Search results for: antibiotic stewardship

Authors: Nazedah Ain Ibrahim, Mohamed Mansor Manan

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Information regarding early onset neonatal sepsis (EONS) pathogens may vary between regions. Global perspectives showed Group B Streptococcal (GBS) as the most common causative pathogens, but the widespread use of intrapartum antibiotics has changed the pathogens pattern towards gram negative microorganisms, especially E. coli. Objective of this study is to describe the pathogens isolated, to assess current treatment and risk of EONS. Records of 899 neonates born in three General Hospitals between 2009 until 2012 were retrospectively reviewed. Proven was found in 22 (3%) neonates. The majority was isolated with gram positive organisms, 17 (2.3%). All grams positive and most gram negative organisms showed sensitivity to the tested antibiotics. Only two rare gram negative organisms showed total resistant. Male was possible risk of proven EONS. Although proven EONS remains uncommon in Malaysia, nonetheless, the effect of intrapartum antibiotics still required continuous surveillance.

Keywords: Early onset neonatal sepsis, neonates, pathogens, gram positive, gram negative.

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Authors: Fatemeh Ardestani, Seyed Safa-ali Fatemi, Bagher Yakhchali, Seyed Morteza Hosseyni, Ghasem Najafpour

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Mycophenolic acid “MPA" is a secondary metabolite of Penicillium bervicompactum with antibiotic and immunosuppressive properties. In this study, fermentation process was established for production of mycophenolic acid by Penicillium bervicompactum MUCL 19011 in shake flask. The maximum MPA production, product yield and productivity were 1.379 g/L, 18.6 mg/g glucose and 4.9 mg/L.h respectively. Glucose consumption, biomass and MPA production profiles were investigated during fermentation time. It was found that MPA production starts approximately after 180 hours and reaches to a maximum at 280 h. In the next step, the effects of methionine and acetate concentrations on MPA production were evaluated. Maximum MPA production, product yield and productivity (1.763 g/L, 23.8 mg/g glucose and 6.30 mg/L. h respectively) were obtained with using 2.5 g/L methionine in culture medium. Further addition of methionine had not more positive effect on MPA production. Finally, results showed that the addition of acetate to the culture medium had not any observable effect on MPA production

Keywords: Penicillium bervicompactum, Methionine, Mycophenolic acid, Submerged culture.

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Authors: R. C. Barbuti, M. N. Oliveira, N. P. Perina, C. Haro, P. Bosch, C. S. Bogsan, J. N. Eisig, T. Navarro-Rodriguez

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The management of Helicobacter pylori (H. pylori) eradication is still a matter of discussion, full effectiveness is rarely achieved, and it has many adverse effects. The use of probiotics may be associated with better eradication rates and possibly prevention of adverse events due to antibiotic therapy. The present clinical study was undertaken to evaluate the efficacy of a specially designed fermented milk product, containing Bifidobacterium lactis B420, on the eradication of H. pylori infection in a prospective, randomized, double-blind, controlled study in humans. Four test fermented milks (FM) were specially designed in which counts of viable cells in all products were 10^10 Log CFU. 100 mL-1 for Bifidobacterium lactis - Bifidobacterium species 420. 190 subjects infected with H. pylori, with previous diagnosis of functional dyspepsia according to Rome III criteria entered the study. Bifidobacterium lactis B420, administered twice a day for 90 days was not able to eradicate H. pylori in Brazilian patients with functional dyspepsia.

Keywords: Antibacterial Therapy, Bifidobacteria Fermented milk, Helicobacter pylori, probiotics.

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Authors: F. O. Omoya

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Food borne illnesses have been reported to be a global health challenge. Annual incidences of food–related diseases involve 76 million cases, of which only 14 million can be traced to known pathogens. Poor hygienic practices have contributed greatly to this. It has been reported that in the year 2000 about 2.1 million people died from diarrheal diseases, hence, there is a need to ensure food safety at all level. This study focused on the sterility examination and inhibitory effect of honey samples on selected gram negative and gram positive food borne pathogen from South West Nigeria. The laboratory examinations revealed the presence of some bacterial and fungal contaminations of honey samples and that inhibitory activity of the honey sample was more pronounced on the gram negative bacteria than the gram positive bacterial isolates. Antibiotic sensitivity test conducted on the different bacterial isolates also showed that honey was able to inhibit the proliferation of the tested bacteria than the employed antibiotics.

