Search results for: resistant microbial strains

Authors: Afroza Khatun, Masuma, Md. Younus Mia, Kamal Kanta Das

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Analysis of the microbial status and physicochemical parameters of some branded and street vended ice cream showed that total viable bacteria in branded ice cream ranged from 4.8×10³ to 1.10×10⁵ cfu/ml, and in street vended ice-cream ranged from 7.5×10⁴ to 1.6×10⁸ cfu/ml. Total coliform bacteria present up to 9.20×10³ cfu/ml in branded ice cream and 5.3×10³ to 9.6×10⁶ cfu/ml observed in street vended ice cream. Total E. coli were found to be present within a range from 0 to 4.5×10³ cfu/ml in branded and 4.1×10² to 7.5×10⁴ cfu/ml in street ice cream. The ranges of Staphylococcus aureus count were 1.8×10² to 2.9×10⁴ cfu/ml (branded) and 3.9×10⁴ to 7.9×10⁶ cfu/ml (street). The pH of both types of ice cream showed acidic to neutral conditions where the concentration of pH for branded ice cream was 5.5 to 6.9, as well as the value of pH in street ice cream, was 6.2 to 7.0. The range of Total soluble solids in several branded ice creams was 26 to 29%, and the value of TSS obtained in street-vended ice-creams ranged from 5 to 10%. The overall results of this research demonstrated that the microbial quality in all street ice creams exceeded the BSTI standard and exhibited lower quality than the industrially produced branded ice creams due to comparatively faulty manufacturing processes and poor hygiene practices. The presence of pathogenic microbes was also observed in branded ice creams which was quite alarming for public health. So it is suggested that the government authorized organization should conduct the proper monitoring system to ensure that both branded and street vended ice-creams are microbiologically safe to prevent public health hazards.

Keywords: food safety, microbiological analysis, physicochemical, ice-cream, E. coli, Staphylococcus aureus

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Authors: Kent Salomonsson, Xuefang Zhao, Sara Kallin

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Human soft tissue is loaded and deformed by any activity, an effect known as a stress-strain relationship, and is often described by a load and tissue elongation curve. Several advances have been made in the fields of biology and mechanics of soft human tissue. However, there is limited information available on in vivo tissue mechanical characteristics and behavior. Confident mechanical properties of human soft tissue cannot be extrapolated from e.g. animal testing. Thus, there is need for non invasive methods to analyze mechanical characteristics of soft human tissue. In the present study, the internal mechanical conditions of the lower limb, which is subject to an external load, is studied by use of the finite element method. A detailed finite element model of the lower limb is made possible by use of MRI scans. Skin, fat, bones, fascia and muscles are represented separately and the material properties for them are obtained from literature. Previous studies have been shown to address macroscopic deformation features, e.g. indentation depth, to a large extent. However, the detail in which the internal anatomical features have been modeled does not reveal the critical internal strains that may induce hypoxia and/or eventual tissue damage. The results of the present study reveals that lumped material models, i.e. averaging of the material properties for the different constituents, does not capture regions of critical strains in contrast to more detailed models.

Keywords: FEM, tissue, indentation, properties

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Authors: S. Sehar, S. Khan, N. Ali, S. Ahmed

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In the last decade, the hunt for cost-effective, eco-friendly and energy sustainable technologies for waste water treatment are gaining much attention due to emerging water crisis and rapidly depleting existing water reservoirs all over the world. In this scenario, constructed wetland being a “green technology” could be a reliable mean for waste water treatment especially in small communities due to cost-effectiveness, ease in management, less energy consumption and sludge production. Therefore, a low cost, lab-scale sub-surface flow hybrid constructed wetland (SS-HCW) was established for domestic waste water treatment.It was observed that not only the presence but also choice of suitable vegetation along with hydraulic retention time (HRT) are key intervening ingredients which directly influence pollutant removals in constructed wetlands. Another important aspect of vegetation is that it may facilitate microbial attachment in rhizosphere, thus promote biofilm formation via microbial interactions. The major factors that influence initial aggregation and subsequent biofilm formation i.e. divalent cations (Ca2+) and extra cellular DNA (eDNA) were also studied in detail. The presence of Ca2+ in constructed wetland demonstrate superior performances in terms of effluent quality, i.e BOD5, COD, TDS, TSS, and PO4- than in absence of Ca2+. Finally, light and scanning electron microscopies coupled with EDS were carried out to get more insights into the mechanics of biofilm formation with or without Ca addition. Therefore, the same strategy can be implemented in other waste water treatment technologies.

Keywords: hybrid constructed wetland, biofilm formation, waste water treatment, waste water

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Authors: Bengi Hakguder Taze, Sevcan Unluturk

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Şalak apricot (Prunus armeniaca L., cv. Şalak) is a specific variety grown in Igdir, Turkey. The fruit has distinctive properties distinguish it from other cultivars, such as its unique size, color, taste and higher water content. Drying is the widely used method for preservation of apricots. However, fresh consumption is preferred for Şalak apricot instead of drying due to its low dry matter content. Higher amounts of water in the structure and climacteric nature make the fruit sensitive against rapid quality loss during storage. Hence, alternative processing methods need to be introduced to extend the shelf life of the fresh produce. Mild heat (MH) treatment is of great interest as it can reduce the microbial load and inhibit enzymatic activities. Therefore, the aim of this study was to evaluate the impact of mild heat treatment on the natural microflora found on Şalak apricot surfaces and some physical quality parameters of the fruit, such as color and firmness. For this purpose, apricot samples were treated at different temperatures between 40 and 60 ℃ for different periods ranging between 10 to 60 min using a temperature controlled water bath. Natural flora on the fruit surfaces was examined using standard plating technique both before and after the treatment. Moreover, any changes in color and firmness of the fruit samples were also monitored. It was found that control samples were initially containing 7.5 ± 0.32 log CFU/g of total aerobic plate count (TAPC), 5.8±0.31 log CFU/g of yeast and mold count (YMC), and 5.17 ± 0.22 log CFU/g of coliforms. The highest log reductions in TAPC and YMC were observed as 3.87-log and 5.8-log after the treatments at 60 ℃ and 50 ℃, respectively. Nevertheless, the fruit lost its characteristic aroma at temperatures above 50 ℃. Furthermore, great color changes (ΔE ˃ 6) were observed and firmness of the apricot samples was reduced at these conditions. On the other hand, MH treatment at 41 ℃ for 10 min resulted in 1.6-log and 0.91-log reductions in TAPC and YMC, respectively, with slightly noticeable changes in color (ΔE ˂ 3). In conclusion, application of temperatures higher than 50 ℃ caused undesirable changes in physical quality of Şalak apricots. Although higher microbial reductions were achieved at those temperatures, temperatures between 40 and 50°C should be further investigated considering the fruit quality parameters. Another strategy may be the use of high temperatures for short time periods not exceeding 1-5 min. Besides all, MH treatment with UV-C light irradiation can be also considered as a hurdle strategy for better inactivation results.

