Assessment of Toxicity Potential of Secnidazole on Reproductive System of Male Wistar Rats

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International Network Organization for Scientific Research

ISSN: 2705 165X Assessment of Toxicity Potential of Secnidazole on Reproductive System of Male Wistar Rats

1,2E.O. Ikuomola, 1O.S Akinsonmisoye, 1R.O. Owolabi and 3M. B. Okon

1Department of Physiological Sciences, Faculty of Basic Medical Sciences, ObafemiAwolowo University,Ile Ife,OsunState,Nigeria.

2Department of Physiological Sciences, Kampala International University, Western Campus, Uganda.

3DepartmentofBiochemistry,KampalaInternationalUniversity,WesternCampus,Uganda.

ABSTRACT

Secnidazole is one of the nitro imidazole drugs, the pharmacological properties of secnidazole have been predicted to be reprotoxic, it is used in the clinical settings to treat verities of reproductive infections. This study provides information on the potential toxicity or otherwise of secnidazole on the reproductive system of male Wistar rats. Forty male Wistar rats aged between (912weeks) with body weight (150 200g) were used in this study. The rats were divided into seven groups, Each groups received various concentrations of secnidazole (14.3 mg/kg (low dose) , 28.6 mg/kg(medium dose ), and 57.2mg/kg(highdose)) andDistilledwater (2mg/kg)for8weeksand werealsoallowedto recover for another 8weeks respectively. It was observed from the results that administration of secnidazole for 8weeks induce male reproductive toxicity, significant decrease in sperm parameters, reproductive hormones and testiscular degeneration following 8 weeks of administration. However, after 8 weeks of recovery it was observed that an improved testicular histology, sperm parameters and reproductive hormones. This studyconcludedthatsecnidazoleshowedtoxiceffectson reproductiveparametersof male wistar rats. However, the toxic effects were dose dependent and reversible after eight weeksofcessationoftreatment.

Keywords: Secnidazole, Toxicity, spermatogenesis, nitro imidazole, Sexual function, infertility,antimicrobial.

INTRODUCTION

Infertility has become an ominous problem in Africa on the average, and it has been reported that about 10% of all couples face difficultyin starting a family and this creates a feeling of great personal failure [1]. Several factors such as extrinsic and environmental factors couple with an increased in the usage of antibiotics and anti effective drugs have been suggested as potential causes of male infertility [2]. Reproductive toxicity is seen as a phenomenon caused by exposure to hazardous chemical substances which interfere or cause detrimental changes in the reproductive system and such substances are called reprotoxic substances. [3] is of the opinion that such substances have adverse effects on sexual function and

fertility in adult male’s reproductive system, as well as developmental toxicity in the offspring. Male reproductive toxicity is also associated with derangement in the structures and functions of the male reproductive system such as sperm parameters, abnormal hormonal secretion and abnormal testicular and epididymal functions. It also induces altered variables indicative of oxidative stress like glutathione (GSH) depletion [3]. [4] are of the view that antimicrobial combination therapy such as metronidazole, quinolones, tetracycline, ketoconazole, fluconazole and other imidazole group of antibiotic drugs are among the most prescribed classes of drugs in medicine and there is a high

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possibility that some of the couples presenting with history of infertility or inability to conceive may be due to these groups of drugs. secnidazole is one of nitroimidazole drugs mostly prescribed in the clinical settings because ofitincreasein patient’scomplianceand most of the infections that it is used to treat are infections of the reproductive system such as urinary trait infection (UTI), Pelvic inflammatory disease (PID), Vaginal infections. Secnidazole drug has not been properly studied and there are limited information about the drug. Studies have shown that some nitro imidazole based anti biotics possess mutagenic activities in bacteria assay in rats and have also been reported to produce toxicity and inhibition of spermatogenesis in them. Secnidazole, an “imidazole” antibiotic, is now frequently used in young population and people in reproductive age bracket. However, the possible hazardous effect of secnidazole on reproductive system has not been properly evaluated. Secnidazole is one of the representatives of nitro imidazole group of drugs. It is highly effective and differs from other compounds in this group by a prolonged serum half life of about 17 29 hours [5]. The prescription of secnidazole have increased due to its pharmacological advantages in treatment of adults and children with intestinal amoebiasis, urinary tract infection (UTI), pelvic inflammatory disease (PID) and vaginal infections mostly of mild or moderate severity [6]. The tolerability profile of secnidazole does not differ markedly from other 5nitroimidazoles [7]. [26] reports an adverse effect of imidazole on male reproductive function and support the hypothesis that imidazoles inhibit male fertility through suppression of two important aspects of testicular function: testosteronesecretion and testicular intestinal fluid TIF formation (Veijola and Rajaneimi, 1986). This mutagenic effect has been proved to reduce a nitro group and attacks the nucleic acid of the micro organism [8]. Furthermore, it

