Volume 2 • Issue 2 • 1000109
Open Access Research Article
Kaur et al., Andrology 2013, 2:2
DOI: 10.4172/2167-0250.1000109
Keywords: Apoptosis; Escherichia coli; Spermagglutinating factor;
Acrosome reaction; Mg
2+
dependent; ATPase activity
Abbreviation: SAF: Spermagglutinating Factor
Introduction
Te signifcance of bacteriospermia has gained increasing
importance in recent years. Acute and chronic infections in the
male reproductive system may compromise the sperm cell function
[1]. Spermatozoa can subsequently be afected by these infections at
diferent points in their development and maturation. A variety of
pathogenic bacterial species are isolated from the semen of fertile and
infertile patients with the ability to interact directly with spermatozoa
[2]. Tese interactions are typifed by attachments between bacteria and
spermatozoa, agglutination phenomena, and morphologic alterations
of spermatozoa [3]. Some investigators have also reported evidence for
soluble spermatotoxic factors.
Among the bacterial species that interact with spermatozoa are well
known causative pathogens of genitourinary infections such as E. coli,
U. urealyticum, M. hominis and C. trachomatis [4,5]. Te most discussed
and tested organism concerning male infertility is Escherichia coli, as
the most important pathogen causing prostatitis and epididymitis.
Several authors have postulated a negative efect of E. coli on sperm
motility. Further, Moretti et al. carried out electron microscopic studies
that revealed the ultrastructural damage of the sperm membrane in all
parts of the spermatozoon, especially in the mid-piece, causing both
swelling of the mid-piece and tail invagination on incubation with E.
coli [1]. Tus, it was suggested that E. coli/spermatozoa-interaction may
be a two-step process: i.e. adhesion to and subsequent destruction of
the sperm membrane [3].
In an earlier work done in our laboratory, we had also isolated a
strain of E. coli from semen sample of male attending infertility clinic,
impeding the motility of spermatozoa by agglutination in vitro. Te ~71
kDa Sperm Agglutinating Factor (SAF), responsible for agglutination
could also be isolated and purifed [6]. Te SAF was found to be
spermicidal at higher concentrations and could lead to adverse efects on
various sperm parameters. Intravaginal administration of SAF in Balb/c
mouse led to infertility [7]. Because SAF is specifcally responsible for
the above mentioned phenomenons, we hypothesized that generation of
anti-SAF antibodies could be exploited as a preventive measure against
SAF induced damage. To experimentally evaluate this hypothesis, anti-
SAF antiserum was raised in Balb/c mouse and examined as an antidote
against SAF mediated efects.
Material and Methods
Animals
Sexually mature 5-6 weeks old male (25 ± 2 g) and 4-5 weeks old
female (22 ± 2 g) Balb/c mice were used in the present study. Te
animals were maintained under standard laboratory conditions (12 h
*Corresponding author: Dr. (Mrs) Vijay Prabha, Professor, Department of
Microbiology, Punjab University, Chandigarh, 160014, India, Tel: + 91-172-
2534140; Fax: +91-172-2541770; E-mail: satishvijay11@yahoo.com
Received October 01, 2013; Accepted October 29, 2013; Published November
06, 2013
Citation: Kaur K, Rishi P, Prabha V (2013) Amelioration of Sperm Agglutinating
Factor (SAF) Induced Sperm Impairment by Anti-SAF Polyclonal Antibody.
Andrology 2: 109. doi: 10.4172/2167-0250.1000109
Copyright: © 2013 Kaur K, et al. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.
Abstract
Background: We have previously isolated a Spermagglutinating Factor (SAF) from Escherichia coli, which was
capable of causing sperm agglutination and impairment of sperm parameters viz. apoptosis, premature acrosome
loss and inhibition of Mg
2+
dependent ATPase activity in vitro. In addition, intravaginal administration of Balb/c
mice with SAF resulted in infertility. To provide evidence that SAF plays an important role in impairment of sperm
parameters and infertility, anti-SAF antiserum was raised and its application as a therapeutic intervention against
SAF induced damage was evaluated.
Methods: Effect of anti-SAF antiserum was evaluated against SAF mediated adverse effects on sperm
parameters. Spermagglutination was observed using light microscopy and Mg
2+
dependent ATPase activity was
estimated in terms of release of inorganic phosphate. Sperm apoptosis and acrosome status were evaluated by
means of fow cytometery and fuorescent microscopy, respectively. Further, the impact of anti-SAF antiserum was
also seen on fertility outcome in mice
Results: Results showed that immunization of mice with SAF lead to the generation of high titer specifc
antibodies. Raised anti-SAF antiserum could neutralize all the biological effects of SAF in contrast to control
antiserum. Furthermore, intravaginal application of anti-SAF antiserum along with SAF rendered mice fertile.
Conclusion: Here we provide evidence that SAF plays an imperative role as all the detrimental effects induced
by SAF whether in vitro or in vivo were blocked on simultaneous incubation with anti-SAF antiserum. Present work
also highlighted the effcacy of the anti-SAF antiserum as a curative measure against SAF.
Amelioration of Sperm Agglutinating Factor (SAF) Induced Sperm
Impairment by Anti-SAF Polyclonal Antibody
Kiranjeet Kaur, Praveen Rishi and Vijay Prabha*
Department of Microbiology, Punjab University, Chandigarh, India
Andrology-Open Access
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ISSN: 2167-0250
Andrology, an open access journal
ISSN: 2167-0250