Keywords: Food borne illness, gram positive and gram negative bacteria, honey, and inhibitory activity.

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Authors: A. Khalique, T. Naseem, N. Haque, Z. Rasool

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The objective of current study was to investigate the effect of Bacillus subtilis PB6 (CloSTAT) as a probiotic in broilers. The corn-soybean based diet was divided into four treatment groups; T1 (basal diet with no probiotic and no Clostridium perfringens); T2 (basal diet challenged with C. perfringens without probiotic); T3 (basal diet challenged with C. perfringens having 0.05% probiotic); T4 (basal diet challenged with C. perfringens having 0.1% probiotic). Every treatment group had four replicates with 24 birds each. Body weight and feed intake were measured on weekly basis, while ileal bacterial count was recorded on day-28 following Clostridium perfringens challenge. The 0.1% probiotic treatment showed 7.2% increase in average feed intake (P=0.05) and 8% increase in body weight compared to T2. In 0.1% treatment body weight was 5% higher than T3 (P=0.02). It was also observed that 0.1% treatment had improved feed conversion ratio (1.77) on 6^th week. No effect of treatment was observed on mortality and ileal bacterial count. The current study indicated that 0.1% use of probiotic had positive response in C. perfringens challenged broilers.

Keywords: Bacillus subtilis PB6, antibiotic growth promoters, Clostridium perfringens, CloSTAT, broilers.

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Authors: F. Javier Benitez, Juan Luis Acero, Francisco J. Real, Gloria Roldan, Francisco Casas

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The degradation of selected pharmaceuticals in some water matrices was studied by using several chemical treatments. The pharmaceuticals selected were the beta-blocker metoprolol, the nonsteroidal anti-inflammatory naproxen, the antibiotic amoxicillin, and the analgesic phenacetin; and their degradations were conducted by using UV radiation alone, ozone, Fenton-s reagent, Fenton-like system, photo-Fenton system, and combinations of UV radiation and ozone with H2O2, TiO2, Fe(II), and Fe(III). The water matrices, in addition to ultra-pure water, were a reservoir water, a groundwater, and two secondary effluents from two municipal WWTP. The results reveal that the presence of any second oxidant enhanced the oxidation rates, with the systems UV/TiO2 and O3/TiO2 providing the highest degradation rates. It is also observed in most of the investigated oxidation systems that the degradation rate followed the sequence: amoxicillin > naproxen > metoprolol > phenacetin. Lower rates were obtained with the pharmaceuticals dissolved in natural waters and secondary effluents due to the organic matter present which consume some amounts of the oxidant agents.

Keywords: Pharmaceuticals, UV radiation, ozone, advancedoxidation processes, water matrices, degradation rates

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Authors: Bahdja Guerfi, N. Hadhoum, I. Azouz, M. Bendoumia, S. Bouafia, F. Z. Hadjadj Aoul

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Background: 'Penicillin V' is a narrow, bactericidal antibiotic of the beta-lactam family of the naturally occurring penicillin group. It is limited to infections due to the germs defined as sensitive. The objective of this work was to identify and to characterize Penicillin V acid and its related compounds by High-performance liquid chromatography (HPLC). Methods: Firstly phenoxymethylpenicillin was identified by an infrared absorption. The organoleptic characteristics, pH, and determination of water content were also studied. The dosage of Penicillin V acid active substance and the determination of its related compounds were carried on waters HPLC, equipped with a UV detector at 254 nm and Discovery HS C18 column (250 mm X 4.6 mm X 5 µm) which is maintained at room temperature. The flow rate was about 1 ml per min. A mixture of water, acetonitrile and acetic acid (65:35:01) was used as mobile phase for phenoxyacetic acid ‘impurity B' and a mixture of water, acetonitrile and acetic acid (650:150:5.75) for the assay and 4-hydroxypenicillin V 'impurity D'. Results: The identification of Penicillin V acid active substance and the evaluation of its chemical quality showed conformity with USP 35th edition. The Penicillin V acid content in the raw material is equal to 1692.22 UI/mg. The percentage content of phenoxyacetic acid and 4-hydroxypenicillin V was respectively: 0.035% and 0.323%. Conclusion: Through these results, we can conclude that the Penicillin V acid active substance tested is of good physicochemical quality.

