Search results for: resistant microbial strains

Authors: Nachawit Tikul

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This study aimed to determine low-income housing adaptations for flooding, which causes living problems and housing damage, and the results from improvement. Three low-income settlements in Chiang Mai which experienced different flood types, i.e. flash floods in Samukeepattana, drainage floods in Bansanku, and river floods in Kampangam, were chosen for the study. Almost all of the residents improved their houses to protect the property from flood damage by changing building materials to flood damage resistant materials for walls, floors, and other parts of the structure that were below the base of annual flood elevation. They could only build some parts of their own homes, so hiring skilled workers or contractors was still important. Building materials which have no need for any special tools and are easy to access and use for construction, as well as low cost, are selected for construction. The residents in the three slums faced living problems for only a short time and were able to cope with them. This may be due to the location of the three slums near the city where assistance is readily available. But the housing and the existence in the slums can endure only the regular floods and residence still have problems in unusual floods, which have been experienced 1-2 times during the past 10 years. The residents accept the need for evacuations and prepare for them. When faced with extreme floods, residence have evacuated to the nearest safe place such as schools and public building, and come back to repair the houses after the flood. These are the distinguishing characteristics of low-income living which can withstand serious situations due to the simple lifestyle. Therefore, preparation of living areas for use during severe floods and encouraging production of affordable flood resistant materials should be areas of concern when formulating disaster assistance policies for low income people.

Keywords: flooding, low-income settlement, housing, adaptation

Procedia PDF Downloads 238

Authors: Manal Suleiman, George Abu-Aqil, Uraib Sharaha, Klaris Riesenberg, Itshak Lapidot, Ahmad Salman, Mahmoud Huleihel

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Klebsiella pneumoniae is one of the most aggressive multidrug-resistant bacteria associated with human infections resulting in high mortality and morbidity. Thus, for an effective treatment, it is important to diagnose both the species of infecting bacteria and their susceptibility to antibiotics. Current used methods for diagnosing the bacterial susceptibility to antibiotics are time-consuming (about 24h following the first culture). Thus, there is a clear need for rapid methods to determine the bacterial susceptibility to antibiotics. Infrared spectroscopy is a well-known method that is known as sensitive and simple which is able to detect minor biomolecular changes in biological samples associated with developing abnormalities. The main goal of this study is to evaluate the potential of infrared spectroscopy in tandem with Random Forest and XGBoost machine learning algorithms to diagnose the susceptibility of Klebsiella pneumoniae to antibiotics within approximately 20 minutes following the first culture. In this study, 1190 Klebsiella pneumoniae isolates were obtained from different patients with urinary tract infections. The isolates were measured by the infrared spectrometer, and the spectra were analyzed by machine learning algorithms Random Forest and XGBoost to determine their susceptibility regarding nine specific antibiotics. Our results confirm that it was possible to classify the isolates into sensitive and resistant to specific antibiotics with a success rate range of 80%-85% for the different tested antibiotics. These results prove the promising potential of infrared spectroscopy as a powerful diagnostic method for determining the Klebsiella pneumoniae susceptibility to antibiotics.

Keywords: urinary tract infection (UTI), Klebsiella pneumoniae, bacterial susceptibility, infrared spectroscopy, machine learning

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Authors: Anoff Anim, Lila A. M. Mahmoud, Maria Katsikogianni, Sanjit Nayak

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Sustained and controlled delivery of antimicrobial drugs have been largely studied recently using metal organic frameworks (MOFs)and different polymers. However, much attention has not been given to combining both MOFs and biodegradable polymers, which would be a good strategy in providing a sustained gradual release of the drugs. Herein, we report a comparative study of the sustained and controlled release of widely used antibacterial drugs, cephalexin and metronidazole, from zinc-based MOF-5 incorporated in biodegradable polycaprolactone (PCL) and poly-lactic glycolic acid (PLGA) membranes. Cephalexin and metronidazole were separately incorporated in MOF-5 post-synthetically, followed by their integration into biodegradable PLGA and PCL membranes. The pristine MOF-5 and the loaded MOFs were thoroughly characterized by FT-IR, SEM, TGA and PXRD. Drug release studies were carried out to assess the release rate of the drugs in PBS and distilled water for up to 48 hours using UV-Vis Spectroscopy. Four bacterial strains from both the Gram-positive and Gram-negative types, Staphylococus aureus, Staphylococuss epidermidis, Escherichia coli, Acinetobacter baumanii, were tested against the pristine MOF, pure drugs, loaded MOFs and the drug-loaded MOF-polymer composites. Metronidazole-loaded MOF-5 composite of PLGA (PLGA-Met@MOF-5) was found to show highest efficiency to inhibit the growth of S. epidermidis compared to the other bacteria strains while maintaining a sustained minimum inhibitory concentration (MIC). This study demonstrates that the combination of biodegradable MOF-polymer composites can provide an efficient platform for sustained and controlled release of antimicrobial drugs and can be a potential strategy to integrate them in biomedical devices.

