Chemistry and Biological Activity of Feed Additive for Poultry Farming

Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.

The Appropriateness of Antibiotic Prescribing within Dundee Dental Hospital

Background: The societal impact of antibiotic resistance is a major public health concern. The increase in incidence of resistant bacteria can ultimately be fatal. Objective: To analyse the appropriateness of antibiotic prescribing in Dundee Dental Hospital, ultimately improving the safety and quality of patient care. Methods: Two examiners independently crosschecked approximately fifty consecutive prescriptions, and corresponding patient case notes, for three data collection cycles between August 2014 – September 2015. The Scottish Dental Clinical Effectiveness Program (SDCEP) Drug Prescribing for Dentistry guidelines was the standard utilised. The criteria: clinical justification, regime justification and review arrangements was measured, and compared to the standard. Results: Cycle one revealed 42% of antibiotic prescriptions were appropriate. Interventions included: multiple staff meetings, introduction of a checklist attached to the prescription pack, and production of patient leaflets explaining indications for antibiotics. Cycle two and three revealed 44%, and 30% compliance, respectively. Conclusion: The results of the audit have yet to meet target standards set out in prescribing guidelines. However, steps are being taken and change has occurred on a cultural level.

Incidence of Acinetobacter in Fresh Carrot (Daucus carota subsp. sativus)

The research aims 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 are analyzed on MacConkey agar and screened by Microbact 24E (Oxoid) and susceptibility of isolates is 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 were similarities in the resistances shown by A. baumannii and A. lwoffii to fluoroquinolones and β- lactame drug families in addition to between sulfonamide and animoglycoside demonstrated by A. lwoffii. Significant correlation in similarities were observed at P < 0.05 to CPX to NA (46.2%), and SXT to AU (52.6%) A. baumannii and A. lwoffii respectively and high multi drug resistance (MDR) of 27.27% and 62.50% by A. baumannii and A. lwoffii respectively. 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.

Microbiological Profile of UTI along with Their Antibiotic Sensitivity Pattern with Special Reference to Nitrofurantoin

Urinary Tract Infections are considered as one of the most common bacterial infections with an estimated annual global incidence of 150 million. Antimicrobial drug resistance is one of the major threats due to wide spread usage of uncontrolled antibiotics. In this study, a total number of 9149 urine samples were collected from R.H Patiala and processed in the Department of Microbiology G. M. C Patiala (January 2013 to December 2013). Urine samples were inoculated on MacConkey’s and blood agar plates and incubated at 370C for 24 hrs. The organisms were identified by colony characters, Gram’s staining, and biochemical reactions. Antimicrobial susceptibility of the isolates was determined against various antimicrobial agents (Hi – Media Mumbai India) by Kirby Bauer DISK diffusion method on Muller Hinton agar plates. Maximum patients were in the age group of 21-30 yrs followed by 31-40 yrs. Males (34%) are less prone to urinary tract infections than females (66%). Culture was positive in 25% of the samples. Escherichia coli was the most common isolate 60.3% followed by Klebsiella pneumoniae 13.5%, Proteus spp. 9% and Staphylococcus aureus 7.6%. Most of the urinary isolates were sensitive to, carbepenems, Aztreonam, Amikacin, and Piperacillin + Tazobactum. All the isolates showed a good sensitivity towards Nitrofurantoin (82%). ESBL production was found to be 70.6% in Escherichia coli and 29.4% in Klebsiella pneumonia. Susceptibility of ESBL producers to Imipenem, Nitrofurantoin and Amikacin were found to be 100%, 76%, and 75% respectively. Uropathogens are increasingly showing resistance to many antibiotics making empiric management of outpatient UTIs challenging. Ampicillin, Cotrimoxazole and Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin could be used in lower urinary tract infection. Knowledge of uropathogens and their antimicrobial susceptibility pattern in a geographical region will help in appropriate and judicious antibiotic usage in a health care setup.

Kinetic and Removable of Amoxicillin Using Aliquat336 as a Carrier via a HFSLM

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.

Bioinformatics and Molecular Biological Characterization of a Hypothetical Protein SAV1226 as a Potential Drug Target for Methicillin/Vancomycin- Staphylococcus aureus Infections

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.

