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The anti-microbial effect of food wrap containing beeswax products


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Food preservation is a vital issue on a global scale. It reflects both the need to prevent food spoilage as well as the necessity for reduction in the occurrence of food-borne pathogens. Progress of technology allowed for the development of various techniques that inhibit the growth or activity of food contaminating microbes. In the last few years we saw creation of various types of antimicrobial food packaging systems. They contain active antimicrobial agents immobilized on a material that acts as a food wrap. In this paper we have tested the efficiency of Abeego, a commercially available food wrap, against representatives of different groups of microbes. The Abeego wrap contains natural products such as waxes and oils, which can display antimicrobial activity. We analyzed the ability of the wrap and its components to affect the growth and replication of various groups of microorganisms. We have discovered that the Abeego wrap was capable of significantly inhibiting the viable cell count of bacterial species - both gram positive and gram negative. However no noticeable antifungal or antiviral activity has been observed. This implies that Abeego wrap can act as an efficient mean of inhibiting the spread of food-borne bacterial pathogens, as well as contribute to preventing food spoilage.
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Crystal T. Pinto1, Jarosław A. Pankowski* 2, Francis E. Nano1
1University of Victoria, Department of Biochemistry and Microbiology, 3800 Finnerty Road, Victoria BC V6P 4W8, Canada.
2University of Nebraska-Lincoln, Department of Chemistry, 1400 R Street, Lincoln, NE 68588, USA.
*Corresponding author:
Keywords: food preservation; antimicrobial; food wrap; Salmonella
The spoilage and contamination of groceries caused by microbes is a serious
concern for many branches in the food industry. It results in food products being
unsuitable for consumption and can account for significant financial losses. It is
estimated that approximately 25% of world’s food supply is lost due to the
activity of microorganisms (Singhet al. 2016). Therefore preventing product
spoilage in food industry is one of the most vital matters. Diseases caused by
food-borne pathogens are another major issue for food distribution. The problem
of food preservation has been known since the beginning of civilization and as a
result multiple techniques of conserving food have been developed. This includes
methods such as drying, salting and pickling. In many cases, especially in
modern food preservation, use of antimicrobial agents plays a significant role.
Especially the chemical compounds that stop the growth and activity of microbes
allow for food to remain consumable for much longer. New possibilities in the
field of food preservation have become available thanks to the use of
antimicrobial food packaging (Malhotra et al. 2015). This type of packaging
material incorporates elements that inhibit the growth of pathogenic bacteria and
fungi. In some cases the active agent can be a natural product.
Abeego is a consumer food wrap material that consists of a coated fabric. The
main constituent of Abeego is beeswax which has been deemed “generally
regarded as safe” (GRAS) (“Select Committee on GRAS Substances (SCOGS)
Opinion: Beeswax (yellow or white),” 1975) by the U.S. Food and Drug
Administration. Lesser components include tree resin and oils. One of the
potential advantages of using beeswax as part of a food wrap material is its anti-
microbial properties (Antúnez et al., 2008; de Andrade Ferreira et al., 2007;
Orsi et al., 2005; Wilsonet al. 2015; Zhang et al. 2013). The anti-microbial
effects of beeswax are attributed to propolis (Marcucci et al., 2001), a sticky
material used by bees to seal holes and cracks in beehives. The major constituents
of propolis are resins derived from the plants that the bees visit while collecting
pollen. The variation in flora in different geographical locations will affect the
constituents of propolis (Wilson et al. 2013; 2015) and, in turn, this will affect
the potency of the anti-microbial effect of each lot of beeswax. Thus, it is
important for manufacturers of food service products using beeswax to have
evidence that each lot of beeswax has a minimal level of anti-microbial activity.
Presumably, much of the anti-bacterial activity of Abeego wrap comes from the
propolis fraction of the beeswax. However there could be contributions from the
resin, oil and fabric components as well.
The aim of this project was to investigate the anti-microbial activity of the
Abeego wrap and its components. The anti-bacterial activity of Abeego is
predominently attributed to the propolis fraction of the beeswax, however there is
a possibility of resin, oil and fabric components to play an important role too.
