Search results for: antibiotic residues

Authors: Ranjith Kumar Kankala, Wei-Zhi Lin, Chia-Hung Lee

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Antibiotic resistance in bacteria has become an emergency situation clinically. To improve the efficacy of antibiotics in resistant strains, advancement of nanoparticles is inevitable than ever. Herewith, we demonstrate a design by immobilizing tetracycline (TET) in copper substituted mesoporous silica nanoparticles (Cu-MSNs) through a pH-sensitive coordination link, enabling its release in the acidic environment. Subsequently, MSNs are coated with silver nanoparticles stabilized polyethyleneimine (PEI-SNP) to act against drug-resistant (MDR) bacterial strains. Silver ions released from SNP are capable of sensitizing the resistant strains and facilitate the generation of free radicals capable of damaging the cell components. In addition, copper ions in the framework are also capable of generating free radicals through Fenton-like reaction. Furthermore, the nanoparticles are well-characterized physically, and various antibacterial efficacious tests against isolated multidrug resistant bacterial strain were highly commendable. However, this formulation has no significant toxic effect on normal mammalian fibroblast cells accounting its high biocompatibility. These MSN trio-hybrids, i.e., SNP, tetracycline, and copper ions result in synergistic effects, and their advancement could bypass resistance and allow synergism for effective treatment of antibiotic clinically.

Keywords: antibiotic resistance, copper, mesoporous silica nanoparticles, Ph-sensitive release, polyethyleneimine, silver, tetracycline

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Authors: Janneth González, George Barreto, Ludis Morales, Angélica Sabogal

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Sea anemone neurotoxins are peptides that interact with Na+ and K+ channels, resulting in specific alterations on their functions. Some of these neurotoxins (1ROO, 1BGK, 2K9E, 1BEI) are important for the treatment of nearly eighty autoimmune disorders due to their specificity for Kv1.3 channel. The aim of this study was to identify the common residues among these neurotoxins by computational methods, and establish whether there is a pattern useful for the future generation of a treatment for autoimmune diseases. Our results showed eight new key common residues between the studied neurotoxins interacting with a histidine ring and the selectivity filter of the receptor, thus showing a possible pattern of interaction. This knowledge may serve as an input for the design of more promising drugs for autoimmune treatments.

Keywords: neurotoxins, potassium channel, Kv1.3, computational methods, autoimmune diseases

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Authors: Jin-Hyeong Park, Hong-Seok Kim, Jin-Hyeok Yim, Young-Ji Kim, Dong-Hyeon Kim, Jung-Whan Chon, Kun-Ho Seo

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Salmonella contamination in chicken samples can cause major health problems in humans. However, not only the effects of antibiotic treatment during growth but also the impacts of poultry slaughter line on the prevalence of Salmonella in final chicken meat sold to consumers are unknown. In this study, we compared the isolation rates and antimicrobial resistance of Salmonella between antibiotic-free, conventional, conventional Korean native retail chicken meat samples and clonal divergence of Salmonella isolates by multilocus sequence typing. In addition, the distribution of extended-spectrum β-lactamase (ESBL) genes in ESBL-producing Salmonella isolates was analyzed. A total of 72 retail chicken meat samples (n = 24 antibiotic-free broiler [AFB] chickens, n = 24 conventional broiler [CB] chickens, and n = 24 conventional Korean native [CK] chickens) were collected from local retail markets in Seoul, South Korea. The isolation rates of Salmonella were 66.6% in AFB chickens, 45.8% in CB chickens, and 25% in CK chickens. By analyzing the minimum inhibitory concentrations of β -lactam antibiotics with the disc-diffusion test, we found that 81.2% of Salmonella isolates from AFB chickens, 63.6% of isolates from CB chickens, and 50% of isolates from CK chickens were ESBL producers; all ESBL-positive isolates had the CTX-M-15 genotype. Interestingly, all ESBL-producing Salmonella were revealed as ST16 by multilocus sequence typing. In addition, all CTX-M-15-positive isolates had the genetic platform of blaCTX-M gene (IS26-ISEcp1-blaCTX-M-15-IS903), to the best of our knowledge, this is the first report in Salmonella around the world. The Salmonella ST33 strain (S. Hadar) isolated in this study has never been reported in South Korea. In conclusion, our findings showed that antibiotic-free retail chicken meat products were also largely contaminated with ESBL-producing Salmonella and that their ESBL genes and genetic platforms were the same as those isolated from conventional retail chicken meat products.

Keywords: antibiotic-free poultry, conventional poultry, multilocus sequence typing, extended-spectrum β-lactamase, antimicrobial resistance

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Authors: Johannes Full, Martin Reisinger, Alexander Sauer, Robert Miehe

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The use of biogenic residues for the biotechnological production of chemical energy carriers for electricity and heat generation as well as for mobile applications is an important lever for the shift away from fossil fuels towards a carbon dioxide neutral post-fossil future. A multitude of promising biotechnological processes needs, therefore, to be compared against each other. For this purpose, a multi-objective target system and a corresponding methodology for the evaluation of the underlying key figures are presented in this paper, which can serve as a basis for decisionmaking for companies and promotional policy measures. The methodology considers in this paper the economic and ecological potential of bio-hydrogen production using the example of hydrogen production from fruit and milk production waste with the purple bacterium R. rubrum (so-called dark photosynthesis process) for the first time. The substrate used in this cost-effective and scalable process is fructose from waste material and waste deposits. Based on an estimation of the biomass potential of such fructose residues, the new methodology is used to compare different scenarios for the production and usage of bio-hydrogen through the considered process. In conclusion, this paper presents, at the example of the promising dark photosynthesis process, a methodology to evaluate the ecological and economic potential of biotechnological production of bio-hydrogen from residues and waste.

