BAOJ Microbiology - Bio Accent

BAOJ Microbiology Asaad Mohammed A Ataa, BAOJ Microbio 2018, 4: 2 4: 034

Research

Study of the Effects of YEMECIN Antibiotic on Bacteria Causing Food Corruption and also as a Preservation for Milk, Meat and Canned Food Asaad Mohammed A Ataa* Department Chemistry, Division Biochemistry, Faculty Applied Science, Thamar University

Abstract

Keywords: YEMECIN antibiotic; Food; Resistant; Sensitivity;

Introduction and Objectives

Escherichiacoli; Salmonellatyphi; Streptococcusmutans; Staphylococcusaureus.

This study aims to identify the effectiveness The YEMECIN antibiotic this is new antibiotic it’s discovery in Yemen. This is studies their effects on bacteria Gram Positive (Streptococcus mutans, Staphylococcus aureus) and Gram Negative (Escherichia. coli, Salmonella typhi), which have been obtained pure in hospitals and others have been isolated at Laboratory . Then we face the foods which are contaminated with such species of these bacteria and their dangers on the public health.

Materials and Methods The study was conducted in two phases, the first of which included knowing the effectiveness of extracts and their effects on some sorts of bacteria which were treated by these extracts through the tests of sensitivity screening. The second phase of this study included the treatment of some types of food which are commonly circulated and speedily damaged – the treatment was with these materials to know their impact in eliminating the microbes that are found in these foods and prevent damage for a long time. We examined for types of bacteria Gram (Positive and Negative): Streptococcus mutans, Staphylococcus aureus, E.coli, Salmonella typhi.

Introduction A rich history Garlic is one of the edible plants, which has generated a lot of interest throughout human history as a medicinal panacea. A wide range of microorganisms including, bacteria, fungi, protozoa and viruses have been shown to be sensitive to crushed garlic preparations. Moreover, garlic has been reported to reduce blood lipids and to have anticancer effects. Chemical analyses of garlic cloves have revealed an unusual concentration of sulphur-containing compounds (1-3%) [1,2]. A quick search of the medical database at the National Library of Medicine in the USA reveals that garlic is top of the league for published research papers that cover a wide variety of disease conditions, the most prevalent of which are its significant antimicrobial properties [1]. Historically, medicinal plants have been a source of novel drug compounds. Plants derived products have made large contributions to human health and well-being. Green pharmacy may become the base for the development of medicines by providing a pharmacophore which could be used for the development of new drug with novel mechanisms of action [2]. Garlic

Results The results of sensitivity screening, which were done by two ways: (CDproliferation plates and confirmation way), showed the sensitivity of these microbes with rates ranging between (100%) for YEMECIN antibiotic to (33.4%) for X, X1 antibiotics. It was high affected for gram positive bacteria, while the less ones were antibiotic gram negative bacteria. Besides, tow samples of food (Milk and Meat), which are commonly circulated in the governorate, were tested and treated with YEMECIN antibiotic.

The inferences The results showed the effectiveness of YEMECIN antibiotic with its quality in eliminating most microbes in meat especially E.coli and its effectiveness in eliminating all microbes in milk with less influence on Salmonella typhi. But it eliminated in the tow samples with no influence on E.coli. BAOJ Microbio, an open access journal

*Corresponding Author: Asaad Mohammed A Ataa, Department Chemistry, Division Biochemistry, Faculty Applied Science, Thamar University, Yemen, E-mail: [email protected] Sub Date: April 5th 2018, Acc Date: April 25th 2018, Pub Date: April 26th 2018. Citation: Asaad Mohammed A Ataa (2018) Study of the Effects of YEMECIN Antibiotic on Bacteria Causing Food Corruption and also as a Preservation for Milk, Meat and Canned Food. BAOJ Microbio 4: 034. Copyright: © 2018 Asaad Mohammed A Ataa. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Volume 4; Issue 2; 034

Citation: Asaad Mohammed A Ataa (2018) Study of the Effects of YEMECIN Antibiotic on Bacteria Causing Food Corruption and also as a Preservation for Milk, Meat and Canned Food. BAOJ Microbio 4: 034.

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(Allium sativum) is a common spice used for flavouring and has been traditionally popular with strong folkloric awareness. It is the edible bulb of lily family, Liliaceae. It contains aromatic sulphur based compounds, which contribute to the characteristic odour and taste. Antimicrobial activity of garlic is attributed to its key component Allicin [3].

