDS (2008) as 11,900 mg/kg body weight. The mercury chloride was the approved laboratory grade chemical by Standard Organization of Nigeria, marketed and sold in Nigeria, while the ascorbic acid tablets was approved by National Agency for Food and Drug Administration and Control to be marketed and used in Nigeria. Before the commencement of the study, ethical approval was sort and obtained from the Ahmadu Bello University Zaria Ethical and Animal Use Committee, Faculty of Veterinary Medicine with reference Number ABU/ FVM/ EAUC/ 2015/12. The animals were dosed as follows: control group was administered with normal saline, group II with 30% mercuric chloride (HgCl2, 49.8 mg/kg) only, group III received HgCl2 with distilled water only, group IV received HgCl2 with 5% low dose ascorbic acid (595 mg/kg), while group V received HgCl2 with 10% high dose ascorbic acid (1,190 mg/kg). However, administrations of distilled water and ascorbic acid from weeks 3-6 were done in order to observe for any possible natural recovery and possible ameliorative potentials of ascorbic acid respectively (Table 1). The administration was by oral route daily and lasted for 3-6 weeks, while animal feed and water were allowed ad libitum.

MOLECULAR STUDIES

Brain tissues were collected in eppendorf tubes containing genomic lysis buffer and were taken to Avian Influenza Laboratory, Veterinary Teaching Hospital, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria for Molecular analysis. The tissues were homogenized, kept in water bath overnight and were washed twice through centrifugation set at 14,000 rpm with 200µl of DNA pre-wash buffer, 200µl DNA wash buffer and DNA elution buffer respectively so as to extract the DNA. This protocol was according to the manufacturer’s instruction (Zymo research; the epigenetics company USA). 1% agarose was added to 100 µl Tris Acetic Acid (TAE) buffer and was dissolved in micro wave oven for 5 minutes and allowed to cool to about 45 ˚C; this was followed by the addition of 1µl ethidium bromide to the gel and was mixed thoroughly and gently poured into the casting tray containing the combs and allowed to solidify. Running buffer (Tris Acetic Acid EDTA) was poured into the tank while 20 μl of the DNA sample was mixed with 5 μl DNA loading dye and was loaded into the wells. The DNA sample was run for 40 minutes at 75 mV while the Gel was observed in gel documentation system and the DNA was documented.

STATISTICAL ANALYSIS

All the results were analyzed using the Statistical package for Social Scientist (SPSS version 21) and the results were expressed as Mean ± SEM. The Statistical significance between means were analyzed using one-way analysis of variance (ANOVA) followed by post HOC test; Turkey’s multiple comparison test to test for statistically significant difference between control and experimental groups. A p-value < 0.05 was considered statistically significant.

RESULTS:

MOLECULAR STUDIES

The results from molecular studies as observed from the Genomic DNA showed that the level of migration of the genomic DNA on agarose gel electrophoresis of the animals in Groups II and III appeared fastest characterized by lighter DNA bands when compared to Groups I (Control), IV and V as shown in Plate I.

MEASUREMENT OF GENOMIC DNA AREA, WIDTH, HEIGHT, PERIMETER AND LENGTH OF MIGRATION OF DNA FROM WELL

The measurement of Genomic DNA area, width, height, perimeter and length of migration of DNA showed significant decrease (p<0.05) in the measured areas and heights of the genomic DNA in Groups II and III when compared to Group one (Control) and Group V, while, measurement of the length of migration of genomic DNA revealed significant increase (p<0.05) in Group II when compared to Groups I (Control), III, IV and V respectively as shown in Table 2.

DISCUSSION

The present study was on whether or not mercuric chloride induced toxicity was accompanied by changes in the Genomic DNA and the role of ascorbic acid. The study revealed that the level of migration of the Genomic DNA as characterized on agarose gel electrophoresis were fastest in Groups treated with mercury only, suggesting lesser molecular weights of the DNA molecules which could be related to the effect of mercury on the Genomic DNA. The study showed significant increase (p<0.05) in the mean Genomic DNA length measured from the well in Groups exposed to mercury only when compared to ascorbic acid treated groups and animals in the Control Group. This study agrees with the work of Hasan et al., (2010) who reported that liver cell DNA of mercury treated fishes appeared further away from the well when compared to the position of the Control Group and this change in position was related to cell death by apoptosis accompanied by DNA fragmentation. In another study, it was reported that DNA damage in mercury exposed individuals suggests that mercury overload induces an imbalance in the redox cycle (Hasan et al., 2013). The present study revealed significant decrease (p<0.05) in height and area of the Genomic DNA of animals exposed to mercury only when compared to animals treated with ascorbic acid and animals in the Control Group suggesting the toxic effects of mercury and the possible ameliorative effects of ascorbic acid against mercury induced DNA damage. However, further study on Polymerase chain reaction (PCR), DNA sequencing and comet assay (Tice et al., 2000) could further reveal any alteration in the Genomic DNA sequence or if any mutation might have occurred following mercury exposure and if this alteration could possibly be ameliorated by ascorbic acid, since it was suggestive that ascorbic acid can protect against DNA damage (Cail et al., 2001; Diatrich et al., 2003).

