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Departments of Medical Microbiology 1, Physiology 3, Pharmacology 6, College of Medicine and
Department of Microbiology 4, Faculty of Natural Sciences, Ambrose Alli University, Ekpoma and
Departments of Obstertrics & Gynaecology2 and Pathology5, Irrua Specialist Teaching Hospital, Irrua.
*Corresponding Author: mcsionelphilrazzy ABSTRACT
A total of 52 semen samples were analysed, following collection through masturbation by males with
history of infertility. Various bacterial strains were isolated with Staphylococcus aureus being the most
common with a prevalent rate of 60%, Streptococcus pyogenes 10%, Proteus mirabilis 23%, Klebsiella
spp 35% and Escherichia coli 23%. Chlamydia trachomatis was diagnosed in 88% of the subjects,
using serological means. A statistical significance (x2= 10.83; P<0.001) revealed Chlamydia
trachomatis as the most probable cause of male infertility compared to Escherichia coli. Mean
spermatozoa motility in the studied subjects reduced with active motile 26.6+17.63, slow motile
23.1+3.01 and non-motile 42.6+4.11. Significant improvement was noted in sperm motility with
accompanying decrease in Leukocytospermia and combined antibiotics therapy also reduced
Leukocytospermia and enhanced spermatozoa motility.

Key Words
: Male infertility, Semen analysis, Leukocytospermia, Combined antibiotic therapy.


cells(RBC), no white blood cells (WBC) nor Male infertility is an important issue, a common problem occurring worldwide. It is a neglected revealed that various microbial infections reproductive health issue in Nigeria (Okonofua reduce the viability of semen. Chlamydia et al., 2005). Reports indicate that the male trachomatis with chronic epididymitis and factor accounts for 20-50% of the causes of epidididymal granuloma formation could lead to blockage of the epidididymis, leading to (Chukwudebelu et al, 1979; Esimai et al., infertility due to azoospermia (Ochsendorf et al, 2002). Male infertility is actually the inability of a 1999). Ness et al (1999) reports an association couple to achieve pregnancy, despite regular between Chlamydia antibodies in semen and unprotected intercourse, usually after a period male infertility. Similarly the presence of of 12 months, with detected factors attributed to antibody to Chlamydia trachomatis is correlated with the presence of white blood cells in semen (Ojengbede et al., 1992) as well as sexually and development of autoimmune response to transmitted diseases may be associated with spermatozoa (Witkin et al., 1995) 2001).Studies also revealed high rates of For Ureaplasma urealyticum infection, induction hyperprolactinemia (Modede, 1994; Adejuwon of leukocytospermia, leads to elevated levels of et al., 1999), antisperm antibodies (Ekwere, leukocyte-derived reactive oxygen species 1995) and genital infections (Onemu and Ibeh, which damage sperms by causing peroxidation 2001; Momoh et el., 2007) in infertile Nigerian of Lipids (Potts et al., 2000). Leukocytospermia men than those in fertile control. Aflatoxin may in infertile males is associated with decreased equally contribute to male infertility (Ibeh et al., sperm numbers and impaired motility with The minimal criteria for normal sperms include a volume of 1.5ml; a minimal count of 20million Another associated factor in male infertility in sperms/ml; motility of 60%, with no red blood Nigeria include sexually transmitted infection
(Imade et al., 1993),while health care seeking trachomatis was assessed and confirmed for all behaviour for STI equally play a vital role in patients using the Chlamydia monoclonal male infertility(Okonofua et al.,2005). antibody spot test kit (Immunocomb) and the Though, pregnancy is the most verifiable means of determining fertility, diagnosis of infertility should not be hurriedly reached even ANTIMICROBIAL SUSCEPTIBILITY TESTING
after1 year of regular unprotected intercourse The disc diffusion method of Bauer et al.,(1996) by a cohabiting couple as the age of the was used. All isolates were subjected to because fertility is optimal in a woman, between ages 18 and 24 years and begin to Staphylococcus aureus Oxford strain NCTC 6751 was used as control for the Gram positive organisms, while Escherichia coli strain NCTC effectiveness of combined antibiotic therapy on 10418 was used as control for Gram negative sperm motility and viability in infertile males in The result of the susceptibility test were MATERIALS AND METHODS
interpreted as sensitive, intermediate or Specimen
resistant following the criteria below as earlier A total of 52 clinical specimens of semen were collected randomly. All the semen samples A zone’s radius equal to or not more than 3mm Specimens were sent to the laboratory for smaller than the control was taken as sensitive, analysis within 15minutes of collection and all a zone’s radius more than 3mm smaller than specimens were from patients with primary the control and not less than 3mm was reported as intermediate and a zone’s radius 2mm or criteria(WHO,1999),ie, collection of semen by less (ie, no zone of inhibition) was reported as specimen bottles, following abstinence from intercourse 2-3 days prior to collection. Following antimicrobial susceptibility testing, Immediate transfer of sample to the laboratory selected antibiotics were prescribed with a four for analysis, a wet preparation made and quick months follow-up period to assess for clinical examination, followed by culture and sensitivity. From a total of 52 samples of semen collected ISOLATION AND IDENTIFICATION
and analysed by standard microbiological All samples were inoculated onto Nutrient agar, methods, different bacterial strains were isolated from the individual sample. Serological method was used to detect Chlamydia spp. The incubated aerobically at 37OC for 24hours, prevalence of Chlamydia spp was 88%, thereafter, discrete colonies were picked from Staphylococcus aureus 60%, Escherichia coli the growth and Gram stained, while further 23%, Klebsiella spp 35%, Streptococcus subculturing was done to obtain pure cultures pyogenes 10% and Proteus mirabilis 23 %.( for biochemical tests. Bacterial isolates were identified using the method described by Bauer et al (1996). The presence of Chlamydia
Table 1: Prevalence of Microorganisms obtained from semen of males with History or Infertility. Microorganism diagnosed Isolated Bacterial strains Via serology Chlamydia trachomatis