Therefore we decided to investigate the effect of the Abeego product and its
individual components on different groups of microbes. Since food-borne
pathogens can be found among bacteria, fungi and viruses, we have investigated
representatives of each group. The individual components of Abeego wrap
analysed during this project have been taken from the lot used to make the final
Preparation of the Abeego wrap and its components
The Whatman™ Filter Paper Circles (12.7 mm) were used as carriers for the oil
and wax. Fragments of Abeego wrap and fabric were cut out to have the same
shape and size as paper circles. Papers and wrap were sterilized in the autoclave.
Oil was prepared by leaving it in boiling water for 1 hour. The resins were
sterilized with 70% ethanol. The sterile paper circles were dipped in oil or molten
wax. For standardization purposes pieces of resins were selected so that their
weight would be within 10% of each other.
Strains and growth conditions
Bacterial strains of Bacillus cereus, Escherichia coli W3110, Pseudomonas
aeruginosa, Salmonella enteritidis and Staphylococcus aureus were used for the
anti-bacterial activity experiments. For positive control discs soaked in
Food preservation is a vital issue on a global scale. It reflects both the need to prevent food spoilage as well as the necessity for
reduction in the occurrence of food-borne pathogens. Progress of technology allowed for the development of various techniques that
inhibit the growth or activity of food contaminating microbes. In the last few years we saw creation of various types of antimicrobial
food packaging systems. They contain active antimicrobial agents immobilized on a material that acts as a food wrap. In this paper we
have tested the efficiency of Abeego, a commercially available food wrap, against representatives of different groups of microbes. The
Abeego wrap contains natural products such as waxes and oils, which can display antimicrobial activity. We analyzed the ability of the
wrap and its components to affect the growth and replication of various groups of microorganisms. We have discovered that the Abeego
wrap was capable of significantly inhibiting the viable cell count of bacterial species both gram positive and gram negative. However
no noticeable antifungal or antiviral activity has been observed. This implies that Abeego wrap can act as an efficient mean of inhibiting
the spread of food-borne bacterial pathogens, as well as contribute to preventing food spoilage.
doi: 10.15414/jmbfs.2017.7.2.145-148
J Microbiol Biotech Food Sci / Pinto et al. 2017 : 7 (2) 145-148
kanamycin (25 µg/ml or 50µg/ml) were used. Two strains of Saccharomyces
cerevisiae CRY1 and MYA3666 (VL6-48) were used as representatives of
yeasts. Bacteriophages M13 (Messing, 1993) and P1vir (Luria et al. 1960) were
used for the viral assays. The M13 phage was amplified by infecting E. coli
JM109 strain, while the P1vir was amplified by infecting E. coli DH10B.
Bacterial broth growth was done in Luria-Bertani (LB) medium (Bertani, 1951).
Yeast broth growth was done in the yeast extract-peptone-dextrose (YPED)
medium supplemented with adenine (YPAD) (Murthy et al. 1975). For solid
media agar was added to concentration of 1.5%.
Zone of inhibition assay
The strains were grown overnight in LB and undiluted samples were spread on
LB plates using sterile cotton swabs. Discs of Abeego wrap, fabric, oil, resin and
beeswax were prepared and sterilely placed on the plates with the above cultures.
The Abeego fabric was used as a negative control (as it contains no beeswax) and
discs soaked in kanamycin were used as positive control. The plates were
incubated at 37C for 18-20 hours until the bacterial lawn of growth was
observed. The zones of inhibition were analyzed by measuring their radius i.e.
the distance from the center of zone to its border.
Bacteria survival assay
For S. enteritidis assay the Abeego discs were left in 1ml LB medium in a
microtiter plate for 18 hours. Then 100µl of S. enteritidis inoculum (~105 cfu/ml)
was added into the wells. The cultures were incubated with shaking at 30°C for
24 hours. After that time the cells in each well were enumerated on dilution
plates. This experiment was performed in quadruplicates. For S. aureus assay the
Abeego discs were left in 1ml LB medium in a microtiter plate for 72 hours.
Then 100µl of S. aureus inoculum (~106 cfu/ml) was added into the wells. The
cultures were incubated with shaking at the room temperature for 40 hours. After
that time the cells in each well were enumerated on dilution plates. This
experiment was performed in pentaplicates. For both experiments the data was
analyzed using unpaired T-test.