inhibits further DNA synthesis and causes degradation of already existing DNA [9], further studies also shown revealed that metronidazole a nitro imidazole based compound at the therapeutic dose of 15mg/kg in rats did nothavesignificantnegativeeffectonthe reproductive parameters assessed while 200mg/kg and 400mg/kg in rats has reduction in testosterone and follicle stimulating hormone. Similarly, luteinizing hormone increased mostly with 400mg/kg in metronidazole treatmentinrats.Itwasalsoreportedthat the body weight of rats significantly reduced when treated with 200mg/kg and 400mg/kg of metronidazole administration for 8 weeks [27] and also [29] also reported that tinidazole (a member of nitro imidazole group) significantlyreducedspermcountinmale Wistarratsfor4weeks

The effect of secnidazole on reproductive system was studied because reports from previous studies [29] which reported the adverse effect of secnidazole in clinical trials and observed 210% adverse effect on the gastrio intestinal tracts of patients alsoheadachedizzinesswereexperienced by about 2% of patient exposed to secnidazole and in recent times has shown increasing number of people using secnidazole in clinical settings especially among the young ones and those in the reproductive age bracket. Also, the pharmacological properties of secnidazole have been predicted to be reprotoxic. The result of this study expressed the toxicity potentials of secnidazole on the male reproductive system. The aim of this study determines the effects of secnidazole on the histology of testis and epididymis of the rats and evaluated the effect on FSH,LH,testosterone and plasma glutathione levels in rats , this were within a view to providing empirical information on the potential toxicity or otherwise of secnidazole on the reproductive system of male wistar rats, the objective of this study investigated the effect of secnidazole on the relative weight of the testis and epididymis of

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male wistar rats ,aslo assessed the effect of secnidazole on sperm parameters like sperm motility, morphology, viability and total sperm count of wistar rats, also determined the histology opf the epididymis and testes of the wistar rats , futher investigated the hormonal assays

Animal Use and Care

(FFSH,LH, testosterone ) and Biochemical assay (GSH). And this studyalso share us relevant toxicity of secnidazole on male reproductive system. The information sharing also help caution or reduce the use of priscription of secnidazole to patientsinclinicalsettings

MATERIALSANDMETHODS

FortyadultmaleWistarratsweighing150 200g were used for this study. The animals were obtained from the Animal House, College of Health Science, ObafemiAwolowo University, Ile Ife, Nigeria. The rats were house and kept in cages under normal environmental conditions and give free access to standard pellet diet and water. They were allowed to acclimatize to the laboratory environment for a weeks before the

commencement of the experiment. Secnidazole was procured by May & Baker Company, SangtoOtta, Ogun State, Nigeria. Ethical clearance for this study was obtained from the Health Research Ethics Committee (HREC) of the Institute of Public Health, College of Health Sciences,ObafemiAwolowoUniversity,Ile Ife, Osun State Nigeria. (HREC Assigned Number:IPHOAU/12/1624).

Experimental Design:

Mode of Sacrifice and Organ Collection

After eight weeks of administration of secnidazole, the rats were sacrificed by cervical dislocation. Their blood was collected through cardiac puncture. The testis and epididymis were harvested and weighed using China 0.1g small laboratory digital weighing scale. The right testis of each rat was immediately fixed in Bouin’s fluid for histological

processing. The semen for sperm parameters was obtained from the caudal epididymis and their sperm parameters were assessed under the microscope. The same sacrificial procedure was repeated forratsintherecoverygroups.

