Comprehensive Literature Review on Curly Kale and Bacterial Infection

This report provides a comprehensive literature review on curly kale (Brassica oleracea var. sabellica), a leafy vegetable known for its nutritional richness and potential health benefits. The review explores the plant's characteristics, including its classification as a cruciferous vegetable, its various types (curly and lacinato), and its cultivation requirements. It delves into the phytochemical analysis of curly kale, highlighting the presence of compounds like flavonoids, ellagic acid, and carotenoids, and their role in antimicrobial properties. The report also examines the interaction between curly kale and Pseudomonas aeruginosa, a gram-negative bacterium, and discusses various treatments for bacterial infections, including the use of antibiotics and the determination of Minimum Inhibitory Concentration (MIC). Furthermore, the report reviews the agar dilution method and qualitative analysis techniques used in assessing the antimicrobial properties of kale extracts and their effects on bacterial growth, emphasizing the importance of sugar content and protein estimation in understanding the plant's response to bacterial infections. The report concludes by highlighting the potential of curly kale as a source of natural antimicrobial agents and the importance of further research in this area.

CHAPTER 2
LITERATURE REVIEW
2.1 Curly Kale (Brassica oleracea var. sabellica)
Curly Kale (Brassica oleracea var. sabellica) is considered to be a leafy vegetable that can be
considered because of its rich nutritious attractiveness, contented, nutraceuticals, in addition to
flavorful perception. Kale is considered as a cruciferous vegetables comprising cauliflower,
broccoli, Brussels sprouts, cabbage as well as bok choy. They come under common family
identified as Brassica oleracea family. Kale originates in diversities in addition to used for
dissimilar purposes. Curly kales have altered colors in addition to have excessively bent leaves in
addition to long stalks. Lacinato kale have dark green-blue leaves along with a imprinted,
horizontal superficial. Curly and lacinato kale are used in special art form. Ornamental kale is the
highly valued amongst the other kales as they cannot be eaten in same way as the others. It
represents vivacious colors like lavender, pink, lavender, blue as well as violet that are highly
appealing for eyes. This is the reason it can be easily used in vegetable and flower bouquets.
Kale growth occurring in the colder climates in addition to lesser temperature seasons associated
to other seasons. Though, it had been observed that the expansively all through the year. It
requires less soil prerequisite nevertheless greater water content for cultivation. Kale developed
in the Middle Eastern region in addition to farmed since 2000 BC (Acikgoz, 2011) in addition
has an imperative place in the diet of the populace in the northern parts of Turkey (Ayaz et al.,
2006). It considered to be one of utmost nutritious foods, rich in dietary fibres, antioxidants,
minerals and proteins (Acikgoz, 2011). Kale is considered to be a opulent source of vitamin C,
vitamin B6, vitamin K, vitamin A, folate, zeaxanthin, lutein, arotenoids, and fiber. It also has all
indispensable minerals like manganese, potassium, iron and copper. Kale in addition to other
cruciferous vegetables comprise bioactive glucosinolates as well as sulfur-containing compounds
when hydrolyzed. It thereby releases elements like isothiocyanates in addition to indole-3-
carbinol. Investigational analysis has shown greater intake of these vegetables impacts
carcinogenesis during commencement in addition to promotion segments of cancer expansion
(Murillo, 2001). An exploration showed by (Ayaz, et al., 2008) established that leaves in
addition to seeds of kale are better foundations of ordinary phenolic antioxidants in addition to
complexes with satisfactory antimicrobial activity. This examination confirmed high efficiency
of antimicrobial action of Kale in contradiction of gram positive bacteria (S. aureus, E. faecalis,
B. subtilis) in addition to gram-negative (M. catarrhalis) bacteria.