Keywords: color, firmness, mild heat, natural flora, physical quality, şalak apricot

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Authors: M. Zafar, S. Rasheed, Imran Hashmi

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Very little is concerned about the bacterial contamination in drinking water biofilm which provide a potential source for bacteria to grow and increase rapidly. So as to understand the microbial density in DWDs, a three-month study was carried out. The aim of this study was to examine biofilm in three different pipe materials including PVC, PPR and GI. A set of all these pipe materials was installed in DWDs at nine different locations and assessed on monthly basis. Drinking water quality was evaluated by different parameters and characterization of biofilm. Among various parameters are Temperature, pH, turbidity, TDS, electrical conductivity, BOD, COD, total phosphates, total nitrates, total organic carbon (TOC) free chlorine and total chlorine, coliforms and spread plate counts (SPC) according to standard methods. Predominant species were Bacillus thuringiensis, Pseudomonas fluorescens , Staphylococcus haemolyticus, Bacillus safensis and significant increase in bacterial population was observed in PVC pipes while least in cement pipes. The quantity of DWDs bacteria was directly depended on biofilm bacteria and its increase was correlated with growth and detachment of bacteria from biofilms. Pipe material also affected the microbial community in drinking water distribution network biofilm while Similarity in bacterial species was observed between systems due to same disinfectant dose, time period and plumbing pipes.

Keywords: biofilm, DWDs, pipe material, bacterial population

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Authors: Recep Keşli, Hayriye Tokay, Cengiz Demir, İsmail Ceyhan

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Objective: In many countries, tuberculosis (TB) is still one of the most important diseases. Tuberculosis is among top 10 causes of death worldwide. The early diagnosis of active tuberculosis still depends on the presence of acid resistant bacilli (ARB) in stained smears. In this study, we aimed to investigate the diagnostic performances of Erlich Ziehl Neelsen (EZN), Kinyoun and two different fluorescent stains. Methods: The specimens were obtained from the patients who applied to Chest Diseases Departments of Ankara Atatürk Chest Diseases and Thoracic Surgery Training and Research Hospital, and Afyon Kocatepe University, ANS Research and Practice Hospital. The study was carried out in the Medical Microbiology Laboratory, School of Medicine, Afyon Kocatepe University. All the non-sterile specimens were homogenized and decontaminated according to the EUCAST instructions. Samples were inoculated onto the Löwenstein-Jensen agars (bio-Merieux Marcy l'Etoile, France) and then incubated at 37˚C, for 40 days. Four smears were prepared from each specimen. Slides were stained with commercial EZN (BD, Sparks, USA), Kinyoun (SALUBRIS Istanbul, Turkey), Auramine (SALUBRIS Istanbul, Turkey) and Rhodamine (SALUBRIS Istanbul, Turkey) kit. While EZN and Kinyoun stainings were examined by light microscope, Auramine and Rhodamine slides were examined by fluorescence microscopy. Results: A total of 158 respiratory system samples (sputum, broncho alveolar lavage fluid…etc) were enrolled into the study. A hundred and two of the samples that processed were found as culture positive. The sensitivity, specificity, positive predictive, and negative predictive values were detected as 100%, 67.5%, 73.5%, and 100% for EZN, 100%, 70.9%, 77.4%, and 100% for Kinyoun, 100%,77.8%, 84.3%, 100% for Auramine, and 100%, 80% , 86.3%, and 100% for Rhodamine respectively. Conclusions: According to our study auramine and rhodamine staining methods showed the best diagnostic performance among the four investigated staining methods. In conclusion, the fluorochrome staining method may be accepted as the most reliable, rapid and useful method for diagnosis of the mycobacterial infections truly.

Keywords: acid resistant bacilli (ARB), auramine, Ehrlich-Ziehl-Neelsen (EZN), Kinyoun, Rhodamine

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Authors: Ajmal Khan Kassi, Humayun Javed, Tariq Mukhtar

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The research was conducted to screen out resistance and susceptibility of okra varieties against Helicoverpa armigera under field conditions 2016. In this experiment, the different management practices viz. release Trichogramma chilonis, hoeing, and weeding, clipping, and lufenuron were tested individually and with all possible combinations for the controlling of American bollworm at 3 diverse localities viz. University research farm Koont, National Agriculture Research Centre (NARC) and farmer field Taxila by using resistant variety Arka Anamika. All the treatment combinations regarding damage of shoot and fruit showed significant results. The minimum fruit infestation, i.e., 3.20% and 3.58% was recorded with combined treatment (i.e., T. chilonis + hoeing + weeding + lufenuron) in two different localities. The minimum shoot infestation, i.e., 7.18%, 7.08%, and 6.85% was also observed with (T. chilonis + hoeing + weeding + lufenuron) combined treatment at all three different localities. The above-combined treatment (T. chilonis + hoeing + weeding + lufenuron) also resulted in maximum yield at NARC and Taxila, i.e., 57.67 and 62.66 q/ha respectively. On the basis of combined treatment (i.e., T. chilonis + hoeing + weeding + lufenuron) in three different localities, Arka Anamika variety proved to be comparatively resistant against H. armigera. So this variety is recommended for the cultivation in Pothwar region to get maximum yield and minimum losses against H. armigera.

Keywords: okra, screening, combine treatment, Helicoverpa armigera

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Authors: Deeba N. Baig, Ateeqa Ijaz, Saloome Rafiq

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Camel is a religious and culturally significant animal in Asian and African regions. In Pakistan Dromedary and Bactrian are common camel breeds. Other than the transportation use, it is a pivotal source of milk and meat. The quality of its milk and meat is predominantly dependent on the geographical location and variety of vegetation available for the diet. Camel milk (CM) is highly nutritious because of its reduced cholesterol and sugar contents along with enhanced minerals and vitamins level. The absence of beta-lactoglobulin (like human milk), makes CM a safer alternative for infants and children having Cow Milk Allergy (CMA). In addition to this, it has a unique probiotic profile both in raw and fermented form. Number of Lactic acid bacteria (LAB) including lactococcus, lactobacillus, enterococcus, streptococcus, weissella, pediococcus and many other bacteria have been detected. From these LAB Lactobacilli, Bifidobacterium and Enterococcus are widely used commercially for fermentation purpose. CM has high therapeutic value as its effectiveness is known against various ailments like fever, arthritis, asthma, gastritis, hepatitis, Jaundice, constipation, postpartum care of women, anti-venom, dropsy etc. It also has anti-diabetic, anti-microbial, antitumor potential along with its robust efficacy in the treatment of auto-immune disorders. Recently, the role of CM has been explored in brain-gut axis for the therapeutics of neurodevelopmental disorders. In this connection, a lot of grey area was available to explore the probiotics and therapeutics latent in the CM available in Pakistan. Thus, current study was designed to explore the predominant probiotic flora and antimicrobial potential of CM from different local breeds of Pakistan. The probiotics have been identified through biochemical, physiological and ribo-typing methods. In addition to this, bacteriocins (antimicrobial-agents) were screened through PCR-based approach. Results of this study revealed that CM from different breeds of camel depicted a number of similar probiotic candidates along with the range of limited variability. However, the nucleotide sequence analysis of selected anti-listerial bacteriocins exposed least variability. As a conclusion, the CM has sufficient probiotic availability and significant anti-microbial potential.

