Search results for: antibiotic resistant

Authors: Rosetta Moshirian Farahi, Ahya Abdi Ali, Sara Gharavi

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The major mechanism of Pseudomonas aeruginosa resistance to fluoroquinolones is the alteration of target enzymes, type II and IV topoisomerases due to mutations in the quinolone resistance-determining regions (QRDR) of the gyrA and parC genes coding A subunits of these enzymes. 37 isolates from patients with burn wounds and 20 isolates from blood, urine and sputum specimen were selected to evaluate mutations involved in antibiotic resistance and were subsequently verified for their resistance to ciprofloxacin. QRDRs regions of gyrA and parC were amplified by polymerase chain reaction (PCR) and were subsequently sequenced. 90% of isolates with MIC≥8 µg/ml to ciprofloxacin had a mutation in gyrA gene in which threonine at position 83 changed to isoleucine. 87.5% of isolates had mutation in parC, Serine 87 changed. 75% had Ser87Leu and 12.5% possessed Serin87Trp. Various silent mutations were also detected such as Val103Val, Ala118Ala, Ala136Ala, His132His in gyrA and Ala115Ala in parC. The data indicates that the common mutation in gyrA is Thr83Ile and in parC is Ser87Leu/Trp. No individual parC mutation was observed while mutations in gyrA and parC occurred simultaneously and appears to be the main reason of high-level resistance to fluoroquinolones in patients with burn wounds and urine infection. The vast majority of P.aeruginosa isolates had mutation in parC which can play a crucial role in increased resistance of these isolates. This is a report of parC mutations from resistant P. aeruginosa isolates from Iran, Tehran.

Keywords: P. aeruginosa, fluoroquinolones, gyrA, parC, antibiotic resistance

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Authors: Komal Fizza

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Irrational and non-judicious use of antibiotics pave the way for an upsurge in antibiotic resistance, diminished effectiveness of different therapeutic regimens and as well as impounding effect on disease management leading to further morbidities. In the backdrop of this the current research is aimed to assess whether antimicrobial prescribing is in accordance with the Infectious Disease Society of America Guidelines in hospitalized patients at Shifa International Hospital, Islamabad, Pakistan. Shifa International Hospital, Islamabad is a 500 bed hospital. With the help of MIS team a form wad developed that gave the information about medical records number, name of the patient, day of start of antibiotic, the day antibiotic is supposed to be stopped and as well as the diagnosis of the patient. A ward pharmacist was employed to generate this report on a daily basis. The therapeutic regiment was reviewed by the pharmacist by monitoring the clinical progress, laboratory report and diagnosis. On the basis of this information, pharmacist made suggestions and forwarded to the hospital doctors responsible for prescribing antibiotics. If desired, changes were made regularly. In the current research our main focus was to implement this action and therefore, started monitoring patients who were on antibiotic regimens for more than 10-15 days. We took this initiative since November, 2013. At the start of the program a maximum 19 patients/day were reported to be on antibiotic regimen for more than 10-15 days. After the implementation of the initiative, the number of patients was decreased to fifteen patients per day in December, further decreased to 7 in the month of January and 9 and 6 in February and March respectively. The average patient census was 350. The current pilot study highlighted the role of pharmacist in initiating antibiotic stewardship programs in hospital settings.

Keywords: stewardship, antibiotics, resistance, clinical process

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Authors: Naima Nur, Safa Islam, Saeema Islam, Faridul Alam

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Background: Drug-resistant pulmonary tuberculosis (DR-PTB), particularly multidrug-resistant tuberculosis (MDR-TB) and pre-extensive drug-resistant (pre-XDR), is a major challenge in effectively controlling TB, especially in developing. This study aimed to identify the strains of M. tuberculosis complex (MTC) and drug resistance patterns among the pulmonary tuberculosis patients. Methods: The study used a cross-sectional design, and 815 patients were recruited randomly in three study periods. In the first-period, 210 treated PTB patients, who were completed their treatment, received their diagnoses using light microscopy, fluorescence microscopy and cultured on Lowenstein-Jensen (L-J) slant, and then strains were identified as MTC by biochemical tests, and then sensitivity test in National Institute of Diseases of the Chest and Hospital. In the second-period, 220 re-treated PTB patients, who were completed their treatment, received their diagnoses using culture on L-J slant, line probe assay (LPA), and GeneXpert in the same hospital. In the last-period, during treatment, 385 MDR-PTB patients received their diagnoses using culture on L-J slant and LPA in the same hospital. Results: Among sixty-two (29.5%) PTB patients, 13% were sensitive to all first-line anti-TB drugs, 26% were MDR-TB patients, and 14.2% were pre-XDR-TB among 14 MDR-TB patients. After three years, 31% were MDR-TB among 220 re-treated PTB patients. After five years, 16.4% was pre-XDR-TB among 385 MDR-TB patients. Compared to females, male patients were significantly higher at all times. Conclusion: The current study demonstrated that in three study periods, the proportions of DR-TB, MDR-TB, and pre-XDR patients were an alarming issue and increasing daily.

Keywords: multi-drug resistant, drug-resistant, pre-extensive drug resistant, pulmonary tuberculosis

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Authors: Garzon V., Bustos R., Salvador J. P., Marco M. P., Pinacho D. G.

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Bacterial resistance to antimicrobial treatment has increased significantly in recent years, making it a public health problem. Large numbers of bacteria are resistant to all or nearly all known antimicrobials, creating the need for the development of new types of antimicrobials or the use of “last line” antimicrobial drug therapies for the treatment of multi-resistant bacteria. Some of the chemical groups of antimicrobials most used for the treatment of infections caused by multiresistant bacteria in the clinic are Glycopeptide (Vancomycin), Polymyxin (Colistin), Lipopeptide (Daptomycin) and Carbapenem (Meropenem). Molecules that require therapeutic drug monitoring (TDM). Due to the above, a methodology based on nanobiotechnology based on an optical and electrochemical biosensor is being developed, which allows the evaluation of the plasmatic levels of some antimicrobials such as glycopeptide, polymyxin, lipopeptide and carbapenem quickly, at a low cost, with a high specificity and sensitivity and that can be implemented in the future in public and private health hospitals. For this, the project was divided into five steps i) Design of specific anti-drug antibodies, produced in rabbits for each of the types of antimicrobials, evaluating the results by means of an immunoassay analysis (ELISA); ii) quantification by means of an electrochemical biosensor that allows quantification with high sensitivity and selectivity of the reference antimicrobials; iii) Comparison of antimicrobial quantification with an optical type biosensor; iv) Validation of the methodologies used with biosensor by means of an immunoassay. Finding as a result that it is possible to quantify antibiotics by means of the optical and electrochemical biosensor at concentrations on average of 1,000ng/mL, the antibodies being sensitive and specific for each of the antibiotic molecules, results that were compared with immunoassays and HPLC chromatography. Thus, contributing to the safe use of these drugs commonly used in clinical practice and new antimicrobial drugs.

