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Microbiologic Review of Seminal Fluids in a Nigerian Tertiary Health Centre

Aims and Objectives: To determine the microbiologic pattern of semenal fluid in male infertility. Subjects and Methods: A retrospective laboratory review of microbiologic analysis of 360 semen samples done in Benue Results: Fifty-one percent
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  Microbiologic Review of Seminal Fluids in a Nigerian Terary  Health Centre Nwadioha SI 1 , Odimayo MS 2 , Jombo GTA 1 , Nwokedi Prince EO 1 , Abba PO 1 , Agaje I 1  and Akor JO 1 1 Department of Medical Microbiology and Parasitology, College of Health Sciences, Benue State University, Makurdi, Nigeria 2 Department of Medical Microbiology and Parasitology, College of Medicine, Eki  State University, Ado Eki,  Nigeria Corresponding author: Nwadioha SI, Department of Medical Microbiology and Parasitology, College of Health Sciences, Benue State University,Makurdi, Nigeria, Tel: +234- 8056838967; E-mail: Rec date: Jun 15, 2016; Acc date: Jul 15, 2016; Pub date: Jul 22, 2016 Copyright: © 2016 Nwadioha SI et al. This is an open-access arcle  distributed under the terms of the Creave  Commons Aribuon  License,which permits unrestricted use, distribuon,  and reproducon  in any medium, provided the srcinal author and source are credited. Citaon:  Nwadioha SI, Odimayo MS, Jombo GTA, Microbiologic Review of Seminal Fluids in a Nigerian Terary  Health Centre. Arch ClinMicrobiol. 2016, 7:4. Abstract Aims and Objecves:   To determine the microbiologic paern  of semenal uid  in male inferlity. Subjects and Methods: A retrospecve  laboratory reviewof microbiologic analysis of 360 semen samples done inBenue State University Teaching Hospital, Makurdi, fromJanuary, 2013 to January, 2016. Results:   Fiy-one  percent (N=184/360) of the semensamples was normospermic, 43% oligospermic (with 3%severe oligospermia) and 6% azoospermic. Sixty per cent(N=216/360) of the semen samples contained sperm cellsthat were mole,  30% sluggish and 10% non-mole.  Nineper cent of the semen samples contained dead sperms(necrospermic). Forty–two percent (N=150/360) of thesemen samples was infected (contained signicant number of pus cells, ≥ 2000 cells per ml). Pathogens wereisolated in 47%(N=70/150) of the infected semen samplesas follows, Staphylococcus aureus  (53%), Staphylococcus saprophytcus   (10%), Escherichia coli (11.4%), Klebsiellaspp (7.1%), Streptococcus pneumoniae (4.4%), Pseudomonas aeruginosa (7.1%) and Candida spp (7.1%). Moxioxacin  was 85% eecve,   Levooxacin  (80%), Ceazidime  (80%), Ceriaxone  (80%), Ciprooxacin  (70%), Ooxacin  (68%) and Gentamicin (60%). While Penicillin(20%) and Tetracycline (10%) showed poor acvity. Conclusion:  Regular semen microbiologic invesgaon  isquite invaluable to male inferlity   intervenon  in poorresource countries. Keywords: Male inferlity;  Microscopy; Culture; Semen introducon Introducon A semen analysis is vital tool to invesgate  male inferlity [1]. It can also be used to validate a successful vasectomy as toabsence of sperm. Fiy  per cent of couples with complaint of inferlity  havebeen associated with male factors. Inferlity,  simply is inabilityof a sexually matured couple to achieve pregnancy in presenceof regular unprotected sexual inmacy  of average of threedays in a week for a minimum of one year [2]. Infecons  of the genito-urinary tracts, hormonal imbalance,age factor, stress, environmental polluon,  some metabolicdisorders among others are contributory to male inferlity  [3].Two separate semen specimens at interval of about seven daysmay be needed to be repeated in three month me  areneeded [4].The picture of semen