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

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Assessment of Toxicity Potential of Secnidazole on Reproductive System of Male Wistar Rats 1,2 E.O. Ikuomola, 1O.S Akinsonmisoye, 1R.O. Owolabi and 3M. B. Okon Department of Physiological Sciences, Faculty of Basic Medical Sciences, ObafemiAwolowo University, Ile-Ife, Osun State, Nigeria.

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

3 Department of Biochemistry, Kampala International University, Western Campus, Uganda.

1

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 (9-12weeks) 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 57.2 mg/kg(high dose)) and Distilled water (2mg/kg) for 8 weeks and were also allowed to 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 study concluded that secnidazole showed toxic effects on reproductive parameters of male wistar rats. However, the toxic effects were dose dependent and reversible after eight weeks of cessation of treatment.

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

INTRODUCTION Infertility has become an ominous fertility in adult male’s reproductive problem in Africa on the average, and it system, as well as developmental toxicity has been reported that about 10% of all in the offspring. Male reproductive couples face difficulty in starting a family toxicity is also associated with and this creates a feeling of great derangement in the structures and personal failure [1]. Several factors such functions of the male reproductive as extrinsic and environmental factors system such as sperm parameters, couple with an increased in the usage of abnormal hormonal secretion and antibiotics and anti-effective drugs have abnormal testicular and epididymal been suggested as potential causes of functions.

It also induces altered male infertility [2]. Reproductive toxicity variables indicative of oxidative stress is seen as a phenomenon caused by like glutathione (GSH) depletion [3]. [4] exposure to hazardous chemical are of the view that antimicrobial substances which interfere or cause combination therapy such as detrimental changes in the reproductive metronidazole, quinolones, tetracycline, system and such substances are called ketoconazole, fluconazole and other reprotoxic substances. [3] is of the imidazole group of antibiotic drugs are opinion that such substances have among the most prescribed classes of adverse effects on sexual function and drugs in medicine and there is a high

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http://www.inosr.net/inosr-applied-sciences/ Ikuomola et al INOSR APPLIED SCIENCES 8(1):120-133, 2022. 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 nitro-imidazole drugs mostly prescribed in the clinical settings because of it increase in patient’s compliance and 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 nitroimidazole 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 halflife 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 5-nitroimidazoles [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 nitroimidazole based compound at the therapeutic dose of 15mg/kg in rats did not have significant negative effect on the 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 treatment in rats. It was also reported that 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) significantly reduced sperm count in male Wistar rats for 4 weeks.

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 2-10% adverse effect on the gastrio intestinal tracts of patients also headache dizziness were experienced 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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http://www.inosr.net/inosr-applied-sciences/ Ikuomola et al INOSR APPLIED SCIENCES 8(1):120-133, 2022. male wistar rats ,aslo assessed the effect (FFSH,LH, testosterone ) and Biochemical of secnidazole on sperm parameters like assay (GSH). And this studyalso share us sperm motility, morphology, viability and relevant toxicity of secnidazole on male total sperm count of wistar rats, also reproductive system. The information determined the histology opf the sharing also help caution or reduce the epididymis and testes of the wistar rats , use of priscription of secnidazole to futher investigated the hormonal assays patients in clinical settings.

MATERIALS AND METHODS Animal Use and Care commencement of the experiment.

Forty adult male Wistar rats weighing 150Secnidazole was procured by May & Baker 200g were used for this study. The Company, Sangto-Otta, Ogun State, animals were obtained from the Animal Nigeria. Ethical clearance for this study House, College of Health Science, was obtained from the Health Research ObafemiAwolowo University, Ile-Ife, Ethics Committee (HREC) of the Institute Nigeria. The rats were house and kept in of Public Health, College of Health cages under normal environmental Sciences, ObafemiAwolowo University, Ileconditions and give free access to Ife, Osun State Nigeria. (HREC Assigned standard pellet diet and water. They were Number: IPHOAU/12/1624). allowed to acclimatize to the laboratory Experimental Design: environment for a weeks before the Table 1 showing parameters division of rats into sub groups and administrations GROUPS GROUP A1 A2 GROUP B