Keywords: antibiotics, electrochemical biosensor, optical biosensor, therapeutic drug monitoring

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Authors: Recep Keşli, Gülşah Aşık, Cengiz Demir, Onur Türkyılmaz

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Objective: Proteus mirabilis (P. mirabilis) is one of Gram-negative pathogens in human and it causes urinary tract and nosocomial infections. P. mirabilis is susceptible to β-lactams, aminoglycosides, fluoroquinolones, and trimethoprim/sulfamethoxazole. It was aimed to investigate the resistance status to antimicrobial agents of Proteus mirabilis strains produced from samples sent to Afyon Kocatepe University, ANS Research and Practice Hospital, Microbiology Laboratory from different clinics and polyclinics during the period of 24 months. Methods: Between October 2013 and September 2015, a total of 30 Proteus were isolated from clinical samples of patients were hospitalized in intensive care units and in various departments of Afyon Kocatepe University, ANS Research and Practice Hospital. Identification of the bacteria was determined by conventional methods and VITEK 2 system (bioMérieux, France) was used additionally. Antibacterial susceptibility tests were performed by Kirby Bauer disc (Oxoid, Hempshire, England) diffusion method following the recommendations of CLSI. Results: Of the total 30 Proteus strains isolated from clinical samples, 19 from urine, 7 from wound, 4 from tracheal aspiration materials were isolated. Antimicrobial resistant for these strains were determined to 24,3% for meropenem, 26.2% for imipenem, 20.2% for amikacin 10.5% for cefepim, 33.3% for ciprofloxacin and levofloxacine, 31.6% for ceftazidime, 20% for ceftriaxone, 15.2% for gentamicin and 26.6% for amoxicillin-clavulanate, 26.2% trimethoprim-sulfamethoxale. Conclusion: In the present study, the highest number of clinical isolates of P. mirabilis were isolated from urine (63,3%), followed by the others (36,6%). The distribution of samples P. mirabilis strains to the clinics were as fallows; 16,8% intensive care unit (ICU), 29,9% polyclinics, 53,3% hospital service units The most effective antibiotic on the total of strains were found to be cefepim, the least effective antibiotics on the total of strains were found to be trimethoprim-sulfamethoxale.

Keywords: proteus mirabilis, antibiotic resistance, intensive care unit, Proteus spp.

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Authors: Maryam Monazzah, Ronak Samadpour, Mehdi Nasr-esfahani, Fatemeh Qalavand, Marziye Motamedi

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Bipolaris sorokiniana, and Heterodera filipjevi, are important wheat diseases that lead to yield losses worldwide. Identifying novel resistant sources helps us combat these devastating diseases. In this study, we studied the role of Cre3 gene and antioxidant enzymes in the immune responses of wheat genotypes to H. filipjevi and B. sorokiniana. Therefore, real-time PCR analysis using Cre3 gene marker, a resistant gene to cereal cyst nematodes, was conducted on leaves and roots, along with changes ‎in the activity of antioxidant enzymes, peroxidase, and catalase. Enzyme activity assay was performed on roots attacked by nematode and in leaves infected with Bipolaris. Wheat accessions including “Bam” (resistant), “Parsi” (moderately-resistant), “Azar2”, “Ohadi”, “Homa” (highly-susceptible) were previously screened against both stresses under greenhouse and field conditions. Results showed that Cre3 expression against cyst nematodes was significantly higher in resistant cultivars compared to susceptible cultivars. Cre3 was used in marker-assisted selection programs to identify genotypes carrying resistant genes to cyst nematodes. Interestingly, Cre3 was also up-regulated in both tissues of resistant cultivars to B. sorokiniana. Therefore, Cre3 in wheat similarly modulates immunity against B. sorokiniana and might be one of the central components of the induced immune system in wheat. The activity of antioxidant enzymes also indicated the highest increase in resistant genotypes upon both stresses that subsequently neutralize oxidative stress in tissues and decrease damage. Further studies on these resistance components may help us gain insight into the molecular basis of resistance and shed new light on the interaction and overlap between different forms of stress.

Keywords: Bipolaris sorokiniana, Heterodera filipjevi, resistant gene expression, wheat

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Authors: Zohra Benfodda, Valentin Duvauchelle, Chaimae Majdi, David Bénimélis, Catherine Dunyach-Remy, Patrick Meffre

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New antibiotics are necessary to treat microbial pathogens, especially ESKAPE pathogens that are becoming increasingly resistant to available treatment. Despite the medical need, the number of newly approved drugs continues to decline. The majority of antibiotics under clinical development are natural products or derivatives thereof. 43 juglone/naphthazarin derivatives were synthesized using Minisci-type direct C–H alkylation and evaluated for their antibacterial properties against various clinical and reference Gram-positive MSSA, clinical Gram-positive MRSA. Different compounds of the synthesized series showed promising activity against clinical and reference MSSA (MIC: 1–8 μg/ml) and good efficacy against clinical MRSA (MIC: 2–8 μg/ml) strains. The synergistic effects of active compounds were evaluated with reference antibiotics (vancomycin and cloxacillin), and it was found that the antibiotic combination with those active compounds efficiently enhanced the antimicrobial activity and consequently the MIC values of reference antibiotics were lowered up to 1/16th of the original MIC. These synthesized compounds did not present hemolytic activity on sheep red blood cells. In addition to the in silico prediction of ADME profile parameter which is promising and encouraging for further development.