2.2 Cruciferous Vegetable
The term ‘cruciferous’ or ‘cruciferae’ is imitative from the distinguishing cruciform growth of
the petals in the plants of this family. They can be filled with vitamins in addition minerals, in
addition to have great fiber content. This have Broccoli (B. oleracea var. italic, Turnip (B. rape
var. rapa), cauliflower (B. oleracea var. botrytis), Cabbage (B. oleracea var. capitate), and
radish (R. sativus) etc. Hu, et al., (2004) did the evaluation of antimicrobial activity of numerous
cruciferous vegetables collections. Water-soluble in addition to methanol-water extracts were
equipped to survey the study in contradiction of dissimilar strains of Gram-positive as well as
gram negative bacteria, and yeast. This examination established that four dissimilar extracts of
all of the six cruciferous vegetables which had a greater inhibitory consequence on Gram-
negative bacteria in comparison to Gram-positive bacteria.
2.3 Pseudomonas aeruginosa
Pseudomonas aeruginosa is a universal, rod-shaped gram-negative, non-spore forming, in
addition to polar-flagellated bacteria, having a diameter range from 1.5-3μm x 0.5μm. It is not
capsulated nonetheless have numerous strains that has a mucoid slime layer. It have outstanding
flexible aptitude to endure in addition to continue in a extensive choice of environmental
circumstances (Singh et al., 2010; Blanc et al., 2007; Dworkin et al., 2006). This bacterium is
accepting of temperatures to be up to 50°C in addition to can cultivate in aerobic circumstances
in addition to in anaerobic situations. P. aeruginosa has an extensive collection of evolution
material, negligible nutrient necessities; non-fastidious microorganisms (Favero et al., 1971). It
is usually recognized to be damaging to plants, insects, as well as animals comprising humans.
P.aeruginosa is a noteworthy source of treacherous communicable diseases such as wounds,
burns irritation, cystic fibrosis, urinary tract infection, along with many other nosocomial
infection, infections otitis external, in addition to causes bacteremia (Hadrich et al., 2014). P.
aeruginosa is the chief foundation of intrusive contaminations in burn patients; there are almost
seventy-five percent of all demises in patients due to the austere scorching are sepsis obtained
due to contamination of offensive hot wounds. It is noticed that the premature wound
expurgation happenings prevented invasive contaminations from the wound site (Barret et al.,
2003). Cystic fibrosis (CF) is instigated by gene mutation that is known as known as CFTR.
Cystic fibrosis is an advanced, protracted, in addition to recurrently incurable genetic
(hereditary) sickness of the body’s secretion glands. CF mostly distresses the respiratory system.
P. aeruginosa laterally with additional bacterial classes that play a part in infection in lung. The
epithelial cells in the respiratory tract converts a stage for colonization of bacteria. In many
cases, colonization of P. aeruginosa of the CF airway caused a protracted antimicrobial resilient

contamination (MacEachran et al., 2007; Döring et al., 2014). The aerosols usage qualifies
remedies in addition to an amount of antibiotics, that includes carbenicillin, tobramycin, colistin,
aztreonam, gentamicin, ceftazidime, in addition to amikacin, that is to be managed straight to the
lung of patients suffering from CF.
2.3.1 Treatments of P.aeruginosa
The greatest challenge is considered to face considering bacterial contamination with
conservative antibiotics is the intensification antibiotic resistance between specie due to the
result of misuse of antibiotic administration. The action develops complex since the outer
membrane of the bacterium is moderately impervious, beside with the action of 12 resistances–
nodulation–division (RND) efflux pumps. These pumps can banish a series of antibiotics. A
broad range of antibiotics are being tested beforehand for the handling of P.aeruginosa.
Amalgamation antibiotic therapy has also been applied to augment P. aeruginosa treatment.
Antibiotics such as Cefotaxime, Gentamycin, Piperacillin, Aztreonam, Streptomycin, Cefepime,
Bacitracin, Ciprofloxacin and Norfloxacin along with Colistin besides Fosfomycin are
recognised to display confrontation to this bacterium. Anti-pseudomonal drugs are like
aminoglycosides, fluoroquinolones and β-lactams. β-lactams are considered to be the most
normally used antimicrobials in the world in addition to account for 60% to 70% of all agreed
antimicrobials in advanced countries. The β-lactam antibiotics performance by constraining the
manufacture of bacterial cell walls from the peptidoglycan layer. Aminoglycosides disturb
amalgamation of proteins by joining to the bacterial 30S ribosome subunit. Fluroquinolones
action by constraining replicative enzymes (DNA gyrase), completing DNA replication.
Confrontation to aminoglycosides in addition to fluoroquinolones considered on the rise
everywhere the globe.
2.4 Minimal Inhibitory Concentration (MIC)
Minimum inhibitory concentration (MIC) is the lowermost absorption of any antimicrobial agent
counting antibiotics in addition to other ingredients precludes microbial progress in addition to
the lowermost absorption of antimicrobial agent necessary to execute microorganisms (Andrews,
2001). The furthermost commonly used approaches for defining the minimum inhibitory
concentration (MIC) where the dilution of agar in addition to broth dilution technique. MIC is
single-minded consuming disk diffusion process demonstrated by Kirby-Bauer method. Bauer et
al., in 1966 resolute association among the scale of disc-inhibition zones in addition to dilution
test results. Kirby-Bauer technique is chosen to be classic systems in microbiology in addition it
is preferred extensively used In case of agar dilution, solutions is given amount of bacterial cells

