ISOLATION OF BACTERIAL PATHOGENS FROM EYE

INFECTION PATIENTS FROM QUETTA

Synopsis

Submitted by

Nida Gul

M.Phil Research Scholar

Zoology Department (2015-2017)

SBKWU Balochistan

Nidagulkhoso@gmail.com

Supervisor

Dr. Shahkeel Babar

Professor

CASVAB

University of Balochistan Quetta

shahkeelbabar@gmail.com

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 Co-supervisor

Dr. Shagufta Sadozai

Assistant Professor

Department of Zoology

SBK Women’s University Quetta

Shagufta_sadozai@yahoo.com

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 Supervisory Committee

Supervisor

Dr. Shahkeel Babar

Professor

CASVAB

University of Balochistan Quetta

Co- Supervisor

Dr. Shagufta Sadozai

Assistant Professor

Department of Zoology

SBK Women’s University Quetta

Shagufta_sadozai@yahoo.com

Head of Department

Dr. Nabeela Tariq

Assistant Professor

Department of Zoology

SBK Women’s University

Nabeela trq @yahoo.com

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 CHAPTER 1

Introduction
An eye is a very thin skinned fragile organ of the body. If the reliability or formation
of the eye is affected somehow, it may cause the severe diseases with the possibilities
to end up in severe disabilities. The external of eye, just like the many other body
parts is liable to contamination by micro-organisms including bacteria, virus, fungi
and parasite. Red eye is one of the most common ophthalmologic conditions in
primary care setting. Inflammation of almost any part of the eye, including lacrimal
glands and eyelids, or faulty tear film can lead to red eye. Primary care physicians
often effectively manage red eye, although knowing when to refer patients to an
ophthalmologist is crucial. The red eyes disease is present everywhere
and there was an epidemic in one of Taif counties, Mahni County in
1998. The department of health in Kingdom of Saudi Arabia
organized many programs for protection from diseases. There are
four different types of conjunctivitis: bacterial conjunctivitis, viral
conjunctivitis, chlamydial conjunctivitis and allergic conjunctivitis
(Leibowitz, 2005). In this communication we will consider the
bacterial conjunctivitis. Bacteria are the
most common pathogens and involved in infections of all the tissues
of the eye. The
most frequently affected part of the eye is conjunctiva, lid and
cornea (Jorgensen et al., 2015; Ubani, 2009) which are external part
of the eye. A healthy conjunctiva is
necessary for the maintenance of a healthy cornea and thus the
visual acuity of the eye. Bacterial infections of the eye are usually
localized but may frequently spread to adjacent tissue due to some
predisposing factors such as during trauma, previous
surgery, ocular surface disease, contact lens wear, ocular adnexal
dysfunction and other exogenous factors, systemic diseases
(Bharathi et al., 2003) and

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immunosuppression may alter the defense mechanisms of the outer
eye and permit
bacteria to spread (Leibowitz, 2005; Seal and Uwe, 2007). Many
opportunistic
pathogenic agents are increasingly encountered in ocular infections
due to
widespread use of topical and systemic immunosuppressive agents,
increasing
numbers of patients with human immunodeficiency virus (HIV)
infection and
with organ transplants who are on immune suppressive therapy.
These opportunistic pathogens also cause ocular infections due to
increased use of contact
lens. The dreaded infections endophthalmitis following cataract
extraction and lens
implantation often is caused by opportunistic pathogens (Leibowitz,
2005;
Caldwell et al., 1992).

Most causes of eye redness can be recognized by taking a careful history and
performing simple examinations with the following main questions:

 Association with pain

 History of preceding trauma

 Seasonal or recurrent occurrence (allergic reactions, iritis)

 Changes in the eye lid (contact dermatitis, ocular rosacea, pemphigoid)

 Use of eye drops (glaucoma, dry eye)

The assessment of clinical signs should include the anatomic location of redness
(eyelids, conjunctiva, cornea, sclera and episclera, or intraocular); symptoms in one or

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both eyes; possible associated symptoms (pain, itching, visual decrease or loss); and
further prominent ocular (mucopurulent discharge, watering, blepharospasm,
lagophthalmus) or systemic (fever, nausea) findings.

In particular, the occurrence of ocular pain is an important indicator. Pain can have
several sources, originating from the eye itself or the periocular region. Painful
symptoms can be projected to the eye from other regions. Typical causes are changes
of the eyelids, ocular muscles, cornea, sclera, uvea, or optical nerve. Frequent pain-
associated syndromes are migraine, cluster headache, arteriitis temporalis
(M. Horton), and zoster or trigeminus neuralgia.