Keywords: juglone, naphthazarin, antibacterial, clinical MRSA, synergistic studies, MIC determination

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Authors: Uraib Sharaha, Ahmad Salman, Eladio Rodriguez-Diaz, Elad Shufan, Klaris Riesenberg, Irving J. Bigio, Mahmoud Huleihel

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Antimicrobial drugs have an important role in controlling illness associated with infectious diseases in animals and humans. However, the increasing resistance of bacteria to a broad spectrum of commonly used antibiotics has become a global health-care problem. Rapid determination of antimicrobial susceptibility of a clinical isolate is often crucial for the optimal antimicrobial therapy of infected patients and in many cases can save lives. The conventional methods for susceptibility testing like disk diffusion are time-consuming and other method including E-test, genotyping are relatively expensive. Fourier transform infrared (FTIR) microscopy is rapid, safe, and low cost method that was widely and successfully used in different studies for the identification of various biological samples including bacteria. The new modern infrared (IR) spectrometers with high spectral resolution enable measuring unprecedented biochemical information from cells at the molecular level. Moreover, the development of new bioinformatics analyses combined with IR spectroscopy becomes a powerful technique, which enables the detection of structural changes associated with resistivity. The main goal of this study is to evaluate the potential of the FTIR microscopy in tandem with machine learning algorithms for rapid and reliable identification of bacterial susceptibility to antibiotics in time span of few minutes. The bacterial samples, which were identified at the species level by MALDI-TOF and examined for their susceptibility by the routine assay (micro-diffusion discs), are obtained from the bacteriology laboratories in Soroka University Medical Center (SUMC). These samples were examined by FTIR microscopy and analyzed by advanced statistical methods. Our results, based on 550 E.coli samples, were promising and showed that by using infrared spectroscopic technique together with multivariate analysis, it is possible to classify the tested bacteria into sensitive and resistant with success rate higher than 85% for eight different antibiotics. Based on these preliminary results, it is worthwhile to continue developing the FTIR microscopy technique as a rapid and reliable method for identification antibiotic susceptibility.

Keywords: antibiotics, E. coli, FTIR, multivariate analysis, susceptibility

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Authors: Choon Seong Ng, P. Petrick, C. L. Lau

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Background: Carbapenem-resistant isolates have been increasingly reported recently. Carbapenem stewardship is designed to optimize its usage particularly among medical wards with high prevalence of carbapenem prescriptions to combat such emerging resistance. Carbapenem stewardship programmes (CSP) can reduce antibiotic use but clinical outcome of such measures needs further evaluation. We examined this in a prospective manner using feedback mechanism. Methods: Our single-center prospective cohort study involved all carbapenem prescriptions across the medical wards (including medical patients admitted to intensive care unit) in a tertiary university hospital setting. The impact of such stewardship was analysed according to the accepted and the rejected groups. The primary endpoint was safety. Safety measure applied in this study was the death at 1 month. Secondary endpoints included length of hospitalisation and readmission. Results: Over the 19 months’ period, input from 144 carbapenem prescriptions was analysed on the basis of acceptance of our CSP recommendations on the use of carbapenems. Recommendations made were as follows : de-escalation of carbapenem; stopping the carbapenem; use for a short duration of 5-7 days; required prolonged duration in the case of carbapenem-sensitive Extended Spectrum Beta-Lactamases bacteremia; dose adjustment; and surgical intervention for removal of septic foci. De-escalation, shorten duration of carbapenem and carbapenem cessation comprised 79% of the recommendations. Acceptance rate was 57%. Those who accepted CSP recommendations had no increase in mortality (p = 0.92), had a shorter length of hospital stay (LOS) and had cost-saving. Infection-related deaths were found to be higher among those in the rejected group. Moreover, three rejected cases (6%) among all non-indicated cases (n = 50) were found to have developed carbapenem-resistant isolates. Lastly, Pitt’s bacteremia score appeared to be a key element affecting the carbapenem prescription’s behaviour in this trial. Conclusions: Carbapenem stewardship program in the medical wards not only saves money, but most importantly it is safe and does not harm the patients with added benefits of reducing the length of hospital stay. However, more time is needed to engage the primary clinical teams by formal clinical presentation and immediate personal feedback by senior Infectious Disease (ID) personnel to increase its acceptance.

Keywords: audit and feedback, carbapenem stewardship, medical wards, university hospital

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Authors: Sumayah Mohammed Asiri A., Aviva Levina B., Elizabeth New C., Peter Lay D.

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Pt compounds have been among the most successful anticancer drugs in the last 40 years, but the development of resistance to them is an increasing problem. Cellular homeostasis of an essential metal, Cu, is known to be involved in Pt resistance, but mechanisms of this process are poorly understood. We used a novel ratiometric Cu(I)-sensitive fluorescent probeInCCu1 dye to detect Cu(I) in the mitochondria. Total Cu and labile Cu pool measured using AAS and InCCu1 dye in A2780 cells and their corresponding resistant cells A2780-cis.R cells treated with Cu and cisplatin. The main difference between both cell lines in the presence and absence of Cu(II) is that resistant cells have lower total Cu content but higher labile Cu levels than cisplatin-sensitive cells. This means that resistant cells can metabolize and export excess Cu more efficiently. Furthermore, InCCu1 has emerged not only as an indicator of labile cellular Cu levels in the mitochondria but as a potentially versatile multi-organelle probe.

Keywords: AAS and ICPMS, A2780 and its resistant cells, ratiometric fluorescent sensors, inCCu1, and total and labile Cu

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Authors: Abdullahi Musa, Yakubu Kukure Enebe Ibrahim, Adeshina Gujumbola

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The emergence of multidrug resistance in clinical Escherichia coli has been associated with plasmid-mediated genes. DNA transfer among bacteria is critical to the dissemination of resistance. Plasmids have proved to be the ideal vehicles for dissemination of resistance genes. Plasmids coding for antibiotic resistance were long being recognized by many researchers globally. The study aimed at determining the antibiotic susceptibility pattern of ESBL E. coli isolates claimed to be multidrug resistance using disc diffusion method. Antibacterial activity of the test isolates was carried out using disk diffusion methods. The results showed that, majority of the multidrug resistance among clinical isolates of ESBL E. coli was as a result of acquisition of plasmid carrying antibiotic-resistance genes. Production of these ESBL enzymes by these organisms which are normally carried by plasmid and transfer from one bacterium to another has greatly contributed to the rapid spread of antibiotic resistance amongst E. coli isolates, which lead to high economic burden, increase morbidity and mortality rate, complication in therapy and limit treatment options. To curtail these problems, it is of significance to checkmate the rate at which over the counter drugs are sold and antibiotic misused in animal feeds. This will play a very important role in minimizing the spread of resistance bacterial strains in our environment.