Keywords: biofuel, hydrogen, R. rubrum, bioenergy

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Authors: Enny Suswati, Supangat Supangat

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Background. Ready-to-eat food products are becoming increasingly popular because consumers are increasingly busy, competitive, and changing lifestyles. Examples of ready-to-eat foods include crispy fried chicken. Escherichia coli is one of the most important causes of food-borne diseases and the most frequent antibiotic-resistant pathogen globally. This study assessed the prevalence and antibiotic resistance profile of E. coli from ready-to-eat crispy fried chicken in Jember city, Indonesia. Methodology. This cross-sectional study was conducted from November 2020 to April 2021 by collecting 81crispy fried chicken samples from 27 food stalls in campus area using a simple random sampling method. Isolation and determination of E. coli use were performed by conventional culture method. An antibiotic susceptibility test was conducted using Kirby Bauer disk diffusion method on the Mueller–Hinton agar. Result. Out of 81crispy fried chicken samples, 77 (95.06%) were positive for E. coli. High E. coli drug resistance was observed on ampicillin, amoxicillin (100%) followed by cefixime (98.72%), erythromycin (97.59%), sulfamethoxazole (93.59%), azithromicin (83.33%), cefotaxime (78.28%), choramphenicol (75.64%), and cefixime (74.36%). On the other hand, there was the highest susceptibility for ciprofloxacin (64.10%). The multiple antibiotic resistance indexes of E. coli isolates varied from 0.4 to 1. The predominant antimicrobial resistance profiles of E. coli were CfmCroAmlAmpAzmCtxSxtCE (n=17), CfmCroAmlCipAmpAzmCtxSxtCE (n=16), and CfmAmlAmpAzmCtxSxtCE (n=5), respectively. Multidrug resistance was also found in the isolates' 76/77 (98.70%). Conclusion. The resistance pattern CfmCroAmlAmpAzmCtxSxtCE was the most common among the E. coli isolates, with 17 showing it. The multiple antibiotic index (MAR index) ranged from 0.4 to 1. Hygienic measures should be rigorously implemented and monitoring resistance of E. coli is required to reduce the risks related to the emergence of multi-resistant bacteria

Keywords: antibacterial drug, ready to eat, crispy fried chicken, escherichia coli

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Authors: Vinod Nair, C. Sadasivan

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Beta-lactam antibiotics are the most frequently prescribed medications in modern medicine. The antibiotic resistance by the production of enzyme beta-lactamase is an important mechanism seen in microorganisms. Resistance to beta-lactams mediated by beta-lactamases can be overcome successfully with the use of beta-lactamase inhibitors. New generations of the antibiotics contain mostly synthetic compounds, and many side effects have been reported for them. Combinations of beta-lactam and beta-lactamase inhibitors have become one of the most successful antimicrobial strategies in the current scenario of bacterial infections. Plant-based drugs are very cheap and having lesser adverse effect than synthetic compounds. The synergistic effect of eugenol acetate with beta-lactams restores the activity of beta-lactams, allowing their continued clinical use. It is reported here the enhanced inhibitory effect of phytochemical, eugenol acetate, isolated from the plant Syzygium aromaticum with beta-lactams on beta-lactamase. The compound was found to have synergistic effect with the antibiotic amoxicillin against antibiotic-resistant strain of S.aureus. The enzyme was purified from the organism and incubated with the compound. The assay showed that the compound could inhibit the enzymatic activity of beta-lactamase. Modeling and molecular docking studies indicated that the compound can fit into the active site of beta-lactamase and can mask the important residue for hydrolysis of beta-lactams. The synergistic effects of eugenol acetate with beta-lactam antibiotics may justify, the use of these plant compounds for the preparation of β-lactamase inhibitors against β-lactam resistant S.aureus.

Keywords: betalactamase, eugenol acetate, synergistic effect, molecular modeling

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Authors: Soukaina Oujana, Peggy Zwolinski

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Fines are heterogeneous residues created during the shredding of non-hazardous waste. They are one of the most challenging issues faced by recyclers, because they are at the present time considered as non-sortable and non-reusable mixtures destined to landfill. However, fines contain a large amount of recoverable materials that could be recycled or reused for the production of solid recovered fuel. This research is conducted in relation to a project named ValoRABES. The aim is to characterize fines and establish a suitable sorting process in order to extract the materials contained in the mixture and define their suitable recovery paths. This paper will highlight the importance of a good sampling and will propose a sampling methodology for fines characterization. First results about the characterization will be also presented.

Keywords: fines, non-hazardous waste, recovery, shredding residues, waste characterization, waste sampling

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Authors: Christina Roman, Deepak Koirala, Joseph A. Piccirilli

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Phage displaying synthetic Fab libraries have been used to obtain Fabs that bind to specific RNA targets with high affinity and specificity. These Fabs have been demonstrated to facilitate RNA crystallization. However, the antibody framework used in the construction of these phage display libraries contains numerous bulky, flexible, and charged residues, which facilitate solubility and hinder aggregation. These residues can interfere with crystallization due to the entropic cost associated with burying them within crystal contacts. To systematically reduce the surface entropy of the Fabs and improve their crystallization properties, a protein engineering strategy termed surface entropy reduction (SER) is being applied to the Fab framework. In this approach, high entropy residues are mutated to smaller ones such as alanine or serine. Focusing initially on Fab BL3-6, which binds an RNA AAACA pentaloop with 20nM affinity, the SER P server (http://services.mbi.ucla.edu/SER/) was used and analysis was performed on existing RNA-Fab BL3-6 co-crystal structures. From this analysis twelve surface entropy reduced mutants were designed. These SER mutants were expressed and are now being measured for their crystallization and diffraction performance with various RNA targets. So far, one mutant has generated 3.02 angstrom diffraction with the yjdF riboswitch RNA. Ultimately, the most productive mutations will be combined into a new Fab framework to be used in a optimized phage displayed Fab library.

