Publication: Aktif İçeriği Sodyum Dimetilditiyokarbamat Olan Antimikrobiyal Maddenin Antibiyotiklere Dirençli Referans Bakteri Suşlarına Karşı Minimum İnhibitör Konsantrasyonlarının Araştırılması
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Tuzlanmış deriler, kaybettiği suyun geri kazandırılarak yeniden yumuşaması ve üzerindeki tuzun, dışkının, mikroorganizmaların temizlenmesiamacıyla ıslatma işlemine sokulmaktadır. Deri endüstrisinde ıslatma işleminin içerdiği organik yük sebebiyle bakteriler gelişebilmektedir.Bakteriyel gelişmenin deride önemli zararlara sebep olduğu bilinmektedir. Islatma işleminde çeşitli antimikrobiyal maddelerkullanılmasına rağmen, ıslatma sıvılarında ve ıslatılmış derilerde antibiyotiklere dirençli Gram-pozitif ve Gram-negatif bakterilerdaha önceki çalışmalarda saptanmıştır. Islatma işleminin verimli ve etkili bir biçimde yapılabilmesi için antimikrobiyal maddenin uygunmiktarda kullanılması oldukça önemlidir. Böylece, derilerde bakteri kaynaklı zararlar kontrol altına alınabilecektir. Bu çalışmada,Gram-negatif ve Gram-pozitif referans bakteri suşlarının farklı antibiyotiklere karşı duyarlılıkları Kirby-Bauer disk difüzyon metodu ilearaştırılmıştır. Ayrıca, ıslatma işleminde kullanılan aktif içeriği sodyum dimetilditiyokarbamat olan antimikrobiyal maddenin farklı konsantrasyonlarıbu bakterilere ve bunların karışık kültürü üzerine uygulanarak minimum inhibisyon konsantrasyonu agar dilüsyon metoduylaaraştırılmıştır. Test bakterileri ampisilin (10 µg), spektinomisin (25 µg), streptomisin (10 µg), meropenem (10 µg) ve penisilin (10 U) antibiyotiklerinekarşı dirençli bulunmuştur. Antimikrobiyal maddenin minimum inhibisyon konsantrasyon değerleri Pseudomonas aeruginosaATCC27853, Staphylococcus aureus ATCC29213, Micrococcus luteus ATCC9341, ve tüm test bakterilerinin karışık kültürü için 1000µg/ml; Escherichia coli ATCC25922 ve Bacillus cereus ATCC11778 için 500 µg/ml; Staphylococcus epidermidis ATCC12228 ve Enterococcusfaecalis ATCC29212 için 15.6 µg/ml; ve Bacillus subtilis ATCC6633 için ise 3.9 µg/ml olarak saptanmıştır. Bu çalışmadan eldeedilen verilere göre, antibiyotiklere dirençli sekiz farklı referans bakteri suşunun ve bunların karışık kültürünün gelişimleri test edilen antimikrobiyalmadde ile inhibe edildiği belirlenmiştir.