Keywords: Escherichia coli, plasmid, multidrug resistance, ESBL, pan drug resistance

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Authors: K. Ali, M. F. Qamar, M. A. Zaman, M. Younus, I. Khan, S. Ehtisham-ul-Haque, R. Tamkeen, M. I. Rashid, Q. Ali

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This study centers on molecular identification of Haemonchus contortus and isolation of Benz-imidazoles (BZ) resistant strains. Different abattoirs’ of two geographic regions of Punjab (Pakistan) were frequently visited for the collection of worms. Out of 1500 (n=1500) samples that were morphologically confirmed as H. contortus, 30 worms were subjected to molecular procedures for isolation of resistant strains. Resistant worms (n=8) were further subjected to DNA gene sequencing. Bio edit sequence alignment editor software was used to detect the possible mutation, deletion, replacement of nucleotides. Genetic diversity was noticed and genetic variation existing in β-tubulin isotype 1 of the H. contortus population of small ruminants of different regions considered in this study. H. contortus showed three different type of genetic sequences. 75%, 37.5%, 25% and 12.5% of the studied samples showed 100% query cover and identity with isolates and clones of China, UK, Australia and other countries, respectively. Interestingly the neighbor countries such as India and Iran haven’t many similarities with the Pakistani isolates. Thus, it suggests that population density of same genetic makeup H. contortus is scattered worldwide rather than clustering in a single region.

Keywords: Haemonchus contortus, Benzimidazole resistant, β-tubulin-1 gene, abattoirs

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Authors: Yousef M. Abouzeed A. Elfahem, F. Zgheel, M. A. Saad, Mohamed O. Ahmed

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The bioactive properties of phytochemicals indicate their potential as natural drug products to prevent and treat human disease; in particular, compounds with antioxidant and antimicrobial activities may represent a novel class of safe and effective drugs. Following desiccation, grapes (Vitis vinifera) become more resistant to microbial-based degradation, suggesting that raisins may be a source of antimicrobial compounds. To investigate this hypothesis, total phenolic extracts were obtained from common raisins, local market-sourced. The acetone extract was tested for antibacterial activity against four prevalent bacterial pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella spp. and Escherichia coli). Antibiotic sensitivity and the Minimum Inhibitory Concentration (MIC) were determined for each bacterium. High performance liquid chromatography was used to identify compounds in the total phenolic extract. The raisin phenolic extract inhibited growth of all the tested bacteria; the greatest inhibitive effect (normalized to cefotaxime sodium control antibiotic) occurred against P. aeruginosa, followed by S. aureus > Salmonella spp.= E. coli. The phenolic extracts contained the bioactive compounds catechin, quercetin, and rutin. Thus, phytochemicals in raisin extract have antibacterial properties; this plant-based extract, or its bioactive constituents, may represent a promising natural preservative or antimicrobial agent for the food industry or anti-infective drug.

Keywords: Vitis vinifera raisin, extraction, phenolic compounds, antibacterial activity

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Authors: M. Dahiru, O. I. Enabulele

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The research wish to investigate the occurrence of multidrug- resistant Acinetobacter, in carrot and estimate the role of carrot in its transmission, in a rapidly growing urban population. Thus, 50 carrot samples were collected from Jakara wastewater irrigation farms and analyzed on MacConkey agar and screened by Microbact 24E (Oxoid) and susceptibility of isolates tested against 10 commonly used antibiotics. Acinetobacter baumannii and A. lwoffii were isolated in 22.00% and 16% of samples respectively. Resistance to ceporex and penicillin of 36.36% and 27.27% in A. baumannii, and sensitivity to ofloxacin, pefloxacin, gentimycin and co-trimoxazole, were observed. However, for A. lwoffii apart from 37.50% resistance to ceporex, it was also resistant to all other drugs tested. There was a similarity in the resistant shown by A. baumannii and A. lwoffii to fluoroquinolones drugs and β- lactame drugs families in addition to between sulfonamide and animoglycoside demonstrated by A. lwoffii. Interestingly, when resistant similarities to different antibiotics were compared for A. baumannii and A. lwoffii as a whole, significant correlation was observed at P < 0.05 to CPX to NA (46.2%), and SXT to AU (52.6%) respectively, and high multi drug resistance (MDR) of 27.27% and 62.50% by A. baumannii and A. lwoffii respectively and overall MDR of 42.11% in all isolates. The occurrence of multidrug-resistance pathogen in carrot is a serious challenge to public health care, especially in a rapidly growing urban population where subsistence agriculture contributes greatly to urban livelihood and source of vegetables.

Keywords: urban agriculture, public health, fluoroquinolone, sulfonamide, multidrug-resistance

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Authors: Muizz Sanni-Anibire

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The 21st century has witnessed growing concerns for the protection of built facilities against natural and man-made disasters. Studies in earthquake resistant buildings, fire, and explosion resistant buildings now dominate the arena. To protect people and facilities from the effects of the explosion, reinforced concrete walls have been designed to be blast resistant. Understanding the performance of these walls is a key step in ensuring the safety of built facilities. Blast walls are mostly designed using simple techniques such as single degree of freedom (SDOF) method, despite the increasing use of multi-degree of freedom techniques such as the finite element method. This study is the first stage of a continuous research into the safety and reliability of blast walls. It presents the SDOF approach applied to the analysis of a concrete wall panel under three representative bomb situations. These are motorcycle 50 kg, car 400kg and also van with the capacity of 1500 kg of TNT explosive.

Keywords: blast wall, safety, protection, explosion

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Authors: Agnese Natali, Francesco Morelli, Walter Salvatore

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Automated Rack Supported Warehouses (ARSWs) are storage buildings whose structure is made of the same racks where goods are placed. The possibility of designing dissipative seismic-resistant ARSWs is investigated. Diagonals are the dissipative elements, arranged as tense-only X bracings. Local optimization is numerically performed to satisfy the over-resistant connection request for the dissipative element, that is hard to be reached due the geometrical limits of the thin-walled diagonals and must be balanced with resistance, the limit of slenderness, and ductility requests.