Isolate of Escherichia coli

Objectives of the study

I took the ready-made seclusion of bacteria Streptococcus mutans then it was reoriented on the Bleed Agar, and then it was activated by putting it in Nutrient broth until it was used.

1.

This study aims to identify the effectiveness of the YEMECIN antibiotic, which is a new antibiotic discovered in Yemen.

2.

Using the YEMECIN antibiotic in maintaining some foods rite (Milk, Meat, Canning food).

I took the ready-made seclusion of bacteria E.coli Then it was activated on MacConcky Agar [6]. Isolate of Streptococcus mutans

Isolate of Staphylococcus aureus

We brought distilled water, YEMECIN antibiotic, Sodium Hydroxide (NaOH), and Bromocresol Purple dye.

We used the replica plate method to identify the recovered aerosolized culturable bacteria by transferring the bacterial colonies onto a selective medium [7]. For this method we used Chapman Stone medium (CSM; Difco Laboratories) for identification of S. aureus. After pressing of the CSM, TSA was used as a final control for the method, being pressed first and last to ensure that the organisms were completely transferred to all plates. All plates were incubated at 35°C and counted at 24 and 48 hr. The presence of S. aureus was further confirmed using Bacto coagulase plasma (Fisher Scientific, Houston, TX). Once counted, the plates were refrigerated in an inverted position at 4°C until used for the Kirby-Bauer disk diffusion method.

Sterilization

Isolate of Salmonella typhi

All work has been carried out under strict sterilization procedures and sterile conditions during the course of isolation, reclamation, bacteriological and dietary tests.

5.5.5.1 Selective Enrichment Phase: Taken from a sample of chicken livers (grilled) by 25 grams, it was developed in 100 ml of the selenite broth Food Center at 37 oC and 24 hours a day. For the purpose of activating bacteria Salmonella [8].

Materials and Methods Culture Media I attended all agricultural communities as instructed by the company processed and sterilized with a temperature of autoclaves (121) oC and under pressure (15) lbs / Wang for period of (15) minutes.

Solutions and Dyes Used

Devices and Tools Devices: Centrifuge, a device measure pH, a delicate balance, refrigerator cooling room, hood, and an incubator. Tools and Glassware: Picrate glass of different sizes, Petri dishes, glass pipettes, test tubes, DH pipes, Loeb, a Needle, a filter paper, and paper punch. YEMECIN extraction We chose garlic with its kinds (Fresh, Fruit) available in the local market in the province, which is known as (garlic male). After Cushing garlic cloves, we extracted the liquid product. The liquid is filtered by filter paper into the clean steriled dry glass pipes and each sample is put separately until use. Saved for YEMCIN antibiotic (7days to three months) under 7 Co [4,5]. Isolate Streptococcus mutans, Staphylococcus aureus, E.coli, Salmonella typhibacteria We obtained them with the help of some Sana’a hospitals like (AlThawra, and Seventy Hospitals), and from (Al-Awolaqi laboratories) in Yemen.

BAOJ Microbio, an open access journal

Working Methods Sensitivity screening test for Streptococcus mutans, Staphylococcus aureus, E.coli, Salmonella typhibacteria Disk Diffusion: It is way in which we use the pre -wetted filter paper in the extracts of (YEMECIN) antibiotic. The circles are prepared which are used to develop the bacteria mentioned earlier. Also preparing the extracts of (YEMECIN) antibiotic and Pharmacy antibiotics to be put in the clean, striated and day pipes. Then patting a certain amount of filter paper which was formed in the form of small disks in a diameter ranging from 0.5 cm; in each tube there is an extracts of (YEMECIN) antibiotic and Pharmacy antibiotics. The disks should be left in the tube for a period ranging between a half an hour to two hours to absorb a solfiaent amount of (YEMECIN) antibiotic and Pharmacy antibiotics [8]. After isolating bacteria, we table the filter paper saturated with the extracts of (YEMECIN) antibiotic and Pharmacy antibiotics and put it in different places by sterilized forceps with alcohol and flame to install on the surface by the light pressure by forceps; each one should be alone [9]. After words, incubating the grath dishes on a temperature of 37°C and for a period ranging between (24-48) hours [10].