CONCLUSION

The findings from the present study justify the ameliorative potential of ascorbic acid against mercury induced molecular changes and DNA alteration; hence the consumption of ascorbic acid should be encouraged to populations exposed to increased risk of mercury poisoning.

Conflict of Interest

None declared.

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3. Animoku AA, Buraimoh AA, Hamman WO, Ibegbu AO, Samuel SB, Uthman AY, Joseph SM, Akpulu, P. (2018). Ameliorative effect of ascorbic acid (vitamin c) on mercury induced temporal lobe damage in rat. Nigerian Journal of Neuroscience. 9(1):9-15.

4. Barregard, L., Hogstedt, B., Schutz, A., Karlsson, A., Sallsten, G. and Thiringer, G. (1991). Effects of occupational exposure to mercury vapor on lymphocyte micronuclei. Scandinavian Journal of Work, Environment and Health, 17(4):263–268

5. Berlin, M., Zalups, R.K. and Fowler, B.A. (2007). “Mercury,” in Handbook on the Toxicology of Metals, G.F. Nordberg, B.A. Fowler, M. Nordberg, and L.T. Friberg, Eds., Chapter 33, Elsevier, New York, NY, USA, 3rd edition.

6. Booth, S. and Zeller, D. (2005). “Mercury, food webs, and marine mammals: implications of diet and climate change for human health,” Environmental Health Perspectives, vol. 113, no pp. 521-526.

7. Burger, J., Jeitne, C. and Gochfeld, M. (2011). Locational differences in mercury and selenium levels in18 species of saltwater fish from New Jersey. Journal of Total Environmental Health;74(13):863- 74.

8. Cail, L., Koropatrick, J. and Cherian, M.G. (2001). Roles of vitamin C in radiation induced DNA damage in the presence and absence of copper. Chem. Biol. Interact., 137: 75-88

9. Choi, M. D., Gao, X., Curhan, G., Xiang Gao, M. D. and Gary Curhan, M. D. (2009). Vitamin C Intake and the Risk of Gout in Men. Archives of Internal Medicine. 169 (5): 502–507.

10. Diatrich, M., Block, G. and Benowitch, N.L. (2003). Vitamin C supplementation decreases oxidative stress biomarker F2-isoprostane in plasma of non-smokers exposed to environmental tobacco smoke. Nutr. Cancer, 45: 176-184.

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12. Farina, M., Rocha, J.B. and Aschner, M. (2011). Mechanisms of methylmercury-induced neurotoxicity: Evidence from experimental studies. Journal of life Sciences 89:555–563

13. Grant, L.D. and Lipman, M. (2009). Human exposure and their effect. Environmental toxicant; 3rded:108-112.

14. Hasan, M.A., Ahmed, M.K., Akhand, A.A., Ahsan, N. and Islam, M.M. (2010). Toxicological effects and molecular changes due to mercury toxicity in fresh water snakehead (Channa punctatus). International Journal of Environmental Research 4(1):91-98.

15. Hasan, F. A., Akram, Umran and Nadhem, H. H. (2013). Oxidative Stress, Antioxidant Status and DNA Damage in Mercury Exposure Workers. British Journal of Pharmacology and Toxicology 4(3): 80-88.

16. Hemila, H. and Louhiala, P. (2007). Vitamin C for preventing and treating pneumonia.Cochrane Database System Rev (1).

18. Ibegbu A.O., Ayuba, M., Animoku, A. A., Brosu, D., Adamu, S. A., Akpulu, P., Hamman, W.O., Umana, U.E. and Musa, S.A. (2014). Effect of Ascorbic Acid on Mercury Induced Changes on the Liver in Adult Wistar Rats. IOSR Journal of Dental and Medical Sciences, Volume 13, Issue 10 Ver. II):10-16.