Heavy Metals and Antibiotic Resistant Bacteria in Green Turtles are Indicators of Environmental Pollution

Freshly laid eggs from green turtles, Chelonia mydas, were randomly collected from Ras Al-Hadd Reserve, Oman. Eggshells taken from eggs and sand collected from the body chamber were analyzed for eight heavy metals (Al, Br, Cd, Co, Cu, Fe, S, and Zn) using inductively coupled plasma mass spectrometry (ICP). Heavy metal concentrations varied significantly (P

Physicochemical Profile of Essential Oil of Daucus carota

Essential oils have a significant antimicrobial activity. These oils can successfully replace the antibiotics. So, the microorganisms show their inefficiencies resistant for the antibiotics. For this reason, we study the physicochemical analysis and antimicrobial activity of the essential oil of Daucus carota. The extraction is done by steam distillation of water which brought us a very significant return of 4.65%. The analysis of the essential oil is performed by GC / MS and has allowed us to identify 32 compounds in the oil of D. carota flowering tops of Bouira. Three of which are in the majority are the α-Pinene (22.3%), the carotol (21.7%) and the limonene (15.8%).

Extracellular Protein Secreted by Bacillus subtilis ATCC21332 in the Presence of Streptomycin Sulfate

The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.

Effects of Ciprofloxacin and Levofloxacin Administration on Some Oxidative Stress Markers in the Rat

Fluoroquinolones are a group of antibiotics widely used because of their broad spectrum activity against both Gram-positive and Gram-negative bacteria. In this study, ciprofloxacin and levofloxacin were administered to rats at therapeutic doses to evaluate their effects on plasma arylesterase activity, as well as, on hepatic advanced oxidized protein products (AOPPs) and malondialdehyde (MDA) levels, as measures of oxidative stress. Ciprofloxacin (80 mg/kg body weight) and levofloxacin (40 mg/kg body weight) were administered to male albino rats for 7 and 14 days. The data obtained demonstrated that plasma arylesterase activity was significantly decreased by both drugs with ciprofloxacin administration inhibiting the activity by 29% and 30% while Levofloxacin treatment resulted in 35% and 30% inhibition, after 7 and 14 days treatment respectively. Hepatic AOPP and MDA levels were both elevated by these antibiotics. This study supplies further evidence that fluoroquinolones at therapeutic doses promote oxidative stress.

Technique for Processing and Preservation of Human Amniotic Membrane for Ocular Surface Reconstruction

Human amniotic membrane (HAM) is a useful biological material for the reconstruction of damaged ocular surface. The processing and preservation of HAM is critical to prevent the patients undergoing amniotic membrane transplant (AMT) from cross infections. For HAM preparation human placenta is obtained after an elective cesarean delivery. Before collection, the donor is screened for seronegativity of HCV, Hbs Ag, HIV and Syphilis. After collection, placenta is washed in balanced salt solution (BSS) in sterile environment. Amniotic membrane is then separated from the placenta as well as chorion while keeping the preparation in BSS. Scrapping of HAM is then carried out manually until all the debris is removed and clear transparent membrane is acquired. Nitrocellulose membrane filters are then placed on the stromal side of HAM, cut around the edges with little membrane folded towards other side making it easy to separate during surgery. HAM is finally stored in solution of glycerine and Dulbecco-s Modified Eagle Medium (DMEM) in 1:1 ratio containing antibiotics. The capped borosil vials containing HAM are kept at -80°C until use. This vial is thawed to room temperature and opened under sterile operation theatre conditions at the time of surgery.