analysis includes as follows; thesemen is expected to liquefy within 30 minutes of ejaculaon with the volume of about 3(±2.5) ml, sperm count per ml of 20million and above; morphology which comprises theappearance, size and shape of the sperms, which must be atleast 50% normal; molity  of the sperm within one hour of  ejaculaon  must be good for at least 50% of the sperms count[5,6]. Infecon  in semen has been incriminated in 15% cases of male inferlity  in ferlity  clinics [7,8]. The infecon  can bethrough sexual transmission or urinary tract. The nidus of  infecon  is usually the prostate [9]. Inferlity  comprises 65% of gynecological consultaons  inAfrica [10]. Male inferlity  is responsible for an average of 50% inferlity  globally [10,11]. The most aected  areas lie withinthe central African region referred to as the “inferlity  belt” of Africa [11].This study therefore, was aimed at the quantave  and qualitave  values of the seminal uid  and the role of microorganisms in male inferlity. Subjects and Method Study populaon Aer  obtaining ethical clearance from the ethics commiee of Benue state university teaching hospital, we collectedsemen samples from three hundred and sixty (360) male Research Article iMedPub Journals Archives of Clinical MicrobiologyISSN 1989-8436  Vol.7 No.4:27 2016 © Copyright iMedPub | This article is available from: 1  subjects. The subjects were aending  the Special TreatmentClinics/ Urology/Reproducve  clinics of Benue State UniversityTeaching Hospital from January, 2013 to January, 2016. Properhygiene was ensured in the collecon  of the samples whichwas done in one of the private rooms in the laboratory. Procedures Aer   liquefacon  of the semen; the duraon  for the liquefacon  was charted, the color noted and volume of thesemen measured. The semen was mounted on a counng chamber (Improved Neubeuer) where sperm count and sperm molity  was read according to WHO standard [1]. Cultures The following culture media plates were used; chocolate for10% carbon dioxide incubaon  and MacConkey and Blood agarfor aerobic incubaon  overnight at 37°C.The 24 hour growth was examined for size, shape, elevaon, hemolysis and other cultural characteriscs.  Gram staining wasdone and pure growth was subjected to biochemical tesng according to standard [5]. Anbiogram Anbioc   suscepbility  of the isolates was tested by agar diusion  method on dry sensivity   tesng  agar (DSTA) usingoxoid muldisc  with standard anbiocs   concentraons according to Clinical and Laboratory Standard Instute  (CLSI),2006 guidelines [12]. Analysis of data The results were analyzed using SPSS 11.0 stascalsoware;  chi-square (x 2 ) was used to compare associaon between proporons  and p-values <0.05 were considered signicant  at 95.0% condence  level. Results A total of 360 seminal uid  samples were collected. Thesperm count was as follows: Fiy-one  percent of the spermcount was normospermic, 25% mildly oligospermic, 15%moderately oligospermic, 3% severely oligospermic and 6%azoospermic (Figure 1) .Forty per cent of the sperm cells were very mole,  20% of moderate progression, 30% sluggish and 10% non-mole (Figure 2).  Viability test revealed 9% dead cells. Forty –two percent (N=150/360) of the seminal uid  samples contained signicant  number of pus cells (≥2000 cells per ml). Pathogenswere isolated in 47% (N=70/150) of this sample. The abovecriteria were distributed according to the percentage molity groups of sperm cells (Figure 3).Figure 1  The sperm count of the sample populaonaending   ferlity  clinics in Benue State University TeachingHospital from Jan, 2013- 2016. Figure 2  The percentage of subjects with their sperm molity  assessment. Figure 3  Percentage Sperm molity  