Yeast survival assay
For the yeast experiments the Abeego discs were left in YPAD medium in a
microtiter plate for 24 hours. Then inoculum from an overnight S. cerevisiae
culture (~10-6 cfu/ml) was used to start the fresh cultures. The cultures with and
without the Abeego wrap were grown at a room temperature for 48 hours. After
that time the cells in each well were enumerated on dilution plates. These
analyses were performed in triplicates. For this experiment the data was analyzed
using unpaired T-test.
Phage activity assay
For the M13 experiment ~103 phage particles were suspended in LB medium in a
microtiter plate. The phages were incubated with and without sterile Abeego
wrap, at a room temperature with shaking. After 50 hours, plaque assay was
carried out by inoculating the phage with the JM109 strain into LB soft agar and
creating an overlay. For the P1/vir experiment ~106 phage particles were
suspended in LB medium in a microtiter plate. The phages were incubated with
and without sterile Abeego wrap, at a room temperature with shaking. After 24
hours, plaque assay was carried out by inoculating the phage with the DH10B
strain into LB soft agar and creating an overlay.
Growth inhibition assay on agar plates
To test the spectrum of anti-bacterial activity of the commercial products, a zone
of inhibition assay was carried out against the bacteria that are some of the most
common food-borne pathogens and environmental contaminants. The zone of
inhibition assay was utilized as it allows for a quick determination of an
inhibitory effect of a given substance. The impact of Abeego wrap and its
components was compared to a positive control in form of antibiotic discs. After
the incubation no zone of inhibition was observed with Abeego fabric, beeswax,
oil and resin against any of the analysed bacteria. At the same time the positive
control generated visible zone of inhibition (Fig 1). The likely explanation for
this lack of effect from the tested products is the limited diffusion into the
surrounding medium. Therefore in order to properly asses the activity we had to
utilize an alternative method.
Figure 1 Zone of inhibition assay to test the antibacterial activity of Abeego
Fabric and its constituent parts. A) Zone of inhibition assay for Abeego wrap
and fabric against B. cereus B) Zone of inhibition assay for Abeego oil and wax
against B. cereus C) Zone of inhibition assay for Abeego resin against B. cereus
D) Zone of inhibition assay for Abeego oil and wax against P. aeruginosa E)
Zone of inhibition assay for Abeego oil and wax against P. aeruginosa F) Zone
of inhibition assay for Abeego oil and wax against S. aureus G) Zone of
inhibition assay for Abeego resin against S. aureus H) Zone of inhibition assay
for Abeego wrap and fabric against E. coli
Anti-bacterial activity of Abeego wrap
We decided that the best way to detect the anti-microbial activity of the Abbego
wrap was to incubate it with bacterial cells in the liquid phase. To represent the
diverse pathogens we have selected both a gram negative bacterium - Salmonella
enteritidis and gram positive bacterium -Staphylococcus aureus. S. enteritidis is
an organism that is commonly associated with gastrointestinal infections (Feasey
et al. 2012). On the other hand S. aureus is a commensal inhabiting skin of
people and animals. It can however act as an opportunistic pathogen. It can also
become a food-borne pathogen when transferred onto dairy products (Fooladi et
al. 2010). During the test bacterial cells of both strains were exposed to Abeego
J Microbiol Biotech Food Sci / Pinto et al. 2017 : 7 (2) 145-148
wrap and its individual components. We discovered that incubation with the wrap
has caused a significantly decline in the number of cells of both S. enteritidis and
S. aureus (Fig 2). At the same time some of the individual components of the
Abeego wrap also had an impact on the number of observed colony forming units
(cfu). However there is no consistency in that manner between both tested
organisms. This could mean that each compound has a major effect on a different
type of bacterial cell. Therefore the effect of wrap could be attributed to a
combined impact of all its components. Overall this confirms the previous
suspicion of the anti-bacterial activity of the tested material.
Anti-yeast activity of Abeego wrap
While the Abeego product shows an effect on bacterial cells it has to be further
investigated if it can inhibit the survival of eukaryotic pathogens. For this
purpose we have used two strains of Saccharomyces cerevisiae CRY1 and
MYA 3666. S. cerevisiae is the most commonly used representative of Eukarya.
It serves as a good approximation of fungal food-borne pathogens like
Aspergillus flavus (Kim et al. 2004). The yeast cells were treated in a similar
manner as S. enteritidis and S. aureus in the survival assay. The analysis of the
number of cfu revealed a small decrease after treatment of CRY1 cells with the
Abeego product. However the observed results were not considered to be
significant (Fig. 3). This strongly suggests that the tested wrap has no impact on
the fitness of eukaryotic cells. It is likely that the composition of yeast membrane
and cell wall gives resistance to the effects that were observed in bacteria.