The leather industry is an important commercial sector for the global economy. The hides and skins are contaminated with microorganisms found in air, soil, water, feces and slaughterhouse. After the animal is flayed, the hide and skin provide an ideal growth condition for these microorganisms due to containing fat and protein. Different chemical agents such as sodium chloride, boric acid, antifungal and antibacterial agents are used to preserve raw hides and skins in order to prevent microbial growth and damage on hides and skins during transportation and storage period. The salted hides are soaked in soaking process in order to re-soften and removing the salt, feces and microorganisms on the hide surface. Bacteria can develop in the leather industry due to the organic load in the soaking process. Bacterial growth is known to cause significant damage to the hides and skins such as discoloration, bad odor, hair slip, grain peeling, fiber disintegration, weakness, looseness and holes. Although various antimicrobial agents have been used in the soaking process, Gram-positive and Gram-negative bacteria resistant to antibiotics in soaking liquids and soaked hides have been identified in previous studies. It is very important to use the antimicrobial agent in the proper amount so that the soaking process can be carried out efficiently and effectively. Thus, bacterial damage can be controlled in the skin. Hence, the aim of the present study was to investigate the susceptibilities of Gram-negative (Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922) and Gram-positive (Staphylococcus epidermidis ATCC 12228, Staphylococcus aureus ATCC 29213, Micrococcus luteus ATCC 9341, Enterococcus faecalis ATCC 29212, Bacillus cereus ATCC 11778, Bacillus subtilis ATCC 6633) reference bacterial strains to five different antibiotics (ampicillin, spectinomycin, streptomycin, meropenem and penicillin) by Kirby-Bauer disc diffusion susceptibility test method on Mueller Hinton Agar. In addition, 18 different concentrations of the antimicrobial agent (4000 µg/ml, 2000 µg/ml, 1000 µg/ml, 500 µg/ ml, 250 µg/ml, 125 µg/ml, 62.5 µg/ml, 31.2 µg/ml, 15.6 µg/ml, 7.8 µg/ml, 3.9 µg/ml, 1.96 µg/ml, 0.98 µg/ml, 0.49 µg/ml, 0.24 µg/ml, 0.12 µg/ml, 0.06 µg/ml and 0.03 µg/ml), containing sodium dimethyldithiocarbamate as the active ingredient and used in the soaking process, were applied to these bacteria and their mixed culture and the minimum inhibitory concentration was investigated by agar dilution method. The series of two-fold dilutions of the test agent, ranging from 4000 µg/ml to 0.03 µg/ml, were prepared in Mueller Hinton Agar. The test strains were grown on Mueller Hinton Agar at 37 °C for 24 hours. Then, overnight cultures of the test strains were inoculated into 10 ml Mueller Hinton Broth with 2 colonies of each test strain. Each bacterial suspension was adjusted to a McFarland standard 0.5 (108 colony forming units per ml). The broth cultures of test strains were incubated at 37 °C for 24 hours. Later, each suspension of test strain was diluted to 107 colony forming units per ml using physiological saline water containing 0.85% sodium chloride. The mixed culture of the test strains was also prepared from these dilutions. An inoculum of 104 colony forming units per ml (1 µl) of each bacterial test strain and their mixed culture was separately transferred to Mueller Hinton Agar plates containing eighteen different concentrations of the antimicrobial agent. The inoculated plates were incubated at 37 °C for 24 hours and the minimum inhibitor concentrations of antimicrobial agent against the test bacteria were determined. According to experimental results, the test bacteria were found to be resistant to ampicillin (10 μg), spectinomycin (25 μg), streptomycin (10 μg), meropenem (10 μg) and penicillin (10 U) antibiotics. The minimum inhibitor concentrations of antimicrobial agent were found as 1000 µg/ml for Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213, Micrococcus luteus ATCC 9341, mixed culture of test bacteria; 500 µg/ml for Escherichia coli ATCC 25922 and Bacillus cereus ATCC 11778; 15.6 µg/ml for Staphylococcus epidermidis ATCC 12228 and Enterococcus faecalis ATCC 29212; and 3.9 µg/ml for Bacillus subtilis ATCC 6633. According to the data obtained from this study, the growths of eight different ATCC reference bacterial strains resistant to antibiotics and their mixed culture were inhibited by the tested antimicrobial agent. It is known that bacterial cells are capable of surviving and growing in the presence of antibacterial agents. Hence, it is generally difficult to control the mixture of different bacterial species with antibacterial agents. In the present study, the mixed culture of eight different ATCC reference bacterial strains were be controlled at 1000 µg/ml concentration of antimicrobial agent. As a result, the use of effective antimicrobial agents at the appropriate concentration in the soaking process is of great importance for the prevention of the development of antibiotic resistant bacteria found in this process and for the production of high quality leather.