Keywords: antimicrobial resistance, biodegradable polymers, cephalexin, drug release metronidazole, MOF-5, PCL, PLGA

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Authors: Tsegay Teklebrhan Gebremariam, Zhiliang, Arjan Jonker

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The study investigated that using corn gluten (CG) instead of cornmeal (CM) increased dietary sulfur shifted H₂ metabolism from methanogenesis to alternative sink and modulated microbiome in the rumen as well as hindgut segments of goats. Ruminal fermentation, CH₄ emissions and microbial abundance in goats (n = 24). The experiment was performed using a randomized block design with two dietary treatments (CM and CG with 400 g/kg DM each). Goats in CG increased sulfur, NDF and CP intake and decreased starch intake as compared with those in CM. Goats that received CG diet had decreased dissolved hydrogen (dH₂) (P = 0.01) and dissolved methane yield and emission (dCH₄) (P = 0.001), while increased dH₂S both in the rumen and hindgut segments than those fed CM. Goats fed CG had higher (p < 0.01) gene copies of microbiota and cellulolytic bacteria, whereas starch utilizing bacterial species were less in the rumen and hindgut than those fed CM. Higher (P < 0.05) methanogenic diversity and abundances of Methanimicrococcus and Methanomicrobium were observed in goats that consumed CG, whilst containing lower Methanobrevibacter populations than those receiving CM. The study suggested that goats fed corn gluten improved the gene copies of microbiota and fibrolytic bacterial species while reducing starch utilizing species in the rumen and hindgut segments as compared with that fed cornmeal. Goats consuming corn gluten had a more enriched methanogenic diversity and reduced Methanobrevibacter, a contributor to CH₄ emissions, as compared with goats fed CM. Corn gluten could be used as an alternative feed to decrease the enteric CH₄ emission in ruminant production.

Keywords: dissolved gasses, methanogenesis, microbial community, metagenomics

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Authors: S. Jaikeaw, S. Chavadej

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The objective of this study was to determine the effects of COD loading rate on hydrogen and methane production and micronutrient transportation using a two-stage upflow anaerobic sludge blanket (UASB) system under mesophilic temperature (37°C) with a constant recycle ratio of 1:1 (final effluent flow rate: feed flow rate). The first (hydrogen) UASB unit having 4 L liquid holding volume was controlled at pH 5.5 but the second (methane) UASB unit having 24 L liquid holding volume had no pH control. The two-stage UASB system operated at different COD loading rates from 8 to 20 kg/m³d based on total UASB working volume. The results showed that, at the optimum COD loading rate of 13 kg/m³d, the produced gas from the hydrogen UASB unit contained 1.5% H₂, 16.5% CH₄, and 82% CO₂ with H₂S of 252 ppm and also provided a hydrogen yield of 1.66 mL/g COD removed (or 0.56 mL/g COD applied) and a specific hydrogen production rate of 156.85 ml H₂/LRd (or 5.12 ml H₂/g MLVSS d). Under the optimum COD loading rate, the produced gas from the methane UASB unit mainly contained methane and carbon dioxide without hydrogen of 74 and 26%, respectively with hydrogen sulfide of 287 ppm and the system also provided a maximum methane yield of 407.00 mL/g COD removed (or 263.23 mL/g COD applied) and a specific methane production rate of 2081.44 ml CH₄/LRd (or 99.75 ml CH₄/g MLVSS d). Under the optimum COD loading rate, all micronutrients markedly dropped by the sulfide precipitation reactions. The reduction of micronutrients mostly appeared in the methane UASB unit. Under the studied conditions, both Co and Ni were found to be greatly precipitated out, causing the deficiency to microbial activity. It is hypothesized that an addition of both Co and Ni can improve the methanogenic activity.

Keywords: hydrogen and methane production, ethanol wastewater, a two-stage upflow anaerobic blanket (UASB) system, mesophillic temperature, microbial concentration (MLVSS), micronutrients

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Authors: Nho-Eul Song, Sang-Ho Baik

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Fermentation characteristics of black raspberry vinegar by using static cultures without any additives were has been investigated to establish of vinegar manufacturing conditions and improve the quality of vinegar by optimization the vinegar manufacturing process. The two vinegar manufacturing conditions were prepared; one-step fermentation condition only using mother vinegar that prepared naturally occurring black raspberry vinegar without starter yeast for alcohol fermentation (traditional method) and two-step fermentation condition using commercial wine yeast and mother vinegar for acetic acid fermentation. Approximately 12% ethanol was produced after 35 days fermentation with log 7.6 CFU/mL of yeast population in one-step fermentation, resulting sugar reduction from 14 to 6oBrix whereas in two-step fermentation, ethanol concentration was reached up to 8% after 27 days with continuous increasing yeast until log 7.0 CFU/mL. In addition, yeast and ethanol were decreased after day 60 accompanied with proliferation of acetic acid bacteria (log 5.8 CFU/mL) and titratable acidity; 4.4% in traditional method and 6% in two-step fermentation method. DGGE analysis showed that S. cerevisiae was detected until 77 days of traditional fermentation and gradually changed to AAB, Acetobacter pasteurianus, as dominant species and Komagataeibacter xylinus at the end of the fermentation. However, S. cerevisiae and A. pasteurianus was dominant in two-step fermentation process. The prepared two-step fermentation showed enhanced total polyphenol and flavonoid content significantly resulting in higher radical scavenging activity. Our studies firstly revealed the microbial community change with chemical change and demonstrated a suitable fermentation system for black raspberry vinegar by the static surface method.

Keywords: bacteria, black raspberry, vinegar fermentation, yeast

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Authors: Afifa Hafidh, Hedia Chaabane

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This work mainly focused on the synthetic strategies and biological activities associated with pyrazoles. Pyrazole derivatives have been successfully synthesized by simple and facile method and studied for their antibacterial activity. These compounds were prepared from pyrazolic difunctional compounds as starting materials, by reaction with salicylic acid, paracetamol and thiosemicarbazide respectively. Structure of all the prepared compounds confirmation were proved using (FT-IR), (1H-NMR) and (13C-NMR) spectra in addition to melting points. The screening of the antimicrobial activity of the pyrazolic derivatives was examined against different microorganisms in the present study. They were screened for their antimicrobial activities against gram positive bacteria, gram negative bacteria and Candida albicans. The synthesized compounds were found to exhibit high antibacterial and antifungal efficiency against several tested bacterial strains, using agar diffusion method and filter paper disc-diffusion method. Ampicillin was used as positive control for all strains except Candida albicans for which Nystatin was used. The obtained results reveal that the antibacterial activity of some pyrazolic derivatives is comparable to that observed for the control samples (Ampicilin and Nystatin), suggesting a strong antibacterial activity. The analysis of these results shows that synthesized products react on the surfaces cell walls that are disrupted. When these products are in contact with the bacteria, they damage the membrane, leading to the perturbation of different cellular processes and then leakage of cytoplasm, resulting in the death of the cells. The results will be presented in details. The obtained products constitute effective antibacterial agents and important compounds for biological systems.