Confirmatory Test to Detect the Inhibitory Ability of the Extract of (YEMECIN) Antibiotic and Pharmacy Antibiotics This way is dove by pouring the extract of (YEMECIN) antibiotic and Pharmacy antibiotic son the growth after transplanting bacteria by planning to observe the inhibition. The way is the some way of (Disk-proliferation), but the difference lies in that after isolation, the process of pouring a certain amount of (YEMECIN) antibiotic and Pharmacy antibiotic son the growth disks and placing the dishes into the incubator to grow at a temperature of 37 ° C for a period, the results are recorded. All work has been carried out under strict sterilization procedures and sterile conditions during the course of isolation, reclamation, bacteriological and dietary tests.

Bacteriological Examination of Food Meat To conduct this process samples of meat (minced meat), pulling 5g of Homogeneous hash in each tube. Then adding one ml of YEMECIN antibiotic with pharmacy antibiotics to each tube separately. The additions are as follows. One ml of YEMECIN antibiotic was added to the first tube; one ml of P antibiotic was added to the second tube; One ml X antibiotic was added to the third fourth tube; one ml of X1 antibiotic was added to fourth tube, except the fifth tube which was not treated by any antibiotics, however, one ml of distilled water was added to it. Then the samples of meat were put in the refrigerator lilt use. Save samples under temperature 7° C for time 2 to 8 days.

Milk The samples of milk were prepared by adding 10 ml of them to each tube [5] tubes. Then one ml of YEMECIN antibiotic extract and P antibiotic, X antibiotic, X1 antibiotic, and mixed X, X1 antibiotics, was added to five tubes, while the milk in the sixth tube was contaminated by the fecal E.coli which contaminates blood. But, the seventh tube was not treated by any sample of the research samples or any bacteria. It was like (a standard sample). All samples were put in the refrigerator lilt using. Save samples under temperature 7° C for time 2 to 8 days.

Prepare the Medium (Lactose Broth) Lactose Broth was prepared after dissolving an amount of 3.25 g in 250 ml of distilled water adding the dye of purple Bromocresol with an amount of 0.112 grams for each 250 ml of distilled water. Then Lactose broth was dissolved adding to it drops of Sodium Hydroxide (NaOH), in order to raise value of pH to [7.8] By the pH Meter, compared to the use of an Buffer solution on purpose to know the charge of the colour in the dye as a result of low pH and at the fermentation of lactose. After that, 10 ml of Lactose Broth were adding to all tube used. The tubes then were placed, down to sterilized Lactose Broth according to instructions of the company [7]. BAOJ Microbio, an open access journal

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Lactose Fermentation Test Part I: Quantitative and Presumptive Test In this part, some samples of meat were prepared and a central centrifuge period of 5 minutes for the purpose of extracting the filtrate. After sterilizing the circle available in the tubes, the vaccination was done by taking a point of milk and meat samples which were added to the tubes separately. After vaccinating the tubes, they were covered by a layer of sterilized was at 121 ° C for a period of 20 minutes. Then cotton was put on each tube for incubating at 37 ° C for 24-48 hours. Directly after the incubation, the results were written down.

Part II: Confirmatory Test After noting the results of lactose fermentation and the composition of the gas, the centres were prepared which are special for isolation of bacteria types which are expected to come out. The media used for that are as follows:

Nutrient, MacConcky, EMB, S.S. Agar After sterilization the oracles, they were poured on the plates and then leaving them to harden. During this process the samples on which the gas was composed were prepared. Also, one tube was taken which represents the samples in which lactose was fermented without composing gas. Loob was taken from the tube after exposing it to the flame to sterilize and vaccinate media planning, and the rest of the work was archer went like wise. After the transplant process by planning on media, the plates were put for incubating and the incubation was at different temperatures between (37- 44.5) ° C. The results were observed and written down.

Part III: Completed Test (Final Test) After the growth of the colonies in the media, loop was taken to the tubes containing Lactose Broth and to the upside down tubes after sterilization. Incubation was at 37° C for a period of 24-48 hours. After the incubation, the results were observed and recorded.

Results The results in Table 1 refer to the sensitivity of YEMECIN antibiotic, P antibiotic , X antibiotic & X1 antibiotic,and their percentages. The results of sensitivity examination state the sensitivity of these microbes for most of these extracts used in this study with rates range between (100%) for YEMECIN antibiotic, (83.4%) for P antibiotic, and (66.6%) for X antibiotic. (50%) for X1 antibiotic, and all kinds of X1,X antibiotics. (33.4%) for antibiotics. The differences can be-seen through the sensitivity of YEMECIN and antibiotics. can be seen that variation in sensitivity through various forms of ID (1, 2, 3)



4

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1 3

1 3

2

4

2

Figure 1: The inhibitory capacity of the extracts on, Staphylococcus aureus, method Disk Diffusion. (1) YEMECIN antibiotic, (2) P antibiotic, (3) X antibiotic,(4) X1 antibiotic. Zone of inhibition (1) 3 Cm, (2) 2.5 Cm, (3) 2 Cm, (4) 1.2 Cm.