Keywords: antibody fragment, crystallography, RNA, surface entropy reduction

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Authors: Nataliya Apanovich, Mark Wright

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Uganda is among the poorest but fastest growing populations in the world. Its annual population growth of 3% puts additional stress through land fragmentation, agricultural intensification, and deforestation on already highly weathered tropical (Ferralsol) soils. All of these factors lead to decreased agricultural yields and consequently diminished food security. The central region of Uganda, Buganda Kingdom, is especially vulnerable in terms of food security as its high population density coupled with mismanagement of natural resources led to gradual loss of its soil and even changes in microclimate. These changes are negatively affecting livelihoods of smallholder farmers who comprise 80% of all population in Uganda. This research focuses on biochar for soil remediation in Masaka District, Uganda. If produced on a small scale from locally sourced materials, biochar can increase the quality of soil in a cost and time effective manner. To assess biochar potential, 151 smallholder farmers were interviewed on the types of crops grown, agricultural residues produced and their use, as well as on attitudes towards biochar use and its production on a small scale. The interviews were conducted in 7 sub-counties, 32 parishes, and 92 villages. The total farmland covered by the study was 606.2 kilometers. Additional information on the state of agricultural development and environmental degradation in the district was solicited from four local government officials via informal interviews. This project has been conducted in collaboration with the international agricultural research institution, Makerere University in Kampala, Uganda. The results of this research can have implications on the way farmers perceive the value of their agricultural residues and what they decide to do with them. The underlying objective is to help smallholders in degraded soils increase their agricultural yields through the use of biochar without diverting the already established uses of agricultural residues to a new soil management practice.

Keywords: agricultural residues, biochar, central Uganda, food security, soil erosion, soil remediation

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Authors: Pajaree Donkhampa, Fuangfa Unob

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Ceftazidime is an antibiotic drug commonly used to treat several human and veterinary infections. However, the presence of ceftazidime residues in the environment may induce microbial resistance and cause side effects to humans. Therefore, monitoring the level of ceftazidime in environmental resources is important. In this work, a melamine foam platform was proposed for simultaneous extraction and colorimetric detection of ceftazidime based on the azo dye formation on the surface. The melamine foam was chemically modified with polyethyleneimine (PEI) and characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Ceftazidime is a sample that was extracted on the PEI-modified melamine foam and further reacted with nitrite in an acidic medium to form an intermediate diazonium ion. The diazotized molecule underwent an azo coupling reaction with chromotropic acid to generate a red-colored compound. The material color changed from pale yellow to pink depending on the ceftazidime concentration. The photo of the obtained material was taken by a smartphone camera and the color intensity was determined by Image J software. The material fabrication and ceftazidime extraction and detection procedures were optimized. The detection of a sub-ppm level of ceftazidime was achieved without using a complex analytical instrument.

Keywords: colorimetric detection, ceftazidime, melamine foam, extraction, azo dye

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Authors: Joseph Clement Akan, Lami Jafiya, Zaynab Muhammad Chellube, Zakari Mohammed, Fanna Inna Abdulrahman

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Five vegetables (spinach, lettuce, cabbage, tomato, and onion) were freshly harvested from the Alau Dam and Gongulong agricultural areas for the determination of some organochlorine pesticide residues (o, p-DDE, p,p’-DDD, o,p’-DDD, p,p’-DDT, α-BHC, γ-BHC, metoxichlor, lindane, endosulfan dieldrin, and aldrin.) Soil samples were also collected at different depths for the determination of the above pesticides. Samples collection and preparation were conducted using standard procedures. The concentrations of all the pesticides in the soil and vegetable samples were determined using GC/MS SHIMADZU (GC-17A) equipped with electron capture detector (ECD). The highest concentration was that of p,p’-DDD (132.4±13.45µg/g) which was observed in the leaf of cabbage, while the lowest concentration was that of p,p’-DDT (2.34µg/g) was observed in the root of spinach. Similar trends were observed at the Gongulong agricultural area, with p,p’-DDD having the highest concentration of 153.23µg/g in the leaf of cabbage, while the lowest concentration was that of p,p’-DDT (12.45µg/g) which was observed in the root of spinach. α-BHC, γ-BHC, Methoxychlor, and lindane were detected in all the vegetable samples studied. The concentrations of all the pesticides in the soil samples were observed to be higher at a depth of 21-30cm, while the lowest concentrations were observed at a depth of 0-10cm. The concentrations of all the pesticides in the vegetables and soil samples from the two agricultural sites were observed to be at alarming levels, much higher than the maximum residue limits (MRLs) and acceptable daily intake values (ADIs) .The levels of the pesticides observed in the vegetables and soil samples investigated, are of such a magnitude that calls for special attention and laws to regulate the use and circulation of such chemicals. Routine monitoring of pesticide residues in these study areas is necessary for the prevention, control and reduction of environmental pollution, so as to minimize health risks.

Keywords: Alau Dam, gongulong, organochlorine, pesticide residues, soil, vegetables

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Authors: Waranya Pongpaiboon, Warangkana Srichamnong, Supat Chaiyakul

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Enzymatic modification of rice flour can produce highly functional derivatives use in food industries. This study aimed to evaluate the physical properties and resistant starch content of rice flour residues hydrolyzed by α-amylase. Rice flour hydrolyzed by α-amylase (60 and 300 u/g) for 1, 24 and 48 hours were investigated. Increasing enzyme concentration and hydrolysis time resulted in decreased rice flour residue’s lightness (L*) but increased redness (a*) and yellowness (b*) of rice flour residues. The resistant starch content and peak viscosity increased when hydrolysis time increased. Pasting temperature, trough viscosity, breakdown, final viscosity, setback and peak time of the hydrolyzed flours were not significantly different (p>0.05). The morphology of native flour was smooth without observable pores and polygonal with sharp angles and edges. However, after hydrolysis, granules with a slightly rough and porous surface were observed and a rough and porous surface was increased with increasing hydrolyzed time. The X-ray diffraction patterns of native flour showed A-type configuration, which hydrolyzed flour showed almost 0% crystallinity indicated that both amorphous and crystalline structures of starch were simultaneously hydrolyzed by α-amylase.