NUMBER RATS 5 5 5

GROUP C

5

GROUP D

5

GROUP E

5

GROUP F

5

GROUP G

5

OF

TREATMENTS OF

NUMBER OF DAYS

2mls of distilled Water (control) 14.3mg secnidazole (low dose) 28.6mg secnidazole (medium dose) 57.2mg secnidazole (high dose) 14.3mg secnidazole (low dose)+ Recovery 28.6mg secnidazole (medium dose) + Recovery 57.2mg secnidazole (high dose)+Recovery

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

8 Weeks treated 8 Weeks Recovery 8 Weeks 8 Weeks 8 Weeks 8 Weeks treated + 8 Weeks Recovery 8 Weeks treated + 8 Weeks Recovery 8 Weeks treated + 8 Weeks Recovery

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 for rats in the recovery groups.

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

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http://www.inosr.net/inosr-applied-sciences/ Ikuomola et al INOSR APPLIED SCIENCES 8(1):120-133, 2022. were done immediately after cervical 4ºC (R-Series Cold Centrifuge, Centurion dislocation. Progressive sperm motility Scientific Ltd., West Sussek, U.K.) to was carried out by puncturing the caudal obtain the serum. The serum luteinizing epididymis to release two drops of the hormone (LH), follicle stimulating semen on a pre-warmed microscope slide. hormone (FSH) and testosterone were Thereafter, two drops of citrate buffer quantified using Enzyme adverse linked solution (pH 7.4) were added and covered Immunosorbent Assay (ELISA) method with a coverslip and examined under the following themanufacturer’s instructions microscope with low magnification (x40) (Inteco diagnostics reagents, UK). It was and assessed. Sperm viability was carried read using a microplate reader (Shanghai out using the eosin/nigrosine stain. Two Yongchuang medical instrument Co., drops of semen were puton a microscope Model SM600, China) at a wavelength of slide and two drops of the stain were 450nm. Reduced glutathione (GSH) was added to it, covered with a coverslip, airmeasured using the method of Beutler et dried and observed using a light al. [14]. This was read using a microplate microscope. The dead sperms were reader (Shanghai Yongchuang Medical stained while the live sperms were not Instrument Co., Model SM600, China) at a stained.

These was counted and wavelength of 412 nm. expressed as percentages [10]. Sperm Histological Analysis morphology was carried out by The left testes and epididymis were placingtwo drops of semen on a carefully excised for the sake of microscopic slide and adding Walls and consistency and prefixed in BouinEwas stain and air dried. The slide was Hollande’s fluid to prevent putrefaction then examined under a microscope with and autolysis.

They were further x100 magnification. A total of 100 sperms processed and the sections (5 microns) were evaluated and the abnormal sperms were then stained with hematoxylin and were expressed as a percentage. Sperm eosin and examined using a light count was evaluated by mincing the microscope [15]. caudal epididymis in a petri dish Statistical Analysis containing 2 ml of distilled water. The Data obtained were presented as mean ± suspension was further diluted with standard error of mean (X± SEM). Data sodium bicarbonate formalin solution ata normality was assessed by Kolmogrovratio of 1:20. The new improved Smirnov test and the statistical Neubauer’shemocytometer counting significance between the control group chamber was filled with the diluted sperm and the treatedgroups was calculated by solution, observed under the microscope the analysis of variance (ANOVA) and post and the numbers of sperm cells in 2mm2 hoc analysis was carried out using were counted, calculated and expressed in Student-Newman-Keuls multiple million/ml [11, 12, 13]. comparison test using the Graph Pad 5.03 Hormonal and Antioxidant Assay statistical software package (Graph Pad Blood was collected by cardiac puncture, Software Inc., CA, USA). A probability transferred into plain sample bottles and value of p less than 0.05 was considered separated by centrifuging in a cold to be significant [16]. centrifuge at 3000 rpm for 10 minutes at RESULTS Body and Relative Organ Weights Table 1: Table showing the effect of Secnidazole on body weight of the male Wistar rats Group Treatment (%) Recovery (%) Control 27.54±3.56 41.98±15.63 Secnidazole at 14.3mg/kg 26.52±1.12 38.24±1.80 Secnidazole at 28.6mg/kg 24.86±2.36 34.83±3.29 Secnidazole at 57.2mg/kg 19.41±2.33 40.03±4.40

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Results are presented as Mean

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 ± 3.29 and 40.03 ± 4.40) (p = 0.94, F = 0.12) at (75%) .