19. Ibegbu, A.O., Animoku, A.A., Ayuba, M., Brosu, D., Adamu, S.A., Akpulu, P., Hamman, W.O., Umana, U.E. and Musa, S.A. (2014). Effect of Ascorbic Acid on Mercuric Chloride Induced Changes on the Histomorphology of the Cerebellum of Wistar Rats. African Journal of Cellular Pathology, Vol.3:9-15.

20. Kacuk, O., Sahin, N., Sahin, K., Gursu, M. F., Gulcu, F., Ozcelik, M. and Issi, M. (2003).Egg production, egg quality and lipid peroxidation status in laying hens maintained at a low ambient temperature (6%) and fed a vitamin C and vitamin E-supplemented diet. Veterinarni Medicina, 1-2: 33-40.

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The post The Role of Ascorbic Acid on Mercury Induced Molecular Changes and DNA Alteration in Rats. appeared first on Nigerian Biomedical Science Journal.

Olufemi SS, Faborode MB, Mustapha RA, Oyegoke TG

Department of Nutrition and Dietetics, Faculty of Applied Sciences, Rufus Giwa,

Polytechnic, P M B 1019, Owo, Ondo State, Nigeria

Adeniran SM

Department of Nutrition and Dietetics, C.S.T, Kaduna Polytechnic, Kaduna

All Corresponding to: Olufemi SS, olufemisamuel2175@gmail.com

ABSTRACT

Background: Breastfeeding is certainly an important child feeding practice, which promotes the wellbeing and survival of children, especially in the first two years of life. Objective: The objective of the study is to assess the impact of maternal education and health facilities on the breastfeeding practices of mothers in Lagos and Ogun State, Nigeria. Material and Methods Three hundred and two (302) breastfeeding mothers and their children were selected using systematic sampling techniques from (5) government and private hospitals in Lagos and Ogun States. Semi-structured interviewer administered questionnaire was administered to obtain information on socio-demographic status, clinical status, knowledge and attitude of mothers towards breastfeeding practices and impact of health facilities on the breastfeeding mothers. Data obtained were analyzed using descriptive statistics and Pearson correlation method. Results: Results showed that more than 60% (198) of the children were between the ages of 0-8 months of which half were males. Majority (76.8%) of the mothers had post-secondary education. More than half 62.3% earning an income of #30,000-#50,000. Clinical characteristics of the breastfeeding mothers showed that majority of the children were delivered through normal delivery and 2.6% had low birth weight. Mothers’ knowledge of breastfeeding practice revealed that half of the population initiated breastfeeding one hour after delivery and majority (94.4%) of mothers gave colostrum to their children. The result also showed that 48.3% of the mothers preferred private hospital and 33.8% of the mothers preferred government hospital. More than half (55.3%) of the mothers gave birth in the private hospital. Conclusion: The study concluded that prevalence of exclusive breastfeeding is low while a high number of mothers preferred private facilities to public facilities during child birth. Recommendation: There is a need for policies which will promote exclusive breastfeeding, encourage mothers and improve quality of health facilities to promote the practice of breastfeeding in both private and public health facilities in Nigeria.

Keywords: Exclusive breastfeeding, Breastfeeding practices, Health facilities, Children , Mothers

INTRODUCTION

Breastfeeding is the feeding of an infant or young child with breast milk directly from female human breasts (i.e.via lactation) rather than using infant formula from a baby bottle or other container [5]. According to a statement by World Health Organization (WHO), Breastfeeding is the ideal method suited for the physiological and psychological needs of an infant [19]. Breastfeeding is an important public health strategy for improving infant and child morbidity and mortality, improving maternal morbidity, and helping to control health care costs [12]. The Nigerian health care has suffered several down-falls. Despite Nigerian’s strategic position in Africa, the country is greatly underserved in the health care sphere. Health facilities (health centers, personnel, and medical equipments) are inadequate in this country, especially in rural areas [10]. According to the 2009 communiqué of the Nigerian national health conference, health care system remains weak as evidenced by lack of coordination, fragmentation of services, dearth of resources, including drug and supplies, inadequate and decaying infrastructure, inequity in resource distribution, and access to care and very deplorable quality of care [18].

Furthermore, It is difficult to ascertain the size of the private health care sector in Nigeria since a sizeable number of existing health facilities are believed to be operating without appropriate licensure by State Ministries of Health (SMOHs). Furthermore, private facilities seldom forward to the SMOH information reflecting health facility case-loads, available personnel and technologies in any given year [3]. In particular, information on private sector personnel resources is compromised by the fact that a considerable proportion of health workers (especially doctors, pharmacists, radiographers and laboratory technicians) working in the private sector hold full-time appointments in the public sector [10]. Faith-based providers are grouped as part of the private, nonprofit sector, and often integrate themselves into national health systems by agreeing with the government to provide specific services. Yet faith-based providers are often more “public” than “private” in both practice and ethos. They often fill gaps left by others, or are designated district hospitals to which other private or lower-level public facilities report. The most obvious example is in Nigeria, where a coalition of churches and faith-based providers manage and operate more than half the national health system [13]. Therefore, this study aimed at assessing maternal knowledge and impact of health facilities on breastfeeding practices of mothers in Lagos and Ogun State.