Keywords: α-Amylase, enzymatic hydrolysis, pasting properties, resistant starch

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Authors: Achiraya Jiraprasertwong, Erdogan Gulari, Sumaeth Chavadej

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Among agricultural residues, sugarcane bagasse is one of the most convincing raw materials for the production of bioethanol due to its availability, and low cost through enzymatic hydrolysis and yeast fermentation. A pretreatment step is needed to enhance the enzymatic step. In this study, sugarcane bagasse (SCB), one of the most abundant agricultural residues in Thailand, was pretreated biologically with various microorganisms of white-rot fungus—Phanerochaete sordid (SK 7), Cellulomonas sp. (TISTR 784), and strain A 002 (Bacillus subtilis isolated from Thai higher termites). All samples with various microbial pretreatments were further hydrolyzed enzymatically by a commercial enzyme obtained from Aspergillus niger. The results showed that the pretreatment with the white-rot fungus gave the highest glucose concentration around two-fold higher when compared with the others.

Keywords: sugarcane bagasse, microorganisms, pretreatment, enzymatic hydrolysis

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Authors: Recep Kesli, Gulsah Asik, Cengiz Demir, Onur Turkyilmaz

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Objective: Pseudomonas aeruginosa (P. aeruginosa) is an important nosocomial pathogen which causes serious hospital infections and is resistant to many commonly used antibiotics. P. aeruginosa can develop resistance during therapy and also it is very resistant to disinfectant chemicals. It may be found in respiratory support devices in hospitals. In this study, the antibiotic resistance of P. aeruginosa strains isolated from bronchial aspiration samples was evaluated retrospectively. Methods: Between October 2013 and September 2015, a total of 318 P. aeruginosa were isolated from clinical samples obtained from various intensive care units and inpatient patients hospitalized at Afyon Kocatepe University, ANS Practice and Research Hospital. Isolated bacteria identified by using both the conventional methods and automated identification system-VITEK 2 (bioMerieux, Marcy l’etoile France). Antibacterial resistance tests were performed by using Kirby-Bauer disc (Oxoid, Hampshire, England) diffusion method following the recommendations of CLSI. Results: Antibiotic resistance rates of identified 318 P. aeruginosa strains were found as follows for tested antibiotics; 32 % amikacin, 42% gentamicin, 43% imipenem, 43% meropenem, 50% ciprofloxacin, 57% levofloxacin, 38% cefepime, 63% ceftazidime, and 85% piperacillin/tazobactam. Conclusion: Resistance profiles change according to years and provinces for P. aeruginosa, so these findings should be considered empirical treatment choices. In this study, the highest and lowest resistance rates found against piperacillin/tazobactam % 85, and amikacin %32.

Keywords: Pseudomonas aeruginosa, antibiotic resistance rates, intensive care unit, Pseudomonas spp.

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Authors: Okafor Joan, Nwodo Uchechukwu

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Klebsiella pneumoniae (K. pneumoniae) is a significant pathogen responsible for opportunistic and nosocomial infection. One of the most significant antibiotic resistance mechanisms in K. pneumoniae isolates is efflux pumps. Our current study identified efflux genes (AcrAB, OqxAB, MacAB, and TolC) and regulatory genes (RamR and RarA) in multidrug-resistant (MDR) K. pneumoniae isolated from hospital effluents and investigated their relationship with antibiotic resistance. The sum of 145 K. pneumoniae isolates was established by PCR and screened for antibiotic susceptibility. PCR detected efflux pump genes, and their link with antibiotic resistance was statistically examined. However, 120 (83%) of the confirmed isolated were multidrug-resistant, with the largest percentage of resistance to ampicillin (88.3%) and the weakest rate of resistance to imipenem (5.5%). Resistance to the other antibiotics ranged from 11% to 76.6%. Molecular distribution tests show that AcrA, AcrB, MacA, oqxB oqxA, TolC, MacB were detected in 96.7%, 85%, 76.7%, 70.8%, 55.8%, 39.1%, and 29.1% respectively. However, 14.3% of the isolates harboured all seven genes screened. Efflux pump system AcrAB (83.2%) was the most commonly detected in K. pneumonia isolated across all the antibiotics class-tested. In addition, the frequencies of RamR and RarA were 46.2% and 31.4%, respectively. AcrAB and OqxAB efflux pump genes were significantly associated with fluoroquinolone, beta-lactam, carbapenem, and tetracycline resistance (p<0.05). The high rate of efflux genes in this study demonstrated that this resistance mechanism is the dominant way in K. pneumoniae isolates. Appropriate treatment must be used to reduce and tackle the burden of resistant Klebsiella pneumonia in hospital wastewater effluents.

Keywords: Klebsiella pneumonia, efflux pumps, regulatory genes, multidrug-resistant, hospital, PCR

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Authors: Sánchez Acuña, Juan Camilo, Granados Gómez, Mildred Magaly, Navarrete Rodríguez, Luisa Fernanda

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Nowadays, the main biofuels source production as bioethanol is food crops. This means a high competition between foods and energy production. For this reason, it is necessary to take into account the use of new raw materials friendly to the environment. The main objective of this paper is to evaluate the potential of the agro-industrial banana crop residues in the production of bioethanol. A factorial design of 2⁴ was used, the design has variables such as pH, time and concentration of hydrolysis, another variable is the time of fermentation that is of 7 or 15 days. In the hydrolysis phase, the pH is acidic (H₂SO₄) or basic (NaOH), the time is 30 or 15 minutes and the concentration is 0.1 or 0.5 M. It was observed that basic media, low concentrations, fermentation, and higher pretreatment times produced better performance in terms of biofuel obtained.