Keywords: bacteriocins, camel milk, probiotics potential, therapeutics

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Authors: Vijay Sodhi, Charanjit Singh, Neelam Sodhi, Puneet P. S. Cheema, Reena Sharma, Mithilesh K. Jha

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The management and secure disposal of the biosludge generated from widely commercialized conventional activated sludge (CAS) treatments become a potential environmental issue. Thus, a sustainable technological upgradation to the CAS for sludge yield minimization has recently been gained serious attention of the scientific community. A number of recently reported studies effectively addressed the remedial technological advancements that in monopoly limited to the municipal wastewater. Moreover, the critical review of the literature signifies side-stream sludge minimization as a complex task to maintain. In this work, therefore, a hybrid moving bed biofilm reactor (MBBR) configuration (named as AMOMOX process) for in-situ minimization of the excess biosludge generated from high organic strength tannery wastewater has been demonstrated. The AMOMOX collectively stands for anoxic MBBR (as AM), aerobic MBBR (OM) and an oxic CAS (OX). The AMOMOX configuration involved a combined arrangement of an anoxic MBBR and oxic MBBR coupled with the aerobic CAS. The AMOMOX system was run in parallel with an identical CAS reactor. Both system configurations were fed with same influent to judge the real-time operational changes. For the AMOMOX process, the strict maintenance of operational strategies resulted about 95% removal of NH4-N and SCOD from tannery wastewater. Here, the nourishment of filamentous microbiota and purposeful promotion of cell-lysis effectively sustained sludge yield (Yobs) lowering upto 0.51 kgVSS/kgCOD. As a result, the volatile sludge scarcity apparent in the AMOMOX system succeeded upto 47% reduction of the excess biosludge. The corroborated was further supported by FE-SEM imaging and thermogravimetric analysis. However, the detection of microbial strains habitat underlying extended SRT (23-26 days) of the AMOMOX system would be the matter of further research.

Keywords: tannery wastewater, moving bed biofilm reactor, sludhe yield, sludge minimization, solids retention time

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Authors: Aniruddha Hore, Saptarshi Mitra, Sandip Ghosh, Sujoy Bose, Avijit Ghosh

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The Indian rupee is the official currency of India. With time, science and technology got advanced, and our society is slowly making its way to a cashless mode of transaction. But as India is still a developing country, a large part of our society still depends on transaction through cash. During times of pandemics, we came to understand that everything that we touch is not safe from microbial contamination. The Indian currency is also not an exception. The Indian currency is the modern-day medium of harmful bacterial as well as other microbial contaminations resulting in diseases in human bodies. Therefore, the need came to make the currency disinfectant to give our people a healthier lifestyle. The main focus of the study is to develop a solution that, when applied to the currency notes, will kill the persisting bacteria or microbes present in the notes. So various natural edible products were used in order to prepare the solution, which is highly effective against the presence of harmful bacteria such as E. coli and S. aureus. The antibacterial activity of these natural ingredients is not unknown to us, so extracts from those products were mixed together to form a solution which was made the Indian currency notes antibacterial for 20min approx. The solution was creating a layer on the surface of currency notes, therefore, making it antibacterial for a given duration of time, i.e., no bacterial growth was seen during the time period of 20 minutes, therefore, making it safe for the usage of human hands.

Keywords: Indian currency, antibacterial property of Indian currency, surface coating, currency disinfectant

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Authors: Lydia Tan, Philippe Lemey, Lieselot Houspie, Marco Viveen, Darren Martin, Frank Coenjaerts

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Introduction: Human respiratory syncytial virus (hRSV) is the leading cause of severe respiratory tract infections in infants under the age of two. Genomic substitutions and related evolutionary dynamics of hRSV are of great influence on virus transmission behavior. The evolutionary patterns formed are due to a precarious interplay between the host immune response and RSV, thereby selecting the most viable and less immunogenic strains. Studying genomic profiles can teach us which genes and consequent proteins play an important role in RSV survival and transmission dynamics. Study design: In this study, genetic diversity and evolutionary rate analysis were conducted on 36 RSV subgroup B whole genome sequences and 37 subgroup A genome sequences. Clinical RSV isolates were obtained from nasopharyngeal aspirates and swabs of children between 2 weeks and 5 years old of age. These strains, collected during epidemic seasons from 2001 to 2011 in the Netherlands and Belgium by either conventional or 454-sequencing. Sequences were analyzed for genetic diversity, recombination events, synonymous/non-synonymous substitution ratios, epistasis, and translational consequences of mutations were mapped to known 3D protein structures. We used Bayesian statistical inference to estimate the rate of RSV genome evolution and the rate of variability across the genome. Results: The A and B profiles were described in detail and compared to each other. Overall, the majority of the whole RSV genome is highly conserved among all strains. The attachment protein G was the most variable protein and its gene had, similar to the non-coding regions in RSV, more elevated (two-fold) substitution rates than other genes. In addition, the G gene has been identified as the major target for diversifying selection. Overall, less gene and protein variability was found within RSV-B compared to RSV-A and most protein variation between the subgroups was found in the F, G, SH and M2-2 proteins. For the F protein mutations and correlated amino acid changes are largely located in the F2 ligand-binding domain. The small hydrophobic phosphoprotein and nucleoprotein are the most conserved proteins. The evolutionary rates were similar in both subgroups (A: 6.47E-04, B: 7.76E-04 substitution/site/yr), but estimates of the time to the most recent common ancestor were much lower for RSV-B (B: 19, A: 46.8 yrs), indicating that there is more turnover in this subgroup. Conclusion: This study provides a detailed description of whole RSV genome mutations, the effect on translation products and the first estimate of the RSV genome evolution tempo. The immunogenic G protein seems to require high substitution rates in order to select less immunogenic strains and other conserved proteins are most likely essential to preserve RSV viability. The resulting G gene variability makes its protein a less interesting target for RSV intervention methods. The more conserved RSV F protein with less antigenic epitope shedding is, therefore, more suitable for developing therapeutic strategies or vaccines.

Keywords: drug target selection, epidemiology, respiratory syncytial virus, RSV

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Authors: Essam A. Makky, Siti H. Mohd Rasdi, J. B. Al-Dabbagh, G. F. Najmuldeen

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The synthesis of silver nano particles (SNPs) extensively studied by using chemical and physical methods. Here, the biological methods were used and give benefits in research field in the aspect of very low cost (from waste to wealth) and safe time as well. The study aims to isolate and exploit the microbial power in the production of industrially important by-products in nano-size with high economic value, to extract highly valuable materials from hazardous waste, to quantify nano particle size, and characterization of SNPs by X-Ray Diffraction (XRD) analysis. Disposal X-ray films were used as substrate because it consumes about 1000 tons of total silver chemically produced worldwide annually. This silver is being wasted when these films are used and disposed. Different bacterial isolates were obtained from various sources. Silver was extracted as nano particles by microbial power degradation from disposal X-ray film as the sole carbon source for ten days incubation period in darkness. The protein content was done and all the samples were analyzed using XRD, to characterize of silver (Ag) nano particles size in the form of silver nitrite. Bacterial isolates CL4C showed the average size of SNPs about 19.53 nm, GL7 showed average size about 52.35 nm and JF Outer 2A (PDA) showed 13.52 nm. All bacterial isolates partially identified using Gram’s reaction and the results obtained exhibited that belonging to Bacillus sp.