Keywords: salicylic acid, antimicrobial activities, antioxidant activity, paracetamol, pyrazole, thiosemicarbazide

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Authors: Vadim Makarov, Stewart T.Cole

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PBTZ169 is novel drug candidate with high efficacy in animals models, and its combination treatment of PBTZ169 with BDQ and pyrazinamide was shown to be more efficacious than the standard treatment for tuberculosis in a mouse model. The target of PBTZ169 is famous DprE1, an essential enzyme in cell wall biosynthesis. The crystal structure of the DprE1-PBTZ169 complex reveals formation of a semimercaptal adduct with Cys387 in the active site and explains the irreversible inactivation of the enzyme. Furthermore, this drug candidate demonstrated during preclinical research ‘drug like’ properties what made it an attractive drug candidate to treat tuberculosis in humans. During first clinical trials several cohorts of the healthy volunteers were treated by the single doses of PBTZ169 as well as two weeks repeated treatment was chosen for two maximal doses. As expected PBTZ169 was well tolerated, and no significant toxicity effects were observed during the trials. The study of the metabolism shown that human metabolism of PBTZ169 is very different from microbial or animals compound transformation. So main pathway of microbial, mice and less rats metabolism connected with reduction processes, but human metabolism mainly connected with oxidation processes. Due to this difference we observed several metabolites of PBTZ169 in humans with antitubercular activity, and now we can conclude that animal antituberculosis activity of PBTZ169 is a result not only activity of the drug itself, but it is a result of the sum activity of the drug and its metabolites. Direct antimicrobial plasma activity was studied, and such activity was observed for 24 hours after human treatment for some doses. This data gets high chance for good efficacy of PBTZ169 in human for treatment TB infection. Second phase of clinical trials was started summer of 2017 and continues to the present day. Available data will be presented.

Keywords: clinical trials, DprE1, PBTZ169, metabolism

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Authors: Muhammad Naeem Chaudhry, Fahim Nawaz, Rana Nauman Shabbir

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New strategies aimed at increasing the resilience of crop plants to the negative effects of climate change represent important research priorities of plant scientists. The use of soil microorganisms to alleviate abiotic stresses like drought has gained particular importance in recent past. A field experiment was planned to investigate the effect of phosphorous solubilizing bacteria on yield and seed quality of Camelina (Camelina sativa L.) under water deficit conditions. The study was conducted at Agronomic Research Farm, University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur, during 4th week of November, 2013. The available seeds of Camelina sativa were inoculated with two bacterial strains (pseudomonas and Bacillus spp.) and grown under various water stress levels i.e. D0, (four irrigations), D3 (three irrigation), D2 (two irrigations), and D1 (one irrigation). The results revealed that drought stress significantly reduced the plant growth and yield, consequently reducing protein contents and oil concentration in camelina. The exposure to drought stress decreased plant height (16%), plant population (27%), number of fertile branches (41-59%), number of pods per plant (35%) and seed per pod (33%). Drought stress also exerted a negative impact on yield characteristics by reducing the 1000-seed weight (65%), final seed yield (52%), biological yield (22%) and harvest index (39%) of camelina. However, the inoculation of seeds with Pseudomonas and Bacillus spp. promoted the plant growth characterized by increased plant height and enhanced plant population. It was noted that inoculation of seeds with Pseudomonas resulted in the maximum plant population (113.4 cm), primary branches (19 plant-1), and number of pods (664 plant-1), whereas Bacillus inoculation resulted in maximum plant height (113.4 cm), seeds per pod (15.9), 1000-seed weight (1.85 g), and seed yield (3378.8 kg ha-1). Moreover, the inoculation with Bacillus also significantly improved the quality attributes of camelina and gave 3.5% and 2.1% higher oil contents than Pseudomonas and control (no-inoculation), respectively. Similarly, the same strain also resulted in maximum protein contents (33.3%). Our results confirmed the hypothesis that inoculation of seeds with phosphorous solubilizing bacterial strains is an effective, viable and environment-friendly approach to improve yield and quality of camelina under water deficit conditions. However, further studies are suggested to investigate the physiological and molecular processes, stimulated by bacterial strains, for increasing drought tolerance in food crops.

Keywords: Camelina, drought stress, phosphate solubilizing bacteria, seed quality

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Authors: Wiqar Hussain Shah

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The Structural, magnetic and transport behavior of La1-xCaxMnO3+ (x=0.48, 0.50, 0.52 and 0.55 and =0.015) compositions close to charge ordering, was studied through XRD, resistivity, DC magnetization and AC susceptibility measurements. With time and thermal cycling (T<300 K) there is an irreversible transformation of the low-temperature phase from a partially ferromagnetic and metallic to one that is less ferromagnetic and highly resistive. For instance, an increase of resistivity can be observed by thermal cycling, where no effect is obtained for lower Ca concentration. The time changes in the magnetization are logarithmic in general and activation energies are consistent with those expected for electron transfer between Mn ions. The data suggest that oxygen non-stoichiometry results in mechanical strains in this two-phase system, leading to the development of irreversible metastable states, which relax towards the more stable charge-ordered and antiferromagnetic microdomains at the nano-meter size. This behavior is interpreted in terms of strains induced charge localization at the interface between FM/AFM domains in the antiferromagnetic matrix. Charge, orbital ordering and phase separation play a prominent role in the appearance of such properties, since they can be modified in a spectacular manner by external factor, making the different physical properties metastable. Here we describe two factors that deeply modify those properties, viz. the doping concentration and the thermal cycling. The metastable state is recovered by the high temperature annealing. We also measure the magnetic relaxation in the metastable state and also the revival of the metastable state (in a relaxed sample) due to high temperature (800 ) thermal treatment.

