Considering the increasing demand for herbal aphrodisiacs, this study investigates the effect of Alomo bitters on thehistology of testis in adult rats. 36 male rats of comparable weight (151.67 ± 2.89 grams) and sizes were involved inthis study. The animals were assigned into four groups; a control group (A) and three test groups (B, C and D). For 3 weeks, group A received growers mash and water, while test groups B1-B3, C1-C3 and D1-D3 representing  experimental durations of 1 week, 2 weeks and 3 weeks respectively; received growers mash and graded doses of  Alomo bitters (7.5, 15, 22.5ml) daily. Histologiclly, micrographs from the test groups presented changes that  included interstitial space exudates, cellular pyknosis cellular degeneration, cell population reduction, and  vacuolations. These changes were dosage-duration dependent suggesting that Alomo bitters can induce testicular damage and by implication, infertility in males. Thus, there is an urgent need to regulate the consumption of Alomo bitters as well as other herbal products considered to possess aphrodisiac potentials.Keywords: Alomo-bitters, Herbs, Male, Testis, Histology

Anyanwu, RA

Ihongbe, JC

Izekor, S

Salisu, AA

Uwuigbe, M

English

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 Anthonio Research Center © 2012 33        Salisu et al., IJHPR; 1(2): 33-39    International Journal of Herbs and Pharmacological Research IJHPR, 2012, 1(2):33 - 39 www.antrescentpub.com  RESEARCH  PAPER  HISTOLOGICAL CHANGES IN THE TESTIS OF RATS TREATED WITH ALOMO BITTER *1Salisu A.A., 1Ihongbe J.C.,1Anyanwu R.A., 1Uwuigbe M., 2Izekor S. Department of 1Medical Laboratory Science, 2Physiology; Faculty of Basic Medical Sciences, College of Medicine, Ambrose Alli University, Ekpoma, Edo-Nigeria.  Corresponding author: akindayommy@gmail.com Received: 21st April, 2012   Accepted: 8th June, 2012   Published: 31st July, 2012  ABSTRACT  Considering the increasing demand for herbal aphrodisiacs, this study investigates the effect of Alomo bitters on the histology of testis in adult rats. 36 male rats of c mparable weight (151.67 ± 2.89 grams) and sizes were involved in this study. The animals were assigned into four groups; a control group (A) and three test groups (B, C and D). For 3 weeks, group A received growers mash and water, while test groups B1-B3, C1-C3 and D1-D3 representing experimental durations of 1 week, 2 weeks and 3 weeks r spectively; received growers mash and graded doses of Alomo bitters (7.5, 15, 22.5ml) daily. Histologiclly, micrographs from the test groups presented changes that included interstitial space exudates, cellular pyknosis cellular degeneration, cell population reduction, and vacuolations. These changes were dosage-duration depen nt suggesting that Alomo bitters can induce testicular damage and by implication, infertility in males. Thus, there is an urgent need to regulate the consumption of Alomo bitters as well as other herbal products considered to possess aphrodisiac potentials.  Keywords: Alomo-bitters, Herbs, Male, Testis, Histology. _____________________________________________________________________ INTRODUCTION Current trends show that the inability to afford modern medical healthcare in developing countries has forced patients to seek traditional medical attentions (Watcho et al., 2007). In this regard, the World Health Organisation (WHO) has estimated that about eighty percent of the world population rely chiefly on traditional medicines (Agaie et al., 2007). Of the 119 plant derived drug listed by WHO study, 74% were discovered as a result of chemical studies to isolate the active compounds responsible for the use of original plant in traditional medicine (Farnsworth et al., 1985). In Nigeria several plants have been claimed traditionally, to have medicinal potentials for the treatment of various ailments in both man and animals (Nwude and Ibrahim, 1980; Akinniyi and Sultanbawa, 1983), although their efficacy and safety remain doubtful as only a few of these have been properly identified and documented (Mbaya et al., 2007; Nwosu et al., 2004).   On the other hand, it is a known fact that healthy sexual functioning contributes significantly to one's sense of well-being and quality of life (Salonia et al., 2004). While early report has revealed a high incidence of sexual problems in the general population (Alain, 1999), recent avail ble records show male impotence (Erectile Dysfunctio , ED) as the common medical condition affecting the sexual life of millions of men worldwide (Montorsi et al., 2003; Shabsigh and Anastasiadis, 2003). To achieve sexual quality, many orthodox therapeutic approaches have been employed for quite a long time (Sagraves et al., 2003). However, they present with limited efficacy, unpleasant side effects and contraindications in certain conditions (Lue et al., 2003).  