Keywords: bioethanol, biofuels, banana waste, hydrolysis

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Authors: Dinesh K. S., D. R. C. V. Jayadevan

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Autism is the most prevalent of a subset of the disorders organized under the umbrella of pervasive developmental disorders. After the publication of Andrew Wakefield's paper in lancet, many critiques deny this connection even without looking in to the matter. The British Medical Journal even put an editorial regarding this issue. BMJ 2010; 340:c1807. But ayurveda has ample of evidences to believe this connectivity. Dysbiosis, yeast growth of the gut, nutritional deficiencies, enzyme deficiencies, essential fatty acid deficiencies, Gastro esophageal reflux disease, indigestion, inflammatory bowel, chronic constipation & its cascade are few of them to note. The purpose of this paper is to present the observed changes in the behavioural symptoms of autistic babies after a gut management protocol which is a usual programme of our autism treatment plan especially after dysbiotic changes after antibiotic administration. Is there any correlation between changes (if significant) in gut symptoms and behavioral problems of autistic babies especially after a dysbiosis induced by antibiotics. Retrospective analysis of the case sheets of autistic patients admitted in Vaidyaratnam P.S.Varier Ayurveda College hospital, kottakkal,kerala, india from September 2010 are taken for the data processing. Autistic patients are used to come to this hospital as a part of their usual course of treatment. We investigated 40 cases diagnosed as autistic by clinical psychologists from different institutions who had dysbiosis induced by antibiotics. Significant change in gut symptoms before and after treatment p<0.05 in most of its components Significant change in behavioral symptoms before and after treatments p<0.05 in most of the components Correlation between gut symptoms change and behavioral symptoms changes after treatment is + 0.86. Conclusion : Selected Polyherbal Ayurveda treatment has significant role to play to make changes abnormal behaviors in autistic babies and has a positive correlation with changes in gut symptoms induced by dysbiosis of antibiotic intake.

Keywords: ayurveda, autism, dysbiosis, antibiotic

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Authors: Mihai-Alexandru Florea, Veronica Drumea, Roxana Nita, Cerasela Gird, Laura Olariu

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The aim of this study was to investigate the residues of pesticide found in tea water infusions. A multi-residues method to determine 147 pesticides has been developed using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe) procedure and dispersive solid phase extraction (d-SPE) for the cleanup the pesticides from complex matrices such as plants and tea. Sample preparation was carefully optimized for the efficient removal of coextracted matrix components by testing more solvent systems. Determination of pesticides was performed using GC-MS/MS (100 of pesticides) and LC-MS/MS (47 of pesticides). The selected reaction monitoring (SRM) mode was chosen to achieve low detection limits and high compounds selectivity and sensitivity. Overall performance was evaluated and validated according to DG-SANTE Guidelines. To assess the pesticide residue transfer rate (qualitative) from dried tea in infusions the samples (tea) were spiked with a mixture of pesticides at the maximum residues level accepted for teas and herbal infusions. In order to investigate the release of the pesticides in tea preparations, the medicinal plants were prepared in four ways by variation of water temperature and the infusion time. The pesticides from infusions were extracted using two methods: QuEChERS versus solid-phase extraction (SPE). More that 90 % of the pesticides studied was identified in infusion.

Keywords: tea, solid-phase extraction (SPE), selected reaction monitoring (SRM), QuEChERS

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Authors: Annie Ramanbhai Mecwan

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Cleaning validation in a pharmaceutical manufacturing facility is documented evidence that a cleaning process has effectively removed contaminants, residues from previous drug products and cleaning agents below a pre-defined threshold from the reusable tools and parts of equipment. In shared manufacturing facilities more than one drug product is prepared. After cleaning of reusable tools and parts of equipment after one drug product manufacturing, there are chances that some residues of drug substance from previously manufactured drug products may be retained on the equipment and can carried forward to the next drug product and thus cause cross-contamination. Health-based limits through the derivation of a safe threshold value called permitted daily exposure (PDE) for the residues of drug substances should be employed to identify the risks posed at these manufacturing facilities. The PDE represents a substance-specific dose that is unlikely to cause an adverse effect if an individual is exposed to or below this dose every day for a lifetime. There are different practices to calculate PDE. Data for all APIs in the public domain are considered to calculate PDE value though, company to company may vary the final PDE value based on different toxicologist’s perspective or their subjective evaluation. Hence, Regulatory agencies should take responsibility for publishing PDE values for all APIs as it is done for elemental PDEs. This will harmonize the PDE values all over the world and prevent the unnecessary load on manufacturers for cleaning validation

Keywords: active pharmaceutical ingredient, good manufacturing practice, NOAEL, no observed adverse effect level, permitted daily exposure

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Authors: S. H. Gasa, L. Singh, B. Pillay, A. O. Olaniran

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Wastewater treatment plants (WWTPs) have been implicated as the leading reservoir for antibiotic resistant bacteria (ARB), including Enterococci spp. and antibiotic resistance genes (ARGs), worldwide. Enterococci are a group of clinically significant bacteria that have gained much attention as a result of their antibiotic resistance. They play a significant role as the principal cause of nosocomial infections and dissemination of antimicrobial resistance genes in the environment. The main objective of this study was to ascertain the role of WWTPs in Durban, South Africa as potential reservoirs for antibiotic resistant Enterococci (ARE) and their related ARGs. Furthermore, the antibiogram and resistance gene profile of Enterococci species recovered from treated wastewater effluent and receiving surface water in Durban were also investigated. Using membrane filtration technique, Enterococcus selective agar and selected antibiotics, ARE were enumerated in samples (influent, activated sludge, before chlorination and final effluent) collected from two WWTPs, as well as from upstream and downstream of the receiving surface water. Two hundred Enterococcus isolates recovered from the treated effluent and receiving surface water were identified by biochemical and PCR-based methods, and their antibiotic resistance profiles determined by the Kirby-Bauer disc diffusion assay, while PCR-based assays were used to detect the presence of resistance and virulence genes. High prevalence of ARE was obtained at both WWTPs, with values reaching a maximum of 40%. The influent and activated sludge samples contained the greatest prevalence of ARE with lower values observed in the before and after chlorination samples. Of the 44 vancomycin and high-level gentamicin-resistant isolates, 11 were identified as E. faecium, 18 as E. faecalis, 4 as E. hirae while 11 are classified as “other” Enterococci species. High-level aminoglycoside resistance for gentamicin (39%) and vancomycin (61%) was recorded in species tested. The most commonly detected virulence gene was the gelE (44%), followed by asa1 (40%), while cylA and esp were detected in only 2% of the isolates. The most prevalent aminoglycoside resistance genes were aac(6')-Ie-aph(2''), aph(3')-IIIa, and ant(6')-Ia detected in 43%, 45% and 41% of the isolates, respectively. Positive correlation was observed between resistant phenotypes to high levels of aminoglycosides and presence of all aminoglycoside resistance genes. Resistance genes for glycopeptide: vanB (37%) and vanC-1 (25%), and macrolide: ermB (11%) and ermC (54%) were detected in the isolates. These results show the need for more efficient wastewater treatment and disposal in order to prevent the release of virulent and antibiotic resistant Enterococci species and safeguard public health.

