Sehr Syed, Umair Azmat,
Shaista Ilyas, Arslan Fazal, Tanzeela Murmim, Fakhra Riaz, Maryam Khan, and Saba Shamim, from
the institute of Pakistan. wrote a Research Article about, "Betel Leaf
Power: Natural Defense Against Acinetobacter Infections. Entitled, Effectiveness
of Piper betel leaf extracts against Acinetobacter species isolated from
bronchitis and pharyngitis patients. This research paper published by the International Journal of Biosciences | IJB. an open access scholarly research journal
on Biosciences. under the affiliation of the International Network
For Natural Sciences| INNSpub. an open access multidisciplinary research
journal publisher.
Abstract
Acinetobacter species
is associated with healthcare-associated infections especially chronic cough
and other related complications. They are becoming increasingly antibiotic
resistant. In the present work, the antimicrobial activity of different extract
(water, ethanol, methanol, and chloroform) of Piper betel leaves were
tested against Acinetobacter sp. SZ-1 and Acinetobacter baumanii TM-1
isolated from clinical samples collected from the patients suffering from
pharyngitis, bronchitis and sore throat. For the antibacterial activity, different
plant extracts were used in which Piper betel leaf stand out to be
the most effective. Six solvents were used for this study in which only
chloroformic extract showed the zones of inhibition. TLC analysis showed five
spots with Rf 0.896,0.973,0.747,0.574 and 0.482. SDS-PAGE revealed
proteins of different molecular weight when the bacterial cells were treated
with ethanolic extract. It was also confirmed by Bradford analysis. GCMS
analysis depicted different bioactive compounds including phytol; phenol,2,2-methylenebis[6-(1,1-dimethylethyl)-4-methyl;
spirost-8-en-11-one,3-hydroxy-,(3β,5α,14β,20β,22β,25R) and
2,2,4-trimethyle-3-(3,8,12,16-tetramethyl-heptadeca-3,7,11,15-tetraenyl)-cyclohexanol.
Furthermore, in silico analysis can enhance the existing knowledge
about establishing the significance of P. betel leaves as an
effective drug to treat the acute and chronic upper respiratory tract
infections.
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Introduction
The genus Acinetobacter
was discovered in the eighteenth century when it was considered as a pathogen
of the soil (Doughari et al., 2011). Since its discovery, the genus and its
originating species have long been characterized and re-characterized under
many genera, before being finally distinguished from other closely affiliated
(on the basis of morphological and genetic grounds) genera (Jung and Park,
2015). The species belonging to this genus are reported to be aerobic, Gram
negative in their Gram morphology and are typically present in the form of
pairs or chains. Their growth characteristics and morphological patterns are
highly dependent upon the type of media used for their isolation. Although the
genus is home to many species, the most commonly known, studied and well
identified species of the genus is Acinetobacter baumanii. It is usually found
to inhabit water bodies and soil, but is also typically found in healthcare
settings, environments and medical equipment (Villegas and Hartstein, 2003).
There have been various studies that report the incidence of multidrug
resistant A. baumanii, which has been the causative agent of many nosocomial
and hospital acquired infections in European countries (Kamolvit et al., 2015).
Over the years, it has emerged as a serious pathogen, as being reported as the
causative agent of many skin, tissue, wound, and blood infections, sepsis,
meningitis, and hospital acquired pneumonia (Howard et al., 2012). A. baumanii
responsible for hospital acquired infections are usually isolated from
different environmental settings like hospital walls, roofs, beddings, curtains,
medical equipment, door knobs and handles, bin stands, as well as dispensers.