ASN-PH-020919     ISSN: 2315-537X   Anthonio Research Center © 2012 34        Salisu et al., IJHPR; 1(2): 33-39   Similarly, many plants have been reported to be employed as remedies for such disorders, while others ave been used as aphrodisiacs (Yakubu et al., 2005; Tajuddin et al., 2005; Dhawan et al., 2003; Sumalata et al., 2010). In fact, a number of plant extracts have being acclaimed to have aphrodisiac potentials (Noumi et al., 1998) and used traditionally to improve sexual performances (Kamtchouing et al, 2002; Carro, 1999). Adjanohoun et al., (1996) and Noumi et al., (1998) listed a variety of plants used in traditional medicine for fertility regulation. Also, several other medicinal plants are known for male contraceptive potentials by suppressing spermatogenesis or by spermicidal action (Paul et al., 2006).  One of such herbal preparations with acclaimed aphrodisiac potentials is Alomo bitters -an alcoholic drink comprising seven herbal ingredients. The ingredients i clude Khaya invorensis, Capparis erythrocarpus, Mondia whitei, Lecaniodiscus-cupanoides, Dialium guineense, Treculia Africana, Crytolepis-sanguinolenta. Considering the rate of consumption, this study therefore, investigates the effects of Alomo Bitters on the histology f the testis.   MATERIALS AND METHODS   Substance of Study: Several bottles of “Alomo bitters” was purchased from main market in Ekpoma, Edo State, Nigeria with NAFDAC registration number: A1-8029; manufactured date: 14/10/2010; and batch number: ALM 287101. No expiry date was found on the bottles. These details were checked to ensure the authenticity of he products purchased.  Experimental Animals: Thirty-two adult male Wister rats of comparable weights and sizes were procured from the animal farm of the Department of Physiology, College of Medicine, Ambrose Alli University, Ekpoma, and moved to the animal house of Anthonio Research Centre, Ekpoma, Edo State, Nigeria, where they were housed in wooden cages.   The animals were assigned into four groups: a control group (A) and three test groups (B, C and D) subdivided into B1-D1, B2-D2, and B3-D3 representing experimental durations of 1 week, 2 weeks and 3 weeks respectively. The rats were allowed to acclimatize for two weeks and were fed ad libitum during this period, with water and grower mesh from Bendel Feeds and Flour Mills, Ewu, Edo State, Nigeria.   Substance Administration: All the animal groups were fed with feed (growers mash) plus water given ad libitum. However, as group A (control) received distilled water only, test groups B to D received water mixed with graded quantities of Alomo Bitters: B (7.5ml/kg), C (15ml/kg) and D (22.5ml/kg). Appropriate quantities of Alomo bitters were administered into the drinking water of the animals in each the groups respectively, using a sterile syringe of 20ml gauge convenience.    Study Duration: This study lasted between October, 2011 - December, 2011. However, the actual experiment lasted for five weeks (2weeks for acclimatization and 3 weeks for substance administration.  During the five weeks study period, the animals were fed and monitored betwe n the hours of 8:00am – 12:00 noon daily.  Sample Collection:  At the end of each week, 3 rats from each of the groups respectively were sacrificed by administering chloroform as anaesthesia. The rats were then dissected to harvest the testes which wereth n fixed immediately in 10% formalin.  For descriptive purposes, the animals sacrificed at the end of week one from the test groups were designated as B1—D1 rats, while B2 – D2 and B3 - D3 represents the rats sacrificed at the end of week two and three respectively.    Tissue Processing: The tissues were processed using automatic tissue processor according to stand histological processing schedule as described by David (2004). Microscopy was done using a binocular light microscope at magnification x400.  RESULTS  Histological observations showed that the issue micrographs obtained from tissue section of the testes in group A (control) presented normal histological architecture (Plate 1). However, those from test group B1- treated with 7.5 ml of alomo bitter for 1 week presented interstitial space exudates (ISE), cellular pyknosis (P), vacuolation (V), cellular degeneration (CD) (Plate 4.2). Micrographs from group B2 -treated with 7.5ml for 2 weeks, presented seminiferous tubule wall degeneration (STD), cellular degeneration (CD) with congested vessels and vacuol tion  Anthonio Research Center © 2012 35        Salisu et al., IJHPR; 1(2): 33-39   (V) (Plate 4.3), while those from group B3 - treated with 7.5 ml for 3 weeks presented interstitial space congestion (ISC) with exudates (E) and vacuolation (V), seminiferous tubule wall degeneration (STD) with mild cellular vacuolations (V) and pale staining cells (Plate 4.4).  