Keywords: antibiogram, enterococci, gentamicin, vancomycin, virulence signatures

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Authors: Tatyane Lopes de Freitas, Antonio Diogo S. Vieira, Susana Marta Isay Saad, Maria Ines Genovese

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The fruit processing industries generate a large volume of residues to produce juices, pulps, and jams. These residues, or by-products, consisting of peels, seeds, and pulps, are routinely discarded. Fruits are rich in bioactive compounds, including polyphenols, which have positive effects on health. Dry residues from two fruits, acerola (M. emarginata D. C.) and orange (C. sinensis), were characterized in relation to contents of ascorbic acid, minerals, total dietary fibers, moisture, ash, lipids, proteins, and carbohydrates, and also high performance liquid chromatographic profile of flavonoids, total polyphenols and proanthocyanidins contents, and antioxidant capacity by three different methods (Ferric reducing antioxidant power assay-FRAP, Oxygen Radical Absorbance Capacity-ORAC, 1,1-diphenyl-2-picrylhydrazil (DPPH) radical scavenging activity). Acerola by-products presented the highest acid ascorbic content (605 mg/100 g), and better antioxidant capacity than orange by-products. The dry residues from acerola demonstrated high contents of proanthocyanidins (617 µg CE/g) and total polyphenols (2525 mg gallic acid equivalents - GAE/100 g). Both presented high total dietary fiber (above 60%) and protein contents (acerola: 10.4%; orange: 9.9%), and reduced fat content (acerola: 1.6%; orange: 2.6%). Both residues showed high levels of potassium, calcium, and magnesium, and were considered sources of these minerals. With acerola by-product, four formulations of probiotics fermented milks were produced: F0 (without the addition of acerola residue (AR)), F2 (2% AR), F5 (5% AR) and F10 (10% AR). The physicochemical characteristics of the fermented milks throughout of storage were investigated, as well as the impact of in vitro simulated gastrointestinal conditions on flavonoids and probiotics. The microorganisms analyzed maintained their populations around 8 log CFU/g during storage. After the gastric phase of the simulated digestion, the populations decreased, and after the enteric phase, no colonies were detected. On the other hand, the flavonoids increased after the gastric phase, maintaining or suffering small decrease after enteric phase. Acerola by-products powder is a valuable ingredient to be used in functional foods because is rich in vitamin C, fibers and flavonoids. These flavonoids appear to be highly resistant to the acids and salts of digestion.

Keywords: acerola, orange, by-products, fermented milk

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Authors: Pratibha Goyal, Nupur Mathur, Anuradha Singh

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Antibiotic resistance is the worldwide threat to human health in this century. Excessive use of antibiotic after their discovery in 1940 makes certain bacteria to become resistant against antibiotics. Most common antibiotic-resistant bacteria include Staphylococcus aureus, Salmonella typhi, E.coli, Klebsiella pneumonia, and Streptococcus pneumonia. Among all Staphylococcus resistant strain called Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for several lives threatening infection in human commonly found in the hospital environment. Our study aimed to isolate bacteriophage against MRSA from the hospital sewage treatment plant and to analyze its efficiency In Vivo in Swiss albino mice model. Sewage sample for the isolation of bacteriophages was collected from SDMH hospital sewage treatment plant in Jaipur. Bacteriophages isolated by the use of enrichment technique and after characterization, isolated phages used to determine phage treatment efficiency in mice. Mice model used to check the safety and suitability of phage application in human need which in turn directly support the use of natural bacteriophage rather than synthetic chemical to kill pathogens. Results show the plaque formation in-vitro and recovery of MRSA infected mice during the experiment. Favorable lytic efficiency determination of MRSA and Salmonella presents a natural way to treat lethal infections caused by Multidrug-resistant bacteria by using their natural host-pathogen relationship.

Keywords: antibiotic resistance, bacteriophages, methicillin resistance Staphylococcus aureus, pathogens, phage therapy, Salmonella typhi

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Authors: Recep Kesli, Huseyin Bilgin, Ela Tasdogan, Ercan Kurtipek