Keywords: Penicillin V acid, characterization, related substances, HPLC.

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Authors: Osama M. Darwesh, Sahar H. Hassan, Abd El-Raheem R. El-Shanshoury, Shawky Z. Sabae

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Nanotechnology as multidisciplinary technology is growing rapidly with important applications in several sectors. Also, nanobiotechnology is known for the use of microorganisms for the synthesis of targeted nanoparticles. The present study deals with the green synthesis of silver nanoparticles using aquatic bacteria and the development of a biogenic nanocomposite for environmental applications. 20 morphologically different colonies were isolated from the collected water samples from eight different locations at the Rosetta branch of the Nile Delta, Egypt. The obtained results illustrated that the most effective bacterial isolate (produced the higher amount of AgNPs after 24 h of incubation time) is isolate R3. Bacillus tequilensis was the strongest extracellular bio-manufactory of AgNPs. Biosynthesized nanoparticles had a spherical shape with a mean diameter of 2.74 to 28.4 nm. The antimicrobial activity of silver nanoparticles against many pathogenic microbes indicated that the produced AgNPs had high activity against all tested multi-antibiotic resistant pathogens. Also, the stabilized prepared AgNPs-SA nanocomposite has greater catalytic activity for the decolourization of some dyes like Methylene blue (MB) and Crystal violet. Such results represent a promising stage for producing eco-friendly, cost-effective, and easy-to-handle devices for the bioremediation of contaminated industrial wastewater.

Keywords: Bioremediation, AgNPs, AgNPs-SA nanocomposite, Bacillus tequilensis, nanobiotechnology.

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Authors: C. Gallottini, W. Di Mari, A. Amaddeo, K. Barbaro, A. Dolci, G. Dolci, L. Gallottini, G. Barraco, S. Eramo

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A group of 10 dogs (group A) with Periodontal Disease in the third stage, were subjected to regenerative therapy of periodontal tissues, by use of nano hydroxy apatite (NHA). These animals induced by general anesthesia, where treated by ultrasonic scaling, root planning, and at the end by a mucogingival flap in which it was applied NHA. The flap was closed and sutured with simple steps. Another group of 10 dogs (group B), control group, was treated only by scaling and root planning. No patient was subjected to antibiotic therapy. After three months, a check was made by inspection of the oral cavity, radiography and bone biopsy at the alveolar level. Group A showed a total restitutio ad integrum of the periodontal structures, and in group B still mild gingivitis in 70% of cases and 30% of the state remains unchanged. Numerous experimental studies both in animals and humans have documented that the grafts of porous hydroxyapatite are rapidly invaded by fibrovascular tissue which is subsequently converted into mature lamellar bone tissue by activating osteoblast. Since we acted on the removal of necrotic cementum and rehabilitating the root tissue by polishing without intervention in the ligament but only on anatomical functional interface of cement-blasts, we can connect the positive evolution of the clinical-only component of the cement that could represent this perspective, the only reason that Periodontal Disease become a Cement Disease, while all other clinical elements as nothing more than a clinical pathological accompanying.

Keywords: Nanoidroxiaphatite, Parodontal Disease, Rigenerative Therapy.

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Authors: Teerapon Pirom, Ura Pancharoen