Figure 1: Clinical picture of red eye

The infectious micro-organisms can spoil the internal structure of the eye, ultimately
resulting in image loss. The bacterial eye disease is a great deal of problem whish has
been reported from all over the world. The familiar fundamental bacterial agents
comprise Streptococcus spp., Staphylococcus spp., Psuedmonas aeroginosa,
Haemophilus influenzae, Moraxella laclunata, Acinetobacter spp., Neisseria
gonorrhoeae, enterobacteria, Chlamydia trachomatis, Branhamella catarrhalis, and a
few anaerobic bacteria.

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The disease is one of the mainly fear complications of view restore subsequent to
intraocular operation, such as waterfall and eye removal, for the reason that the
beginning and spoil can arise quickly. Bacterial keratitis is individual of the mainly
visually aggressive optical catching pathologies, anywhere corneal perforation can
arise in not as much of than 24 hours. Endophthalmitis is an intraocular disease
through severe trouble that can consequence in stable failure of image. Trachoma is
initiate universal cause through chlamydia trachomatis and is the most only cause of
loss of sight. Bacterial conjunctivitis is a global frequent trouble, it occurs frequently
in Sudan, anywhere the best division of belongings were between infants and kids
(Nabila ,1985). High commonness and huge complications of the eye disease were
reported in several studies (Schaefer et al., 2001). still extra do research work is
necessary in this grassland in Sudan, attractive in thought the meaning of
multigraviton strains with any monotherapy. Staphylococcus spp. are a face up to for
the new day medication due to the difficulty of infection process and presence and
appearance patterns of their personal virulence factors. The members of this group
own many known toxins, many immune evasion mechanisms and devotion factors,
most of which show fleeting, timed, and disease specific appearance. They cause
different types of infections in a host that are also planktonic, biofilm mediated or
together. Sepsis and pneumonia are largely caused by planktonic forms whereas, a
entire range of diseases, namely, endophthalmitis, osteomyelitis, endocarditis,
constant skin infections, indwelling medical gadget infections, chronic rhino-sinusitis,
and dental implantits are caused by the biofilmic form of the bacteria. swelling can be
caused by equally of the forms (Harro et al., 2010). Staphylococci are human
pathogen, known for their capability to become unwilling to antibiotics. They have
been connected, as well causing ophthalmic infections, with skin infections and
sepsis. Methicillin resistant S. aureus (MRSA), in adding together to resistance to
supplementary drugs, have emerged as a prevalent cause of society disease as fine. In
this section, we explain the epidemiology and antibiotic resistance amongst S. aureus
and additional genus through individual location to ophthalmic infection and focal
point lying on newer approach for behavior of staphylococcus infection resembling
phage treatment and vaccines.

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Antibiotics can be purchased without prescription in less
development countries,
which leads to misuse of antibiotics. This may lead to the
emergence and spread of
antimicrobial resistance (Caldwell et al. 1992, Anagaw et al. 2011;
Tewelde et al.
2013). Also, poor hygienic and infection control practice in the area
may play an
important role in an increased prevalence of resistant bacteria in a
community. Therefore, periodic controlling of etiology and
reevaluation of antibiotics is essential to make a rational choice of
initial antimicrobial therapy.

The increasing use of PCR, over the last few years, rapid template
purification, and
automated DNA sequencing has dramatically reduced the time
necessary to
yield a high-quality sequence. The use of 16S- rRNA gene
sequencing to study the
relatedness of prokaryotic species is well established and has led to
increased
availability of 16S- rRNA databases. The convergence of these
technical and
computational advances has also enhanced the application of 16S-
rRNA gene sequence analysis to bacterial identification
(Rantakokko-Jalava et al., 2000). It was
recently reported that subtle sequence differences in the 16S rRNA
gene could be
used for species identification (Sacchi et al., 2002) and for subtyping
and identifying
hyper virulent bacterial clones (Nilsson et al., 2003).

1. Aims and Objectives

The aims and objectives of the research are listed here.

1.1 Aim of the research

 Isolation of B. subtilis strep Pyogens and staph aureus from eye infection patients.

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1.2 Objectives of the research
 Bio Chemical Identification of isolated Bacillius subtilis, S.pyogems and staph
aureus.
 PCR confirmation of biochmically identified S.aureus, S,pyogens and B.subtitis.