MATERIALS AND METHOD

Study Design

A cross-sectional descriptive study was used to assess the impact of health facilities on the breastfeeding practices of mothers in selected health facilities in Lagos and Ogun state.

Study Location

The study was carried out in some selected Local Government Areas (L.G.A) in Lagos and Ogun State

Study Population

The study focused on breastfeeding mothers and their children that are between age 0-24months.

Ethical consideration

A written permission to carry out this research was obtained from each of the hospitals within the study area, verbal consent were also received from the mothers after the purpose and procedure of the study was read to the participants, giving the mothers a fair understanding into the study aims and the chance to freely choose to participate in the study.

Sampling Technique

Five Local Government Areas (LGAs) 5 health facilities were purposively selected for the study (three (3) public hospitals and five (5) private health facilities). One hundred and fifty-two (152) mothers with their children were further selected from government hospitals and One hundred and fifty (150) mothers with their children from private and faith-based hospitals systematic sampling. A total number of Three hundred and two (302) breastfeeding mothers with

Statistical Analysis

Data analysis was carried out with the aid of Statistical Package for the Social Sciences (SPSS) version 21.0. Questionnaires were scrutinized for completeness. Relevant descriptive and inferential statistics was used to analyze data.

RESULTS

Socio-demographic characteristics of the respondent

Table 2 shows that104 (34.4%) of the children were between the age 0-5months, 94 (31.1%) were between the ages of 6-8 months, 43 (14.2%) were between the ages of 9-11 months while 61 (20.2%) were between12-24 months. Result also showed that 153 (50.7%) of the children were males and 146(48.3%) were females. The results also showed that majority (78.6%) had tertiary education and 17.3% obtained secondary education. Estimated family monthly income shows that, 39 (12.9%) earned below N10,000, 61 (20.2%) earned between N10,000- N30,000 while 67 (22.2%) earned

above N50,000.

Clinical Characteristics of the respondents

Table 2 revealed the clinical data of the respondents. It shows that 8 (2.6%) of the children were less than 2.5kg in weight at birth, 248 (82.1%) weighed between 2.5kg-5.0kg at birth while 14(4.6%) weighed above 5.0kg. The results showed 295(97.7%) had no birth defects or malformation as at the time of the study and 3(1.0%) had birth defects. The clinical data also shows that 11 (3.6%) of the children suffered illness and 283 (93.7%) had no illness.

Maternal Knowledge of Mothers on the Practice of Exclusive Breastfeeding

The results in table 3 shows that thirty six (11.9%) of the mothers introduced breastfeeding immediately after delivery, (31.8%) introduced it 30minutes after delivery while (53.6%) of mothers introduced one hour after delivery. In the practice of exclusive breastfeeding, 49.0% of the mothers fed their children with only breast milk, 70 (23.2%) fed their children with breast milk and water. Colostrum was given to the children immediately after birth by 285 (94.4%) of mothers while 12(4.0%) didn’t give colostrum.

Table 4 shows that two hundred and seventy nine (92.4%) revealed that the health facilities were good and encouraged them to adequately breastfeed, 16 (5.3%) indicated average, 2 (0.7%) of the mothers reported that the health facilities were poor. Majority (97.7%) agreed that the health workers are competent and practice good hygiene, 98.3% of the respondents reported that there is no nutritionist in the facilities.

Mothers preference of health facilities as a result of breastfeeding practices

Figure 1 shows that one hundred and forty six (48.3%) preferred the private hospitals for their breastfeeding practices, 102 (33.8%) preferred Public/Government hospital and 48 (15.9%) preferred faith-based hospital.

Figure 1: Mothers preference of health facilities as a result of breastfeeding practices

Health facilities by child delivery

Figure 2 shows that one hundred and sixty-one (53.3%) gave birth to their children in private hospitals, 106 (35.1%) Public/Government hospital hospitals, 21 (7.0%) for faith-based and 6 (2.0%) mothers specify other places.