Keywords: steel racks, thin-walled cold-formed elements, structural optimization, hierarchy rules, dog-bone philosophy

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Authors: Ramesh Subramani, Mathivanan Narayanasamy, William Aalbersberg

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It is well accepted by last 35 years of research and on-going programmes that marine environment harbours abundant and unique biodiversity, which is currently playing as an important source in bioprospecting. It has become apparent that marine microorganisms are lead in the biodiscovery. Among marine organisms, actinobacteria are a target phylum for discovering novel antibiotics against increasing the multi-drug resistant human pathogens because of these taxa representing for novel genera and species. Marine actinomycetes are a proven source of new antibiotic leads and novel enzymes with important industrial applications. A total of 183 streptomycete and 25 non-streptomycete strains were isolated from different marine samples collected from north-eastern part of the Indian Ocean. Among them, 111 isolates displayed antibacterial activity against human pathogens and 151 exhibited antifungal activity against phytopathogens. Importantly, most of them produced various extracellular enzymes and 58 of them produced exopolysaccharides. Totally eight small bioactive compounds and a thermostable alkaline protease have been purified from a selected strain, Streptomyces fungicidicus. Besides, our on-going studies on non-streptomycete strains (rare actinomycetes) are most likely promising resource for new and unique compounds against current emerging drug-resistant pathogens. We have just recognised the chemical diversity in marine microorganisms. Therefore it is worthwhile to continue the exploration of marine microorganisms for new drug leads, novel enzymes and other bioprospecting research.

Keywords: bioactive compounds, industrial enzymes, marine actinobacteria, microbial metabolites, marine natural products

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Authors: Khushbu Priyadarshi, Chandramani Pathak

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Cancer cells can develop resistance to conventional therapies especially chemotherapeutic drugs. Resistance to chemotherapy is another challenge in cancer therapeutics. Therefore, it is important to address this issue. Gallic acid (GA) is a natural plant compound that exhibits various biological properties including anti-proliferative, anti-inflammatory, anti-oxidant and anti-bacterial. Despite of the wide spectrum biological properties GA has cytotoxic response and low bioavailability. To overcome this problem, GA was conjugated with the Polyamidoamine(PAMAM) dendrimer for improving the bioavailability and efficient delivery in drug-resistant HCT-116 Colon Cancer cells. Gallic acid was covalently linked to 4.0 G PAMAM dendrimer. PAMAM dendrimer is well established nanocarrier but has cytotoxicity due to presence of amphiphilic nature of amino group. In our study we have modified surface of PAMAM dendrimer with Gallic acid and examine their anti-proliferative effects in drug-resistant HCT-116 cells. Further, drug-resistant colon cancer cells were established and thereafter treated with different concentration of PAMAM-GA to examine their anti-proliferative potential. Our results show that PAMAM-GA conjugate induces apoptotic cell death in HCT-116 and drug-resistant cells observed by Annexin-PI staining. In addition, it also shows that multidrug-resistant drug transporter P-gp protein expression was downregulated with increasing the concentration of GA conjugate. After that we also observed the significant difference in Rh123 efflux and accumulation in drug sensitive and drug-resistant cancer cells. Thus, our study suggests that conjugation of anti-cancer agents with PAMAM could improve drug resistant property and cytotoxic response to treatment of cancer.

Keywords: drug resistance, gallic acid, PAMAM dendrimer, P-glycoprotein

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Authors: Ilknur Tuncer, Nihayet Bizsel

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The scarcity of research in bacterial diversity and antimicrobial resistance in coastal environments as in Turkish coasts leads to difficulties in developing efficient monitoring and management programs. In the present study, biogeochemical analysis of sediments and antimicrobial susceptibility analysis of bacteria in Izmir Bay, eastern Aegean Sea under high anthropogenic pressure were aimed in summer period when anthropogenic input was maximum and at intertidal zone where the first terrigenious contact occurred for aquatic environment. Geochemical content of the intertidal zone of Izmir Bay was firstly illustrated such that total and organic carbon, nitrogen and phosphorus contents were high and the grain size distribution varied as sand and gravel. Bacterial diversity and antibiotic resistance were also firstly given for Izmir Bay. Antimicrobially assayed isolates underlined the multiple resistance in the inner, middle and outer bays with overall 19% high MAR (multiple antibiotic resistance) index. Phylogenetic analysis of 16S rRNA gene sequences indicated that 67 % of isolates belonged to the genus Bacillus and the rest included the families Alteromonadaceae, Bacillaceae, Exiguobacteriaceae, Halomonadaceae, Planococcaceae, and Staphylococcaceae.

Keywords: bacterial phylogeny, multiple antibiotic resistance, 16S rRNA genes, Izmir Bay, Aegean Sea

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Authors: Zunnu Raen Akhtar, U. Irshad, M. Majid

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Due to the widespread cultivation of transgenic cotton, intense selection pressure resulted in resistant allele in pink bollworm, Pectinophora gossypiella (Gelechiidae: Lepidoptera). A resistant strain of pink bollworm against transgenic cotton has become a challenge to Integrated Resistance Management (IRM) in the World. Laboratory and field studies were conducted to determine the resistant strains of pink bollworm by performing bioassay, extracting the DNA, conducting PCR of both laboratory as well as field collected pink bollworms to observe the developed resistance. In all of the studies, two Bt varieties FH-142 and FH-118 expressing Cry1Ac compared to non-Bt (Control) were tested against pink bollworm. In the laboratory, bioassay results showed that there was no significant mortality difference between Bt and non-Bt varieties. Similar mortality percentage was observed in transgenic and non-transgenic (control) variety. Insects which were survived after bioassay, as well as those collected from the Bt cotton fields, were selected for further molecular studies. DNA extraction followed by PCR was conducted to check the resistant strains in pink bollworm. In field studies, we also observed the population dynamics of pink boll worms on Bt as compared to non-Bt varieties. Laboratory and field studies confirmed that resistant strains occurs in Pakistani Bt cotton fields. Different strategies should be adopted to combat that serious prevailing resistance issues.