Keywords: nanotechnology, bioremediation, disposal X-ray film, nanoparticle, waste, XRD

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Authors: Radheshyam Yadav, Ramakrishna Wusirika

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Plant Growth Promoting Bacteria (PGPB) and Arbuscular Mycorrhizal (AM) Fungi are ubiquitous in soil and often very critical for crop yield and agriculture sustainability, and this has motivated the agricultural practices to support and promote PGPB and AM Fungi in agriculture. PGPB can be involved in a range of processes that affect Nitrogen (N) and Phosphorus (P) transformations in soil and thus influence nutrient availability and uptake to the plants. A field study with two wheat cultivars, HD-3086, and HD-2967 was performed in Malwa region, Bathinda of Punjab, India, to evaluate the effect of native and non-native PGPB alone and in combination with AM fungi as an inoculant on wheat grain yield, nutrient uptake and soil health parameters (dehydrogenase, urease, β‐glucosidase). Our results showed that despite an early insignificant increase in shoot length, plants treated with PGPB (Bacillus sp.) and AM Fungi led to a significant increase in shoot growth at maturity, aboveground biomass, nitrogen (45% - 40%) and phosphorus (40% - 34%) content in wheat grains relative to untreated control plants. Similarly, enhanced grain yield and nutrients uptake i.e. copper (27.15% - 36.25%) iron (43% - 53%) and zinc (44% - 47%) was recorded in PGPB and AM Fungi treated plants relative to untreated control. Overall, inoculation with native PGPB alone and in combination with AM Fungi provided benefits to enhance grain yield, wheat biofortification, and improved soil fertility, despite this effect varied depending on different PGPB isolates and wheat cultivars. These field study results provide evidence of the benefits of agricultural practices involving native PGPB and AM Fungi to the plants. These native strains and AM Fungi increased accumulations of copper, iron, and zinc in wheat grains, enhanced grain yield, and soil fertility.

Keywords: AM Fungi, biofortification, PGPB, soil microbial enzymes

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Authors: Afsoun Nikravan, Parisa Babaei, Gulen Gullu

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We aimed to study the association between house dust endotoxin, β-(1→3)-D-glucan, and asthma in a sample representative of the Turkish population. We analyzed data from 240 participants. The house dust was collected from the homes of 110 asthmatics and 130 control (without asthma) school-aged children (6-11 years old). House dust from the living room and from bedroom floors were analyzed for endotoxin and beta-glucan contents. House dust was analyzed for endotoxin content by the kinetic limulus amoebocyte lysate assay and for β-(1→3)-D-glucan by the inhibition enzyme immunoassay. The parents answered questions regarding potential determinants. We found geometric means 187.5 mg/m² for dust. According to statistical values, the endotoxin geometric mean was 13.86×103 EU/g for the control group and 6.16×103 EU/g for the asthma group. As a result, the amount of bacterial endotoxin was measured at a higher level in the homes of children without asthma. The geometric mean for beta-glucan was 46.52 µg/g and 44.39 µg/g for asthma and control groups, respectively. No associations between asthma and microbial agents were observed in Turkish children. High correlations (r > 0.75) were found between floor dust and endotoxin loads, while endotoxin and β-(1→3)-D-glucan concentrations were not correlated. The type of flooring (hard-surface or textile) was the strongest determinant for loads of floor dust and concentrations of endotoxin. Water damage and dampness at home were determinants of β-(1→3)-D-glucan concentrations. Endotoxin and β-(1→3)-D-glucan concentrations in Turkish house dust might lower than concentrations seen in other European countries.

Keywords: indoor air quality, asthma, microbial pollutants, case-control

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Authors: Cengiz Demir, Recep Keşli, Gülşah Aşık

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Objective: This study was carried out with the purpose of defining by using the E test method and determining the antibiotic resistance profiles of Gram-negative anaerobic bacilli isolated from various clinical specimens obtained from patients with suspected anaerobic infections and referred to Medical Microbiology Laboratory of Afyon Kocatepe University, ANS Application and Research Hospital. Methods: Two hundred and seventy eight clinical specimens were examined for isolation of the anaerobic bacteria in Medical Microbiology Laboratory between the 1st November 2014 and 30th October 2015. Specimens were cultivated by using Scheadler agar that 5% defibrinated sheep blood added, and Scheadler broth. The isolated anaerobic Gram-negative bacilli were identified conventional methods and Vitek 2 (ANC ID Card, bioMerieux, France) cards. Antibiotic resistance rates against to penicillin G, clindamycin, cefoxitin, metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem were determined with E-test method for each isolate. Results: Of the isolated twenty-eight anaerobic gram negative bacilli fourteen were identified as the B. fragilis group, 9 were Prevotella group, and 5 were Fusobacterium group. The highest resistance rate was found against penicillin (78.5%) and resistance rates against clindamycin and cefoxitin were found as 17.8% and 21.4%, respectively. Against to the; metronidazole, moxifloxacin, imipenem, meropenem, ertapenem and doripenem, no resistance was found. Conclusion: Since high rate resistance has been detected against to penicillin in the study penicillin should not be preferred in empirical treatment. Cefoxitin can be preferred in empirical treatment; however, carrying out the antibiotic sensitivity testing will be more proper and beneficial. No resistance was observed against carbapenem group antibiotics and metronidazole; so that reason, these antibiotics should be reserved for treatment of infectious caused by resistant strains in the future.

Keywords: anaerobic gram-negative bacilli, anaerobe, antibiotics and resistance profiles, e-test method

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Authors: S. S. R. Shojaei, S. Kord Afshari

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The study was initiated to compare the production and defecation of Eimerial oocysts of two internationally reputed broiler strains under the local environmental and management conditions. 40 one-day old male chickens of Arbor Acres strain and ROSS strain (20 chicks from each strain) used in this study and were divided randomly into four control and challenge groups. Feed and water were provided for ad libitum consumption. At 15 d of age, chickens of challenge groups (from each strain) were individually inoculated with a mixture of 50000 of sporulated oocysts of 4 species including of E. acervulina (20%), E. maxima (40%), E. tenella (25%) and E. necatrix (15%) and also from the fourth day after Eimerial challenge, faecal droppings (litter samples) were collected 10 days consecutively for counting oocyst per gram (OPG). The results indicated that in the challenge groups, there was an increasing process of OPG in days of 4 to 7 post challenging and the pick level of OPG was seen at seventh day after challenging. From day 8 to 9, decreasing of OPG was happened. This decreasing continues with mild, fast and mild process to day of 13. Respectively and totally the average of OPG in the Arbor Acres group was lower than the group Ross in all days post inoculation and this difference was significant according to t-test. According to the obtained results in this study and since oocyst index almost always is considered as one of the most important indicators for coccidiosis evaluation, it can be realized that in the same surveillance condition the regarding the severity evaluation of coccidiosis, Arbor Acres strain broilers shed less oocysts than Ross strain broilers.