Keywords: Rare-earth maganites, nano-structural materials, doping effects on electrical, magnetic properties, competing interactions

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Authors: Shivankar Agrawal, Sunil Kumar Deshmukh, Colin Barrow, Alok Adholeya

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A variety of genetic and environmental factors cause various cosmetics and dermatological problems. There are already claimed drugs available in market for treating these problems. However, the challenge remains in finding more potent, environmental friendly, causing minimal side effects and economical cosmeceuticals. This leads to an increased demand for natural cosmeceutical products in the last few decades. Plant derived ingredients are limited because plants either contain toxic metabolites, grow too slow or seasonal harvesting is a problem. The research work carried out in this project aims at isolation, characterization of marine fungal secondary metabolite and evaluating their potential use in future cosmetic skin care products. We have isolated and purified 35 morphologically different fungal isolates from various marine habitats of the India. These isolates have been functionally characterized for anti-tyrosinase, antioxidant and anti-acne activities. For molecular characterization, the Internal Transcribed spacer (ITS) region of 15 functionally active marine fungal isolates was amplified using universal primers, ITS1 and ITS4 and sequenced. Out of 15 marine fungal isolates crude extract of strains D4 (Aspergillus terreus) and P2 (Talaromyces stipitatus) showed 70% and 57% tyrosinase inhibition at 1mg/mL respectively. Strain D5 (Simplicillium lamellicola) has showed significant inhibition against Propionibacterium acnes and Staphylococcus epidermidis. In addition, all these strains also displayed DPPH- radical scavenging activity and may be utilized as skin cosmeceutical applications. Purification and characterization of crude extracts for identification of active lead molecule is under process.

Keywords: anti-acne, anti-tyrosinase, cosmeceutical, marine fungi

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Authors: Gina Leisching, Ray-Dean Pietersen, Carel Van Heerden, Paul Van Helden, Ian Wiid, Bienyameen Baker

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The distinguishing factors that characterize the host response to infection with virulent Mycobacterium tuberculosis (M.tb) are largely confounding. We present an infection study with two genetically closely related M.tb strains that have vastly different pathogenic characteristics. The early host response to infection with these detergent-free cultured strains was analyzed through RNAseq in an attempt to provide information on the subtleties which may ultimately contribute to the virulent phenotype. Murine bone marrow-derived macrophages (BMDMs) were infected with either a hyper- (R5527) or hypovirulent (R1507) Beijing M. tuberculosis clinical isolate. RNAseq revealed 69 differentially expressed host genes in BMDMs during comparison of these two transcriptomes. Pathway analysis revealed activation of the stress-induced and growth inhibitory Gadd45 signaling pathway in hypervirulent infected BMDMs. Upstream regulators of interferon activation such as and IRF3 and IRF7 were predicted to be upregulated in hypovirulent-infected BMDMs. Additional analysis of the host immune response through ELISA and qPCR included the use of human THP-1 macrophages where a robust proinflammatory response was observed after infection with the hypervirulent strain. RNAseq revealed two early-response genes (IER3 and SAA3) and two host-defence genes (OASL1 and SLPI) that were significantly upregulated by the hypervirulent strain. The role of these genes under M.tb infection conditions are largely unknown but here we provide validation of their presence with use of qPCR and Western blot. Further analysis into their biological role under infection with virulent M.tb is required.

Keywords: host-response, Mycobacterium tuberculosis, RNAseq, virulence

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Authors: Prajna Paramita Naik, Sujit Kumar Bhutia

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Background: Regardless of the development multimodal treatment strategies, oral squamous cell carcinoma (OSCC) is often associated with a high rate of recurrence, metastasis and chemo- and radio- resistance. The present study inspected the relevance of CD44, ABCB1 and ADAM17 expression as a putative stem cell compartment in oral squamous cell carcinoma (OSCC) and deciphered the role of autophagy in regulating the expression of aforementioned proteins, stemness and chemoresistance. Methods: A retrospective analysis of CD44, ABCB1 and ADAM17 expression with respect to the various clinicopathological factors of sixty OSCC patients were determined via immunohistochemistry. The correlation among CD44, ABCB1 and ADAM17 expression was established. Sphere formation assay, flow cytometry and fluorescence microscopy were conducted to elucidate the stemness and chemoresistance nature of established cisplatin-resistant oral cancer cells (FaDu). The pattern of expression of CD44, ABCB1 and ADAM17 in parental (FaDu-P) and resistant FaDu cells (FaDu-CDDP-R) were investigated through fluorescence microscopy. Western blot analysis of autophagy marker proteins was performed to compare the status of autophagy in parental and resistant FaDu cell. To investigate the role of autophagy in chemoresistance and stemness, sphere formation assay, immunofluorescence and Western blot analysis was performed post transfection with siATG14 and the level of expression of autophagic proteins, mitochondrial protein and stemness-associated proteins were analyzed. The statistical analysis was performed by GraphPad Prism 4.0 software. p-value was defined as follows: not significant (n.s.): p > 0.05;*: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001; ****: p ≤ 0.0001 were considered statistically significant. Results: In OSCC, high CD44, ABCB1 and ADAM17 expression were significantly correlated with higher tumor grades and poor differentiation. However, the expression of these proteins was not related to the age and sex of OSCC patients. Moreover, the expression of CD44, ABCB1 and ADAM17 were positively correlated with each other. In vitro and OSCC tissue double labeling experiment data showed that CD44+ cells were highly associated with ABCB1 and ADAM17 expression. Further, FaDu-CDDP-R cells showed higher sphere forming capacity along with increased fraction of the CD44+ population and β-catenin expression FaDu-CDDP-R cells also showed accelerated expression of CD44, ABCB1 and ADAM17. A comparatively higher autophagic flux was observed in FaDu-CDDP-R against FaDu-P cells. The expression of mitochondrial proteins was noticeably reduced in resistant cells as compared to parental cells indicating the occurrence of autophagy-mediated mitochondrial degradation in oral cancer. Moreover, inhibition of autophagy was coupled with the decreased formation of orospheres suggesting autophagy-mediated stemness in oral cancer. Blockade of autophagy was also found to induce the restoration of mitochondrial proteins in FaDu-CDDP-R cells indicating the involvement of mitophagy in chemoresistance. Furthermore, a reduced expression of CD44, ABCB1 and ADAM17 was also observed in ATG14 deficient cells FaDu-P and FaDu-CDDP-R cells. Conclusion: The CD44+ ⁄ABCB1+ ⁄ADAM17+ expression in OSCC might be associated with chemoresistance and a putative CSC compartment. Further, the present study highlights the contribution of mitophagy in chemoresistance and confirms the potential involvement of autophagic regulation in acquisition of stem-like characteristics in OSCC.