Keywords: transgenic cotton, resistance, pectinophora gossypiella, , integrated resistance management (IRM), polymerase chain reaction (PCR)

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Authors: Omar Nasani, Chaza Kouchaji, Muznah Alkhani, Maisaa Abd-alkareem

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Objective: To evaluate the influence of sweetening agents used in pediatric medications on the growth of Streptococcus mutans colonies and its effect on the cariogenic activity in the oral cavity. No previous studies are registered yet in Syrian children. Methods: Specimens were isolated from the oral cavity of pediatric patients, then in-vitro study is applied on locally manufactured liquid pediatric antibiotic drugs, containing natural or synthetic sweeteners. The selected antibiotics are Ampicillin (sucrose), Amoxicillin (sucrose), Amoxicillin + Flucloxacillin (sorbitol), Amoxicillin+Clavulanic acid (Sorbitol or sucrose). These antibiotics have a known inhibitory effect on gram positive aerobic/anaerobic bacteria especially Streptococcus mutans strains in children’s oral biofilm. Five colonies are studied with each antibiotic. Saturated antibiotics were spread on a 6mm diameter filter disc. Incubated culture media were compared with each other and with the control antibiotic discs. Results were evaluated by measuring the diameter of the inhibition zones. The control group of antibiotic discs was resourced from Abtek Biologicals Ltd. Results: The diameter of inhibition zones around discs of antibiotics sweetened with sorbitol was larger than those sweetened with sucrose. The effect was most important when comparing Amoxicillin + Clavulanic Acid (sucrose 25mm; versus sorbitol 27mm). The highest inhibitory effect was observed with the usage of Amoxicillin + Flucloxacillin sweetened with sorbitol (38mm). Whereas the lowest inhibitory effect was observed with Amoxicillin and Ampicillin sweetened with sucrose (22mm and 21mm). Conclusion: The results of this study indicate that although all selected antibiotic produced an inhibitory effect on S. mutans, sucrose weakened the inhibitory action of the antibiotic to varying degrees, meanwhile antibiotic formulations containing sorbitol simulated the effects of the control antibiotic. This study calls attention to effects of sweeteners included in pediatric drugs on the oral hygiene and tooth decay.

Keywords: pediatric, dentistry, antibiotics, streptococcus mutans, biofilm, sucrose, sugar free

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Authors: A. Kumar, D. Himanshu, Ak Vaish, K. Usman , A. Singh, R. Misra, V. Atam, S. P. Verma, S. Singhal

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Introduction: Resistant and uncontrolled hypertension offer great challenge, in terms of higher risk of morbidity, mortality and not the least, difficulty in diagnosis and management. Our study tries to identify the importance of two crucial aspects of hypertension management, i.e. drug compliance and optimum dosing and also the effect of spironolactone on blood pressure in cases of resistant hypertension. Methodology: A prospective study was carried out among patients, who were referred as case of resistant hypertension to Hypertension Clinic at Gandhi memorial and associated hospital, Lucknow, India from August, 2013 to July 2014. A total of 122 Subjects having uncontrolled BP with ≥3 antihypertensives were selected. After ruling out secondary resistance and with appropriate lifestyle modifications, effect of adherence and optimum doses was seen with monitoring of BP. Only those having blood pressure still uncontrolled were true resistant. These patients were given spironolactone to see its effect on BP over next 12 weeks. Results: Mean baseline BP of all (n=122) patients was 150.4±7.2 mmHg systolic and 92.1±5.7 mmHg diastolic. After promoting adherence to the regimen, there was reduction of 4.20±3.65 mmHg systolic and 2.08±4.74 mmHg Diastolic blood pressure, with 26 patients achieving target blood pressure goal. Further reduction of 6.66±5.99 mmHg in systolic and 2.59±3.67 mmHg in diastolic BP was observed after optimizing the drug doses with another 66 patients achieving target blood pressure goal. Only 30 patients were true resistant hypertensive and prescribed spironolactone. Over 12 weeks, mean reduction of 20.62±3.65 mmHg in systolic and 10.08 ± 6.46 mmHg in diastolic BP was observed. Out of these 30, BP was controlled in 24 patients. Side effects observed were hyperkalemia in 2 patients and breast tenderness in 2 patients. Conclusion: Improper adherence and suboptimal regimen appear to be the important reasons for uncontrolled hypertension. By virtue of maintaining proper adherence to an optimum regimen, target BP goal can be reached in many without adding much to the regimen. Spironolactone is effective in patients with resistant hypertension, in terms of blood pressure reduction with minimal side effects.

Keywords: resistant, hypertension, spironolactone, blood pressure

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Authors: Kristina Karapetyan, Flora Tkhruni, Tsovinar Balabekyan, Arevik Israyelyan, Tatyana Khachatryan

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The ability of the lactic acid bacteria (LAB) to prevent and cure a variety of diseases, their protective role against infections and colonization of pathogenic microorganisms in the digestive tract, has lead to the coining of the term probiotics or pro-life. LAB inhibiting the growth of pathogenic and food spoilage microorganisms, maintaining the nutritive quality and improving the shelf life of foods. They have also been used as flavor and texture producers. Enterococcus strains have been used for treatment of diseases such as diarrhea or antibiotic associated diarrhea, inflammatory pathologies that affect colon such as irritable bowel syndrome, or immune regulation, diarrhea caused by antibiotic treatments. The obtaining and investigation of biological properties of proteinoceous antibiotics, on the basis of probiotic LAB shown, that bacteriocins, metabiotics, and peptides of LAB represent bactericides have a broad range of activity and are excellent candidates for development of new prophylactic and therapeutic substances to complement or replace conventional antibiotics. The genotyping by 16S rRNA sequencing for LAB were used. Cell free culture broth (CFC) broth was purified by the Gel filtration method on the Sephadex Superfine G 25 resin. Antimicrobial activity was determined by spot-on-lawn method and expressed in arbitrary units (AU/ml). The diversity of multidrug-resistance (MDR) of pathogenic strains to antibiotics, most widely used for treatment of human diseases in the Republics of Armenia and Nagorno Karabakh were examined. It was shown, that difference of resistance of pathogens to antibiotics depends on their isolation sources. The influences of partially purified antimicrobial preparations (AMP), obtained from the different strains of Enterococcus genus on the growth of MDR pathogenic bacteria were investigated. It was shown, that bacteriocin containing partially purified preparations, obtained from different strains of Enterococcus faecium and durans species, possess bactericidal or bacteriostatic activity against antibiotic resistant intestinal, spoilage and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus, E. coli, and Salmonella. Endemic strains of LAB, isolated from Matsoni made from donkey, buffalo and goat milk, shown broad spectrum of activity against food spoiling microorganisms, moulds and fungi, such as Salmonella sp., Esherichia coli, Aspergillus and Penicillium species. Highest activity against MDR pathogens shown bacteria, isolated from goat milk products. High stability of the investigated strains of the genus Enerococcus, isolated from samples of matsun from different regions of Nagorno-Karabakh (NKR) to the antibiotics was shown. The obtained data show high stability of the investigated different strains of the genus Enerococcus. The high genetic diversity in Enterococcus group suggests adaptations for specific mutations in different environments. Thus, endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms isolated from different sources and belong to different taxonomic group. Prospect of the use of certain antimicrobial preparations against pathogenic strains is obvious. These AMP can be applied for long term use against different etiology antibiotic resistant pathogens for prevention or treatment of infectional diseases as an alternative to antibiotics.