Keywords: arbor acres, ross, coccidiosis, OPG

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Authors: Jou-An Chen, Julio Castillo, Errol Duncan Cason, Gabre Kemp, Leana Esterhuizen, Angel Valverde Portal, Esta Van Heerden

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Emerging contaminants (EC) such as pharmaceutical and personal care products have been accumulating for years in water bodies all over the world. However, very little is known about the occurrences, levels, and effects of ECs in South African water resources. This study provides an initial assessment of the distribution of eight ECs (Acetaminophen, Atrazine, Terbuthlyazine, Carbamazepine, Phenyton, Sulfmethoxazole, Nevirapine and Fluconozole) in fifteen water sources from the Free State and Easter Cape provinces of South Africa. Overall, the physiochemical conditions were different in surface and groundwater samples, with concentrations of several elements such as B, Ca, Mg, Na, NO3, and TDS been statistically higher in groundwater. In contrast, ECs levels, quantified at ng/mL using the LC/MS/ESI, were much lower in groundwater samples. The ECs with higher contamination levels were Carbamazepine, Sulfmethoxazole, Nevirapine, and Terbuthlyazine, while the most widespread were Sulfmethoxazole and Fluconozole, detected in all surface and groundwater samples. Fecal and E. coli tests indicated that surface water was more contaminated than groundwater. Microbial communities, assessed using NGS, were dominated by the phyla Proteobacteria and Bacteroidetes, in both surface and groundwater. Actinobacteria, Planctomycetes, and Cyanobacteria, were more dominant in surface water, while Verrucomicrobia were overrepresented in groundwater. In conclusion, ECs contamination is closely associated with human activities (human wastes). The microbial diversity identified can suggest possible biodegradation processes.

Keywords: emerging contaminants, EC, personal care products, pharmaceuticals, natural attenuation process

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Authors: Bahareh Asefi, Fereidoun Farzaneh, Ghazaleh Asefi

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Continuous increasing food waste produced on a global as well as national scale may lead to burgeoning environmental and economic problems. Simultaneously, decreasing the use efficiencies of natural resources such as land, water, and energy is occurring. On the other hand, food waste has a high-energy content, which seems ideal to achieve dual benefits in terms of energy recovery and the improvement of resource use efficiencies. Therefore, to decrease the environmental impacts of food waste and resource conservation, the researcher has focused on traditional methods of using food waste as a resource through different approaches such as anaerobic digestion, composting, incineration, and landfill. The adverse environmental effects of growing food waste make it difficult for traditional food waste treatment and management methods to balance social, economic, and environmental benefits. The old technology does not need to develop, but several new technologies such as microbial fuel cells, food waste disposal, and bio-converting food waste technology still need to establish or appropriately considered. It is pointed out that some new technologies can take into account various benefits. Since the information about food waste and its management method is critical for executable policy, a review of the latest information regarding the source of food waste and its management technology in some counties is provided in this study.

Keywords: food waste, management technology, innovative method, bio converting food waste, microbial fuel cell

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Authors: Magdalena Smoktunowicz, Renata Wawrzyniak, Malgorzata Waleron, Krzysztof Waleron

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Pectobacterium spp. were previously classified into the Erwinia genus founded in 1917 to unite at that time all Gram-negative, fermentative, nonsporulating and peritrichous flagellated plant pathogenic bacteria. After work of Waldee (1945), on Approved Lists of Bacterial Names and bacteriology manuals in 1980, they were described either under the species named Erwinia or Pectobacterium. The Pectobacterium genus was formally described in 1998 of 265 Pectobacterium strains. Currently, there are 21 species of Pectobacterium bacteria, including Pectobacterium betavasculorum since 2003, which caused soft rot on sugar beet tubers. Based on the biochemical experiments carried out for this, it is known that these bacteria are gram-negative, catalase-positive, oxidase-negative, facultatively anaerobic, using gelatin and causing symptoms of soft rot on potato and sugar beet tubers. The mere fact of growing on sugar beet may indicate a metabolism characteristic only for this species. Metabolomics, broadly defined as the biology of the metabolic systems, which allows to make comprehensive measurements of metabolites. Metabolomics, in combination with genomics, are complementary tools for the identification of metabolites and their reactions, and thus for the reconstruction of metabolic networks. The aim of this study was to apply the GC-MS-based untargeted metabolomics to study the metabolism of P. betavasculorum in different growing conditions. The metabolomic profiles of biomass and biomass media were determined. For sample preparation the following protocol was used: extraction with 900 µl of methanol: chloroform: water mixture (10: 3: 1, v: v) were added to 900 µl of biomass from the bottom of the tube and up to 900 µl of nutrient medium from the bacterial biomass. After centrifugation (13,000 x g, 15 min, 4oC), 300µL of the obtained supernatants were concentrated by rotary vacuum and evaporated to dryness. Afterwards, two-step derivatization procedure was performed before GC-MS analyses. The obtained results were subjected to statistical calculations with the use of both uni- and multivariate tests. The obtained results were evaluated using KEGG database, to asses which metabolic pathways are activated and which genes are responsible for it, during the metabolism of given substrates contained in the growing environment. The observed metabolic changes, combined with biochemical and physiological tests, may enable pathway discovery, regulatory inference and understanding of the homeostatic abilities of P. betavasculorum.

Keywords: GC-MS chromatograpfy, metabolomics, metabolism, pectobacterium strains, pectobacterium betavasculorum

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Authors: Thu Thuy Pham, Thi Chau Loan Tran, Van Duy Nguyen

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Marine fungi play an important role in the marine ecosystems. Marine fungi also supply biomass and metabolic products of industrial value. Currently, the biodiversity of marine fungi along the coastal areas of Vietnam has not yet been studied fully. The objective of this study is to assess the spatial and temporal diversity of planktonic fungi from the coastal waters of Nha Trang Bay and Van Phong Bay in Central Vietnam using culture-dependent and independent approach. Using culture-dependent approach, filamentous fungi and yeasts were isolated on selective media and then classified by phenotype and genotype based on the sequencing of ITS (internal transcribed spacers) regions of rDNA with two primer pairs (ITS1F_KYO2 and ITS4; NS1 and NS8). Using culture-independent approach, environmental DNA samples were isolated and amplified using fungal-specific ITS primer pairs. A total of over 160 strains were isolated from 10 seawater sampling stations at 50 cm depth. They were classified into diverse genera and species of both yeast and mold. At least 5 strains could be potentially novel species. Our results also revealed that planktonic fungi were molecularly diverse with hundreds of phylotypes recovered across these two bays. The results of the study provide data about the distribution and taxonomy of mycoplankton in this area, thereby allowing assessment of their positive role in the biogeochemical cycle of coastal ecosystems and the development of new bioactive compounds for industrial applications.