SEM, n =

5 (p < 0.05). The results showed that there 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 ± 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 Results are presented as Mean 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 Results are presented as Mean 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 in the recovery groups it was reversible to (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 and 97.40 ± 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 weeks of recovery.

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Sperm Motility (% Motile Sperm)

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100

*

GROUP A1-CONTROL GROUP B-LOW DOSE GROUP C-MEDIUM DOSE GROUP D-HIGH DOSE

*

80

 60



GROUP A2-CONTROL GROUP E-LD + RECOVERY GROUP F-MD+RECOVERY GROUP G-HD+RECOVERY

40 20 0

Figure 4: Effect of Secnidazole on sperm motility Graph showing the effect of Secnidazole on sperm motility Results are presented as Mean SEM, n = 5

Sperm Morphology ( % of Abnormal Sperm)

*= significantly different from group A (control) (p < 0.05) α= significantly different from group B (low dose) (p < 0.05) β= significantly different from group C (Medium dose) (p < 0.05)

GROUP A1-CONTROL GROUP B-LOW DOSE GROUP C-MEDIUM DOSE GROUP D-HIGH DOSE

20 15

GROUP A2-CONTROL GROUP E-LD + RECOVERY GROUP F-MD+RECOVERY GROUP G-HD+RECOVERY

10 5 0

Figure 5. Effect of Secnidazole on sperm morphology Graph showing the effect of Secnidazole on sperm morphology Results are presented as Mean SEM, n = 5 (p < 0.05)

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Sperm Viability (%Live Sperm)

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GROUP A1-CONTROL GROUP B-LOW DOSE GROUP C-MEDIUM DOSE GROUP D-HIGH DOSE

150

100

GROUP A2-C0NTROL GROUP E-LD + RECOVERY GROUP F-MD+RECOVERY GROUP G-HD+RECOVERY

50

0

Sperm Count (x106/ml)

Figure 6: Effect of Secnidazole on sperm viability Graph showing the effect of Secnidazole on sperm viability Results are presented as Mean SEM, n = 5 (p < 0.05)

150

100

GROUP A1-CONTROL GROUP B-LOW DOSE GROUP C-MEDIUM DOSE GROUP D-HIGH DOSE

* 



GROUP A2-CONTROL GROUP E-LD + RECOVERY GROUP F-MD+RECOVERY GROUP G-HD+RECOVERY

50

0

Figure 7: Effect of Secnidazole on total sperm count Graph showing the effect of Secnidazole on total sperm count Results are presented as Mean SEM, n = 5 *= significantly different from group A (control) (p < 0.05) α= significantly different from group B (low dose) (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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FSH concentration (mlU/ml)

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0.4

*

0.3

* 0.2



GROUP A1-CONTROL GROUP B-LOW DOSE GROUP C-MEDIUM DOSE GROUP D-HIGH DOSE GROUP A2-CONTROL GROUP E-LD + RECOVERY GROUP F-MD+RECOVERY GROUP G-HD+RECOVERY

0.1 0.0

Figure 8: Effect of Secnidazole on FSH concentration Graph showing the effect of Secnidazole on FSH concentration Results are presented as Mean SEM, n = 5 *= significantly different from group A (control) (p < 0.05) α= significantly different from group B (low dose) (p < 0.05) β= significantly different from group C (high dose) (p < 0.05) δ= significantly different from group E (low dose + recovery) (p < 0.05) ᵡ= significantly different from group F (medium dose + recovery) (p < 0.05) Table 4: Table showing the Wistarrats Group Control Secnidazole at 14.3mg/kg Secnidazole at 28.6mg/kg Secnidazole at 57.2mg/kg Results are presented as Mean

effect of Secnidazole on LH concentration of the male Treatment (%) 0.18±0.00 0.20±0.01 0.22±0.01* 0.23±0.01* SEM, n = 5 (p < 0.05)