Keywords: antimicrobial biopreparation, endemic lactic acid bacteria, intra-species diversity, multidrug resistance of pathogens

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Authors: Warda Fatima, Hasnain Javed

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The emerging trend of Drug resistance in childhood Tuberculosis is increasing worldwide and now becoming a priority challenge for National TB Control Programs of the world. Childhood TB accounts for 10-15% of total TB burden across the globe and same proportion is quantified in case of drug resistant TB. One third population suffering from MDR TB dies annually because of non-diagnosis and unavailability of appropriate treatment. However, true Childhood MDR TB cannot be estimated due to non-confirmation. Diagnosis of Pediatric TB by sputum Smear Microscopy and Culture inoculation are limited due to paucibacillary nature and difficulties in obtaining adequate sputum specimens. Diagnosis becomes more difficult when it comes to HIV infected child. New molecular advancements for early case detection of TB and MDR TB in adults have not been endorsed in children. Multi centered trials are needed to design better diagnostic approaches and efficient and safer treatments for DR TB in high burden countries. The aim of the present study is to sketch out the current situation of the childhood Drug resistant TB especially in the developing world and to highlight the classic and novel methods that are to be implemented in high-burden resource-limited locations.

Keywords: drug resistant TB, childhood, diagnosis, novel methods

Procedia PDF Downloads 374

Authors: L. D. Upegui, Isabel Alvarez-Solorza, Karina Garduno-Ulloa, Maren Boecker

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Introduction: The increasing prevalence rate of resistant and multiresistant bacterial strains to antibiotics is a threat to public health and requires a rapid multifunctional answer. Individuals that are affected by resistant strains present a higher morbidity and mortality than individuals that are infected with the same species of bacteria but with sensitive strains. There have been identified risk factors that are related to the misuse and overuse of antibiotics, like socio-demographic characteristics and psychological aspects of the individuals that have not been explored objectively due to a lack of valid and reliable instruments for their measurement. Objective: To validate a questionnaire for the evaluation of the levels of knowledge related to the use of antibiotics in a Mexican population. Materials and Methods: Analytical cross-sectional observational study. The questionnaire consists of 12 items to evaluated knowledge (1=no, 2=not sure, 3=yes) regarding the use of antibiotics, with higher scores corresponding to a higher level of knowledge. Data are collected in a sample of students. Data collection is still ongoing. In this abstract preliminary results of 30 respondents are reported which were collected during pilot-testing. The validation of the instrument was done using the Rasch model. Fit to the Rasch model was tested checking overall fit to the model, unidimensionality, local independence and evaluating the presence of Differential Item Functioning (DIF) by age and gender. The software Rumm2030 and the SPSS were used for the analyses. Results: The participants of the pilot-testing presented an average age of 32 years ± 12.6 and 53% were women. The preliminary results indicated that the items showed good fit to the Rasch model (chi-squared=12.8 p=0.3795). Unidimensionality (number of significant t-tests of 3%) could be proven, the items were locally independent, and no DIF was observed. Knowledge was the smallest regarding statements on the role of antibiotics in treating infections, e.g., most of the respondents did not know that antibiotics would not work against viral infections (70%) and that they could also cause side effects (87%). The knowledge score ranged from 0 to 100 points with a transformed measurement (mean of knowledge 27.1 ± 4.8). Conclusions: The instrument showed good psychometric proprieties. The low scores of knowledge about antibiotics suggest that misinterpretations on the use of these medicaments were prevalent, which could influence the production of antibiotic resistance. The application of this questionnaire will allow the objective identification of 'Hight risk groups', which will be the target population for future educational campaigns, to reduce the knowledge gaps on the general population as an effort against antibiotic resistance.

Keywords: antibiotics, knowledge, misuse, overuse, questionnaire, Rasch model, validation

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Authors: Nichole Haag, Kimberly Velk, Tyler McCune, Chun Wu

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Methicillin/multiple-resistant Staphylococcus aureus (MRSA) are infectious bacteria that are resistant to common antibiotics. A previous in silico study in our group has identified a hypothetical protein SAV1226 as one of the potential drug targets. In this study, we reported the bioinformatics characterization, as well as cloning, expression, purification and kinetic assays of hypothetical protein SAV1226 from methicillin/vancomycin-resistant Staphylococcus aureus Mu50 strain. MALDI-TOF/MS analysis revealed a low degree of structural similarity with known proteins. Kinetic assays demonstrated that hypothetical protein SAV1226 is neither a domain of an ATP dependent dihydroxyacetone kinase nor of a phosphotransferase system (PTS) dihydroxyacetone kinase, suggesting that the function of hypothetical protein SAV1226 might be misannotated on public databases such as UniProt and InterProScan 5.

Keywords: Methicillin-resistant Staphylococcus aureus, dihydroxyacetone kinase, essential genes, drug target, phosphoryl group donor