Sperm Analysis

Sperm analysis was carried out as described by W.H.O (1999). Briefly, progressive sperm motility and viability

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were done immediately after cervical dislocation. Progressive sperm motility was carried out by puncturing the caudal epididymis to release two drops of the semen on a pre warmed microscope slide. Thereafter, two drops of citrate buffer solution (pH 7.4) were added and covered with a coverslip and examined under the microscope with low magnification (x40) and assessed. Sperm viability was carried out using the eosin/nigrosine stain. Two drops of semen were puton a microscope slide and two drops of the stain were added to it, covered with a coverslip, air dried and observed using a light microscope. The dead sperms were stained while the live sperms were not stained. These was counted and expressed as percentages [10]. Sperm morphology was carried out by placingtwo drops of semen on a microscopic slide and adding Walls and Ewas stain and air dried. The slide was then examined under a microscope with x100magnification. A totalof 100sperms were evaluated and the abnormal sperms were expressed as a percentage. Sperm count was evaluated by mincing the caudal epididymis in a petri dish containing 2 ml of distilled water. The suspension was further diluted with sodium bicarbonate formalin solution ata ratio of 1:20. The new improved Neubauer’shemocytometer counting chamberwasfilledwiththedilutedsperm solution, observed under the microscope and the numbers of sperm cells in 2mm2 werecounted,calculatedandexpressedin million/ml[11,12,13].

Hormonal and Antioxidant Assay

Blood was collected by cardiac puncture, transferred into plain sample bottles and separated by centrifuging in a cold centrifuge at 3000 rpm for 10 minutes at

4ºC (RSeries Cold Centrifuge, Centurion Scientific Ltd., West Sussek, U.K.) to obtain the serum. The serum luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone were quantified using Enzyme adverse linked Immunosorbent Assay (ELISA) method following themanufacturer’s instructions (Inteco diagnostics reagents, UK). It was read using a microplate reader (Shanghai Yongchuang medical instrument Co., Model SM600, China) at a wavelength of 450nm. Reduced glutathione (GSH) was measured using the method of Beutler et al. [14]. This was read using a microplate reader (Shanghai Yongchuang Medical Instrument Co., Model SM600, China) at a wavelengthof412nm.

Histological Analysis

The left testes and epididymis were carefully excised for the sake of consistency and prefixed in Bouin Hollande’s fluid to prevent putrefaction and autolysis. They were further processed and the sections (5 microns) were then stained with hematoxylin and eosin and examined using a light microscope[15].

Statistical Analysis

Data obtained were presented as mean ± standard error of mean (X± SEM). Data normality was assessed by Kolmogrov Smirnov test and the statistical significance between the control group and the treatedgroups was calculated by theanalysisofvariance (ANOVA)andpost hoc analysis was carried out using Student Newman Keuls multiple comparison test using the Graph Pad 5.03 statistical software package (Graph Pad Software Inc., CA, USA). A probability value of p less than 0.05 was considered tobesignificant[16].

RESULTS

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Results are presented as Mean SEM, n = 5(p<0.05).Theresultsshowedthatthere was no significant change in the body weight of the rats in the treated groups (26.52 ± 1.12, 24.86 ± 2.36 and 19.41 ±

2.33) at (60% ) when compared with the control group(27.54 ±3.56) (p =0.19, F = 1.79) and also the recovery groups (38.24 ±1.80,34.83±3.29and40.03±4.40)(p= 0.94,F=0.12)at(75%)

Table 2. Table showing the effect of Secnidazole on the relative weight of testis of the male Wistar rats

Group

Treatment (%) Recovery (%)

Control 0.53±0.02 0.49±0.03

Secnidazole at 14.3mg/kg 0.52±0.00 0.51±0.02

Secnidazole at 28.6mg/kg 0.52±0.02 0.49±0.01 Secnidazole at 57.2mg/kg 0.53±0.01 0.44±0.04

ResultsarepresentedasMean SEM,n=5(p<0.05)

Table 3: Table showing the effect of Secnidazole on the relative weight of epididymis of the male Wistar rats