Keywords: ABCB1, ADAM17, autophagy, CD44, chemoresistance, mitophagy, OSCC, stemness

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Authors: M. Imran Hamid, Muzammil Hussain, Yunpeng Wu, Meichun Xiang, Xingzhong Liu

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The rhizosphere microbiome elucidate multiple functioning in the soil suppressiveness against plant pathogens. Soybean rhizosphere microbial communities may involve in the natural suppression of soybean cyst nematode (SCN) populations in disease suppressive soils. To explore these ecological mechanisms of microbes, a long term monoculture suppressive soil were taken into account for further investigation to test the disease suppressive ability by using different treatments. The designed treatments are as, i) suppressive soil (S), ii) conducive soil (C), iii) conducive soil mixed with 10% (w/w) suppressive soil (CS), iv) suppressive soil treated at 80°C for 1 hr (S80), and v) suppressive soil treated with formalin (SF). By using an ultra-high-throughput sequencing approach, we identified the key bacterial and fungal taxa involved in SCN suppression. The Phylum-level investigation of bacteria revealed that Actinobacteria, Bacteroidetes, and Proteobacteria in the rhizosphere soil of soybean seedlings were more abundant in the suppressive soil than in the conducive soil. The phylum-level analysis of fungi in rhizosphere soil indicated that relative abundance of Ascomycota was higher in suppressive soil than in the conducive soil, where Basidiomycota was more abundant. Transferring suppressive soil to conducive soil increased the population of Ascomycota in the conducive soil by lowering the populations of Basidiomycota. The genera, such as, Pochonia, Purpureocillium, Fusarium, Stachybotrys that have been well documented as bio-control agents of plant nematodes were far more in the disease suppressive soils. Our results suggested that the plants engage a subset of functional microbial groups in the rhizosphere for initial defense upon nematode attack and protect the plant roots later on by nematodes to response for suppression of SCN in disease-suppressive soils.

Keywords: disease suppressive soil, high-throughput sequencing, rhizosphere microbiome, soybean cyst nematode

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Authors: Kubra Dogan, Fatih Tornuk

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Due to increasing consumer demand for the high-quality food products and awareness regarding the health benefits of different nutrients in food minimal processing becomes more popular in modern food preservation. To date, heat treatment is often used for inactivation of spoilage microorganisms in foods. However, it may cause significant changes in the quality and nutritional properties of food. In order to overcome the detrimental effects of heat treatment, several alternatives of non-thermal microbial inactivation processes have been investigated. Ultraviolet (UV) inactivation is a promising and feasible method for better quality and longer shelf life as an alternative to heat treatment, which aims to inhibit spoilage and pathogenic microorganisms and to inactivate the enzymes in vegetable juice production. UV-C is a sub-class of UV treatment which shows the highest microcidal effect between 250-270 nm. The wavelength of 254 nm is used for the surface disinfection of certain liquid food products such as vegetable juice. Effects of UV-C treatment on microbiological load and quality parameter of vegetable juice which is a mix of celery, carrot, lemon and orange was investigated. Our results showed that storing of UV-C applied vegetable juice for three months, reduced the count of TMAB by 3.5 log cfu/g and yeast-mold by 2 log cfu/g compared to control sample. Total phenolic content was found to be 514.3 ± 0.6 mg gallic acid equivalent/L, and there wasn’t a significant difference compared to control. The present work suggests that UV-C treatment is an alternative method for disinfection of vegetable juice since it enables adequate microbial inactivation, longer shelf life and has minimal effect on degradation of quality parameters of vegetable juice.

Keywords: heat treatment, phenolic content, shelf life, ultraviolet (UV-C), vegetable juice