DISCUSSION

This study assessed breastfeeding practices among nursing mothers attending selected government and faith-based health facilities and the impact of health facilities on breastfeeding practices of mothers in the selected facilities. Proper and Adequate health facility with good nutrition practices during infancy and early childhood is very essential to ensure the optimum growth, healthy lifestyle and development of children to their full potential.

The study revealed that a high number of the children were between the ages of 0-8 years, this is because the study focused on children of age 0-24 months

Majority of the mothers who participated in the study (92.4%) were married and a high number of them were highly educated, with 75.2% having of tertiary education. The impact and support from husband in breastfeeding practices cannot be underestimated. In line with other studies [15], this study showed that educated mothers will have a fair knowledge about the meaning of exclusive breastfeeding and its recommendation as proposed by WHO and UNICEF that infant should be exclusively breastfed for six months. Educated mothers are more likely to expose to health information and make use of such information especially that which concern their children. The study shows that 42.2% of the mothers were trader/business woman; this tends to be higher compared to findings of Onah et al., [17] which reveals that 9.1% of lactating women were traders/business women. The clinical profile revealed that majority of the children (82.1%) had normal birth weight This supports the findings of Akinyemi et al., [1] that 81.7% of live births in urban Nigeria between 2008 and 2018 reported normal birth weight (2,500g and 4,000g. The prevalence of HBW, estimated to be 10.7%, was higher than the LBW estimated at 7.5%. Birth weight is a strong predictor of child’s health and development. A low birth weight (LBW) of less than 2,500g is associated with adverse health outcomes and is a leading risk factor for neonatal morbidity and mortality.

Some of the mothers considered breast milk as the best form of food and nutrition for infants, they agreed to the concept of six months exclusive breastfeeding, however, feeding children with only breast milk, exclusive breastfeeding (EBF) rate of 49% recorded in this studies is far below the expected 90% set by WHO [20] for the EBF to produce a reasonable impact in the population. The introduction of colostrum had increased among the breastfeeding mothers in this study with the rate 94.4%, This is higher than what has been documented by other studies in Nigeria [2].

In this study, timely initiation of breastfeeding (within one hour of birth) found among majority of the respondents in this study is commendable with53.6% of the mothers initiated breastfeeding within the first hour after delivery while 31.8% initiated breastfeeding thirty minutes after delivery. This result is higher compare to the findings of Ọládòyìnbó et al.,[16] which revealed that 28.6% of the infants who initiates breast milk in less than 1 hour. Early breastfeeding initiation has been found to increase mother to child bonding, regulate infant temperature, breathing and enhance breastfeeding longevity and findings on the time of breastfeeding initiation on nutritional status of mother and infant shows that within 24 hours is the best [4].

The results of this survey indicated that 49.0% of the mothers had good knowledge and positive attitude towards exclusive breastfeeding. This study also found out that more than half of the mothers obtained their recommended length of breastfeeding information from health workers, 16.6% from media and 2.0% from family/husband. This is again almost similar to the findings of Bolanle [7], in which 64.5% of the mothers obtained such information from health workers, 9.0% from the media and 7.3% from their husbands. The finding of this study shows that decreased value which was 35.1% of the mothers gave birth at public (government hospitals). This is lower when compared to the reports of Nigeria Demographic and health survey [14], that 43.2% of the mothers in Nigeria give birth in public/government hospitals across the nation. The impacts of health workers cannot be underestimated in breastfeeding, even their knowledge also contribute to the effective practice of breastfeeding [11]. Mothers assessed in this study were satisfied with services of the health workers in terms of competency and good hygiene practices (97.7%). Successful breastfeeding is dependent on an array of factors related to the mother, infant, and supportive environment. Although the majority of the women in the study cited support from the hospital staff and partner or family members. The presence of a support system, whether it is personal or professional, has a positive influence on the duration of exclusive breastfeeding. Research has shown that peers, partners, and families play a major role in helping and supporting mothers to continue EBF for six months, thereby significantly decreasing the risk of early is continuation of EBF before six months [6], [[9].

Conclusion

This study brings to the conclusion that there is awareness on exclusive breastfeeding among women attending public (government owned hospitals) and private hospitals bur the practice of exclusive breastfeeding is still very low in urban areas. In addition, the study gave account of almost no presence of nutritionist could deliver adequate information about diet and nutritional intake of the breastfeeding mothers in all the health facilities

Recommendations

Based on the result findings of the study, the following recommendations were made:

Exclusive breastfeeding counseling should be more elaborative with emphasis on its advantages in both private and government hospitals in the study are. The government should to employ the services of nutrition experts in all the health facilities to improve breastfeeding practices and the consumption of adequate diets that will support growth and development of mothers and their babies.

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