Increase of Peroxidase Activity of Haptoglobin (2-2)-Hemoglobin at Pathologic Temperature and Presence of Antibiotics

Free Hemoglobin promotes the accumulation of hydroxyl radicals by the heme iron, which can react with endogenous hydrogen peroxide to produce free radicals which may cause severe oxidative cell damage. Haptoglobin binds to Hemoglobin strongly and Haptoglobin-Hemoglobin binding is irreversible. Peroxidase activity of Haptoglobin(2-2)-Hemoglobin complex was assayed by following increase of absorption of produced tetraguaiacol as the second substrate of Haptoglobin-Hemoglobin complex at 470 nm and 42°C by UV-Vis spectrophotometer. The results have shown that peroxidase activity of Haptoglobin(2-2)-Hemoglobin complex is modulated via homotropic effect of hydrogen peroxide as allostric substrate. On the other hand antioxidant property of Haptoglobin(2- 2)-Hemoglobin was increased via heterotropic effect of the two drugs (especially ampicillin) on peroxidase activity of the complex. Both drugs also have mild effect on quality of homotropic property of peroxidase activity of Haptoglobin(2-2)-Hemoglobin complex. Therefore, in vitro studies show that the two drugs may help Hp-Hb complex to remove hydrogen peroxide from serum at pathologic temperature ature (42 C).

Some Peculiarities of Growth and Functional Activity of Escherichia coli Strain from Probiotic Formula “ASAP“

It has been shown that pH 7,3 and 37 0C are the optimal condition for the growth of E. coli “ASAP". The cells grow well on Glucose, Lactose, D-Mannitol, D-Sorbitol, (+)-Xylose, L- (+)-Arabinose and Dulcitol. No growth has been observed on Sucrose, Inositol, Phenylalanine, and Tryptophan. The strain is sensitive to a range of antibiotics. The present study has demonstrated that E. coli “ASAP" inhibit the growth of S. enterica ATCC #700931 in vitro. The studies on conjugating activity has revealed no conjugant of E. coli “ASAP" with plasmid strains E. coli G35#59 and S. enterica ATCC #700931. On the other hand, the conjugants with low frequencies were obtained from E. coli “ASAP" with E. coli G35#61, and E. coli “ASAP" with randomly chosen isolate from healthy human gut microflora: E. coli E6. The results of present study have demonstrated improvements in gut microflora condition of patients with different diseases after the administration of “ASAP"

Protein Production by Bacillus Subtilis Atcc 21332 in the Presence of Cymbopogon Essential Oils

Proteins levels produced by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antimicrobial agents or antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics or natural compounds in nature as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was focusing on the effect of essential oils from Cymbopogon flexuosus and C. nardus in regulating proteins production by Bacillus subtilis ATCC 21332. The Minimum Inhibition Concentrations (MICs) of both essential oils on B. subtilis were determined by using microdilution assay, resulting 0.2% and 1.56% for each C. flexuosus and C. nardus subsequently. The bacteria were further exposed to each essential oils at concentration of 0.01XMIC for 2 days. The proteins were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Protein profile showed that a band with approximate size of 250 kD was appeared for the treated bacteria with essential oils. Thus, Bacillus subtilis ATCC 21332 in stressful condition with the presence of essential oils at low concentration could induce the protein production.

Antibiotic Resistance Profile of Bacterial Isolates from Animal Farming Aquatic Environments and Meats in a Peri-Urban Community in South Korea

The increasing usage of antibiotics in the animal farming industry is an emerging worldwide problem contributing to the development of antibiotic resistance. The purpose of this work was to investigate the prevalence and antibiotic resistance profile of bacterial isolates collected from aquatic environments and meats in a peri-urban community in Daejeon, Korea. In an antibacterial susceptibility test, the bacterial isolates showed a high incidence of resistance (~ 26.04 %) to cefazolin, tetracycline, gentamycin, norfloxacin, erythromycin and vancomycin. The results from a test for multiple antibiotic resistance indicated that the isolates were displaying an approximately 5-fold increase in the incidence of multiple antibiotic resistance to combinations of two different antibiotics compared to combinations of three or more antibiotics. Most of the isolates showed multi-antibiotic resistance, and the resistance patterns were similar among the sampling groups. Sequencing data analysis of 16S rRNA showed that most of the resistant isolates appeared to be dominated by the classes Betaproteobacteria and Gammaproteobacteria in the phylum Proteobacteria.

Sterility Examination and Comparative Analyses of Inhibitory Effect of Honey on Some Gram Negative and Gram Positive Food Borne Pathogens in South West Nigeria

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.

Early Onset Neonatal Sepsis Pathogens in Malaysian Hospitals: Determining Empiric Antibiotic

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.