Figure 2: The inhibitory capacity of the extracts on, E.coli, method Disk Diffusion. (1) YEMECIN antibiotic, (2) P antibiotic, (3) X antibiotic,(4) X1 antibiotic. Zone of inhibition (1) 2.3 Cm, (2) 2 Cm, (3) 1.4 Cm, (4) 1 Cm.

3 2

4 1 3

1 2

4

Figure 3: The inhibitory capacity of the extracts on, Streptococcus mutans, method Disk Diffusion. (1) YEMECIN antibiotic, (2) P antibiotic, (3) X antibiotic,(4) X1 antibiotic.Zone of inhibition (1) 3.9 Cm, (2) 3.2 Cm, (3) 2.6 Cm, (4) 2.4 Cm

Figure 4: The inhibitory capacity of the extracts on, Salmonella typhi, method Disk Diffusion. (1) YEMECIN antibiotic, (2) P antibiotic, (3) X antibiotic,(4) X1 antibiotic. Zone of inhibition (1) 1.8 Cm, (2) 2.4 Cm, (3) 0.6 Cm, (4) 0 Cm.

Table 1: The Percentage of Sensitivity and Resistance of Bacteria Isolated Extract (YEMECIN, Antibiotics) Extracted

Number

The Test Number of Times

Sensitivity

%

The Number of Resistance

%

YEMECIN

1

3

3

100

-

-

P antibiotic

2

6

3

83.4

3

16.6

X1 antibiotic

3

6

5

50

1

50

X antibiotic

4

6

2

66.6

4

33.4

X, X1 antibiotics

5

6

5

33.4

1

66.6




Graph 1: Sensitive Bacteria (YEMECIN and Antibiotics) (Method of Disks Spread)

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The affected bacterium was Streptococcus mutans, Staphylococcus aureus treated by YEMECIN antibiotic, but it was not affected by P antibiotic, X antibiotic and X1 antibiotic. Regarding the effectiveness of P antibiotic was high on Salmonella typhi and no the plates treated. When mixing X antibiotic and X1 antibiotic both, these is growth. Regarding E.coli the results stated that this bacteria was affected by YEMECIN antibiotic with less affected by all Antibiotics (Disk diffusion way) [7]. For Salmonella bacteria, the result stated the effectiveness of P antibiotic on Salmonella typhi, with less affected by YEMECIN antibiotic, however these was no growth for this bacteria when using the X1 antibiotic, those is no growth when mixing X antibiotic and X1 antibiotic both, these is growth. These mean that the mixture of X and X1 antibiotic does not give effectiveness. Some results did not state the effectiveness of YEMECIN extracts by using them separately or mixing them together on Salmonella typhi. Through the results, it was stated that there is on effect of YEMECIN antibiotic on bacteria E.coli, Streptococcus mutans, &Staphylococcus aureus.These results correspond with [8], which state that is an effect on bacteria E.coli. Lysozyme effect on bacteria Salmonella was similar to (8). study which stated that there is effectiveness on Salmonella.

Bacteriological Examination of Food Meat Through the experiments conducted on YEMECIN and Antibiotics with their phases (first and second), the experiment states that YEMECIN antibiotic has an effect on E. Coli, Staphylococcus aureus and limited on Streptococcus mutans. On the other hard, it was rot rated that it has an effect on Salmonella typhi, although, it has strong effectiveness in eliminating most microbes found in most. This corresponds [3]. More over it has strong effectiveness in eliminating all microbes found in milk when treating with these extracts (local and foreign).