Procedia PDF Downloads 377

Authors: Avirup Chowdhury, Angus K. McFadyen, Linsey Batchelor

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Blood cultures (BCs) are an important aspect of management of the septic patient, identifying the underlying pathogen and its antibiotic sensitivities. However, while the current literature outlines indications for initial BCs to be taken, there is little guidance for repeat sampling in the following 5-day period and little information on how antibiotic use can affect the usefulness of this investigation. A retrospective cohort study was conducted using inpatients who had undergone 2 or more BCs within 5 days between April 2016 and April 2017 at a 400-bed hospital in the west of Scotland and received antibiotic therapy between the first and second BCs. The data for BC sampling was collected from the electronic microbiology database, and cross-referenced with data from the hospital electronic prescribing system. Overall, 283 BCs were included in the study, taken from 92 patients (mean 3.08 cultures per patient, range 2-10). All 92 patients had initial BCs, of which 83 were positive (90%). 65 had a further sample within 24 hours of commencement of antibiotics, with 35 positive (54%). 23 had samples within 24-48 hours, with 4 (17%) positive; 12 patients had sampling at 48-72 hours, 12 at 72-96 hours, and 10 at 96-120 hours, with none positive. McNemar’s Exact Test was used to calculate statistical significance for patients who received blood cultures in multiple time blocks (Initial, < 24h, 24-120h, > 120h). For initial vs. < 24h-post BCs (53 patients tested), the proportion of positives fell from 46/53 to 29/53 (one-tailed P=0.002, OR 3.43, 95% CI 1.48-7.96). For initial vs 24-120h (n=42), the proportions were 38/42 and 4/42 respectively (P < 0.001, OR 35.0, 95% CI 4.79-255.48). For initial vs > 120h (n=36), these were 33/36 and 2/36 (P < 0.001,OR ∞). These were also calculated for a positive in initial or < 24h vs. 24-120h (n=42), with proportions of 41/42 and 4/42 (P < 0.001, OR 38.0, 95% CI 5.22-276.78); and for initial or < 24h vs > 120h (n=36), with proportions of 35/36 and 2/36 respectively (P < 0.001, OR ∞). This data appears to show that taking an initial BC followed by a BC within 24 hours of antibiotic commencement would maximise blood culture yield while minimising the risk of false negative results. This could potentially remove the need for as many as 46% of BC samples without adversely affecting patient care. BC yield decreases sharply after 48 hours of antibiotic use, and may not provide any clinically useful information after this time. Further multi-centre studies would validate these findings, and provide a foundation for future health policy generation.

Keywords: antibiotics, blood culture, efficacy, inpatient

Procedia PDF Downloads 144

Authors: Alazar Nebyou, Sujata Pandit

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Lactic acid bacteria (LAB) are essential ingredients in probiotic foods, intestinal microflora, and dairy products that are capable of coping up with harsh gastrointestinal tract conditions and are available in a variety of environments. The objective of this study is to evaluate the probiotic property of LAB isolated from bovine milk. Milk samples were collected from local dairy farms. Samples were obtained using sterile test tubes and transported to a laboratory in the icebox for further biochemical characterization. Preliminary physiological and biochemical identification of LAB isolates was conducted by growing on MRS agar after ten-fold serial dilution. Seven of the best isolates were selected for the evaluation of the probiotic property. The LAB isolates were checked for resistance to antibiotics and their antimicrobial activity by disc diffusion assay and agar well diffusion assay respectively. Bile salt hydrolase activity of isolates was studied by growing isolates in a BSH medium with bile salt. Cell surface property of isolates was assayed by studying their autoaggregation and coaggregation percentage with S. aerues. All isolates were found BSH positive. In addition, BCM2 and BGM1 were susceptible to all antibiotic disks except BBM1 which was resistant to all antibiotic disks. BCM1 and BGM1 had the highest autoaggregation and coaggregation potential respectively. Since all LAB isolates showed gastrointestinal tolerance and good cell surface property they could be considered as good potential probiotic candidates for treatment and probiotic starter culture preparation.

Keywords: probiotic, aggregation, lactic acid bacteria, antimicrobial activity

Procedia PDF Downloads 182

Authors: V. V. Lakshmi, Y. V. S. Annapurna

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Urinary tract infection (UTI) is one of the most common infectious diseases seen in the community. Susceptible individuals experience multiple episodes, and progress to acute pyelonephritis or uro-sepsis or develop asymptomatic bacteriuria (ABU). Ability to cause extraintestinal infections depends on several virulence factors required for survival at extraintestinal sites. Presence of virulence phenotypes enhances the pathogenicity of these otherwise commensal organisms and thus augments its ability to cause extraintestinal infections, the most frequent in urinary tract infections(UTI). The present study focuses on detection of the virulence characters exhibited by the uropathogenic organism and most common factors exhibited in the local pathogens. A total of 700 isolates of E.coli and Klebsiella spp were included in the study. These were isolated from patients from local hospitals reported to be suffering with UTI over a period of three years. Isolation and identification was done based on Gram character and IMVIC reactions. Antibiotic sensitivity profile was carried out by disc diffusion method and multi drug resistant strains with MAR index of 0.7 were further selected.. Virulence features examined included their ability to produce exopolysaccharides, protease- gelatinase production, hemolysin production, haemagglutination and hydrophobicity test. Exopolysaccharide production was most predominant virulence feature among the isolates when checked by congo red method. The biofilms production examined by microtitre plates using ELISA reader confirmed that this is the major factor contributing to virulencity of the pathogens followed by hemolysin production

Keywords: Escherichia coli, Klebsiella sp, Uropathogens, Virulence features.

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Authors: Sreejani Barua, P. P. Srivastav

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Formulation of new functional food products for diabetes patients and obsessed people is a challenge for food industries till date. Starch is a certainly happening, ecological, reasonable and profusely obtainable polysaccharide in plant material. In the present scenario, there is a great interest in modifying starch functional properties without destroying its granular structure using different modification techniques. Resistant starch (RS) contains almost zero calories and can control blood glucose level to prevent diabetes. The current study focused on modification of mung bean starch which is a good source of legumes carbohydrate for the production of functional food. Heat moisture treatment (HMT) of mung starch was conducted at moisture content of 10-30%, temperature of 80-120 °C and time of 8-24 h.The content of resistant starch after modification was significantly increased from native starches containing RS 7.6%. The design combinations of HMT had been completed through Central Composite Rotatable Design (CCRD). The effects of HMT process variables on the yield of resistant starch was studied through Rapid Surface Methodology (RSM). The highest increase of resistant starch was found up to 34.39% when treated the native starch with 30% m.c at 120 °C temperature for 24 h.The functional properties of both native and modified mung bean starches showed that there was a reduction in the swelling power and swelling volume of HMT starches. However, the solubility of the HMT starches was higher than that of untreated native starch and also observed change in structural (scanning electron microscopy), X-Ray diffraction (XRD) pattern, blue value and thermal (differential scanning calorimetry) properties. Therefore, replacing native mung bean starch with heat-moisture treated mung bean starch leads to the development of new products with higher resistant starch levels and functional properties.

Keywords: Mung bean starch, heat moisture treatment, functional properties, resistant starch

Procedia PDF Downloads 172