Citation
Zulkiflee, Nur Izzatie
(2019)
Assessment of pathogenicity of community-acquired methicillin-resistant Staphylococcus aureus isolates in peritonitis-induced mice.
Masters thesis, Universiti Putra Malaysia.
Abstract
Staphylococcus aureus strain with distinct genetic backgrounds may exhibit
different virulence in animal models as well as associations with different
clinical outcomes. Methicillin-Resistant S. aureus (MRSA) is known to be more
problematic as it cannot be treated with common antibiotics. Traditionally,
MRSA infections have been limited to hospitals and predisposed immunecompromised
individuals. Recently, MRSA infections have been reported to
occur outside of hospital setting. There has been an alarming increase in the
incidence of Community-Acquired MRSA (CA-MRSA) infections. CA-MRSA
can be a serious threat to public health, which spreads in community. In
addition, the present of Panton-Valentine Leucocidin (PVL), pore-forming gene,
has also been reported to be epidemiologically associated with CA-MRSA and
may cause aggressive infections. Peritonitis is one of the results of S. aureus
infection which caused potentially fatal inflammation of the peritoneum. S.
aureus contributed for the greatest number of positive peritonitis cultures from
patients with continuous ambulatory peritoneal dialysis and end-stage renal
disease. Since the presence of CA-MRSA is continuously emerging, it is
important to continue monitoring the distribution pattern and the pathogenic
status of CA-MRSA in the community. In this study, the pathogenicity of CAMRSA
isolates were assessed using in vivo model of peritonitis with
comparison to a clinical isolate, ATCC 700699 MRSA (ATCC-MRSA). Two
different CA-MRSA isolates (CA-MRSA1 and CA-MRSA2) were previously
isolated from a healthy population were studied. Mice were assigned into 4
groups and intraperitoneally injected with 200 μl of 109 CFU/ml of ATCCMRSA,
CA-MRSA1 and CA-MRSA2, respectively. Control group was injected
with sterile DPBS. After inoculation, mice were observed twice daily until 72
hours post-infection and any distress signs were recorded. Mice were
euthanized at 72 hours post-inoculation or had severe symptoms. Interested
organs and peritoneal lavage were collected for bacterial load and histopathological analysis. All mice inoculated with MRSA showed clear signs
of illness and developed symptoms of peritonitis (p<0.001). In addition, CAMRSA
caused significant mortality rate and higher Peritonitis Severity Scoring
(PSS) scores compared to un-infected mice, but comparable to reference
isolate, ATCC-MRSA, with 80% and 100% of mortality recorded in CA-MRSA2
and CA-MRSA1, respectively. There is no significant different in mortality rate
between CA-MRSA and ATCC-MRSA-infected mice. Bacterial count in liver,
lung and spleen tissues isolated from CA-MRSA-infected mice were
comparable to ATCC-MRSA group. Although it was not statistically different,
CA-MRSA2 showed slightly higher bacterial counts compared to CA-MRSA1
and ATCC-MRSA in culture samples. In addition, histopathological comparison
had showed mild increase in various indicators including inflammation,
necrosis, edema, and haemorrhage, in tissue samples isolated from all MRSAinfected
groups. Tissues isolated from CA-MRSA- and ATCC-MRSA-infected
mice showed consistent histopathological scores. Abscess was only observed
in CA-MRSA2 tissues samples. These results indicate that nasal carriage CAMRA
isolated from a healthy population has potential to cause peritonitis, with
comparable severity as clinical isolate, ATCC-MRSA. Together, informations
obtained from this study provide new insight on the virulence status of MRSA
nasal carriage isolates. Without appropriate intervention, CA-MRSA is likely to
be a continuous threat to public health for the foreseeable future. Thus, it is
important to ensure consistent practices are adopted towards early diagnosis,
appropriate intervention and widespread surveillance to increase awareness
that needed in terms of information or education on the risks, management and
treatment of MRSA.
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