Therefore similar reaction might be expected from food-borne fungal
Activity on viral capsid
The last relevant group of food-borne pathogen consists of viruses. The common
examples could be noroviruses, rotaviruses as well as hepatitis A and E viruses
(Atreya, 2004; Carter, 2005). All of these pathogens possess non-enveloped
capsids. On the other hand there are much fewer enveloped viruses that can cause
food-borne infections i.e. astroviruses have enveloped viroids and are associated
with gastroenteritis (Fleet et al. 2000). Additionally coronaviruses can cause
gastrointestinal symptoms, however these viruses are mostly associated with
pneumonic infections (Jevšnik et al., 2013). To investigate the effect of Abeego
wrap on viral capsids we tested representatives of non-enveloped viruses. For
quick and easy assay we decided to analyse the phage particles. These viruses can
be replicated in short time and assayed in a reproducible manner. As viral
particles they can act as an approximation of the eukaryotic viruses. The
commonly used bacteriophages M13 and P1 were chosen for this experiment.
The phage particles were tested using similar method as the one used for bacterial
and yeast survival assay. Each phage was then enumerated on two-layer plates by
infecting E. coli cells. Results show decrease in the overall number of active
phage particles (Fig 4). However these changes are not significant enough to
support conclusion that the Abeego wrap can inactivate viral particles. A possible
explanation for this result would be that the effect of beeswax components is
primarily targeting the membrane and therefore has no strong impact on the
protein structure of viroid.
Figure 2 Anti-bacterial activity of Abeego product. The Abeego wrap and its components (wax, resins, oil and fabric) were tested for their anti-microbial effect on
cells of Salmonella enteridis and Staphylococcus aureus. Pure LB medium and antibiotic were used as negative and positive control, respectively. Significant decrease
in cell number has been observed for S. enteridis when comparing LB medium with wrap (P=0.002) or resins (P=0.043). Other individual elements of Abeego wrap
(wax, oil and fabric) had no significant impact on cell number. Significant decrease in cell number has been observed for S. aureus when comparing LB medium with
wax (P=0.02), oil (P=0.04) or wrap (P=0.02). The exposure to antibiotic reduced number of cells to zero in both cases. The significance level of 0.05>P>0.01 is
represented by * symbol on the graph. The significance level of P<0.01 is represented by *** symbol on the graph. Bars represent standard deviation.
Figure 3 Effect of Abeego product on yeast cells. The Abeego wrap was tested for its anti-microbial effect on cells of Saccharomyces cerevisiae strains CRY1 and
MYA 3666. The yeast cells grown in regular YPAD medium were compared to the ones grown in the presence of the wrap. After enumerating cells in both populations
no significant differences were observed (P>0.05, not significant indicated as ns). Bars represent standard deviation.
J Microbiol Biotech Food Sci / Pinto et al. 2017 : 7 (2) 145-148
Figure 4 Effect of Abeego product on activity of phage particles. The Abeego wrap was tested for its effect on suspended particles of P1vir phage and M13 phage.
The phages exposed to the wrap and the negative control were enumerated on two-layer plates with appropriate strains of E.coli. No significant differences were
observed between the two populations (P>0.05, not significant indicated as ns). Bars represent standard deviation.
The presented results show that Abeego wrap possesses anti-bacterial activity.
This effect can be observed when the microorganisms are exposed to the surface
of the wrap, since the active substance cannot diffuse into the environment. The
tested product displayed activity against both gram negative and gram positive
cells. At the same time this effect does not extend to yeast cells and viral capsids.
Acknowledgments: The work was supported by grant from the Natural Sciences
and Engineering Council of Canada, EGP 477804-14. We woud also like to thank
Toni Desrosiers for providing resources necessary for this project.
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... A recent novel study showed that the wraps also possess antibacterial activity, therefore enhancing their food preservation properties (Pinto, Pankowski and Nano 2017). Although the research did not determine the individual contribution of each wrap component to its bactericidal properties, separate studies have been carried out on those substances. ...