Graph 2: The Sensitivity of Bacteria to (YEMECIN and Antibiotics) Sensitive bacteria method of disks spread and confirmed test YEMECIN antibiotic 87.5% P antibiotic 56.25% X antibiotic 50% X1 antibiotic 37.5% Strep: Streptococcus mutans E.coli: Escherichia. Coli Sal: Salmonella typhi Staph: Staphylococcus aureus

Milk Results have shown a sample of Milk, which is known to milk from the appropriate sources for the growth of various microorganisms and which ones cause damaged. The results that YEMECIN antibiotic is highly effective not prove ferment lactose or gas in the pipeline that treated milk which YEMECIN antibiotic remained colored as color

Table 2: The Fermentation of Lactose, And the Product Gas in a Sample Of Meat Number Tube

Meat-Treatment Sample

The Result Fermentation of Lactose

1 2 3 4 5 6

YEMECIN P antibiotic X antibiotic X1 antibiotic X, X1 antibiotic Control

+ + + + + +


The Result Be Gas − − + + + + Volume 4; Issue 2; 034


pigment blue Dark, and was also confirmation of the results through the implant on different media which did not show any colonies in central EMB, MacConcky Agar and S.S. Agar. The P antibiotic has demonstrated the effectiveness of the approach of YEMECIN antibiotic it of P antibiotic and X antibiotic has effective in the inhibition did not show any planting colonies when the central S.S. Agar, X1 antibiotics ,and mixed X, X1 antibiotics did not have an impact notes ferment lactose with gas formation and colonies appeared in various media used. Through studies conducted on extracts Lysozyme and YEMECIN antibiotic stages I and II experience has shown the influence of these

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substances uneven and that was YEMECIN antibiotic has an effect on E.coli effectively and limited Streptococcusmutans was observed effect on Salmonellatyphi, and effectiveness of high eliminate most microbes found in meat This corresponds to what is stated in [8], and its effectiveness concluded in eliminating all of the microbes in the milk counter when treated in this local and external quality, compared with the standard sample. And effectiveness in eliminating bacteria Salmonella found in milk and this corresponds to what Lord’s [8,10], and is identical to a previous study conducted for [4], which explained all to the effectiveness of YEMECIN antibiotic Salmonella typhi.

Table 3: The Fermentation of Lactose, and the Product Gas in a Sample of Milk Number Tube

Meat-Treatment Sample

The Result Fermentation of Lactose

The Result Be Gas

1

YEMECIN

−

−

2

P antibiotic

−

−

3

X antibiotic

−

−

4

X1 antibiotic

+

+

5

X, X1 antibiotic

+

+

6

Control

−

−

Microbiological Food Quantitative and Presumptive tests

Conclusions and Recommendations Conclusions 1- High efficiency of the YEMECIN antibiotic with P antibiotic on Streptococcusmutans and Salmonellatyphi&Staphylococcus aureus. 2- The effectiveness of some food; the local one was more effective among them. 3- The effectiveness of YEMECIN antibiotic in eradicating E.coli in all transactions. 4- The effectiveness of YEMECIN antibiotic and its ineffectiveness in the other treatments with its local and foreign types on bacteria Streptococcusmutans and Salmonellatyphi. 5- The possibility of using YEMECIN in some foods, quickly damaged because of their impact on many microbe existing in foods causing damage, as well as food poisoning. 6- The effectiveness of X antibiotic on microbes when treating foods which it.




Recommendations 1- Studying the possibility of using YEMECIN antibiotic in the field, of food processing such as improving conservation meat with its kinds. 2- Extraction and purifying the components of both YEMECIN and P, X,X1 antibiotics and studying their effect on microbes which cause food damage and diseases widely.

9.

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Klancnik A, Piskernik S, Jersek B, Mozina SS, (2010) Evaluation of diffusion and dilution methods to determine the antibacterial activity of plant extracts. J Microbiol Methods 81: 121-126.

10. Gandara A, Mota LC, Flores C, Perez HR, Green CF, et al. (2006) Isolation of Staphylococcus aureus and antibiotic-resistant Staphylococcus aureus from residential indoor bioaerosols 114: 1859-64.

3- Using YEMECIN antibiotic pure in treating meat, milk and other foods, especially those which Coli form increases and those that are kept for a long time because YEMECIN antibiotic is effective against microbes. It is of high therapeutic and healthy, importance with using P antibiotic with foods in which largely Salmonellatyphi is found especially meat and us derivatives for X1 antibiotic has strong effectiveness on Salmonella typhi.

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Koch HP, Lawson LD (1996) Garlic, the science and therapeutic application of Allium sativum L. and related species. Williams & Wilkins 1-233.

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Kim JS, Kim YH (2007) The inhibitory effect of natural bioactives on the growth of pathogenic bacteria. Nutrition Research and Practice 1: 273-278.

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Ataa Asaad Mohammed (2017) Use of Enzyme Lysozyme & antibacterial Allicin for food preservation and prevent at damage. World Journal of Biology and Biotechnology 02: 169-173.