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Authors: Abdul Haq

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The goal of this research is to better understand how wind loads affect bridges and develop strategies for designing bridges that are more stable and resistant to wind. The effect of wind on bridges is essential to their safety and functionality, especially in areas that are prone to high wind speeds or violent wind conditions. The study looks at the aerodynamic forces and vibrations caused by wind and how they affect bridge construction. Part of the research method involves first understanding the underlying ideas influencing wind flow near bridges. Computational fluid dynamics (CFD) simulations are used to model and forecast the aerodynamic behaviour of bridges under different wind conditions. These models incorporate several factors, such as wind directionality, wind speed, turbulence intensity, and the influence of nearby structures or topography. The results provide significant new insights into the loads and pressures that wind places on different bridge elements, such as decks, pylons, and connections. Following the determination of the wind loads, the structural response of bridges is assessed. By simulating their dynamic behavior under wind-induced forces, Finite Element Analysis (FEA) is used to model the bridge's component parts. This work contributes to the understanding of which areas are at risk of experiencing excessive stresses, vibrations, or oscillations due to wind excitations. Because the bridge has inherent modes and frequencies, the study considers both static and dynamic responses. Various strategies are examined to maximize the design of bridges to withstand wind. It is possible to alter the bridge's geometry, add aerodynamic components, add dampers or tuned mass dampers to lessen vibrations, and boost structural rigidity. Through an analysis of several design modifications and their effectiveness, the study aims to offer guidelines and recommendations for wind-resistant bridge design. In addition to the numerical simulations and analyses, there are experimental studies. In order to assess the computational models and validate the practicality of proposed design strategies, scaled bridge models are tested in a wind tunnel. These investigations help to improve numerical models and prediction precision by providing valuable information on wind-induced forces, pressures, and flow patterns. Using a combination of numerical models, actual testing, and long-term performance evaluation, the project aims to offer practical insights and recommendations for building wind-resistant bridges that are secure, long-lasting, and comfortable for users.

Keywords: wind effects, aerodynamic forces, computational fluid dynamics, finite element analysis

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Authors: Islam Zeb

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Water of poor quality has a potential of probable contamination and a way to spread pollutant in the field and surrounding environment. A number of comprehensive reviews articles have been published which highlight irrigation water as a source of pathogenic microorganisms and heavy metals toxicity that leads to chronic diseases in human. Here a study was plan to determine the microbial status of irrigation water collected from various location of district Swat in various months. The analyses were carried out at Environmental Horticulture Laboratory, Department of Horticulture, The University of Agriculture Peshawar, during the year 2018 – 19. The experiment was laid out in Randomized Complete Block Design (RCBD) with two factors and three replicates. Factor A consist of different locations, and factor B represent various months. The results of microbial status for various locations in irrigation water showed the highest value for Total Bacterial Count, Enterobacteriacea, E. coli, Salmonella, and Listeria (9.05, 8.54, 6.01, 5.84, and 5.03 log cfu L-1 respectively) for samples collected from mingora location, whereas the lowest values for Total Bacterial Count, Enterobacteriacea, E. coli, Salmonella and Listeria (6.70, 6.38, 4.47, 4.42 and 3.77 log cfu L-1 respectively) were observed for matta location. Data for various months showed maximum Total Bacterial Count, Enterobacteriacea, E. coli, Salmonella, and Listeria (12.01, 11.70, 8.46, 8.41, and 6.88 log cfu L-1, respectively) were noted for the irrigation water samples collected in May/June whereas the lowest range for Total Bacterial Count, Enterobacteriacea, E. coli, Salmonella and Listeria (4.41, 4.08, 2.61, 2.55 and 3.39 log cfu L-1 respectively) were observed in Jan/Feb. A significant interaction was found for all the studied parameters it was concluded that maximum bacterial groups were recorded in the months of May/June from Mingora location, it might be due to favorable weather condition.

Keywords: contamination, irrigation water, microbes, SWAT, various months

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Authors: Asmuddin Natsir, A. Mujnisa, Syahriani Syahrir, Marhamah Nadir, Nurul Purnomo

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Bacteria, protozoa, Archaea, and fungi are the dominant microorganisms found in the rumen ecosystem that has an important role in converting feed ingredients into components that can be digested and utilized by the livestock host. This study was conducted to assess the diversity of rumen bacteria of bali cattle raised under traditional farming condition. Three adult bali cattle were used in this experiment. The rumen fluid samples from the three experimental animals were obtained by the Stomach Tube method before the morning feeding. The results of study indicated that the Illumina sequencing was successful in identifying 301,589 sequences, averaging 100,533 sequences, from three rumen fluid samples of three cattle. Furthermore, based on the SILVA taxonomic database, there were 19 kinds of phyla that had been successfully identified. Of the 19 phyla, there were only two dominant groups across the three samples, namely Bacteroidetes and Firmicutes, with an average percentage of 83.68% and 13.43%, respectively. Other groups such as Synergistetes, Spirochaetae, Planctomycetes can also be identified but in relatively small percentage. At the genus level, there were 157 sequences obtained from all three samples. Of this number, the most dominant group was Prevotella 1 with a percentage of 71.82% followed by 6.94% of Christencenellaceae R-7 group. Other groups such as Prevotellaceae UCG-001, Ruminococcaceae NK4A214 group, Sphaerochaeta, Ruminococcus 2, Rikenellaceae RC9 gut group, Quinella were also identified but with very low percentages. The sequencing results were able to detect the presence of 3.06% and 3.92% respectively for uncultured rumen bacterium and uncultured bacterium. In conclusion, the results of this experiment can provide an opportunity for a better understanding of the rumen bacterial diversity of the bali cattle raised under traditional farming condition and insight regarding the uncultured rumen bacterium and uncultured bacterium that need to be further explored.