are positioned straight on the agar plate with precise antimicrobial agent concentrations
(Wiegand, 2008). The bacterial growth over the agar plate is inspected for the MIC. Broth
dilution method procedures liquid growth medium comprising cumulative absorptions of the
antimicrobial agent which is injected with a particular number of bacterial cells. The occurrence
of turbidity or a residue after incubation designates the growth of the microorganism (Wiegand,
2008).
2.5 Agar Dilution Method
The possible of microorganisms to accomplish observable growth on a variety of agar plates
(agar dilution) encompassing antimicrobial dilutions is established. The lowest antimicrobial
agent absorption that will avoid a microorganism's measureable growth is defined as the MIC.
Ohikhena et al., (2017) conducted a research estimating of the antibacterial in addition to
antifungal possessions of Phragmanthera capitate where they combined the plant extract inside
the molten Mueller Hinton agar (MHA). Klancnik (2010), for the agar dilution method serial
dilutions of plant extracts were made in molten MHA when accompanying an experimentation
assessing of diffusion in addition to dilution methods. The examination concluded that agar
dilution method is more operative in comparison to any other method employed for finding MIC.

2.5 Phytochemical analysis
The qualitative and quantitative phytochemical need to be done for evaluating the in vitro
antioxidant properties for numerous aqueous along with alcoholic leaf extracts and root parts
extract for Curly kale. The phytochemical analysis is done as this substance is naturally present
in the Curly Kale and have shown biological significance (Szutowska et al., 2020). It plays
several essential roles in plants like to protect themselves in contradiction of different pathogenic
microbes by showing different antimicrobial analysis features thereby inhibiting or killing
procedures. The four different classifications of phytochemicals are flavonoids, ellagic acid,
allium compound and carotenoids. The caretonoids present in plant is lycopene, lutein and beta-
carotenoids prevent red blood cells and fat cell along with other body fluids from free radicles.
2.5.1 Qualitative analysis
Different qualitative analysis was done with methanol, ethanol and aqueous extract to identify
the phytochemicals. This analysis is required for recognizing the phytochemical The Qualitative
analysis is very essential to identify the phytochemical constituents present in plants. Kale is a
medicinal plant, and the value to this plant is because it contains bioactive elements. Kale
contains tannis, aponins, starch, sugar, aponins, tannins, phenols, flavonoids, glycosoides,
ascorbic acid and alkaloids, for qualitative analysis (Šamec et al., 2018). Terpenoids as well as
starch were available in all plant species other than Nicotiana tobaccum. Saponins are
present in curly kale. Hence, this Qualitative analysis is very vital to organize the phytochemical
elements present in medicinal plants such as Kale. The medicinal value of kale is because the
bioactive elements are present.
2.5.2 Total protein estimation
Total protein estimate for curly kale is required as it helps in determining the bacterial protein.
The plant itself have some specific proteins and bacterial infection can be identified if any extra
proteins are preset. This would help in interpreting the effect of the bacterial infection and