Recovery (%) 0.18±0.00 0.18±0.00 0.15±0.01 0.12±0.01*α

*=significantly different from group A (control) (p<0.05) α= significantly different from group E (low dose + 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 Results are presented as Mean SEM, n = 5 (p < 0.05) *=significantly different from group A (control) (p<0.05) α= significantly different from group C (medium dose) (p < 0.05) β= significantly different from group E (low dose + recovery) (p < 0.05)

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Glutathione mMol/mL

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GROUP A1-CONTROL GROUP B-LOW DOSE GROUP C-MEDIUM DOSE GROUP D-HIGH DOSE

0.6

0.4

GROUP A2-CONTROL GROUP E-LD + RECOVERY GROUP F-MD+RECOVERY GROUP G-HD+RECOVERY

0.2

0.0

Figure 11: Effect of Secnidazole on plasma glutathione Graph showing the effect of Secnidazole on serum glutathione Results are presented as Mean SEM, n = 5 exposed to secnidazole but were reversed after 8 weeks of recovery. However, No visible damage was observed in the histoarchitecture of the epididymis.

Histology There were alterations in the histoarchitecture of the testis of rats

PLATE 1

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

GE- germinal epithelium, l- lumen,LC- leydig cell ,s- spermatogonia , * exfoliated germ cells LEGEND, A1– 2ml/kg distilled water only, B – 14.3mg/kg secnidazole, C – 28.6 mg/kg secnidazole, D- 57.2 mg/kg secnidazole, A2- Recovery control ,E,F and G – 14.3 mg/kg, 28.6 mg/kg, 57.2 mg/kg + 8 weeks withdrawal of secnidazole administration

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PLATE 2

S- spermatogonia , * exfoliated germ cells EP- epithelium , CT- connective tissue, BS- Basal cells , P- Principal cells.

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- 57.2 mg/kg secnidazole, A2- Recovery control ,E,F and G – 14.3 mg/kg, 28.6 mg/kg, 57.2 mg/kg + 8 weeks withdrawal of secnidazole administration DISCUSSION This study assessed the toxicity potential surrounding the seminiferous tubules in of secnidazole on body weights, relative the mammalian testis has been document testicular weight, relative epididymal in literature [18]. The absence of a weight, sperm parameters, male significant change in the relative reproductive hormones, plasma testicular weight suggests that when glutathione, and histology of the testis as compared to its counterpart in the nitrowell as the epididymis in Wistar Rats. imidazole drugs, secnidazole may not Three doses were administered orally. induce its toxicity through permeation of The lowest dose was 14.3mg/kg and it the blood-testis barrier. Also, it has been was a sub therapeutic dose; 28.6mg/kg found that ordinazole a group of nitrorepresented the average daily imidazole drug cause of no significant recommended dose; and 57.2mg/kg was change in relative testicular weight of the maximum daily dose. There was no male Wistar rats when treated with significant change in body weight of the 400mg/kg/day for four weeks [19]. Also, rats in the treated groups when compared there was no significant difference in the with the control following treatment and relative epididymal weight between the even after recovery. This may be due to drug treated rats and the control and after the fact that secnidazole inhibits recovery. This suggests that the blood intestinal absorption thereby preventing epididymis barrier restricted the lipid deposition (which may promote permeability of epididymal tissues to weight gain). [17] reported a significant secnidazole. However, [5] reported a reduction in body weight when treated significant change in the relative with 200mg/kg and 400mg/kg of epididymal weight when treated with metronidazole (a member of nitro metronidazole a group of nitro-imidazole imidazole group) for eight weeks. There drug treated with 400mg/kg and was no significant difference in the 500mg/kg for eight weeks. There was a relative testicular weight between the significant reduction in percentage sperm drug treated groups and recovery groups. motility in the treated rats when The existence of blood-testis barrier compared with the control. The reduction

