Antibacterial Effect of Thiazole Derivatives on Rhodoccocus Equi, Brucella Abortus, and Pasteurella Multocida
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Iranian Journal of Veterinary Medicine Antibacterial effect of thiazole derivatives on Rhodoccocus equi, Brucella abortus, and Pasteurella multocida Ghasemi B.1, Najimi, M.2* 1Gratuated from the Faculty of Veterinary Medicine, University of Zabol, Zabol, Iran 2Department of Pathobiology, Faculty of Veterinary Medicine, University of Zabol, Zabol, Iran Key words: Abstract: antibacterial activity, B. abrotus, BACKGROUND: Rhodoccocus equi, Brucella abortus, and Pas- P. multocida, R. equi, thiazole teurella multocida are important veterinary bacterial pathogens derivatives that in recent years have been resisted to current antibiotics, and this problem threats the livestock industry. To control this Correspondence resistant microorganisms, use of new antibacterial compounds, Najimi, M. such as thiazole derivatives, in veterinary is necessary. OBJEC- Department of Pathobiology, TIVES: In this study, antibacterial effects of thiazole derivatives Faculty of Veterinary Medicine, on Rhodoccocus equi, Brucella abortus, and Pasteurella mul- University of Zabol, Zabol, Iran tocida were evaluated. METHODS: Synthesized thiazole deriv- Tel: +98(54) 31232250 atives were prepared in DMSO, then the disk diffusion meth- Fax: +98(54) 31232251 od was used to measure growth inhibition zone diameter and Email: najimi.mohsen@gmail. the broth microdilution method was applied to determine the com minimum inhibitory concentration (MIC). RESULTS: Results showed that thiazole derivatives had no significant inhibitory Received: 21 October 2015 effects on B. abrotus, while they had inhibitory effects on R. Accepted: 11 January 2016 equi and P. multocida with inhibition zone 12.7±0.4 -30.1±0.2 mm and MICs 32- 256 μg/ml. CONCLUSIONS: Results of this study indicate that thiazole derivatives have considerable in- hibitory effects on R. equi and P. multocida as veterinary bac- terial pathogens. Introduction derivatives. These derivatives perceive to have multi-therapeutic effects and they have been Rhodoccocus equi, Brucella abrotus, and utilized in treatment of cancer, blood fat, blood Pasteurella multocida are important veterinary pressure, and HIV infection (Chementi et al., bacterial pathogens that cause high mortality 2011). Strong antioxidant and anti-inflam- rates and economic losses in the livestock in- matory effects of thiazoles have been proven dustry (Adesiyun et al., 2011; Bakavoli et al., (Cheng et al., 2013; Coleman et al., 2010). 2009; Bakavoli et al; 2011). Development of In laboratory, thiazoles have shown the bacterial resistant in these microorganisms to ability to kill anopheles insects (Helul et al., current antibiotics, such as ciprofloxacin, tri- 2013). Thiazole derivatives have inhibitory methoprim, and tetracycline have caused se- effects on Trypanosoma and Candida spp. rious problems in veterinary in recent years (Horohov et al., 2011; Jaishree et al., 2013). and use of new antibacterial compounds is the Thiazole derivatives can inhibit the growth of best solution for this problem (Bondock et al., a variety of gram-positive and gram-negative 2010; Bondock et al., 2013; Brvar et al., 2010). bacteria, including Escherichia coli, Staphy- One of these novel antimicrobials is thiazole lococcus epidermidis, Staphylococcus aureus, IJVM (2016), 10 (1):47-52 47 Antibacterial effect of thiazole Ghasemi B. Streptococcus pyogenes, Enterococcus faeca- tometer and standard McFarland tube number lis and Pseudomonas fluorescens. Strong and 0.5 from each bacterium which is assigned as a wide range of antibacterial properties of thi- stock solution (Kofteridis et al., 2009). azole derivatives has generally made the an- Minimum inhibition concentration tibacterial test to be among the initial exper- (MIC): The MIC test was done in a sterile 96- iments that is studied after synthesizing these well plates by broth micro dilution as CLSI agents in many countries (Juspin et al., 2010; standard. First, 100 µl of Muller-Hinton broth Katsuda et al., 2013. Very few studies on thi- medium (Merck®, Germany) was added to azole antibacterial effects against these veteri- each well. Then, 100 µl of thiazole derivatives nary bacterial pathogens have been published. (in control groups, 100 µl of penicillin and In the current study, the antibacterial effects of gentamycin antibiotics (Sigma®)) were added thiazole derivatives on R. equi, P. multocida, to the first well and, after mixing, 100 µl of this and B. abortus were assessed. mixture was embedded into the second well. Similarly, dilution procedure was done in other Materials and Methods wells. 