necessary treatment can be done with the plant to prevent this effect. Bacterial infection mainly
kills the plant slowly. Glutamic is present in large amount in Kale, which is around 33.2mg/dry
weight. Aspartic acid is also available in larger amount which includes 27.6mg/dry weight which
further accounts for 10.2% of the total amino acid content of kale leaf. The content of amino acid
can be analyzed with amount of amino acids present on kale leaf in comparison to that bacterial
infected kale plant (Podda et al., 2019). Hence, microbial activity can be analyzed and protein
can be recognized causing the disease.
2.5.3 Sugar content in plant
To prevent the microbial activity of Pseudomonas aeruginosa which is gram negative bacteria.
The antifungal drug such as ciclopirox, have the capability to correspondingly blocks
susceptibility of antibiotic or having multidrug resistant (Krishnamachari & V, 2017). There are
several bacterial which shows that it is not infected by any prevailing resistance procedures.
Drug such as ciclopirox have the capability to prevent the growth of Pseudomonas aeruginosa.
Hence in order to prove it strain from similar gram negative bacteria chosen that showed
interruption of genes coding products involving in sugar substances such as galactose salvage,
antigen synthesis which is an enterobacterial in addition to transport of the iron binding
enterobactin, siderophore, and by lowering the MIC thereby blocking the mutant growth in
comparison to other isogenic strain from parents.
2.6 Curley kale and Pseudomonas aeruginosa
The medium of action can be received by complete spoilage. The organic acids reduce the pH
leading to inhibition of different mechanisms. However, the antimicrobial impact of organic
acids is linked with undisscociated style. It can easily diffuse out through the cell membrane as
well as internalized through the pH of the cytoplasm of the cell and hence getting dissociated
into protons and anions. Proton release leads to the reduction in the internal pH thereby
disrupting the proton force and preventing the transport of substrate. There are some elements

which can prevent the pathogen growth by a modest prohibition of the environmental vocation
by substrate preferences and production of bacteriocins. The antibacterial impact drugs reduce
the phenolic component activity as there are many sites of action involved in the reaction. Hence,
antimicrobial activity prevents the bacterial infection and saves plants from dyeing.

References
Krishnamachari, H., & V, N. (2017). Phytochemical Analysis and Antioxidant Potential of
Cucumis Melo Seeds. International Journal Of Life-Sciences Scientific Research, 3(1).
https://doi.org/10.21276/ijlssr.2017.3.1.19
Omondi, E., Engels, C., Nambafu, G., Schreiner, M., Neugart, S., Abukutsa-Onyango, M., &
Winkelmann, T. (2017). Nutritional compound analysis and morphological
characterization of spider plant ( Cleome gynandra ) - an African indigenous leafy
vegetable. Food Research International, 100, 284-295.
https://doi.org/10.1016/j.foodres.2017.06.050
Podda, A., Pollastri, S., Bartolini, P., Pisuttu, C., Pellegrini, E., & Nali, C. et al. (2019). Drought
stress modulates secondary metabolites in Brassica oleracea L. convar. acephala (DC)
Alef, var. sabellica L. Journal Of The Science Of Food And Agriculture, 99(12), 5533-
5540. https://doi.org/10.1002/jsfa.9816
Šamec, D., Urlić, B., & Salopek-Sondi, B. (2018). Kale (Brassica oleracea var. acephala) as a
superfood: Review of the scientific evidence behind the statement. Critical Reviews In
Food Science And Nutrition, 59(15), 2411-2422.
https://doi.org/10.1080/10408398.2018.1454400
Szutowska, J., Rybicka, I., Pawlak‐Lemańska, K., & Gwiazdowska, D. (2020). Spontaneously
fermented curly kale juice: Microbiological quality, nutritional composition, antioxidant,
and antimicrobial properties. Journal Of Food Science. https://doi.org/10.1111/1750-
3841.15080