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Amoxicillin is an antibiotic which is widely used to treat various infections in both human beings and animals. However, when amoxicillin is released into the environment, it is a major problem. Amoxicillin causes bacterial resistance to these drugs and failure of treatment with antibiotics. Liquid membrane is of great interest as a promising method for the separation and recovery of the target ions from aqueous solutions due to the use of carriers for the transport mechanism, resulting in highly selectivity and rapid transportation of the desired metal ions. The simultaneous processes of extraction and stripping in a single unit operation of liquid membrane system are very interesting. Therefore, it is practical to apply liquid membrane, particularly the HFSLM for industrial applications as HFSLM is proved to be a separation process with lower capital and operating costs, low energy and extractant with long life time, high selectivity and high fluxes compared with solid membranes. It is a simple design amenable to scaling up for industrial applications. The extraction and recovery for (Amoxicillin) through the hollow fiber supported liquid membrane (HFSLM) using aliquat336 as a carrier were explored with the experimental data. The important variables affecting on transport of amoxicillin viz. extractant concentration and operating time were investigated. The highest AMOX- extraction percentages of 85.35 and Amoxicillin stripping of 80.04 were achieved with the best condition at 6 mmol/L [aliquat336] and operating time 100 min. The extraction reaction order (n) and the extraction reaction rate constant (kf) were found to be 1.00 and 0.0344 min-1, respectively.

Keywords: Aliquat336, amoxicillin, HFSLM, kinetic.

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Authors: Fereshteh Shacheraghi, Mohammad Reza Shakibaie, Hanieh Noveiri

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Fourty one strains of ESBL producing P.aeruginosa which were previously isolated from burn patients in Kerman University general hospital, Iran were subjected to PCR, RFLP and sequencing in order to determine the type of extended spectrum β- lactamases (ESBL), the restriction digestion pattern and possibility of mutation among detected genes. DNA extraction was carried out by phenol chloroform method. PCR for detection of bla genes was performed using specific primer for each gene. Restriction Fragment Length Polymorphism (RFLP) for ESBL genes was carried out using EcoRI, NheI, PVUII, EcoRV, DdeI, and PstI restriction enzymes. The PCR products were subjected to direct sequencing of both the strands for identification of the ESBL genes.The blaCTX-M, blaVEB-1, blaPER-1, blaGES-1, blaOXA-1, blaOXA-4 and blaOXA-10 genes were detected in the (n=1) 2.43%, (n=41)100%, (n=28) 68.3%, (n=10) 24.4%, (n=29) 70.7%, (n=7)17.1% and (n=38) 92.7% of the ESBL producing isolates respectively. The RFLP analysis showed that each ESBL gene has identical pattern of digestion among the isolated strains. Sequencing of the ESBL genes confirmed the genuinety of PCR products and revealed no mutation in the restriction sites of the above genes. From results of the present investigation it can be concluded that blaVEB-1 and blaCTX-M were the most and the least frequently isolated ESBL genes among the P.aeruginosa strains isolated from burn patients. The RFLP and sequencing analysis revealed that same clone of the bla genes were indeed existed among the antibiotic resistant strains.

Keywords: ESBL genes, PCR, RFLP, Sequencing, P.aeruginosa

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Authors: Nadia El Alami El Hassani, Soukaina Motia, Benachir Bouchikhi, Nezha El Bari

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Doxycycline (DXy) is a cycline antibiotic, most frequently prescribed to treat bacterial infections in veterinary medicine. However, its broad antimicrobial activity and low cost, lead to an intensive use, which can seriously affect human health. Therefore, its spread in the food products has to be monitored. The scope of this work was to synthetize a sensitive and very selective molecularly imprinted polymer (MIP) for DXy detection in honey samples. Firstly, the synthesis of this biosensor was performed by casting a layer of carboxylate polyvinyl chloride (PVC-COOH) on the working surface of a gold screen-printed electrode (Au-SPE) in order to bind covalently the analyte under mild conditions. Secondly, DXy as a template molecule was bounded to the activated carboxylic groups, and the formation of MIP was performed by a biocompatible polymer by the mean of polyacrylamide matrix. Then, DXy was detected by measurements of differential pulse voltammetry (DPV). A non-imprinted polymer (NIP) prepared in the same conditions and without the use of template molecule was also performed. We have noticed that the elaborated biosensor exhibits a high sensitivity and a linear behavior between the regenerated current and the logarithmic concentrations of DXy from 0.1 pg.mL⁻¹ to 1000 pg.mL⁻¹. This technic was successfully applied to determine DXy residues in honey samples with a limit of detection (LOD) of 0.1 pg.mL⁻¹ and an excellent selectivity when compared to the results of oxytetracycline (OXy) as analogous interfering compound. The proposed method is cheap, sensitive, selective, simple, and is applied successfully to detect DXy in honey with the recoveries of 87% and 95%. Considering these advantages, this system provides a further perspective for food quality control in industrial fields.