Keywords: biodiversity, ITS, marine fungi, Nha Trang Bay, Van Phong Bay

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Authors: Dmitriy Berillo, Areej K. A. Al-Jwaid, Jonathan L. Caplin, Andrew Cundy, Irina Savina

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Chlorophenols (CPs) are used as a precursor in the production of higher CPs and dyestuffs, and as a preservative. Contamination by CPs of the ground water is located in the range from 0.15-100mg/L. The EU has set maximum concentration limits for pesticides and their degradation products of 0.1μg/L and 0.5μg/L, respectively. People working in industries which produce textiles, leather products, domestic preservatives, and petrochemicals are most heavily exposed to CPs. The International Agency for Research on Cancers categorized CPs as potential human carcinogens. Existing multistep water purification processes for CPs such as hydrogenation, ion exchange, liquid-liquid extraction, adsorption by activated carbon, forward and inverse osmosis, electrolysis, sonochemistry, UV irradiation, and chemical oxidation are not always cost effective and can cause the formation of even more toxic or mutagenic derivatives. Bioremediation of CPs derivatives utilizing microorganisms results in 60 to 100% decontamination efficiency and the process is more environmentally-friendly compared with existing physico-chemical methods. Microorganisms immobilized onto a substrate show many advantages over free bacteria systems, such as higher biomass density, higher metabolic activity, and resistance to toxic chemicals. They also enable continuous operation, avoiding the requirement for biomass-liquid separation. The immobilized bacteria can be reused several times, which opens the opportunity for developing cost-effective processes for wastewater treatment. In this study, we develop a bioremediation system for CPs based on macroporous materials, which can be efficiently used for wastewater treatment. Conditions for the preparation of the macroporous material from specific bacterial strains (Pseudomonas mendocina and Rhodococus koreensis) were optimized. The concentration of bacterial cells was kept constant; the difference was only the type of cross-linking agents used e.g. glutaraldehyde, novel polymers, which were utilized at concentrations of 0.5 to 1.5%. SEM images and rheology analysis of the material indicated a monolithic macroporous structure. Phenol was chosen as a model system to optimize the function of the cryogel material and to estimate its enzymatic activity, since it is relatively less toxic and harmful compared to CPs. Several types of macroporous systems comprising live bacteria were prepared. The viability of the cross-linked bacteria was checked using Live/Dead BacLight kit and Laser Scanning Confocal Microscopy, which revealed the presence of viable bacteria with the novel cross-linkers, whereas the control material cross-linked with glutaraldehyde(GA), contained mostly dead cells. The bioreactors based on bacteria were used for phenol degradation in batch mode at an initial concentration of 50mg/L, pH 7.5 and a temperature of 30°C. Bacterial strains cross-linked with GA showed insignificant ability to degrade phenol and for one week only, but a combination of cross-linking agents illustrated higher stability, viability and the possibility to be reused for at least five weeks. Furthermore, conditions for CPs degradation will be optimized, and the chlorophenol degradation rates will be compared to those for phenol. This is a cutting-edge bioremediation approach, which allows the purification of waste water from sustainable compounds without a separation step to remove free planktonic bacteria. Acknowledgments: Dr. Berillo D. A. is very grateful to Individual Fellowship Marie Curie Program for funding of the research.

Keywords: bioremediation, cross-linking agents, cross-linked microbial cell, chlorophenol degradation

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Authors: Rudzani Manafe, Ezekiel Green

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Brucella, has many different virulence factors that act as a causative agent of brucellosis, depending on the environment and other factors, some factors may play a role more than others during infection and as a result, play a role in becoming a causative agent for pathogenesis. Brucella melitensis and Brucella abortus are considered to be pathogenic to humans. The genetic regularity of nine potential causes of virulence of two Brucella species in Eastern Cape livestock have been examined. A hundred and twenty isolates obtained from Molecular Pathogenesis and Molecular Epidemiology Research Group (MPMERG) were used for this study. All isolates were grown on Brucella agar medium. Nine primer pairs were used for the detection of virB2, virB5, vceC, btpA, btpB, prpA, betB, bpe275, and bspB virulence factors using Polymerase chain reaction (PCR). Approximately 100% was observed for genes BecC and BetB from B. arbotus. While the lowest gene observed was PrpA at 4.6% from B. arbotus. BetB was detected in 34.7%, while virB2 and prpA (0%) were not detected in B. melitensis. The results from this research suggest that most isolates of Brucella have virulence-related genes associated with disease pathogenesis. Finally, our findings showed that Brucella strains in the Eastern Cape Province are extremely virulent as virulence characteristics exist in most strains investigated.

Keywords: putative virulence genes, brucella, polymerase chain reaction, milk

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Authors: Alsulami H., Alghamdi N., Alasker A., Almohen N., Shome D.

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Most conventional chemotherapeutic agents which are used for the treatment of cancers not only eradicate cancer cells but also affect normal hematopoietic Stem cells (HSCs) that leads to severe pancytopenia during treatment. Therefore, a need exists for novel approaches to treat cancer without or with minimum effect on normal HSCs. Parthenolide (PTL), a herbal product occurring naturally in the plant Feverfew, is a potential new chemotherapeutic agent for the treatment of many cancers such as acute myeloid leukemia (AML) and chronic lymphocytic leukemia (CLL). In this study we investigated the effect of different PTL concentrations on the viability of normal HSCs and also on the ability of these cells to form colonies after they have been treated with PTL in vitro. Methods: In this study, 24 samples of bone marrow and cord blood were collected with consent, and mononuclear cells were separated using density gradient separation. These cells were then exposed to various concentrations of PTL for 24 hours. Cell viability after culture was determined using 7ADD in a flow cytometry test. Additionally, the impact of PTL on hematopoietic stem cells (HSCs) was evaluated using a colony forming unit assay (CFU). Furthermore, the levels of NFҝB expression were assessed by using a PE-labelled anti-pNFκBP65 antibody. Results: this study showed that there was no statistically significant difference in the percentage of cell death between untreated and PTL treated cells with 5 μM PTL (p = 0.7), 10 μM PTL (p = 0.4) and 25 μM (p = 0.09) respectively. However, at higher doses, PTL caused significant increase in the percentage of cell death. These results were significant when compared to untreated control (p < 0.001). The response of cord blood cells (n=4) on the other hand was slightly different from that for bone marrow cells in that the percentage of cell death was significant at 100 μM PTL. Therefore, cord blood cells seemed more resistant than bone marrow cells. Discussion &Conclusion: At concentrations ≤25 μM PTL has a minimum or no effect on HSCs in vitro. Cord blood HSCs are more resistant to PTL compared to bone marrow HSCs. This could be due to the higher percentage of T-lymphocytes, which are resistant to PTL, in CB samples (85% in CB vs. 56% in BM. Additionally, CB samples contained a higher proportion of CD34+ cells, with 14.5% of brightly CD34+ cells compared to only 1% in normal BM. These bright CD34+ cells in CB were mostly negative for early-stage stem cell maturation antigens, making them young and resilient to oxidative stress and high concentrations of PTL.