in the infected Seminal uid  of some subjects aending   ferlity  clinics in BenueState University Teaching Hospital, Jan 2013-2016Total number of isolates was 70 as follows, S. aure us (53%), S. saprophytcus   (10%), E. coli   (11.4%), Klebsiella spp (7.1%), S. Archives of Clinical MicrobiologyISSN 1989-8436  Vol.7 No.4:27 2016 2 This article is available from:   pneumonia (4.4%), P. aeruginosa (7.1%) and Candida spp (7.1%) (Table 1).Table 1  Microbial Isolates from semen of some subjects aending   ferlity  clinics in Benue State University Teaching Hospital,January, 2013 to January, 2016. Isolates Number Percentage (%) Staphylococcus aureus 37/7052.9 Staphylococcus saprophyticus 7/7010 Escherichia coli  8/7011.4 Klebsiella species 5/707.1 Streptococcus pneumoniae 3/704.4 Pseudomonas aeruginosa 5/707.1 Candida species 5/707.1Total Isolates7019Sterile cultures (of samples with Pus Cells ≥ 2000/ml)8022Total sterile cultures29081Total cultures360100 Moxioxacin  (85%), levooxacin  (80%), ceazidime  (80%), ceriaxone  (80%) ciprofoxacin (70%), ooxacin  (68%) andgentamicin (60%) are eecvely   acve  across the spectrum of bacteria tested. While penicillin (20%) and tetracycline (10%)are low in acvity   (Table 2). Discussion The study shows a 6% azoospermia, 43% oligospermia and51% normospermic. Some studies in Nigeria showed thefollowing results such as 8.8 % [11] azoospermia in Ogun and14.9% [13] in Osun both in South West Nigeria; oligospermiccases were reported as 51% [11], 46.5% [13], 45.3% in Ilorin,North-Central Nigeria [14], 60% [11] in Enugu ,Eastern Nigeriaare in synchrony with the present study. However, in Tunisia,North Africa was a 13% [15] lower in rate of oligospermia.Azoospermia categorically can be responsible for any couple's inferlity,  since it is complete sterility, while oligospermia onlydecreases the chancesof ferlizaon.  This  condion  mayresult from autoimmune natural sperm agglunanganbodies  in the serum of sub ferle  men [10].Only 40% of the sperm cells were mole  within one hour of  ejaculaon,  ten per cent non mole,  while remaining 50%were sluggish. Drake et al. [11] documented that bacterial infecons  could be responsible for decline in sperm molity.  Inearly infecon,  there may be normal sperm count and sperm molity  before the complicaon  sets in. Role of bacterial infecon  may include change in the chemical milieu of thesemen, inammatory  damage of the passage, destrucon  of the sperm cells and impede their molity.  The consequence istherefore, reduced sperm count, teratospermia andasthenospermia. Escherichia coli have been found to decreasesperm molity  and cause agglunaon  [11,16].The male reproducve  tract is essenally  sterile except thelower urethra. Hence, greater than 2000 pus cells per ml of semen may be signicant  for diagnosis of infecon  [10]. Thepresence of certain organisms in semen or high number of theorganisms is associated with inferlity  [17]. In the presentstudy, 42% (N=150/360) of the semen samples had signicant pus cells (≥2000 cells per ml) and pathogens were isolated inonly 47% (N=70/150) of these samples. The y-  three percent (N=80/150) rate of sterile culture samples with signicant pus cells is a pointer to reckless anbioc  usage in Nigeria [13,18]. This ‘culture-negave  semen' challenge can be solved bythe introducon  of non-culture diagnoscs. The 19% (N=70/360) of the semen contained total numberof isolates was as follows, S. aureus  (53%), E. coli   (11.4%), S. saprophytcus  (10%), Klebsiella spp  (7.1%), P. aeruginosa (7.1%), Candida spp  (7.1%) and S. pneumoniae  ( 4.4%) indescending order. The above data are comparable to studiesby Orji et al. [16] and Olajubu et al. [11] which idened   S.aureus  as the