Keywords: Electrochemical sensor, molecular imprinted polymer, doxycycline, food control.

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Authors: Lubis A., Widayanti R., Hikmah Z., Endaryanto A., Harsono A., Harianto A., Etika R., Handayani D. K., Sampurna M.

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Neonatal lupus erythematous (NLE) is a rare disease marked by clinical characteristic and specific maternal autoantibody. Many cutaneous, cardiac, liver, and hematological manifestations could happen with affect of one organ or multiple. In this case, both babies were premature, low birth weight (LBW), small for gestational age (SGA) and born through caesarean section from a systemic lupus erythematous (SLE) mother. In the first case, we found a baby girl with dyspnea and grunting. Chest X ray showed respiratory distress syndrome (RDS) great I and echocardiography showed small atrial septal defect (ASD) and ventricular septal defect (VSD). She also developed anemia, thrombocytopenia, elevated C-reactive protein, hypoalbuminemia, increasing coagulation factors, hyperbilirubinemia, and positive blood culture of Klebsiella pneumonia. Anti-Ro/SSA and Anti-nRNP/sm were positive. Intravenous fluid, antibiotic, transfusion of blood, thrombocyte concentrate, and fresh frozen plasma were given. The second baby, male presented with necrotic tissue on the left ear and skin rashes, erythematous macula, athropic scarring, hyperpigmentation on all of his body with various size and facial haemorrhage. He also suffered from thrombocytopenia, mild elevated transaminase enzyme, hyperbilirubinemia, anti-Ro/SSA was positive. Intravenous fluid, methyprednisolone, intravenous immunoglobulin (IVIG), blood, and thrombocyte concentrate transfution were given. Two cases of neonatal lupus erythematous had been presented. Diagnosis based on clinical presentation and maternal auto antibody on neonate. Organ involvement in NLE can occur as single or multiple manifestations.

Keywords: Neonatus lupus erythematous, maternal autoantibody, clinical characteristic.

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Authors: Jatindra N. Bhakta, Kouhei Ohnishi, Yukihiro Munekage, Kozo Iwasaki

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The growing health hazardous impact of arsenic (As) contamination in environment is the impetus of the present investigation. Application of lactic acid bacteria (LAB) for the removal of toxic and heavy metals from water has been reported. This study was performed in order to isolate and characterize the Asresistant LAB from mud and sludge samples for using as efficient As uptaking probiotic. Isolation of As-resistant LAB colonies was performed by spread plate technique using bromocresol purple impregnated-MRS (BP-MRS) agar media provided with As @ 50 μg/ml. Isolated LAB were employed for probiotic characterization process, acid and bile tolerance, lactic acid production, antibacterial activity and antibiotic tolerance assays. After As-resistant and removal characterizations, the LAB were identified using 16S rDNA sequencing. A total of 103 isolates were identified as As-resistant strains of LAB. The survival of 6 strains (As99-1, As100-2, As101-3, As102-4, As105-7, and As112-9) was found after passing through the sequential probiotic characterizations. Resistant pattern pronounced hollow zones at As concentration >2000 μg/ml in As99-1, As100-2, and As101-3 LAB strains, whereas it was found at ~1000 μg/ml in rest 3 strains. Among 6 strains, the As uptake efficiency of As102-4 (0.006 μg/h/mg wet weight of cell) was higher (17 – 209%) compared to remaining LAB. 16S rDNA sequencing data of 3 (As99- 1, As100-2, and As101-3) and 3 (As102-4, As105-7, and As112-9) LAB strains clearly showed 97 to 99% (340 bp) homology to Pediococcus dextrinicus and Pediococcus acidilactici, respectively. Though, there was no correlation between the metal resistant and removal efficiency of LAB examined but identified elevated As removing LAB would probably be a potential As uptaking probiotic agent. Since present experiment concerned with only As removal from pure water, As removal and removal mechanism in natural condition of intestinal milieu should be assessed in future studies.