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In recent years, wraps made with beeswax, jojoba oil, and tree resin have emerged both in the form of commercial and home-made products as a sustainable alternative to clingfilm. This study evaluated the antimicrobial properties of their components and explored a potential enhancement with propolis. Food grade beeswax and propolis samples were extracted by reflux, and jojoba oil and pine resin by Soxhlet, and solvent extraction respectively. The antimicrobial effects of the extracts against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) were evaluated using the agar disc diffusion method. A novel method using extract-impregnated discs in bacterial suspensions was also trialled. Combinations of extracts were tested to detect interaction effects (IE) between components. Beeswax and jojoba oil showed no antimicrobial activity (P>0·05), resin reduced the growth of S. aureus (P<0.001), and propolis inhibited both S. aureus (P=0·011) and E. coli. Synergistic interactions were detected between the wrap components (IE=-59·73%) and between beeswax and propolis (IE=-27·88%), but only against S. aureus. The results showed that beeswax food wraps possess antimicrobial properties, and that propolis may enhance these by additivity or synergy. Overestimation of bacterial concentrations likely occurred due to method flaws, therefore possible improvements were identified to increase method reliability. Further studies including larger number of replicates are required to confirm the described results.
... There are some similarities with the studies of Pinto et al. (2017), who tested the efficiency of Abeego (a commercially available food wrap) against representatives of different groups of microorganisms. Abeego consists of a fabric coated with beeswax, with minor components such as tree resin and oils. ...
... The authors demonstrated the antibacterial activity of Abeego against Salmonella enteridis and Staphylococcus aureus, which they attributed to all its ingredients (wax, resin, oil and fabric). They found no antifungal activity against Saccharomyces cerevisiae yeast [40]. ...
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... The use of other mixing ingredients such as oil for the addition of beeswax mixtures typically brings some benefits. Jojoba has anti-bacterial and antifungal properties, or VCO coconut oil has antibacterial properties [6]. 3.the feeling of the beeswax-wrap surface is tolerable, in particular the stickiness of the beeswax coating with natural oils, which can be decreased by the addition of resin or gondorukem. ...
Conference Paper
Abstrak Pandemi covid-19 mengakibatkan terbatasnya ruang gerak bagi pelaksanaan pengabdian masyarakat oleh para dosen. Universitas Trilogi menawarkan fasilitas pengabdian kepada masyarakat yang diintegrasikan dengan acara seminar daring yang dikelola oleh mahasiswa KKN Materi seminar adalah beeswax-wrap. Tujuannya menginspirasi pebisnis pada saat pandemi, yaitu menghasilkan produk penunjang penyimpanan makanan dan sosialisasi penghentian penggunaan single use plastic. Pelatihan dilaksanakan dalam bentuk ceramah, demo, dan tanya jawab. Seminar berbasis web mengggunakan aplikasi Zoom, koordinasi melalui WhatsApp, dan pengelolaan pendaftaran melalui Google Formulir. Peserta yang mendaftar sebanyak 422, sedangkan yang hadir 348 orang. Peserta yang hadir tersebut terdiri atas 64% perempuan dan 36% laki-laki; 90% berdomisili di pulau Jawa dan sisanya di luar pulau Jawa. Jenjang pendidikan peserta 74% masih mahasiswa, 14%lulusan S-1, dan 3% lulusan S-3. Dalam rangka mengetahui respons peserta, dilaksanakan survei lewat Google Formulir. Hasilnya 58% merespons seluruh kegiatan secara positif dan 36% kecewa.. Respons negatif itu disebabkan oleh buruknya koordinasi panitia, waktu yang buruknya koordinasi panitia, waktu yang terlalu pendek, kendala sinyal internet, dan kualitas suara yang tidak memuaskan.Secara umum peserta memahami pengembangan produk kreatif yang berkaitan dengan isu global pandemik dan darurat sampah. Kata kunci : seminar daring, zero waste, beeswax-wrap, abdimas era pandemi Covid19 Abstract The Covid-19 pandemic resulted in minimal wiggle space for lecturers to implement community service. Trilogy University provides community service services combined with online workshops organized by KKN students. The seminar materials are beeswax wraps. During pandemics, the aim is to encourage business people, namely to create food storage support goods and socialize the cessation of the use of plastic for single use. The instruction is conducted in the form of seminars, demonstrations, and q & a. Web-based seminars use the Zoom app, organize via WhatsApp, and administer registration via Google Forms. Participants who signed up were 422, 348 of whom attended. Participants were 64 % female and 36 % male; 90 % live on the island of Java and the rest outside the island of Java. The educational standard of participants is now 74 % of candidates, 14 % of respondents of undergraduate, and 3 percent of graduates of graduate. A survey is conducted via Google Forms. To find out the answers of the participant a survey is administered through Google Forms. As a result, 58% reacted favourably to all events, and 36% were dissatisfied. The negative response was due to weak management of the committee, poor coordination of the committee, very little time, internet signal limitations, and inadequate sound quality. In general, participants recognize the creation of innovative goods relevant to global pandemics and waste emergencies.
