Download Characterization of Mycoplasma Isolated

Haneen Mosleh
Professor Lata Moses
Biology lab 1615
February 27, 2014
Characterization of Mycoplasma Isolated
Introduction
There are many pathogens have been reported to cause eye disease in wild ruminants.
Among the bacterial factors, a Mycoplasma conjunctiva is the most known and studied
disease. A Mycoplasma conjunctiva causes infectious called keratoconjunctivitis (IKC). IKC
is highly contagious ocular disease. This particular etiologic factor was first report by Surman
(1968) from an Australian sheep suffering from phlogosis of the ocular mucosa, and was
further identified and classified by Barile (1972). In northern Italy, infections caused by
Mycoplasma species have not been investigate in wild animals. However, an M. conjunctiva
has reported previously in wild animals in the Apline regions of Italy. Genomic
characterization has performed on only six isolates from chamois from the provinces of
Bergamo and Sondrio, Lombardia Region. They have reported that the diagnostic
examinations performed on an adult male ibex (number 130104/2005), which was euthanized
in 2005 by forest rangers in Val Veny (Valle d’Aosta Region) due to severe blindness. They
also reported the subsequent isolation and characterization of Mycoplasma species.
Materials/ Methods
At this point, they used Anatomopathology in order to diagnose the disease more. The total
disease injuries were determined at postmortem examination and histological procedures
completed the anatomopathologic investigations. They took samples from the cornea and
conjunctiva and cut them to 4 mm thickness, washed, dehydrated, cleared, and placed in
paraffin using an automated system. The Samples embedded in paraffin wax for 14 hr. After
cooling at 4 C, the paraffin blocks cut into 5-mm sections using a microtome.
Results
From the monitoring program for IKC undertaken by CeRMAS in the northern Italian
region of Valle d’Aosta, several cases of clinical disease have reported during the 3-yr
survey. They submitted all cases for the total disease, histological, and bacteriologic
examination. A confirmed diagnosis of Mycoplasma species was only successful in one of
the 86 samples tested. Bacteriologic examination on selective PPLO agar showed growth of
small ‘‘fried egg’’–shaping colonies, ascribable to Mycoplasma sp. Colonies showed slightly
darker granulations, which varied depending on culture conditions. After several days of
growth, the fully developed colonies were 1 mm in diameter. There were not any other
bacterial agents isolated on blood agar or McConkey agar plates. The Mycoplasma diseases
are not diagnosed because they are solely based on clinical signs, pathologic lesions, or
serologic tests because of the close relatedness between Mycoplasma species. Identification
of the causative organism is therefore required to confirm diagnosis. However, it requires
specialized laboratories with experience with these fastidious organisms.
Discussion
Mycoplasma species isolation was only successful in one of the 86 samples tested. In
this case, the animal euthanized and method is a sensitive single generic test capable of
detecting and differentiating Mycoplasmas to the species level. By DGGE, the ibex isolate
observed to be an Mmc serovar. The use of PCR/DGGE represents a significant improvement
on current tests as diagnosis of Mycoplasma infection could be made directly from clinical
samples in less than 24 hr. Mycoplasma infections might also detect and identified in this
single test. The use of molecular testing has limitation because sometimes it is essential to
obtain live organisms for research and other purposes, such as antibiotic sensitivity testing.
Conclusion
The study demonstrated that infection with MmcLC serovar occurs in wild animals in
the northern Italian region of Valle d’Aosta and may be associated with clinical
keratoconjunctivitis. This observation provides a novel facet of the study on the Mycoplasma
spp. infection in the wild animals of the Alpine regions of northern Italy, considering that
several outbreaks of IKC have reported, particularly in chamois and ibex, evidencing the
circulation of M. conjunctivae (Grattarola et al., 1999; Belloy et al., 2003). This indicates the
need for regular and intensive laboratory investigations for the differential diagnosis of
clinical cases in wild ruminants suffering from ocular lesions, which are not always
ascribable to M. conjunctivae.