Table 2: shows some selected antibiotics and the in-vitro antimicrobial properties. Augmentin showed
similar activity as Ciprofloxacin when its efficacy against Proteus mirabilis, Klebsiella spp as well as
Escherichia coli was assessed. Perfloxacin and Ofloxacin had a MIC of 0.125µg/ml against Proteus
mirabilis, with the same concentration of Ofloxacin being bacteriocidal with respect to Streptococcus
Pyogenes and Klebsiella spp.
Table 2 In Vitro MICs/Antimicrobial Activity of selected Antibiotics Against Isolated Microorganisms
Chlamydia trachomatis N\A N\A N\A N\A
Proteus mirabilis 0.250 0.125 0.125 0.250
Streptococcus pyogenes 0.250 0.250 0.125 0.500
Klebsiella spp 0.250 0.250 0.125 0.250
Escherichia coli 0.250 0.125 0.250 0.250
Staphylococcus aureus 0.250 0.250 0.250 0.500
CRP = Ciprofloxacin
PEF = Perfloxacin
OFX= Ofloxacin
AUG= Augmentin
N/A = Not Applicable.
Table 3 show actively motile sperms, slow motile sperms and non- motile sperms of male with
infertility, have associated white blood cells. While there are numerous leukocytes associated with non-
motile sperms and slowly motile sperms, about 0-1 white blood cells were seen among the active
motile sperms.
Table 3: Viability of spermatozoa using mean motility and Leukocytospermia.
Motility Mean +sd Leukocytospermia
Active motile 26.6+_17.63 + Slow motile 23.1 +_ 3.0 +++ Non-motile 42.6 _+4.11 +++ Table 4 shows various degree of improvement in sperm viability when patients are placed on antibiotics. However, viability is greatly enhanced when combined therapy (Ofloxacin and doxycycline) are used and leukocytospermia was eliminated among active motile sperms.

Table 4 Leukocytospermia in combined and mono-Antibiotic therapy
Sperm motility Leukocytospermia in Leukocytospermia in Ofloxacin
Ofloxacin therapy + doxycycline combined therapy
Treatment Duration in months Treatment Duration in months
1 2 3 4 1 2 3 4
Active motile + _ _ _ _ _ _ _
Slow motile +++ +++ ++ + + + _ _
Non motile +++ +++ ++ + +++ + + +

+ = 0-5 WBC \HPF
++ = 6-9 WBC \HPF
+++ = > 10 (Numerous) WBC \HPF
phosphate, fructose). Leukocytes are the main Chlamydia trachomatis and Staphylococcus radicals) which can decrease sperm function organisms associated with male infertility in this (acrosome reaction and zona-binding).MAGI is work, with percentage prevalence of 88% and also associated with an increased prevalence 31% respectively. Other isolated organisms of sperm antibodies (Witkin,1995). The role of were Streptococcus spp (10%), Escherichia coli different micro-organism such as Mycoplasma (23%), Proteus mirabilis (23%) and Klebsiella or Chlamydia in prostatitis and infertility and the role of leukocytes in seminal fluid are also responsible for male accessory gland infection (MAGI) occurs twice as often in the male partner of infertile couples than in infertile men. However, the role of infection and antibiotic spontaneously, however, they tend to reoccur treatment in male infertility is still controversial and only frequent ejaculation together with antibiotics treatment have a long-lasting effect Escherichia coli, an organism isolated in this study has previously been reported as the most implicated microorganism in urinary tract The protocol in some cases, is to treat with infection (Momoh et al, 2007) MAGI, can be prostatitis, if it is associated with positive diagnosed when 2 or more criteria are fulfilled: urethral Chlamydia, Mycoplasma or bacterial (1.) history of recurrent UTI or prostatitis, (2.) culture in semen, with positive Mar test or expressed prostatic fluid with more than 40 associated genital tract infection in the female leukocytes or urinary sediment with more than 15 leukocytes after prostatic massage and\or positive bacterial culture, (3.) more than 1 In this study, the quinolones, particularly million\ml leukocytes in the ejaculate , (4.) Ofloxacin is effective in the treatment of growth of 1,000 or more pathogens (E coli, bacterial associated with infertility, as this study Streptococcus faecalis, Proteus spp) in seminal 0.125µg/ml -0.25µg\ml, Ofloxacin has inhibitory (Staphylococcus epidermis, Corynebacterium effects on all the bacterial isolates. The spp, Acinetobacter)(Comhaire,1986). implication is that Ofloxacin can be relatively relied upon to decrease the bacterial load in MAGI could affect male infertility by decreasing these condition. Augmentin at concentrations of sperm count or motility and accessory gland 0.250µg\ml-0.050µg\ml, also has inhibitory function (decreased levels of zinc, acid effects on the isolated bacterial strains.
Yanushpolsky(1996),reports that if no germ is Sperm motility is an index of sperm viability and isolated, suspected cases are treated with male infertility while leukocytospermia is an important critical index for assessing male metronidazole (2x400mg) and AINS (diclofenac 100mg) in the male partner in case of severe The use of a combined therapy of doxycyline and Ofloxacin proved to be better in the management of these patients, leading to improved sperm motility and reduction and elimination of white blood cells from semen. associated with the semen samples. While active motile sperms has between 0-1 white Finally, this study has shown that the role of blood cells with a mean value of 26.6+17.63, Chlamydia in prostatitis and infertility as well as the role of leukocytes in seminal fluids are not numerous white blood cells (>10) associated with their mean value being 23.1+3.01 and 42.6 REFERENCES
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