... Depending on the composition, some of these waxes can bear antimicrobial activity and one of the most representative is the beeswax, its antimicrobial activity is being assigned to propolis. Pinto et al. [60] presented the antimicrobial activity of Abeego food wrap (a beeswax based product along with tree resin and oils, both as minor components), the coating is being realized using a propolis-rich fraction of the beeswax. According to their study, Abeego food wrap exhibited a strong antibacterial activity against some representative Gram-positive and Gram-negative bacteria (Salmonella entritidis and Staphyloccocus aureus) but not against fungal or viral strains. ...
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This paper offers a general view of the solutions that are able to confer bioactivity to the packaging materials, especially antimicrobial and antioxidant activity. These properties can be induced by the nature of the polymers blend or due to the addition of ternary components from natural agents (essential oils or other extracts) to synthetic organic and inorganic agents, including nanoparticles with a broad antimicrobial activity such as metals (e.g., Ag, Au, Cu) or metal oxide (e.g., TiO2, ZnO) nanoparticles, and even bacterial cells such as probiotics. Many times, these components are synergistically used, each of them assuring a specific role or potentiating the role of the other components. The antimicrobial activity can be induced due to the applied coatings or due to the whole bulk material. Along with an increasing food stability which means a longer shelf-life some smart packaging can be exploited in order to highlight the freshness of the food. These act as a sensor (usually pH sensitive but also other mechanisms can be exploited such as aggregation/agglomeration of AuNPs leading to color change or even aldehyde-specific reactions such as the Cannizzaro reaction), and thus, consumers can be confident about the freshness of the food, especially perishable food such as seafood or fish.
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Microorganisms are the major cause of spoilage in most seafood products; however, only few microbes, called the specific spoilage organisms (SSOs), contribute to the offensive off-flavors associated with seafood spoilage. In food, microbial degradation manifests itself as spoilage, or changes in the sensory properties of a food product, rendering it unsuitable for human consumption. The use of antimicrobial substances can control the general microflora as well as specific microorganisms related to spoilage to provide products with higher safety and better quality. Many antimicrobial compounds have been evaluated in film structures for use in seafood, especially organic acids and their salts, enzymes, bacteriocins; some studies have considered inorganic compounds such as AgSiO2, zinc oxide, silver zeolite, and titanium oxide. The characteristics of some organic antimicrobial packaging systems for seafood and their antimicrobial efficiency in film structures are reviewed in this article.
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Nowadays food preservation, quality maintenance, and safety are major growing concerns of the food industry. It is evident that over time consumers’ demand for natural and safe food products with stringent regulations to prevent food-borne infectious diseases. Antimicrobial packaging which is thought to be a subset of active packaging and controlled release packaging is one such promising technology which effectively impregnates the antimicrobial into the food packaging film material and subsequently delivers it over the stipulated period of time to kill the pathogenic microorganisms affecting food products thereby increasing the shelf life to severe folds. This paper presents a picture of the recent research on antimicrobial agents that are aimed at enhancing and improving food quality and safety by reduction of pathogen growth and extension of shelf life, in a form of a comprehensive review. Examination of the available antimicrobial packaging technologies is also presented along with their significant impact on food safety. This article entails various antimicrobial agents for commercial applications, as well as the difference between the use of antimicrobials under laboratory scale and real time applications. Development of resistance amongst microorganisms is considered as a future implication of antimicrobials with an aim to come up with actual efficacies in extension of shelf life as well as reduction in bacterial growth through the upcoming and promising use of antimicrobials in food packaging for the forthcoming research down the line.