Keywords: stem cell, parthenolide, NFKB, CLL

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Authors: Iyiola Olusola Oluwaleye, Samson A. Adeleye, Omojola Awogbemi

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This paper reports an investigation of the comparative effects of drying on the quality of some leafy vegetables at three different temperatures namely: 50ᵒC, 60ᵒC and 70ᵒC. The vegetables investigated are spinach (Amaranthus cruentus); water leaf (Talinum triangulare); lettuce (Lactuca satuva); and fluted pumpkin (Telfaria occidentalis). These vegetables are available in abundance during raining season and are commonly consumed by average Nigerians. A convective dryer was used for the drying process at the stipulated temperatures which were maintained with the aid of a thermostat. The vegetable samples after washing was cut into smaller sizes of 0.4 cm-0.5 cm and loaded into the drying cage of the convective dryer. The daily duration of the drying is six hours from 9:00 am to 3:00 pm. The dried samples were thereafter subjected to microbial and proximate analyses. The result of the tests shows that the microbial load decreases as the drying temperature increases. As temperature increases, the moisture content and carbohydrate of all the samples decreases while the crude fiber, ash and protein increases. Percentage fat content decreases as drying temperature increases with the exception of fluted pumpkin. The shelf life of the vegetable samples increase with drying temperature, Spinach has the lowest shelf life followed by Fluted Pumpkin, followed by lettuce while Water Leaf has the highest shelf life at the three drying temperatures of 50ᵒC, 60ᵒC and 70ᵒC respectively.

Keywords: convective drying, leafy vegetables, quality, shelf life

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Authors: Simin Khodabakhshi, Hossein Rassi

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Hyperlipidemia refers to any of several acquired or genetic disorders that result in a high level of lipids circulating in the blood. Helicobacter pylori infection is a contributing factor in the progression of hyperlipidemia with serum lipid changes. The aim of this study was to detect of Helicobacter pylori by PCR and serological methods in patients with hyperlipidemia. In this case-control study, 174 patients with hyperlipidemia and 174 healthy controls were studied. Also, demographics, physical and biochemical parameters were performed in all samples. The DNA extracted from blood specimens was amplified by H pylori cagA specific primers. The results show that H. pylori cagA positivity was detected in 79% of the hyperlipidemia and in 56% of the control group by ELISA test and 49% of the hyperlipidemia and in 24% of the control group by PCR test. Prevalence of H. pylori infection was significantly higher in hyperlipidemia as compared to controls. In addition, patients with hyperlipidemia had significantly higher values for triglyceride, total cholesterol, LDL-C, waist to hip ratio, body mass index, diastolic and systolic blood pressure and lower levels of HDL-C than control participants (all p < 0.0001). Our result detected the ELISA was a rapid and cost-effective detection and considering the high prevalence of cytotoxigenic H. pylori strains, cag A is suggested as a promising target for PCR and ELISA tests for detection of infection with toxigenic strains. In general, it can be concluded that molecular analysis of H. pylori cagA and clinical parameters are important in early detection of hyperlipidemia and atherosclerosis with H. pylori infection by PCR and ELISA tests.

Keywords: Helicobacter pylori, hyperlipidemia, PCR, ELISA

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Authors: Bastian Vollrath, Hartwig Hubel

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In case of over-elastic and cyclic loading, strain may accumulate due to a ratcheting mechanism until the state of shakedown is possibly achieved. Load history dependent numerical investigations by a step-by-step analysis are rather costly in terms of engineering time and numerical effort. In the case of multi-parameter loading, where various independent loadings affect the final state of shakedown, the computational effort becomes an additional challenge. Therefore, direct methods like the Simplified Theory of Plastic Zones (STPZ) are developed to solve the problem with a few linear elastic analyses. Post-shakedown quantities such as strain ranges and cyclic accumulated strains are calculated approximately by disregarding the load history. The STPZ is based on estimates of a transformed internal variable, which can be used to perform modified elastic analyses, where the elastic material parameters are modified, and initial strains are applied as modified loading, resulting in residual stresses and strains. The STPZ already turned out to work well with respect to cyclic loading between two states of loading. Usually, few linear elastic analyses are sufficient to obtain a good approximation to the post-shakedown quantities. In a multi-dimensional load domain, the approximation of the transformed internal variable transforms from a plane problem into a hyperspace problem, where time-consuming approximation methods need to be applied. Therefore, a solution restricted to structures with four stress components was developed to estimate the transformed internal variable by means of three-dimensional vector algebra. This paper presents the extension to cyclic multi-parameter loading so that an unlimited number of load cases can be taken into account. The theoretical basis and basic presumptions of the Simplified Theory of Plastic Zones are outlined for the case of elastic shakedown. The extension of the method to many load cases is explained, and a workflow of the procedure is illustrated. An example, adopting the FE-implementation of the method into ANSYS and considering multilinear hardening is given which highlights the advantages of the method compared to incremental, step-by-step analysis.

Keywords: cyclic loading, direct method, elastic shakedown, multi-parameter loading, STPZ

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Authors: Ilze Dimanta, Zane Rutkovska, Vizma Nikolajeva, Janis Kleperis, Indrikis Muiznieks

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Majority of word’s steadily increasing energy consumption is provided by non-renewable fossil resources. Need to find an alternative energy resource is essential for further socio-economic development. Hydrogen is renewable, clean energy carrier with high energy density (142 MJ/kg, accordingly – oil has 42 MJ/kg). Biological hydrogen production is an alternative way to produce hydrogen from renewable resources, e.g. using organic waste material resource fermentation that facilitate recycling of sewage and are environmentally benign. Hydrogen gas is produced during the fermentation process of bacteria in anaerobic conditions. Bacteria are producing hydrogen in the liquid phase and when thermodynamic equilibrium is reached, hydrogen is diffusing from liquid to gaseous phase. Because of large quantities of available crude glycerol and the highly reduced nature of carbon in glycerol per se, microbial conversion of it seems to be economically and environmentally viable possibility. Such industrial organic waste product as crude glycerol is perspective for usage in feedstock for hydrogen producing bacteria. The process of biodiesel production results in 41% (w/w) of crude glycerol. The developed lab-scale test system (experimental bioreactor) with hydrogen micro-electrode (Unisense, Denmark) was used to determine hydrogen production yield and rate in the liquid phase. For hydrogen analysis in the gas phase the RGAPro-100 mass-spectrometer connected to the experimental test-system was used. Fermentative bacteria strains were tested for hydrogen gas production rates. The presence of hydrogen in gaseous phase was measured using mass spectrometer but registered concentrations were comparatively small. To decrease the hydrogen partial pressure in liquid phase reactor with a system for continuous bubbling with inert gas was developed. H2 production rate for the best producer in liquid phase reached 0,40 mmol H2/l, in gaseous phase - 1,32 mmol H2/l. Hydrogen production rate is time dependent – higher rate of hydrogen production is at the fermentation process beginning when concentration increases, but after three hours of fermentation, it decreases.