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Aim: The aim of this study was to compare resin based Bactec plus aerobic/anaerobic blood culture bottles (Becton Dickinson, MD, USA) and polymeric beads based BacT/Alert FA/FN plus blood culture bottles (bioMerieux, NC, USA) in terms of microorganisms recovery rates and time to detection (TTD) in the patients receiving antibiotic treatment. Method: Blood culture samples were taken from the patients who admitted to the intensive care unit and received antibiotic treatment. Forty milliliters of blood from patients were equally distributed into four types of bottles: Bactec Plus aerobic, Bactec Plus anaerobic, BacT/Alert FA Plus, BacT/Alert FN Plus. Bactec Plus and BacT/Alert Plus media were compared to culture recovery rates and TTD. Results: Blood culture samples were collected from 382 patients hospitalized in the intensive care unit and 245 patients who were diagnosed as having bloodstream infections were included in the study. A total of 1528 Bactec Plus aerobic, Bactec Plus anaerobic, BacT/Alert FA Plus, BacT/Alert FN Plus blood culture bottles analyzed and 176, 144, 154, 126 bacteria or fungi were isolated, respectively. Gram-negative and gram-positive bacteria were significantly more frequently isolated in the resin-based Bactec Plus bottles than in the polymeric beads based BacT/Alert Plus bottles. The Bactec Plus and BacT/Alert Plus media recovery rates were similar for fungi and anaerobic bacteria. The mean TTDs in the Bactec Plus bottles were shorter than those in the BacT/Alert Plus bottles regardless of the microorganisms. Conclusion: The results of this study showed that resin-containing media is a reliable and time-saving tool for patients who are receiving antibiotic treatment due to sepsis in the intensive care unit.

Keywords: Bactec Plus, BacT/Alert Plus, blood culture, antibiotic

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Authors: Chioma Nwakanma, Onyeka Okoh Irene, Emmanuel Eze

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Pesticide residues left in agricultural soils after cropping are always accumulative, difficult to degrade and harmful to animals, plants, soil and human health in general. The biodegrading potential of pesticides- resistant PGPB on soil pollution was investigated using in situ remediation technique following recommended standards. In addition, screening for insecticide utilization, maximum insecticide concentration tolerance, insecticide biodegradation and insecticide residues analyses via gas chromatographic/electron column detector were determined. The location of bacterial degradation genes was also determined. Three plant growth-promoting rhizophere (PGPR) were isolated and identified according to 16S rRNA as Paraburkholderia tropica, Burkolderia glumae and Achromobacter insolitus. From the results, all the three isolates showed phosphate solubilizing traits and were able to grow on nitrogen free medium. The isolates were able to utilize the insecticide as sole carbon source and increase in biomass. They were statistically significantly tolerant to all the insecticide concentrations screened. The gas chromatographic profiles of the insecticide residues showed a reduction in the peak areas of the insecticides, indicating degradation. The bacterial consortium had the lowest peak areas, showing the highest degradation efficiency. The genes responsible for degradation were found to be in the plasmids of the isolates. Therefore, the use of PGPR is recommended for bioremediation of agricultural soil insecticide polluted areas and can also enhance soil fertility.

Keywords: biodegradation, rhizosphere, insecticides utilization, agricultural soil

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Authors: Stefan Nicolae, Cristina Cirtoaje, Emil Petrescu, Florin-Razvan Duca

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In the context of the accelerated expansion of urban agglomerations and the exponential development of industry, a huge amount of water is used, and a crisis of drinking water may occur any time. Classic wastewater treatment removes most pollutants but, for some chemical residues, special methods are needed. Carbon nanotubes and other carbon materials might be used in many cases [1-2], especially for heavy metals removal but also on pharmaceutical products such as paracetamol [3]. Our research has confirmed the better efficiency of nanotubes compared to graphene on paracetamol removal from water, but even better results were obtained on single-walled nanotubes (SWCNTs) and graphene nanoplatelets. This can be due to their better dispersion in water which leads to an increased contact surface, so we propose a filtration system of membranes and carbon materials that can be used for paracetamol removal from wastewater but also for other drugs that affect the aquatic life as well as terrestrial animals and people who use this contaminated water.

Keywords: applied physics, wastewater, nanomaterials, enviromental science

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Authors: K. Ighilahriz, M. Taleb Ahmed, R. Maachi

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Oil industries are responsible for most cases of contamination of our ecosystem by oil and heavy metals. They are toxic and considered carcinogenic and dangerous even when they exist in trace amounts. At Algiers refinery, production, transportation, and refining of crude oil generate considerable waste in storage tanks; these residues result from the gravitational settling. The composition of these residues is essentially a mixture of hydrocarbon and lead. We propose in this work the application of electrooxidation treatment for the leachate of the leaden sludge. The effect of pH, current density and the electrolysis time were studied, the effectiveness of the processes is evaluated by measuring the chemical oxygen demand (COD). The dissolution is the best way to mobilize pollutants from leaden mud, so we conducted leaching before starting the electrochemical treatment. The process was carried out in batch mode using graphite anode and a stainless steel cathode. The results clearly demonstrate the compatibility of the technique used with the type of pollution studied. In fact, it allowed COD removal about 80%.

Keywords: electrooxidation, leaching, leaden sludge, oil industry

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Authors: Margita Čanigová, Jana Račková, Miroslav Kročko, Viera Ducková, Vladimíra Kňazovická

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The aim of the study was to test the milk samples in terms of enterococci presence and their counts. Tested samples were as follows: raw cow milk, raw cow milk stored at 10°C for 16 hours and milk pasteurised at 72°C for 15 seconds. The typical colonies were isolated randomly and identified by classical biochemical test - EN-COCCUS test (Lachema, CR) and by PCR. Isolated strains were tested in terms of antibiotic resistance by well diffusion method. Examined antibiotics were: vancomycin (30 μg/disc), gentamicin (120 μg/disc), erythromycin (15 μg/disc), teicoplanine (30 μg/disc), ampicillin (10 μg/disc) and tetracycline (30 μg/disc). Average value of enterococci counts in raw milk cistern samples (n=30) was 8.25 ± 1.37 ×103 CFU/cm3. Storage tank milk samples (n=30) showed an increase (P > 0.05) and average value was 9.16 ± 1.49 × 103 CFU/cm3. Occurrence of enterococci in pasteurized milk (n=30) was sporadic and their counts were mostly below 10 CFU/cm3. Overall, 96 enterococci strains were isolated. In samples of raw cow milk and stored raw cow milk, Enterococcus faecalis was a dominant species (58.1% and 71.7%, respectively), followed by E. faecium (16.3% and 0%, respectively). Enterococcus mundtii, E. casseliflavus, E. durans and E. gallinarum were isolated, too. Resistances to ampicillin, erythromycin, gentamicin, tetracycline and vancomycin were found in 7.29%, 3.13%, 4.00%, 13.54% and 10.42% of isolated enterococci strains, respectively. Resistance to teicoplanine was not found in any isolated strain. All Vancomycin-Resistant Enterococci (VRE) belonged to E. faecalis. Obtained results confirmed that raw milk is a potential risk of enterococci resistant to antibiotics transmission into the food chain.