Group Treatment (%) Recovery (%)

Control

0.24±0.01 0.25±0.02

Secnidazole at 14.3mg/kg 0.26±0.00 0.30±0.02

Secnidazole at 28.6mg/kg 0.27±0.01 0.27±0.00

Secnidazole at 57.2mg/kg 0.30±0.01 0.24±0.02

ResultsarepresentedasMean SEM,n=5(p<0.05)

There was also no significant difference in the relative testicular and epididymal weight in the treated rats (0.52 ± 0.00, 0.52 ± 0.02, 0.53 ± 0.01 and 0.26 ± 0.00, 0.27 ± 0.01, 0.30 ± 0.01) at (0.5%) when compared with the control (0.53 ± 0.02 and 0.24 ± 0.01) (p = 0.99, F = 0.04), (p = 0.15,F=2.03)at(0.6%)

Sperm parameters

There was a significant decrease in the sperm motility of the treated groups (68.00 ± 2.00, 62.00 ± 2.00 and 54.00 ± 2.44) at (50%, 60%, and 65%) when compared with the control (93.00 ± 1.22) (p = 0.001, F = 73.183) at (95%). However intherecoverygroupsitwasreversibleto (85%, 87% and 90%) after 8 weeks of

recovery. There was no significant difference in sperm morphology and viability of the treated rats (13.70 ± 0.55, 14.00 ± 0.37, 14.00 ± 0.50 and 94.20 ± 2.39, 95.20 ± 1.46, 96.80 ± 0.73) when compared with the control (12.26 ± 0.56 and97.40±0.60) (p=0.42,F=0.98),(p= 0.080, F = 2.71). There was a significant reduction in the sperm count of the treated groups (109.60 ± 4.47, 95.40 ± 3.28 and 88.40 ± 3.01) at (60%, 50%, and 40%) when compared with the control (138.00 ± 4.08) (p = 0.05, F = 3.03) (96%) However in the recovery groups it was reversible to (85%, 87% and 90%) after 8 weeksofrecovery

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GROUPA1CONTROL GROUPBLOWDOSE GROUPCMEDIUMDOSE GROUPDHIGHDOSE

GROUPA2CONTROL GROUPELD+RECOVERY GROUPFMD+RECOVERY GROUPGHD+RECOVERY

Figure 4: Effect of Secnidazole on sperm motility

Graph showing the effect of Secnidazole on sperm motility

ResultsarepresentedasMean SEM,n=5

*=significantlydifferentfromgroupA(control)(p<0.05)

α=significantlydifferentfromgroupB(lowdose)(p<0.05)

β=significantlydifferentfromgroupC(Mediumdose)(p<0.05)

GROUPA1CONTROL GROUPBLOWDOSE GROUPCMEDIUMDOSE GROUPDHIGHDOSE

GROUPA2CONTROL GROUPELD+RECOVERY GROUPFMD+RECOVERY GROUPGHD+RECOVERY

Figure5.EffectofSecnidazoleonspermmorphology

GraphshowingtheeffectofSecnidazoleonspermmorphology

ResultsarepresentedasMean SEM,n=5(p<0.05)

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GROUPA1CONTROL GROUPBLOWDOSE GROUPC-MEDIUMDOSE GROUPDHIGHDOSE

GROUPA2C0NTROL GROUPE-LD+RECOVERY GROUPF-MD+RECOVERY GROUPGHD+RECOVERY

Figure 6: Effect of Secnidazole on sperm viability

GraphshowingtheeffectofSecnidazoleonspermviability

ResultsarepresentedasMean SEM,n=5(p<0.05)

GROUPA1CONTROL GROUPBLOWDOSE GROUPCMEDIUMDOSE GROUPD-HIGHDOSE

GROUPA2CONTROL GROUPELD+RECOVERY GROUPFMD+RECOVERY GROUPG-HD+RECOVERY

Figure 7: Effect of Secnidazole on total sperm count

GraphshowingtheeffectofSecnidazoleontotalspermcount

ResultsarepresentedasMean SEM,n=5

*=significantlydifferentfromgroupA(control)(p<0.05)