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The deposition of antimicrobial plant resins in honey bee, Apis mellifera, nests has important physiological benefits. Resin foraging is difficult to approach experimentally because resin composition is highly variable among and between plant families, the environmental and plant-genotypic effects on resins are unknown, and resin foragers are relatively rare and often forage in unobservable tree canopies. Subsequently, little is known about the botanical origins of resins in many regions or the benefits of specific resins to bees. We used metabolomic methods as a type of environmental forensics to track individual resin forager behavior through comparisons of global resin metabolite patterns. The resin from the corbiculae of a single bee was sufficient to identify that resin's botanical source without prior knowledge of resin composition. Bees from our apiary discriminately foraged for resin from eastern cottonwood (Populus deltoides), and balsam poplar (P. balsamifera) among many available, even closely related, resinous plants. Cottonwood and balsam poplar resin composition did not show significant seasonal or regional changes in composition. Metabolomic analysis of resin from 6 North American Populus spp. and 5 hybrids revealed peaks characteristic to taxonomic nodes within Populus, while antimicrobial analysis revealed that resin from different species varied in inhibition of the bee bacterial pathogen, Paenibacillus larvae. We conclude that honey bees make discrete choices among many resinous plant species, even among closely related species. Bees also maintained fidelity to a single source during a foraging trip. Furthermore, the differential inhibition of P. larvae by Populus spp., thought to be preferential for resin collection in temperate regions, suggests that resins from closely related plant species many have different benefits to bees.
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Background Human coronaviruses (HCoVs) are a well-known cause of respiratory infections but their role in gastrointestinal infections is unclear. The objective of our study was to assess the significance of HCoVs in the etiology of acute gastroenteritis (AGE) in children <6 years of age. Methods Stool samples and nasopharyngeal (NP) swabs collected from 260 children hospitalized for AGE (160 also had respiratory symptoms) and 157 otherwise healthy control children admitted for elective surgery were tested for the presence of four HCoVs using real time RT-PCR. Registered at (reg. NCT00987519). Results HCoVs were more frequent in patients with AGE than in controls (23/260, 8.8% versus 4/151, 2.6%; odds ratio, OR 3.3; 95% confidence interval, CI 1.3–10.0; P = 0.01). Three of four HCoV-positive members in the control group, asymptomatic when sampled, recalled gastrointestinal or respiratory symptoms within the previous 14 days. In patients with AGE, HCoVs were present in NP samples more often than in stools (22/256, 8.6%, versus 6/260, 2.3%; P = 0.0004). In 5/6 children with HCoVs detected in stools, the viruses were also detected in NP swabs. Patients had a significantly higher probability of HCoV detection in stool (OR 4; 95% CI 1.4–15.3; P = 0.006) and also in stool and/or NP (OR 3.3, 95% CI 1.3–10.0; P = 0.01) than healthy controls. All four HCoVs species were detected in stool and NP samples. Conclusions Although HCoVs were more frequently detected in patients with AGE than in the control group, high prevalence of HCoVs in NP swabs compounded by their low occurrence in stool samples and detection of other viruses in stool samples, indicate that HCoVs probably play only a minor role in causing gastrointestinal illness in children <6 years old.
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Invasive strains of non-typhoidal salmonellae have emerged as a prominent cause of bloodstream infection in African adults and children, with an associated case fatality of 20-25%. The clinical presentation of invasive non-typhoidal salmonella disease in Africa is diverse: fever, hepatosplenomegaly, and respiratory symptoms are common, and features of enterocolitis are often absent. The most important risk factors are HIV infection in adults, and malaria, HIV, and malnutrition in children. A distinct genotype of Salmonella enterica var Typhimurium, ST313, has emerged as a new pathogenic clade in sub-Saharan Africa, and might have adapted to cause invasive disease in human beings. Multidrug-resistant ST313 has caused epidemics in several African countries, and has driven the use of expensive antimicrobial drugs in the poorest health services in the world. Studies of systemic cellular and humoral immune responses in adults infected with HIV have revealed key host immune defects contributing to invasive non-typhoidal salmonella disease. This emerging pathogen might therefore have adapted to occupy an ecological and immunological niche provided by HIV, malaria, and malnutrition in Africa. A good understanding of the epidemiology of this neglected disease will open new avenues for development and implementation of vaccine and public health strategies to prevent infections and interrupt transmission.