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http://www.inosr.net/inosr-applied-sciences/ Ikuomola et al INOSR APPLIED SCIENCES 8(1):120-133, 2022. was dose dependent, which may be 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 serum FSH level in the recovery groups 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 require more prolong time for the animals to recover fully. [19] also reported tinidazole (a member of nitro imidazole group) significantly reduce sperm motility 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 partly due to reduction in serum FSH level 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 difference in abnormal sperm morphology 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 that secnidazole 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 evidenced in the sperm count as observed in this study, or the deleterious effect of the sertoli cells as it was also an evidence in the histo-architecture 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 shows secnidazole toxic effect, and may require a longer time for the animals to recover fully. This is in line with [5], who documented earlier that intra-peritoneal 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

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http://www.inosr.net/inosr-applied-sciences/ Ikuomola et al INOSR APPLIED SCIENCES 8(1):120-133, 2022. hormone (LH) and testosterone levels in testis is regarded as the most sensitive the treated group when compared with end point in the evaluation of male the control group. This increase is reproductive toxicity [24]. In this study, possibly due to a probable negative histological observation of testicular feedback effect of secnidazole primarily sections subjected to hematoxylin and on the anterior pituitary to cause increase eosin stain in the rats subjected to in the secretion of LH and testosterone as secnidazole treatments groups showed observed. However, [17] reported that alterations in the germinal epithelium, that intra-peritoneal administration of seminiferous tubules and the depletion metronidazole (400 mg kg-1 day-1), for 30 of spermatogonic cells, large interstitium, days (a member of nitro-imidazole group) partial depletion of elongated spermatids showed reduction in the levels of in the lumen, when compared with the luteinizing hormone and testosterone. control. The drug treatment was highly The significant increase in LH levels and effective in the rats treated with medium testosterone in the treated rats was dose(C) and high dose(D), however the reversed in the recovery group when alterations observed in histo architectural compared with the control.

After structure of the testis was reversed in the recovery, the LH level in the rats that recovery groups after the rats were received high dose of secnidazole was allowed to recovered for 8weeks. The significantly reduced far below the degenerated histo architectural structure control group, this showed that more time observed in group F and G were partially might be required for full restoration for reversed in the recovery group when the animal that receive high dose of compared with the control. This study is secnidazole. The significant reduction in in line with that reported by serum LH level after recovery showed [25,26,27,28,29,30,31], who reported toxic effect of secnidazole, and may sloughing of germinal epithelium, require more time for the animals to seminiferous tubules and the recover fully. In the present study, there spermatogonic cells, large interstitium, was no significant difference in the partial depletion of elongated spermatids plasma glutathione between treated rats in the lumen in male white mice, treated and the control and after recovery. This with 0.37mg/kg/mouse/30 days of suggested that, when compared with its metronidazole(a member of nitrocounterparts in the nitro-imidazole drug, imidazole group). This might be due to secnidazole is not associated with the fact that secnidazole breaches the oxidative stress, it also suggested a better blood testis barrier and gains access to pharmacological profile of secnidazole is the germ cells and seminiferous tubules. better than other imidazole group.

Whereas, there was no damage recorded However, [23] reported a significant in the epididymis for both drug-treated reduction in plasma GSH level in rats and recovery groups when compared with exposed to metronidazole 500mg/kg/day the control groups.

This shows (a member of imidazole group) for 30 secnidazole does not affect the days. Histopathological evaluation of epididymis and suggested secnidazole to animal reproductive tissue is useful in be better than other nitro-imidazole male reproductive risk assessment. The group of drugs.

CONCLUSION This study concluded that secnidazole these toxic effects were dose dependent showed toxic effects on reproductive and reversible after eight weeks of parameters of male Wistar rats. However, cessation of treatment.

RECOMENDATIONS

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

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