α=significantlydifferentfromgroupB(lowdose)(p<0.05)

Hormonal concentration and glutathione levels

There was a dose dependent significant increase in testosterone (0.26± 0.01, 0.34 ± 0.01 and 0.21 ± 0.01) (p = 0.001, F = 5.78) and LH levels (0.22 ± 0.01 and 0.23 ± 0.01) (p = 0.001, F = 6.92). While There was a significant decrease in FSH of the treated rats (0.12 ± 0.01, 0.14 ± 0.01 and 0.09 ± 0.00) (p= 0.001, F= 6.4905) at

(0.12%, 0.14% and 0.08%) when compared with the control (0.24 ± 0.00) However in the recovery groups it was reversible to (0.25%, 0.3% and 0.35%) after 8 weeks of recovery. There was no significant difference in the plasma glutathione of the treated rats (0.13 ± 0.01, 0.15 ± 0.01 and 0.14 ± 0.01) when compared with the control(0.13±0.01)(p=0.15,F=2.30).

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GROUPA1CONTROL GROUPBLOWDOSE GROUPCMEDIUMDOSE GROUPDHIGHDOSE

GROUPA2CONTROL GROUPELD+RECOVERY GROUPFMD+RECOVERY GROUPGHD+RECOVERY

Figure 8: Effect of Secnidazole on FSH concentration

GraphshowingtheeffectofSecnidazoleonFSHconcentration

ResultsarepresentedasMean SEM,n=5

*=significantlydifferentfromgroupA(control)(p<0.05)

α=significantlydifferentfromgroupB(lowdose)(p<0.05)

β=significantlydifferentfromgroupC(highdose)(p<0.05)

δ=significantlydifferentfromgroupE(lowdose+recovery)(p<0.05)

ᵡ=significantlydifferentfromgroupF(mediumdose+recovery)(p<0.05)

Table 4: Table showing the effect of Secnidazole on LH concentration of the male Wistarrats

Group

Control

Treatment (%)

Recovery (%)

0.18±0.00 0.18±0.00

Secnidazole at 14.3mg/kg 0.20±0.01 0.18±0.00

Secnidazole at 28.6mg/kg 0.22±0.01* 0.15±0.01

Secnidazole at 57.2mg/kg 0.23±0.01* 0.12±0.01*α

ResultsarepresentedasMean SEM,n=5(p<0.05)

*=significantlydifferentfromgroupA(control)(p<0.05)

α=significantlydifferentfromgroupE(lowdose+recovery)(p<0.05)

Table 5: Table showing the effect of Secnidazole on serum testosterone of the male Wistar rats

Group Treatment (%) Recovery (%)

Control

0.17±0.02 0.17±0.02

Secnidazole at 14.3mg/kg 0.26±0.01*α 0.20±0.01 Secnidazole at 28.6mg/kg 0.34±0.01* 0.13±0.00β

Secnidazole at 57.2mg/kg 0.21±0.01α 0.10±0.00β

ResultsarepresentedasMean SEM,n=5(p<0.05)

*=significantlydifferentfromgroupA(control)(p<0.05)

α=significantlydifferentfromgroupC(mediumdose)(p<0.05)

β=significantlydifferentfromgroupE(lowdose+recovery)(p<0.05)

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There were alterations in the histoarchitecture of the testis of rats

GROUPA1CONTROL GROUPBLOWDOSE GROUPCMEDIUMDOSE GROUPDHIGHDOSE

GROUPA2CONTROL GROUPELD+RECOVERY GROUPFMD+RECOVERY GROUPGHD+RECOVERY

exposed to secnidazole but were reversed after 8 weeks of recovery. However, No visible damage was observed in the histoarchitectureoftheepididymis. PLATE 1

LEGEND,

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S spermatogonia,*exfoliatedgermcellsEP epithelium, CT connectivetissue,BS Basal cells,P Principalcells.

Photomicrograph of the epididymis of the rats (H & E x 400 Magnification).