commonest isolated bacteria with 37.1% and51% respecvely  unlike a study [10] which found E. coli   as themost prevalent with 70.4%. The isolaon  of S. aureus  mighthave resulted from the skin ora  of the couples. However,cross- infecon  might be a factor in some partners “tryingoutside marriage” [16]. Sex preferences may as well inuence the nature and types of isolates in the semen [17,18]. Forinstance, anal sex may result in the transmission of bacteriawhich are normal ora  of the intesne. The most acve   anbiocs  tested against the various typesof bacterial isolates includes, Moxioxacin  (85%), Levooxacin (80%), Ceazidime  (80%), Ceriaxone  (80%). However, someolder and commonly used anbiocs,  Penicillin (20%) andTetracycline (10%) have been noted to be crying orphans withlow acvity.  Some usually acve   anbiocs,   Ciprooxacin (70%), Ooxacin  (68%) and Gentamicin (60%) have turned to Archives of Clinical MicrobiologyISSN 1989-8436  Vol.7 No.4:27 2016 © Copyright iMedPub  3  be moderately eecve.  The newer generaon   anbiocs  are relavely  expensive and are mostly given in hospitals.Therefore, they are not easily reached by the people. Table 2   Anbioc   suscepbility   paern  of the isolates from infected semen of the subjects Drugs Bacterial Isolates ** S. aureus 37 (%) S. saprophyticus 7 (%) E. coli  8 (%) Klebsiella spp. 5 (%) P. aeruginosa 5 (%) S. pneumonia 3 (%) *  AMP10 (27)--3 (60)1(20)1 (33)PEN8 (22)0 (0)2 (25)1 (20)-1 (33) AMX----1 (20)-SXT11 (30)3 (43)2 (25)--2 (66)TCN---0 (0)1 (20)-GENT25 (68)5 (71)4 (50)3 (60)2 (40)2 (66)ERYTH12 (32)1 (14)3 (38)-0 (0)-MOXI35 (95)7 (100)-4 (80)--CEFTR33 (89)-6 (75)--3 (100)CEFT-7 (100)-4 (80)3 (60)-CIPRO32 (87)6 (86)5 (63)3 (60)-2 (66)OFL30 (81)6 (86)--3 (60)-LEV34 (92)-7 (88)4 (80)3 (60)3 (100) *  AMP=Ampicillin; PEN=Penicillin; AMX=Amoxicillin; MOXI=Moxifloxacin; TCN=Tetracycline; GENT=Gentamicin; ERYTH=Erythromycin; SXT=Trimethoprim-Sulphamethoxazole; CEFTR=Ceftriaxone; CEFT=Ceftazidime; CIPRO=Ciprofloxacin; OFL=Ofloxacin; LEV=Levofloxacin; ** S. aureus = Staphylococcus aureus ; S. pneumoniae = Streptococcus pneumoniae ; E.coli  = Escherichia coli  ; spp=species Male inferlity  has largely been aributed  to nonspecic seminal tract infecon  [17,18]. Alausa and Osoba for inferlity [9], discovered that 40% of oligospermia and azoospermiacases presenng  to ferlity  clinics in Lagos Nigeria gave ahistory of two or more aack  of urethris.   Ferlity  is jeopardized by these infecons  in several ways by decreasingsperm molity,  changing the chemical milieu of the seminal uid  or illicit some inammatory  changes in seminal tract.Bacterial presence may cause tescular  damage or the obstrucon  of the sperm ducts leading to low sperm count.Chronicity of some urinary tract infecon  is traceable toseminal infecon  by acng  as a reservoir of infecon  [19].In excepon  to infecons,  other condions  can beincriminated in male inferlity.  Such condions  like, exposureto heavy metals e.g. lead, zinc and arsenic. Others include;heat, chemical, X-ray exposure, smoking, nutrionaldeciencies  and use of alcohol, tobacco and caeine.  Also,many recreaonal  and prescripon  drugs (e.g. nitro furans, cimedine),  some herbal medicines (such as St. John's wort)and specic  herbicides and pescides  have been found to betoxic to spermatogenesis. However, the correlaons  are dicult  to develop, as supporng  evidence is sketchy[9,18,19].An automated system example, Computer-Assisted SpermAnalysis (CASA), would have given us a beer  analysis of sperm molity  and other parameters may be a limitaon  tothe study. Intervenons  should be focused at prevenon  since surgicaltreatment of inferlity  and Assisted Reproducve  Techniques(ARTs) are beyond the reach of many in Africa. Therefore,regular microbiologic screening and invesgaon  for inferlity is advised in this sub region. References 1. World Health Organizaon  (2010) WHO Laboratory Manual forthe Examinaon  and Processing of Human Semen. (5thEdn),Cambridge University Press, Cambridge, UK. 2. Jimoh AA (2002) The Management of Inferlity  in Africa.Medilorin Journal 1: 3-9. 