The leather industry is an important commercial sector for the global economy. The hides and skins are contaminated with microorganisms found in air, soil, water, feces and slaughterhouse. After the animal is flayed, the hide and skin provide an ideal growth condition for these microorganisms due to containing fat and protein. Different chemical agents such as sodium chloride, boric acid, antifungal and antibacterial agents are used to preserve raw hides and skins in order to prevent microbial growth and damage on hides and skins during transportation and storage period. The salted hides are soaked in soaking process in order to re-soften and removing the salt, feces and microorganisms on the hide surface. Bacteria can develop in the leather industry due to the organic load in the soaking process. Bacterial growth is known to cause significant damage to the hides and skins such as discoloration, bad odor, hair slip, grain peeling, fiber disintegration, weakness, looseness and holes. Although various antimicrobial agents have been used in the soaking process, Gram-positive and Gram-negative bacteria resistant to antibiotics in soaking liquids and soaked hides have been identified in previous studies. It is very important to use the antimicrobial agent in the proper amount so that the soaking process can be carried out efficiently and effectively. Thus, bacterial damage can be controlled in the skin. Hence, the aim of the present study was to investigate the susceptibilities of Gram-negative (Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922) and Gram-positive (Staphylococcus epidermidis ATCC 12228, Staphylococcus aureus ATCC 29213, Micrococcus luteus ATCC 9341, Enterococcus faecalis ATCC 29212, Bacillus cereus ATCC 11778, Bacillus subtilis ATCC 6633) reference bacterial strains to five different antibiotics (ampicillin, spectinomycin, streptomycin, meropenem and penicillin) by Kirby-Bauer disc diffusion susceptibility test method on Mueller Hinton Agar. In addition, 18 different concentrations of the antimicrobial agent (4000 µg/ml, 2000 µg/ml, 1000 µg/ml, 500 µg/ ml, 250 µg/ml, 125 µg/ml, 62.5 µg/ml, 31.2 µg/ml, 15.6 µg/ml, 7.8 µg/ml, 3.9 µg/ml, 1.96 µg/ml, 0.98 µg/ml, 0.49 µg/ml, 0.24 µg/ml, 0.12 µg/ml, 0.06 µg/ml and 0.03 µg/ml), containing sodium dimethyldithiocarbamate as the active ingredient and used in the soaking process, were applied to these bacteria and their mixed culture and the minimum inhibitory concentration was investigated by agar dilution method. The series of two-fold dilutions of the test agent, ranging from 4000 µg/ml to 0.03 µg/ml, were prepared in Mueller Hinton Agar. The test strains were grown on Mueller Hinton Agar at 37 °C for 24 hours. Then, overnight cultures of the test strains were inoculated into 10 ml Mueller Hinton Broth with 2 colonies of each test strain. Each bacterial suspension was adjusted to a McFarland standard 0.5 (108 colony forming units per ml). The broth cultures of test strains were incubated at 37 °C for 24 hours. Later, each suspension of test strain was diluted to 107 colony forming units per ml using physiological saline water containing 0.85% sodium chloride. The mixed culture of the test strains was also prepared from these dilutions. An inoculum of 104 colony forming units per ml (1 µl) of each bacterial test strain and their mixed culture was separately transferred to Mueller Hinton Agar plates containing eighteen different concentrations of the antimicrobial agent. The inoculated plates were incubated at 37 °C for 24 hours and the minimum inhibitor concentrations of antimicrobial agent against the test bacteria were determined. According to experimental results, the test bacteria were found to be resistant to ampicillin (10 μg), spectinomycin (25 μg), streptomycin (10 μg), meropenem (10 μg) and penicillin (10 U) antibiotics. The minimum inhibitor concentrations of antimicrobial agent were found as 1000 µg/ml for Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213, Micrococcus luteus ATCC 9341, mixed culture of test bacteria; 500 µg/ml for Escherichia coli ATCC 25922 and Bacillus cereus ATCC 11778; 15.6 µg/ml for Staphylococcus epidermidis ATCC 12228 and Enterococcus faecalis ATCC 29212; and 3.9 µg/ml for Bacillus subtilis ATCC 6633. According to the data obtained from this study, the growths of eight different ATCC reference bacterial strains resistant to antibiotics and their mixed culture were inhibited by the tested antimicrobial agent. It is known that bacterial cells are capable of surviving and growing in the presence of antibacterial agents. Hence, it is generally difficult to control the mixture of different bacterial species with antibacterial agents. In the present study, the mixed culture of eight different ATCC reference bacterial strains were be controlled at 1000 µg/ml concentration of antimicrobial agent. As a result, the use of effective antimicrobial agents at the appropriate concentration in the soaking process is of great importance for the prevention of the development of antibiotic resistant bacteria found in this process and for the production of high quality leather.