LEGEND, A1 2ml/kg distilled water only, B 14.3mg/kg secnidazole, C 28.6 mg/kg secnidazole,D 57.2mg/kgsecnidazole,A2 Recoverycontrol,E,FandG 14.3mg/kg,28.6 mg/kg,57.2mg/kg+8weekswithdrawalofsecnidazoleadministration

DISCUSSION

This study assessed the toxicity potential of secnidazole on body weights, relative testicular weight, relative epididymal weight, sperm parameters, male reproductive hormones, plasma glutathione, and histology of the testis as well as the epididymis in Wistar Rats. Three doses were administered orally. The lowest dose was 14.3mg/kg and it was a sub therapeutic dose; 28.6mg/kg represented the average daily recommended dose; and 57.2mg/kg was the maximum daily dose. There was no significant change in body weight of the rats in the treatedgroups when compared with the control following treatment and even after recovery. This may be due to the fact that secnidazole inhibits intestinal absorption thereby preventing lipid deposition (which may promote weight gain). [17] reported a significant reduction in body weight when treated with 200mg/kg and 400mg/kg of metronidazole (a member of nitro imidazole group) for eight weeks. There was no significant difference in the relative testicular weight between the drug treated groups and recovery groups. The existence of blood testis barrier

surrounding the seminiferous tubules in the mammalian testis has been document in literature [18]. The absence of a significant change in the relative testicular weight suggests that when compared to its counterpart in the nitro imidazole drugs, secnidazole may not induce its toxicity through permeation of the blood testis barrier. Also, it has been found that ordinazole a group of nitro imidazole drug cause of no significant change in relative testicular weight of male Wistar rats when treated with 400mg/kg/day for four weeks [19]. Also, there was no significant difference in the relative epididymal weight between the drugtreatedratsandthecontrolandafter recovery. This suggests that the blood epididymis barrier restricted the permeability of epididymal tissues to secnidazole. However, [5] reported a significant change in the relative epididymal weight when treated with metronidazole a group of nitro imidazole drug treated with 400mg/kg and 500mg/kg for eight weeks. There was a significant reduction in percentage sperm motility in the treated rats when compared with the control. The reduction

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was dose dependent, which maybe partly due to reduction in serum FSH level as observed in this study. [20] also reported a reduction in sperm motility in the groups treated with either 200 or 400mg/kg of metronidazole for 4 weeks in male Wistar rats. However after 8 weeks of drug free recovery period, the sperm motility significantly recovers when compared with the control. This may be due to significant reverse in the serumFSH level in the recoverygroups as observed in this study. Even after recovery, the rats that received high dose of secnidazole significantly reduced below the recovery control group. This showed secnidazole toxic effect, and may requiremoreprolongtimefortheanimals to recover fully. [19] also reported tinidazole (a member of nitro imidazole group)significantlyreducespermmotility in male Wistar rats for 4 weeks Furthermore, there was a significant reduction in sperm count in the treated rats when compared with the control. The effect was dose dependent. This may be partlyduetoreductioninserumFSHlevel as observed in this study. [20] also reported a reduction in sperm count in the groups treated with either 200 or 400mg/kg of metronidazole for 4 weeks in male Wistar rats. However, after 8 weeks of drug free recovery period, the sperm count significantly recovered when compared with the control. This may be due to significant reversal in the serum FSH level in the recovery groups as observed in this study. [19] also reported that tinidazole (a member of nitro imidazole group) significantly reduced sperm count in male Wistar rats for 4 weeks. Furthermore, there was no significant difference in the sperm viability between the treated rats and the control and after recovery. This may be due to the fact that sperm viability is the least affected among the sperm characteristics (sperm count, motility, morphology and sperm viability) after the administration of secnidazole drug. [21] reported that sperm viability is the least affected among sperm parameters after the administration of xenobiotic. This