Keywords: 16S rRNA sequencing, bali cattle, rumen microbial diversity, uncultured rumen bacterium

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Authors: Naser Valipour Motlagh, Majid Aliabadi, Elnaz Rahmani, Samira Ghorbanpour

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Strawberry is one of the most favored fruits all along the world. But due to its vulnerability to microbial contamination and short life storage, there are lots of problems in industrial production and transportation of this fruit. Therefore, lots of ideas have tried to increase the storage life of strawberries especially through proper packaging. This paper works on efficient packaging as well. The primary material used is produced through simple mixing of low-density polyethylene (LDPE) and silver nanoparticles in different weight fractions of 0.5 and 1% in presence of dicumyl peroxide as a cross-linking agent. Final packages were made in a twin-screw extruder. Then, their effect on the quality maintenance of strawberry is evaluated. The SEM images of nano-silver packages show the distribution of silver nanoparticles in the packages. Total bacteria count, mold, yeast and E. coli are measured for microbial evaluation of all samples. Texture, color, appearance, odor, taste and total acceptance of various samples are evaluated by trained panelists and based on 9-point hedonic scale method. The results show a decrease in total bacteria count and mold in nano-silver packages compared to the samples packed in polyethylene packages for the same storage time. The optimum concentration of silver nanoparticles for the lowest bacteria count and mold is predicted to be around 0.5% which has attained the most acceptance from the panelist as well. Moreover, organoleptic properties of strawberry are preserved for a longer period in nano-silver packages. It can be concluded that using nano-silver particles in strawberry packages has improved the storage life and quality maintenance of the fruit.

Keywords: antimicrobial properties, polyethylene, silver nanoparticles, strawberry

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Authors: Idan Chiyanzu, Maruping Mangena

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Crude glycerol, a major by-product from the transesterification of triacylglycerides with alcohol to biodiesel, is known to have a broad range of applications. For example, its bioconversion can afford a wide range of chemicals including alcohols, organic acids, hydrogen, solvents and intermediate compounds. In bacteria, the 2-oxoglutarate dioxygenase (2-OGD) enzymes are widely found among the Pseudomonas syringae species and have been recognized with an emerging importance in ethylene formation. However, the use of optimized enzyme function in recombinant systems for crude glycerol conversion to ethylene is still not been reported. The present study investigated the production of ethylene from crude glycerol using engineered E. coli MG1655 and JM109 strains. Ethylene production with an optimized expression system for 2-OGD in E. coli using a codon optimized construct of the ethylene-forming gene was studied. The codon-optimization resulted in a 20-fold increase of protein production and thus an enhanced production of the ethylene gas. For a reliable bioreactor performance, the effect of temperature, fermentation time, pH, substrate concentration, the concentration of methanol, concentration of potassium hydroxide and media supplements on ethylene yield was investigated. The results demonstrate that the recombinant enzyme can be used for future studies to exploit the conversion of low-priced crude glycerol into advanced value products like light olefins, and tools including recombineering techniques for DNA, molecular biology, and bioengineering can be used to allowing unlimited the production of ethylene directly from the fermentation of crude glycerol. It can be concluded that recombinant E.coli production systems represent significantly secure, renewable and environmentally safe alternative to thermochemical approach to ethylene production.

Keywords: crude glycerol, bioethylene, recombinant E. coli, optimization

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Authors: Anoff Anim, Lila Mahmound, Maria Katsikogianni, Sanjit Nayak

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Sustained and controlled delivery of antimicrobial drugs have been largely studied recently using metal organic frameworks (MOFs)and different polymers. However, much attention has not been given to combining both MOFs and biodegradable polymers which would be a good strategy in providing a sustained gradual release of the drugs. Herein, we report a comparative study of the sustained and controlled release of widely used antibacterial drugs, cephalexin and metronidazole, from zinc-based MOF-5 incorporated in biodegradable polycaprolactone (PCL) and poly-lactic glycolic acid (PLGA) membranes. Cephalexin and metronidazole were separately incorporated in MOF-5 post-synthetically, followed by their integration into biodegradable PLGA and PCL membranes. The pristine MOF-5 and the loaded MOFs were thoroughly characterized by FT-IR, SEM, TGA and PXRD. Drug release studies were carried out to assess the release rate of the drugs in PBS and distilled water for up to 48 hours using UV-Vis Spectroscopy. Four bacterial strains from both the Gram-positive and Gram-negative types, Staphylococus aureus, Staphylococuss epidermidis, Escherichia coli, Acinetobacter baumanii, were tested against the pristine MOF, pure drugs, loaded MOFs and the drug-loaded MOF-polymer composites. Metronidazole-loaded MOF-5 composite of PLGA (PLGA-Met@MOF-5) was found to show highest efficiency to inhibit the growth of S. epidermidis compared to the other bacteria strains while maintaining a sustained minimum inhibitory concentration (MIC). This study demonstrates that the combination of biodegradable MOF-polymer composites can provide an efficient platform for sustained and controlled release of antimicrobial drugs, and can be a potential strategy to integrate them in biomedical devices.

Keywords: antimicrobial resistance, biodegradable polymers, cephalexin, drug release metronidazole, MOF-5, PCL, PLGA

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Authors: N. Valderrama, W. Albarracín, N. Algecira

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It was studied the effect of the inclusion of thyme and rosemary essential oils into chitosan films, as well as the microbiological and physical properties when storing chitosan film with and without the mentioned inclusion. The film forming solution was prepared by dissolving chitosan (2%, w/v), polysorbate 80 (4% w/w CH) and glycerol (16% w/w CH) in aqueous lactic acid solutions (control). The thyme (TEO) and rosemary (REO) essential oils (EOs) were included 1:1 w/w (EOs:CH) on their combination 50/50 (TEO:REO). The films were stored at temperatures of 5, 20, 33°C and a relative humidity of 75% during four weeks. The films with essential oil inclusion did not show an antimicrobial activity against strains. This behavior could be explained because the chitosan only inhibits the growth of microorganisms in direct contact with the active sites. However, the inhibition capacity of TEO was higher than the REO and a synergic effect between TEO:REO was found for S. enteritidis strains in the chitosan solution. Some physical properties were modified by the inclusion of essential oils. The addition of essential oils does not affect the mechanical properties (tensile strength, elongation at break, puncture deformation), the water solubility, the swelling index nor the DSC behavior. However, the essential oil inclusion can significantly decrease the thickness, the moisture content, and the L* value of films whereas the b* value increased due to molecular interactions between the polymeric matrix, the loosing of the structure, and the chemical modifications. On the other hand, the temperature and time of storage changed some physical properties on the chitosan films. This could have occurred because of chemical changes, such as swelling in the presence of high humidity air and the reacetylation of amino groups. In the majority of cases, properties such as moisture content, tensile strength, elongation at break, puncture deformation, a*, b*, chrome, ΔE increased whereas water resistance, swelling index, L*, and hue angle decreased.