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Staphylococcus aureusis a one of THE most frequent causes of food poisoning (FP) in dairy products. The main etiologic agents of FP are staphylococcal enterotoxins (SE). There are different types of SE; types A (SEA) and B (SEB) are the most clinically important enterotoxins. Traditional dairy products are still produced in small batches and sold by some vendors without a permit from the Ministry of Health. This study focuses on the molecular and serological detection of enterotoxigenic Staphylococcus aureus SEA and SEB genes and its products, respectively from samples of such traditional products. 100 samples from dairy products were produced under sterile conditions via traditional methods and were transported to the laboratory. The samples were cultured and identified by routine bacteriological methods. The isolated bacteria were evaluated by PCR tests for detection of the genes encoding SEA and SEB. Subsequently, the ability of these strains to produce enterotoxin was examined by Sac's culture method and was confirmed by Sigel Radial Immounodiffussion (SRID). The results indicated that 32% of the dairy products were contaminated by S. aureus (cream 18%, cheese 10%, milk 4%). The PCR results showed that 15.6% of the S. aureus isolates possessed the SEA gene, 9.3% had the SEB gene, and 6.2% possessed both genes. The evaluation of enterotoxin production indicated that 80% of SEA and 33% of SEB genes were expressed. Enterotoxins SEA and SEB are heat stable and consequently; heating has no effect on dairy products contaminated by entertoxins. Subsequently, gastritis may occur within several hours after consumption. Our findings suggest that PCR is a rapid, sensitive, specific, and inexpensive method for detecting SE and can replace the traditional assays.
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Propolis antibiotic action has been widely investigated. This assay was carried out in order to observe the in vitro antibacterial activity of propolis against Salmonella enteritidis isolated from food and Salmonella typhimurium isolated from human infections. Propolis was collected by Apis mellifera in two regions of Brazil (Mossoró, Rio Grande do Norte State; and Urubici, Santa Catarina State). Both strains survival percentage decreased with time of incubation in Ethanolic Extracts of Propolis (EEP), demonstrating bactericidal effect after 24 hours. It was also observed that EEP from Mossoró was more effective than that from Urubici. The control of the propolis solvent - 70% ethanol - was less effective than EEP, showing only a bacteriostatic effect. We can conclude that propolis shows an activity against Gram-negative bacteria that varies according to the geographical region where it was collected by bees.
Propolis is a substance derived from antimicrobial plant resins that honey bees use in the construction of their nests. Propolis use in the hive is an important component of honey bee social immunity and confers a number of positive physiological benefits to bees. The benefits that bees derive from resins are mostly due to their antimicrobial properties, but it is unknown how the diversity of antimicrobial activities among resins might impact bee health. In our previous work, we found that resins from different North American Populus spp. differed in their ability to inhibit in vitro growth of the bee bacterial pathogen Paenibacillus larvae. The goal of our current work was to characterize the antimicrobial activity of propolis from 12 climatically diverse regions across the US against the bee pathogens P. larvae and Ascosphaeraapis and compare the metabolite profiles among those samples using LC–MS-based metabolomic methods. Samples differed greatly in their ability to inhibit both bacterial and fungal growth in vitro, but propolis from Nevada, Texas, and California displayed high activity against both pathogens. Interestingly, propolis from Georgia, New York, Louisiana, and Minnesota were active against A. apis, but not very active against P. larvae. Metabolomic analysis of regional propolis samples revealed that each sample was compositionally distinct, and LC–FTMS profiles from each sample contained a unique number of shared and exclusive peaks. Propolis from Aspen, CO, Tuscon, AZ, and Raleigh, NC, contained relatively large numbers of exclusive peaks, which may indicate that these samples originated from relatively unique botanical sources. This is the first study to characterize how the diversity of bee preferred resinous plants in the US may affect bee health, and could guide future studies on the therapeutic potential of propolis for bees.
The guanidine-based antimicrobial polymers were grafted onto the surface of beeswax latex particles stabilized with or without amphoteric surfactant, n-dodecyl-N,N-dimethylglycine. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) was used as a coupling reagent to introduce the covalent bonding or ensure sufficient binding force. The resulting novel beeswax latexes were utilized as a dual-functional paper additive, which improved both water-vapor resistance and antimicrobial activities of the paper. Compared with control sample, the water vapor transmission rate of the paper was reduced significantly. The antimicrobial activity of paper against E. coli. was also improved substantially even at a low dosage of the beeswax latex derivatives (20 mg/g fibre).