10 µl of bacterial suspension was added to each well. For negative control 100 µl of Preparation of thiazole derivatives: The Muller-Hinton broth, 100 µl DMSO and 10 µl number 6a-d of thiazole derivative was incor- of bacterial suspension were added to the last porated in a three-phase process and its chem- well in each row. The result of incubation was ical structure was verified with monocrystal- read after 24 hours incubation in 37oC. The lu- line X-ray diffraction, HNMR, CNMR, IR, cidity and turbidity in each well indicated lack element decomposition, and spectrometry. Af- or existence of bacterial growth, respectively. terwards, this derivative was dissolved in the The last well that did not show any turbidity DMSO solvent with the concentration of 8192 was reported as MIC (Kofteridis et al., 2009). µg/ml (7). Inhibition zone diameter: First, in Synthesis of 2-[(E)-(benzo[d]thiazol-2(3H)- Muller-Hinton agar medium, the superficial ylidene)(cyano)methyl]thiazoles (10a-c); the bacterial culture was performed with a swab number 10a-c of thiazole derivative was incor- impregnated to bacterial suspension. 20 µl of porated in a three-phase process and its chem- obtained MIC thiazole derivatives and also an- ical structure was verified with monocrystal- tibiotics was shed on blank sterile disks. For line X-ray diffraction, HNMR, CNMR, IR, negative control, the DMSO-impregnated disk element decomposition, and spectrometry. Af- was used. Then, after 24 hours incubation in terwards, this derivative was dissolved in the 37oC, the growth inhibition zone diameter was DMSO solvent with the concentration of 8192 measured with particular ruler. The results of µg/ml (Khalil et al., 2009). growth inhibition zone diameter have been Bacterial suspensions: Rhodococcus equi provided as average ± standard deviation and ATCC 33701, P. multocida ATCC 12948, and for the aim of analyzing the data, the SPSS B. abortus ATCC 23448 were provided by the statistical software (version 22) was used Faculty of Veterinary Medicine, University of (Kofteridis et al., 2009). Shiraz. Each bacterium was cultured in Muel- ler-Hinton agar medium in 37oC for 24 hours. Results Henceforth, in sterile conditions of Muel- ler-Hinton medium and in logarithmic growth Thiazole derivatives showed no significant phase, a concentration of 0.5 McFarland (1.5 inhibitory effects on B. abortus; also 6a-c, 10a × 108 CFU/ml) was obtained with spectropho- and 10c did not have inhibitory effects on any 48 IJVM (2016), 10 (1):47-52 Ghasemi B. Iranian Journal of Veterinary Medicine Scheme 1. Synthesis of 2-(E)-cyano(thiazolidin-2-ylidene)thiazoles (6a-d) (derivative from reference No. 7). Scheme2. Synthesis of 2-[(E)-(benzo[d]thiazol-2(3H)-ylidene)(cyano)methyl]thiazoles (10a-c) (Bakavoli et al., 2015). bacterial tested. Just the two derivatives 6d and compounds which promise good replacements 10b showed inhibitory effects on R. equi and for some antibacterial drugs. In the current P. multocida. The maximum inhibitory effects study, inhibitory effects of eight thiazole de- on R. equi and P. multocida belonged to deriv- rivatives have been assessed on three veteri- atives 6d and 10b with MIC of 64 and 32 µg/ nary bacterial pathogens. Results show that ml respectively. Ampicillin and neomycin had the maximum inhibitory effect against R. equi the maximum and minimum inhibitory effects belongs to derivative 6d. The structural study on R. equi, respectively and penicillin and na- of this derivative shows that this compound lidixic acid had the maximum and minimum includes thiazolidine ring as well as thiazole inhibitory effects on P. multocida, respective- ring. This thiazolidine ring is similar to that ly. In antibiogram test, the most and the least of penicillin family of antibiotics. However, susceptibility were recorded for P. multocida this derivative is expected to affect beta-lact- to penicillin with MIC of 0.5 µg/ml and for amase producing bacteria due to the lack of a B. abortus to penicillin with MIC of 16 µg/ml beta-lactam ring (Lv et al., 2009). (Tables 1 and 2). Thiazolidines are a novel class of antibac- terial agents which include inhibitory effects Discussion on a broad-spectrum of gram-positive bacteria, such as streptococci and staphylococci. The in- Thiazole derivatives are novel antibacterial hibitory effect of thiazolidine derivatives on S. IJVM (2016), 10 (1):47-52 49 Antibacterial effect of thiazole Ghasemi B. Table 1. Bacterial growth inhibitory zone (mm) of the thiazole to significantly inhibit the growth of Pseudo- derivatives and antibiotics on studied bcteria. - absence of in- monas aeruginosa, S. aureus, and B. subtilis, hibition effect. compared to penicillin G and kanamycin. Deriva- R. equi P. multocida B. abortus tives/Drugs ATCC ATCC 12948 ATCC 23448 Results have also shown that these deriva- 33701 tives have higher inhibition effects with MICs 6a-c - - - of 12.5-100 µg/ml. Derivatives within the cur- 6d 25.6±0.1 26.3±0.0 - rent study possibly include excited Cl and Br 10a - - - in thiazole ring and, therefore, show intensi- 10b 12.7±0.4 30.1±0.2 - fied inhibitory effects (Venugolpa et al., 2013). 10c - - - In a study by Zaky and Yousef (2011), MIC Gentamicin 25.4±0.3 21.2±0.0 16.3±0.2 and inhibition zone of thiazole derivatives on Penicillin 27.2±0.5 30.5±0.3 22.1±0.1 S. aureus and P. aeruginosa were assessed and Table 2. MICs (μg/ml) of thiazole derivatives and antibiotics showed high antibacterial effects, compared on studied bcteria. - absence of inhibition effect.