Keywords: antibiotic resistance, enterococci, milk, biosystems engineering

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Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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Authors: Muhammad Shahzad Hussain

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Two major problems are facing generally by conventional poultry farming that is disease outbreaks and poor performance, which results due to improper management. To enhance the growth performance and efficiency of feed and reduce disease outbreaks, antibiotic growth promoters (AGPs) which are antibiotics at sub-therapeutic levels, are extensively used in the poultry industry. European Union has banned the use of antibiotics due to their presence in poultry products, development of antibiotic-resistant pathogens, and disturbance of normal gut microbial ecology. These residues cause serious health concerns and produce antibiotic resistance in pathogenic microbes in human beings. These issues strengthen the need for the withdrawal of AGPs from poultry feed. Nowadays, global warming is a major issue, and it is more critical in tropical areas like Pakistan, where heat stress is already a major problem. Heat stress leads to poor production performance, high mortality, immuno-suppression, and concomitant diseases outbreak. The poultry feed industry in Pakistan, like other countries of the world, has been facing shortages and high prices of local as well as imported feed ingredients. Prebiotics are potential replacer for AGP as prebiotics has properties to enhance the production potential and reduce the growth of harmful bacteria as well as stimulate the growth/activity of beneficial bacteria. The most commonly used prebiotics in poultry includes mannan oligosaccharide (MOS). MOS is an essential component of the yeast cell wall (YCW) (Saccharomyces cerevisiae); therefore, the YCW wall possesses prebiotic properties. The use of distillery yeast wall (YCW) has the potential to replace conventional AGPs and to reduce mortality due to heat stress as well as to bind toxins in the feed. The dietary addition of YCW has not only positive effects on production performance in poultry during normal conditions but during stressful conditions. A total of 168-day-old broilers were divided into 6 groups, each of which has 28 birds with 4 replicates (n=7).Yeast cell wall (YCW) supplementation @ 0%, 1%, 1.5%, 2%, 2.5%, 3% from day 0 to 35. Heat stress was exposed from day 21 to 35 at 30±1.1ᵒC with relative humidity 65±5%. Zootechnical parameters like body weight, FCR, Organ development, and histomorphometric parameters were studied. A significant weight gain was observed at group C supplemented @ 1.5% YCW during the fifth week. Significant organ weight gain of Gizzard, spleen, small intestine, and cecum was observed at group C supplemented @ 1.5% YCW. According to morphometric indices Duodenum, Jejunum, and Ileum has significant villus height, while Jejunum and Ileum have also significant villus surface area in the group supplemented with 1.5% YCW. IEL count was only decreased in 1.5% YCW-fed group in jejunum and ileum, not in duodenum, that was less in 2% YCW-supplemented group. Dietary yeast cell wall of saccharomyces cerevisiae partially reduced the effects of high ambient temperature in terms of better growth and modified gut histology and components of mucosal immune response to better withstand heat stress in broilers.

Keywords: antibiotics, AGPs, broilers, MOS, prebiotics, YCW

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Authors: Fatima Bouazza, Rachida Hassikou, Lamiae Amallah, Jihane Ennadir, Khadija Khedid

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This study evaluated the in vitro resistance and susceptibility of Enterobacteriaceae (Escherichia coli and Klebsiella oxytoca strains) and Staphylococci strains, isolated from sheep’s milk, against antibiotics and essential oils from Thymus satureioides and Mentha pulegium. Antibiotic resistance tests were done using disc diffusion while essential oils were extracted by steam distillation, and yields were calculated relative to plant dry matter. Gas chromatography-mass Spectrometry (GC-MS) was used to analyze each oil's chemical composition. The AMC, CTX, FOX, NA, CN, CIP, and OFX were very effective against the E. coli strains tested. Half of the strains were resistant to AMC, 60% to TIC, and 80% to TE. The K. oxytoca was resistant against AMC, FOX, and TIC (100%). Antibiotic-resistant testing on Staphylococci strains indicated Staphylococcus capitis and Staphylococcus chromogenes as the most sensitive. Staphylococcus aureus, Staphylococcus xylosus, and Staphylococcus cohnii ureal exhibited less resistance to OX, TE, PT, E, and P. The M. pulegium resulted in a higher yield of essential oil of 3.2% oil compared to T. satureioides with only 1.85% yield. Staphylococcus aureus, Staphylococcus xylosus, and Staphylococcus cohnii ureal had lower OX, TE, PT, E, and P resistance. M. pulegium yielded 3.2% essential oil compared to 1.85% for T. satureioides. The monoterpene oxygenated derivatives, monoterpene hydrocarbons, and phenols are found in essential oil extracts. T. satureioides essential oil had high antibacterial activity even at low concentrations (0.2; 0.55 g/mL). The Minimal Bactericidal Concentration (MBC) values indicate that the essential oils from the plants analyzed had bactericidal effects on all strains tested and are similar to the Minimal Inhibitory Concentration (MIC) values. The high antibacterial properties of these medicinal plants, against bacteria isolated from sheep’s milk, provide an opportunity to use these medicinal plants in the breeding sector as additives and preservatives in the dairy industry.

Keywords: antibiotic resistance, medicinal plants, essential oils, enterobacteriaceae, staphylococci, sheep milk

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