Keywords: bio-hydrogen, fermentation, experimental bioreactor, crude glycerol

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Authors: Hamada Ahmed, Mervat A. Abdel-Latif, Alaa. A. Ghoraba, Samah A. Ganna

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An experiment was conducted to determine the effect of microbial phytase (Quantum Blue®) supplementation on layer chickens fed diets with required or low phosphorous level in corn-soybean based diets. One hundred and sixteen 23-week-old Lohman brown laying hens were used in 8-week feeding trial. Hens were randomly allotted into four treatments where the group (1) (control group) was fed basal diet without phytase, group (2) fed basal diet supplemented with phytase, group (3) fed diet supplemented with phytase as a replacement of 25% of monocalcium phosphate and group (4) fed diet supplemented with phytase as a replacement of 50% of monocalcium phosphate. Records on daily egg production, egg mass, egg weight and body weight of hens at the end of experimental period were recorded. Results revealed no significant (p ≥ 0.05) differences were observed among the other dietary treatments in BW, egg production, egg mass, feed intake or feed conversion when these parameters were evaluated over the duration of the experiment while egg weight showed significant (p < 0.05) increase in all phytase supplemented groups. There was no significant (p ≥ 0.05) differences in egg quality including egg length, egg width, egg shape index, yolk height, yolk width, yolk index, yolk weight and yolk albumin ratio while egg albumin was significantly increased (p < 0.05) in group (2) and group (3). Egg shell weight increased significantly (p < 0.05) in all phytase supplemented groups when compared with the control group also shell thickness increased significantly (p < 0.05) in both group (2 &3). No significant (P ≥ 0.05) difference was observed in serum Ca, P level while alkaline phosphatase was significantly (P ˂ 0.05) increased in group (3). Egg shell analysis showed increase in egg shell ash% in all phytase supplemented groups when compared with the control group, egg shell calcium % was higher in group (3) and group (4) than the control group while group (2) showed lower egg shell calcium% than the other experimental groups, egg shell phosphorous% was higher in all phytase supplemented groups than the control group. Phosphorous digestability was significantly (P ˂ 0.05) increased in all phytase supplemented groups than the control group and the highest p digestability was in group (4). Calcium digestability showed significant (P ˂ 0.05) increase in all phytase supplemented groups when compared with the control group and the highest digetability was in group (4).

Keywords: layers, microbial phytase, Ca and P availability, egg production, egg characteristics

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Authors: Young Ah Kim, Hyunsoo Kim, Eun-Jeong Yoon, Young Hee Seo, Kyungwon Lee

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Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli from food animals are considered as a reservoir for transmission of ESBL genes to human. The aim of this study is to assess the prevalence and molecular epidemiology of ESBL-producing E. coli colonization in pigs, farm workers, and farm environments to elucidate the transmission of multidrug-resistant clones from animal to human. Nineteen pig farms were enrolled across the country in Korea from August to December 2017. ESBL-producing E. coli isolates were detected in 190 pigs, 38 farm workers, and 112 sites of farm environments using ChromID ESBL (bioMerieux, Marcy l'Etoile, France), directly (stool or perirectal swab) or after enrichment (sewage). Antimicrobial susceptibility tests were done with disk diffusion methods and blaTEM, blaSHV, and blaCTX-M were detected with PCR and sequencing. The genomes of the four CTX-M-55-producing E. coli isolates from various sources in one farm were entirely sequenced to assess the relatedness of the strains. Whole genome sequencing (WGS) was performed with PacBio RS II system (Pacific Biosciences, Menlo Park, CA, USA). ESBL genotypes were 85 CTX-M-1 group (one CTX-M-3, 23 CTX-M-15, one CTX-M-28, 59 CTX-M-55, one CTX-M-69) and 60 CTX-M-9 group (41 CTX-M-14, one CTX-M-17, one CTX-M-27, 13 CTX-M-65, 4 CTX-M-102) in total 145 isolates. The rectal colonization rates were 53.2% (101/190) in pigs and 39.5% (15/38) in farm workers. In WGS, sequence types (STs) were determined as ST69 (E. coli PJFH115 isolate from a human carrier), ST457 (two E. coli isolates PJFE101 recovered from a fence and PJFA1104 from a pig) and ST5899 (E. coli PJFA173 isolate from the other pig). The four plasmids encoding CTX-M-55 (88,456 to 149, 674 base pair), whether it belonged to IncFIB or IncFIC-IncFIB type, shared IncF backbone furnishing the conjugal elements, suggesting of genes originated from same ancestor. In conclusion, the prevalence of ESBL-producing E. coli in swine farms was surprisingly high, and many of them shared common ESBL genotypes of clinical isolates such as CTX-M-14, 15, and 55 in Korea. It could spread by horizontal transfer between isolates from different reservoirs (human-animal-environment).

Keywords: Escherichia coli, extended-spectrum β-lactamase, prevalence, whole genome sequencing

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Authors: Amr Saeb, Khalid Al-Rubeaan, Mohamed Abouelhoda, Manojkumar Selvaraju, Hamsa Tayeb

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Background: P. mirabilis is a common uropathogenic bacterium that can cause major complications in patients with long-standing indwelling catheters or patients with urinary tract anomalies. In addition, P. mirabilis is a common cause of chronic osteomyelitis in diabetic foot ulcer (DFU) patients. Methodology: P. mirabilis SCDR1 was isolated from a diabetic ulcer patient. We examined P. mirabilis SCDR1 levels of resistance against nano-silver colloids, the commercial nano-silver and silver containing bandages and commonly used antibiotics. We utilized next generation sequencing techniques (NGS), bioinformatics, phylogenetic analysis and pathogenomics in the identification and characterization of the infectious pathogen. Results: P. mirabilis SCDR1 is a multi-drug resistant isolate that also showed high levels of resistance against nano-silver colloids, nano-silver chitosan composite and the commercially available nano-silver and silver bandages. The P. mirabilis-SCDR1 genome size is 3,815,621 bp with G+C content of 38.44%. P. mirabilis-SCDR1 genome contains a total of 3,533 genes, 3,414 coding DNA sequence genes, 11, 10, 18 rRNAs (5S, 16S, and 23S), and 76 tRNAs. Our isolate contains all the required pathogenicity and virulence factors to establish a successful infection. P. mirabilis SCDR1 isolate is a potential virulent pathogen that despite its original isolation site, wound, it can establish kidney infection and its associated complications. P. mirabilis SCDR1 contains several mechanisms for antibiotics and metals resistance including, biofilm formation, swarming mobility, efflux systems, and enzymatic detoxification. Conclusion: P. mirabilis SCDR1 is the spontaneous nano-silver resistant bacterial strain. P. mirabilis SCDR1 strain contains all reported pathogenic and virulence factors characteristic for the species. In addition, it possesses several mechanisms that may lead to the observed nano-silver resistance.

Keywords: Proteus mirabilis, multi-drug resistance, silver nanoparticles, resistance, next generation sequencing techniques, genome analysis, bioinformatics, phylogeny, pathogenomics, diabetic foot ulcer, xenobiotics, multidrug resistance efflux, biofilm formation, swarming mobility, resistome, glutathione S-transferase, copper/silver efflux system, altruism

Procedia PDF Downloads 335