It has the ability to sustain on living and non-living sources for long periods
of time, and its persistence in hospitals and healthcare facilities is the
leading factor behind its resistance to antibiotics and other disinfectants
(Evans et al., 2013). It mainly targets exposed tissues and organs like the
mucous membranes and those areas which have been exposed open by any trauma or
wound injury (Sebeny et al., 2008). These infections can take a turn for the
worse if these are left untreated, by leading to septicemia and eventual death
(Howard et al., 2012). The other reason may be the exposure to the pathogen and
its acquiring from environment of the infected hospital as well as the exposure
and contact of the healthcare personnel with an infected patient (RodríguezBaño
et al., 2009). The other species of Acinetobacter such as A. pittii and A.
nosocomialis also cause infection in the patients of intensive care unit that
is reported around the world while A. calcoaceticus causes bacteremia and
pneumonia. The other species like A. lwoffii, and A. junii also have been
observed to cause infections in patients with compromised immune systems
(AlAtrouni et al., 2016). The affected groups of people can range from all age
periods and ranges, but A. baumannii can particularly affect those people who
are hospitalized and are immunocompromised, which may or may not be
hospitalized. The patients who have a protracted stay at the hospitals are also
susceptible to infection. Interestingly enough, the peculiar group with an
increased risk of its infection are the soldiers and the armed forces who have
been sent to conflicted war zones, especially those environments which are dry
and humid. The desert areas provide an ideal setting for its growth which is
reported to be the main causative agent of infection in wounded armed personnel
(CDC, 2004).
The incidence of
multiple drug resistant A. baumannii is usually reported in patients which are
kept in the ICU of hospitals, where the estimated fatality rate is often high
(Seifert et al., 1995). Therefore, it is often difficult to ascertain the
fatality strictly related to it, and not depending upon the patients‟ other
root causes of disease. However, it would not be wrong to say that the presence
of A. baumannii certainly elevates the risk of high fatality (Abbo et al.,
2007).
The other probable
cause in the case of A. baumannii is that it acts as a biomarker or a precursor
of an increased rate of mortality in the case of bacterial infections
associated with patients admitted under critical care in hospitals (Eliopoulos
et al., 2008).
The use of plants in
the form of drugs has been in use since the advent and enlightenment of
disease. The origin of this phenomenon is most probably the result of basic
instinct and interest, as the case with animals and their first interaction
with humans as a food source. The knowledge was beginning to take shape at the
time, in the light of inadequate information regarding the pathogenesis of
disease and the use of plants as their cure. The passage of time resulted in
the discovery of specific plants which were solely used for the treatment of
specific diseases (Petrovska, 2012). The medicinal plants may be defined as those
plants that have been associated with herbal treatments since ancient and
recent times, for the treatment of established and new infections and diseases,
as well as for use in simple herbal and therapeutic concoctions that provide
relief against simple ailments like insect bites, headache, nausea etc (Schulz
et al., 2001).
There have been many
plants that are reported for their beneficial properties, countless therapeutic
and commercial applications, due to which they have been in use since ancient
times, long before their beneficial properties were scientifically proven and
researched about. Piper betel (Piper betle Linn.) plant is one such example. It
is one of the most widely known and used plants worldwide. It belongs to the
family of Piperaceae, which is known by various names across many countries of
the world. It is known as „ikmo‟ in the Philippines, where it is largely
cultivated (Quisumbing, 1978). It is also grown in many other Asian countries
like China, Taiwan, India, Pakistan, Indonesia and Malaysia (Guha, 2006). It
has been reported to be effective against various bacterial strains like:
Bacillus cereus, Enterococcus faecalis, Listeria monocytogenes, Micrococcus
luteus, Staphylococcus aureus, Aeromonas hydrophila, Escherichia coli, Salmonella
Enteritidis, Pseudomonas aeruginosa, Streptococcus mutans (Khan and Kumar,
2011), Enterococcus faecium, Actinomycetes viscosus, Streptococcus sanguis,
Fusobacterium nucleatum as well as Prevotella intermedia and Streptococcus
pyogenes (Datta et al., 2011).
The aims of the current
study were to find the Acinetobacter species associated with pharyngitis,
bronchitis and sore throat. The biochemical and molecular characterization of
Acinetobacter sp., screening of the medicinal plants against it, selection of
the medicinal plant on the basis of its bioactivity, thin layer chromatography
(TLC), qualitative and quantitative assays of proteins, estimation of
antioxidative enzymes including superoxide dismutase (SOD), peroxidase (POX),
ascorbate peroxidase (APOX), glutathione reductase (GR), catalase (CAT) and gas
chromatography mass spectrophotometry (GC-MS) will be ascertained.
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