Similarly, tissue micrographs from group C1-treated with 15.0ml of alomo bitter for 1 week presented severe cellular degeneration (CD) with cell-population reduction, interstitial space enlargement (ISE) with exudates (E), seminiferous tubule degeneration (STD) with vacuolati n (V), cellular pyknosis (P) (Plate 4.5). Microgaphs from group C2 -treated with 15.0ml of alomo bitter for 2 weeks presented cellular degeneration (CD), interstitial space enlargement (ISE) and sminiferous tubular wall degen ration (STD) with pale staining cells, cellular pyknosis (P) and vacuolation (V) (Plate 4.6). Those from group C3 -treated with 15.0ml of alomo bitter for 3 weeks presented cellular degeneration (CD), interstitial space enlargement and congestion (ISC), cellular necrosis (N)with vacuolation (V), seminiferous tubules wall degeneration (STD), and reduction in cell population (Plate 4.7).  Also, micrographs from group D1-treated with 22.5ml of alomo bitter for 1 week presented severe cellular necrosis (N) with vacuolations (V), exudations (E), interstitial space enlargement (N), and seminiferous tubule wall degeneration (Plate 4.8). Micrographs from group D2 -treated with 22.5ml of alomo bitter for 2 weeks presented vacuolation (V), pyknosis (P), and interstitial space exudation (E) with cells staining pale and “ghost-like” in appearance (Plate 4.9). Group D3; treated with 22.5ml of alomo bitter for 3 weeks (Plate 4.10) presented cellular necrosis (N), interstitial space congestion (ISC), vacuolation (V), seminiferous tubules degeneration (STD), pale staining cells with “ghost-like” appearance.  Generally, the histological findings suggests alomo-bitters-induced testicular damage and the observed changes were dosage-duration dependent.             Plate 4: Testis (H&E X400) showing Interstitial space congestion (ISC), Seminiferous tubule degeneration (STD), Vacuolation (V)  Plate 1: Testis (H&E X400) showing normal histological features of the Testis  Plate 2: Testis (H&E X400) showing cellular degeneration (CD), interstitial space exudate (E), and Vacuolation (V) Plate 3: Testis (H&E X400) showing Seminiferous tububles degeneration (STD), Pyknosis (P), Vascular congestion (VC)  Anthonio Research Center © 2012 36        Salisu et al., IJHPR; 1(2): 33-39                        Plate 5: Testis (H&E X400) showing Pyknosis (P), Interstitial space enlargement (ISE), Seminiferous tubules degeneration (STD)  Plate 6: Testis (H&E X400) showing Pyknosis (P), Interstitial space enlargement (ISE), Seminiferous tubules degeneration (STD)  Plate 7: Testis (H&E X400) showing Cellular degeneration (CD), Necrosis (N), Vacuolation (V)  Plate 8: Testis (H&E X400) showing Pyknosis (P), Seminiferous tubule degeneration (STD), Vacuolation (V)  Plate 9: Testis (H&E X400) showing Pyknosis (P), interstitial space exudate (E), and Vacuolation (V)  Plate 10: Testis (H&E X400) showing Pyknosis (P), interstitial space congestion (E), and cell degeneration (CD)  Anthonio Research Center © 2012 37        Salisu et al., IJHPR; 1(2): 33-39   DISCUSSION   The observed histological changes in the testis, following the administration of Alomo Bitters, suggest that Alomo Bitters can induce male sterility. In fact, the findi gs reported by Lampio (2009), Watcho et al., (2007), Patman et al., (2005) that some of the constituents of Alomo Bitters - Mondia whitei and Khaya ivorensis, can cause several testicular damages in a dose depended manner surely pports this assertion.  Furthermore, the presence of interstitial space exudates, shrinkage of the seminiferous tubules, complete disorganization of the testicular tissue with destruction of spermatogonial cells and germinal cells, cellular pyknosis, vacuoles in the cytoplasm of the Sertoli cells, necrosis and degenerating cells in the present study, implies that Alomo Bitters is capable of inducing testicular cell damage like snake venom as previously reported by Penna-Videaú, et al. (2000).  Similarly, Russell (1999) had reported that such testicular lesions are the early morphological sign of testicular injury which was considered by Nolte et al. (1995) and Penna-Videaú et al. (2000) as the main testicular response to harmful stimuli. Thus, the histological findings in this study contradict the general belief that Alomo Bitters boost male fertility.  From the foregoing, it can be concluded that the structural integrity of the testes was compromised uner the influence of Alomo Bittters in a dosage-duration dependent manner. Definitely, the observed changes would adversely affect the function of the testes and might particularly become worse, if the dosage is high and consumption prolonged. It is our opinion therefore, that prolonged or repeated ingestion of alomo bitters should be discouraged considering the increasing cases of male sterility hitherto tagged idiopathic.  ACKNOWLEDGEMENT  We express our appreciation to all those who in one way or the other contributed to the success of this research work.  REFERENCES  Adjanohoun, J. C., Aboubaker, N., Dramane, K., Ebot, M. E., Ekpere, J.A., Enow-Orock, E.G.,Focho, D., Gbile, Z.O., Kamanyi, A., Kamsu Kom, J., Keita, A., Mbenkum, T., Mbi, C. N., Mbiele, A.L., Mbome, L. 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Histological changes in the testis of rats treated with Alomo Bitter

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Histological changes in the testis of rats treated with Alomo Bitter

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