Keywords: Lactic acid bacteria, As-resistant, characterization, Pediococcus sp., As removal probiotic.

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Authors: Goksen Arik, Mihriban Korukluoglu

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Lactic acid bacteria (LAB) and some yeast species are common microorganisms found in dairy products and most of them are responsible for the fermentation of foods. Such cultures are isolated and used as a starter culture in the food industry because of providing standardisation of the final product during the food processing. Choice of starter culture is the most important step for the production of fermented food. Isolated LAB and yeast cultures which have the ability to create a biofilm layer can be preferred as a starter in the food industry. The biofilm formation could be beneficial to extend the period of usage time of microorganisms as a starter. On the other hand, it is an undesirable property in pathogens, since biofilm structure allows a microorganism become more resistant to stress conditions such as antibiotic presence. It is thought that the resistance mechanism could be turned into an advantage by promoting the effective microorganisms which are used in the food industry as starter culture and also which have potential to stimulate the gastrointestinal system. Development of the biofilm layer is observed in some LAB and yeast strains. The resistance could make LAB and yeast strains dominant microflora in the human gastrointestinal system; thus, competition against pathogen microorganisms can be provided more easily. Based on this circumstance, in the study, 10 LAB and 10 yeast strains were isolated from various dairy products, such as cheese, yoghurt, kefir, and cream. Samples were obtained from farmer markets and bazaars in Bursa, Turkey. As a part of this research, all isolated strains were identified and their ability of biofilm formation was detected with two different methods and compared with each other. The first goal of this research was to determine whether isolates have the potential for biofilm production, and the second was to compare the validity of two different methods, which are known as “Tube method” and “96-well plate-based method”. This study may offer an insight into developing a point of view about biofilm formation and its beneficial properties in LAB and yeast cultures used as a starter in the food industry.

Keywords: Biofilm, dairy products, lactic acid bacteria, yeast.

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Authors: Wisam H. Benamer, Ehab A. Elfallah, Mohamed A. Elshaari, Farag A. Elshaari

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A challenge for laboratories is to provide bacterial identification and antibiotic sensitivity results within a short time. Hence, advancement in the required technology is desirable to improve timing, accuracy and quality. Even with the current advances in methods used for both phenotypic and genotypic identification of bacteria the need is there to develop method(s) that enhance the outcome of bacteriology laboratories in accuracy and time. The hypothesis introduced here is based on the assumption that the chromosome of any bacteria contains unique sequences that can be used for its identification and typing. The outcome of a pilot study designed to test this hypothesis is reported in this manuscript. Methods: The complete chromosome sequences of several bacterial species were downloaded to use as search targets for unique sequences. Visual basic and SQL server (2014) were used to generate a complete set of 18-base long primers, a process started with reverse translation of randomly chosen 6 amino acids to limit the number of the generated primers. In addition, the software used to scan the downloaded chromosomes using the generated primers for similarities was designed, and the resulting hits were classified according to the number of similar chromosomal sequences, i.e., unique or otherwise. Results: All primers that had identical/similar sequences in the selected genome sequence(s) were classified according to the number of hits in the chromosomes search. Those that were identical to a single site on a single bacterial chromosome were referred to as unique. On the other hand, most generated primers sequences were identical to multiple sites on a single or multiple chromosomes. Following scanning, the generated primers were classified based on ability to differentiate between medically important bacterial and the initial results looks promising. Conclusion: A simple strategy that started by generating primers was introduced; the primers were used to screen bacterial genomes for match. Primer(s) that were uniquely identical to specific DNA sequence on a specific bacterial chromosome were selected. The identified unique sequence can be used in different molecular diagnostic techniques, possibly to identify bacteria. In addition, a single primer that can identify multiple sites in a single chromosome can be exploited for region or genome identification. Although genomes sequences draft of isolates of organism DNA enable high throughput primer design using alignment strategy, and this enhances diagnostic performance in comparison to traditional molecular assays. In this method the generated primers can be used to identify an organism before the draft sequence is completed. In addition, the generated primers can be used to build a bank for easy access of the primers that can be used to identify bacteria.

Keywords: Bacteria chromosome, bacterial identification, sequence, primer generation.

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