3. Hauser R, Paz G, Botchan A, Yoger L, Yaretz H (2001) Varicocele: Eect  on Sperm Funcons.  Human Reproducon  Update 7:482-485. 4. Kenkel S, Rolf C, Nieschlag E (2001) Occupaonal  Risks for Male Ferlity:  An Analysis of Paents   Aending  a Terary  ReferralCenter. Internaonal  Journal of Andrology 24: 318-326. 5. Jeyendra RS (2001) Semen analysis: method and interpretaon. In:Gynaecology and Obstetrics. Sciarra JJ, Lipincot Williams andWilkins 5: 4-20. Archives of Clinical MicrobiologyISSN 1989-8436  Vol.7 No.4:27 2016 4 This article is available from:  6. Budia A, Luis PJ, Broseta E, Tejadios S, Benedicto A, et al. (2006)Value of Semen Culture in the Diagnosis of Chronic Bacterial Prosas  : A Simplied  Method. Scandinavian Journal of Urology &. Nephrology 40: 326-331. 7.  Pella  D, Mylonakis I, Bertoloni G, Fiore C, Andrisani A, et al.(2008) Genital Tract Infecons  and Inferlity.  European Journalof Obstetrics& Gyneacologic Reproducve  Biology 140: 3-11. 8. Sanocka-Maciejewsky D, Ciupinska M, Kurpisz M (2005)Bacterial Infecon  and Semen Quality. Journal of Reproducve immunology 67: 51-56. 9. Ekhaise FO, Richard FR (2008) Common Bacterial IsolatesAssociated with Semen of Men Complaining of Inferlity  inUniversity of Benin Teaching Hospital (U.B.T.H), Benin City,Nigeria. World Journal of Medical Sciences 3: 28-33. 10. Alo MN, Ugah UI, Elom MO (2013) Semen Culture: A Comparave  Analysis between Solid Media and Liquid Media Supplementaon  . IOSR Journal of Pharmacy and BiologicalSciences 5: 67-72. 11. Festus AO, Deji-Agboola M, Olubunmi AO, Olusoji EJ (2013)Seminal Fluid Characteriscs  of Men Aending   Inferlity  Clinicof a Teaching Hospital. Open Journal of Medical Microbiology 3:1-4. 12. Clinical Laboratory Standard Instute  (2007) Performancestandard for anmicrobial  disk suscepbility  tests. 13. Kosamat YA, Adebolu TT (2005) Bacteriological study of isolatesfrom seminal uids  of inferle  men in Ila and Ifedore localgovernment areas, Osun State, South-Western Nigeria. Journalof Food, Agriculture & Environment 3: 73-74. 14. Nwabuisi C, Onile BA (2001) Male Inferlity  among Sexually Transmied  Disease Clinic Aendees  in Ilorin. Nigerian Journalof Medicine 10: 68-71. 15. Gdoura R, Keskes-Ammar L, Bouzid F (2001) Chlamydia trachomas  and Male Inferlity  in Tunisia. European Journal of  Contracepon  and Reproducve  Health Care 6: 102-107. 16. Orji I, Ezeifeka G, Amadi ES, Okafor F (2007) Role of Enrichedmedia in Bacterial Isolaon  from Semen and Eect  of Microbial Infecon  on Semen Quality: A Study on 100 Inferle  men.Pakistan Journal of Medical Sciences 23: 855-888. 17.  Coell  E, Harrison RF, McCarey  M, Walsh T, Mallon E (2000)Microorganisms are commonly found in insignicant   quanes in the semen of asymptomac  men. African Journal of Ferlity and Sterility 74: 465-470. 18. Onemu SO, Ibeh IN (2001) Studies on the Signicance  of Posive Bacterial Semen Cultures in Male Ferlity  in Nigeria. Internaonal  Journal of Ferlity  and Women’s Medicine 46:210-214. 19. Audu BM, Massa AA, Bukar M, Melah GS, Kudi A (2010). Eect of bacterial isolates on the seminal indices of men invesgated for inferlity  in Gombe. Nigerian Journal of Clinical Pracce  13:16-19.   Archives of Clinical MicrobiologyISSN 1989-8436  Vol.7 No.4:27 2016 © Copyright iMedPub  5
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