may partly explain why there was no significant change in sperm viability after treatment in this study. However, there was significant reduction in sperm count and sperm motility respectively. In addition, there was no significant differenceinabnormalspermmorphology in the drug treated rats when compared with the control and after recovery. This may be due to the fact that secnidazole does not cause mutagenic effect in the development of sperm cells, also may be due to the facts thatsecnidazole does not induce its toxicity through permeation of the blood epididymis barrier [22]. It has been reported that tinidazole (600mg/kg/d) and ornidazole (400mg/kg/d) does not cause abnormal sperm morphology in respect to hook shaped head, mid piece and tail, after 4 and 8 weeks administration [19]. This may partly explain why secnidazole did not affect sperm morphology in this study. Also there was a significant decrease in the serum FSH of the rats in the treated groups when compared with the control. The effect was dose dependent. The reduction in FSH may be due to the significant increase in testosterone levels as observed in this study, also it was also evidencedinthespermcountasobserved in this study, or the deleterious effect of the sertoli cells as it was also an evidence in the histoarchitecture of the testis as observed in this study. However it was significantly reversed in recovery group after 8 weeks of recovery. Even after recovery the serum FSH in the rats that receive high dose of secnidazole significantly reduce below control. This shows that more longer time might be required for full restoration for the animal that receive high dose of secnidazole,this showssecnidazoletoxic effect, and may require a longer time for theanimalstorecoverfully.Thisisinline with [5], who documented earlier that intraperitoneal administration of metronidazole (400 mg kg 1 day 1), for 30 days, reduced the hormone levels of follicle stimulating hormone. There was also a significant increase of luteinizing

al

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hormone (LH) and testosterone levels in the treated group when compared with the control group. This increase is possibly due to a probable negative feedback effect of secnidazole primarily on the anteriorpituitaryto cause increase in the secretion of LH and testosterone as observed. However, [17] reported that that intraperitoneal administration of metronidazole (400 mg kg 1 day 1), for 30 days (a member of nitro imidazole group) showed reduction in the levels of luteinizing hormone and testosterone. The significant increase in LH levels and testosterone in the treated rats was reversed in the recovery group when compared with the control. After recovery, the LH level in the rats that received high dose of secnidazole was significantly reduced far below the controlgroup,thisshowedthatmoretime might be required for full restoration for the animal that receive high dose of secnidazole. The significant reduction in serum LH level after recovery showed toxic effect of secnidazole, and may require more time for the animals to recover fully. In the present study, there was no significant difference in the plasma glutathione between treated rats and the control and after recovery. This suggested that, when compared with its counterparts in the nitro imidazole drug, secnidazole is not associated with oxidative stress,italso suggesteda better pharmacological profile of secnidazole is better than other imidazole group. However, [23] reported a significant reduction in plasma GSH level in rats exposed to metronidazole 500mg/kg/day (a member of imidazole group) for 30 days. Histopathological evaluation of animal reproductive tissue is useful in male reproductive risk assessment. The

testis is regarded as the most sensitive end point in the evaluation of male reproductive toxicity [24]. In this study, histological observation of testicular sections subjected to hematoxylin and eosin stain in the rats subjected to secnidazole treatments groups showed alterations in the germinal epithelium, seminiferous tubules and the depletion of spermatogonic cells, large interstitium, partial depletion of elongated spermatids in the lumen, when compared with the control. The drug treatment was highly effective in the rats treated with medium dose(C) and high dose(D), however the alterations observed in histo architectural structure of the testis was reversed in the recovery groups after the rats were allowed to recovered for 8weeks. The degenerated histo architectural structure observed in group F and G were partially reversed in the recovery group when compared with the control. This study is in line with that reported by [25,26,27,28,29,30,31], who reported sloughing of germinal epithelium, seminiferous tubules and the spermatogonic cells, large interstitium, partial depletion of elongated spermatids in the lumen in male white mice, treated with 0.37mg/kg/mouse/30 days of metronidazole(a member of nitro imidazole group). This might be due to the fact that secnidazole breaches the blood testis barrier and gains access to the germ cells and seminiferous tubules. Whereas, there was no damage recorded in the epididymis for both drug treated andrecoverygroups when compared with the control groups. This shows secnidazole does not affect the epididymis and suggested secnidazole to be better than other nitro imidazole groupofdrugs.

CONCLUSION

This study concluded that secnidazole showed toxic effects on reproductive parameters of male Wistar rats. However,

these toxic effects were dose dependent and reversible after eight weeks of cessationoftreatment.

RECOMENDATIONS

I recommend this imidazole group of drugs should also be investigated on other relevant organs in the body.

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