Keywords: chitosan, food additives, modified films, polymers

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Authors: Soukaina Ounss, Hamid Mounir, Abdellatif El Marjani

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Sandwich materials with composite reinforced skins are mostly required in advanced construction applications with a view to ensure resistant structures. Their lightweight, their high flexural stiffness and their optimal thermal insulation make them a suitable solution to obtain efficient structures with performing rigidity and optimal energy safety. In this paper, the mechanical behavior of a sandwich beam with composite skins reinforced by unidirectional carbon fibers is investigated numerically through analyzing the impact of reinforcements specifications on the longitudinal elastic modulus in order to select the adequate sandwich configuration that has an interesting rigidity and an accurate convergence to the analytical approach which is proposed to verify performed numerical simulations. Therefore, concerned study starts by testing flexion performances of skins with various fibers orientations and volume fractions to determine those to use in sandwich beam. For that, the combination of a reinforcement inclination of 30° and a volume ratio of 60% is selected with the one with 60° of fibers orientation and 40% of volume fraction, this last guarantees to chosen skins an important rigidity with an optimal fibers concentration and a great enhance in convergence to analytical results in the sandwich model for the reason of the crucial core role as transverse shear absorber. Thus, a resistant sandwich beam is elaborated from a face-sheet constituted from two layers of previous skins with fibers oriented in 60° and an epoxy core; concerned beam has a longitudinal elastic modulus of 54 Gpa (gigapascal) that equals to the analytical value by a negligible error of 2%.

Keywords: fibers orientation, fibers volume ratio, longitudinal elastic modulus, sandwich beam

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Authors: N. G. Vepkhvadze, T. Enukidze

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Anthrax is a fatal disease caused by strains of Bacillus anthracis, a spore-forming gram-positive bacillus that causes the disease anthrax in animals and humans. Anthrax is a zoonotic disease that is also well-recognized as a potential agent of bioterrorism. Infection in humans is extremely rare in the developed world and is generally due to contact with infected animals or contaminated animal products. Testing of this zoonotic disease began in 1907 in Georgia and is still being tested routinely to provide accurate information and efficient testing results at the State Laboratory of Agriculture of Georgia. Each clinical sample is analyzed by RT-PCR and bacteriology methods; this study used Real-Time PCR assays for the detection of B. anthracis that rely on plasmid-encoded targets with a chromosomal marker to correctly differentiate pathogenic strains from non-anthracis Bacillus species. During the period of 2015-2022, the State Laboratory of Agriculture (SLA) tested 250 clinical and environmental (soil) samples from several different regions in Georgia. In total, 61 out of the 250 samples were positive during this period. Based on the results, Anthrax cases are mostly present in Eastern Georgia, with a high density of the population of livestock, specifically in the regions of Kakheti and Kvemo Kartli. All laboratory activities are being performed in accordance with International Quality standards, adhering to biosafety and biosecurity rules by qualified and experienced personnel handling pathogenic agents. Laboratory testing plays the largest role in diagnosing animals with anthrax, which helps pertinent institutions to quickly confirm a diagnosis of anthrax and evaluate the epidemiological situation that generates important data for further responses.

Keywords: animal disease, baccilus anthracis, edp, laboratory molecular diagnostics

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Authors: Obied A. Alwan, Elgerbi, M. Ali

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This study was conducted on the cow milk which is used in the local milk factories of Zawia. This was completely random sampling the unscheduled samples. The microbiologic result have approved that the count of bacteria and the count of E.Coli are very high and all the manufacturing places which were included in the study have lacked the health conditions.

Keywords: raw milk, dairy factories, Libya, microbiologic

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Authors: Hidayah Triana, Mahir S. Gani, Asmi Citra Malina, Hamka

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This study was conducted to determine genetic diversity of tiger groupers that are resistant to V. parahaemolyticus and tolerant to low extreme salinities. This research is useful to obtain superior broodstock of fish. Tiger grouper used were 6 to 8 cm obtained from Brackish Water Aquaculture Research Center Gondol (Bali). This study consists of four stages: preliminary stage was adaptation of fish exposed to several concentrations of V. parahaemolyticus (103, 104, 105, 106, and 107 CFU / ml); second stage was test of Lethal Concentration (LC50) of bacteria to fish; third stage was salinity tolerance test (low salinity 12, 14 and 16 ppt) and fourth stage was analysis of DNA profiles. For DNA profiles analysis, genomic DNA of fish were extracted for PCR using primers YNZ-22 and UBC-122 and visualized by electrophoresis method. The results showed that Lethal concentration of bacteria (LC50) to fish was 1,56x106 CFU/ml. Furthermore, survival rate of groupers exposed with low salinities (12, 14, 16 ppt) survival rates were found to be 54,17 %, 66,67 % and 79,16 % respectively. Average of DNA fragment (5 fragments) generated from primer UBC-122 in the group of fish resistant to V.parahaemolyticus and tolerant to low salinities was similar to group of susceptible to low salinities. Primer YNZ-22 generated more diverse of DNA fragments (8,0 and 5,8 fragments) both in the group of fish tolerant and susceptible to low salinities compared to primer UBC-122 (5,0 fragments). Size of DNA 1.5 kb resulted from primer YNZ-22. Primer YNZ-22 generated 4 (50 %) and 3 (42,8 %) polymorfic fragments in the group of fish tolerant and susceptible to low salinities, respectively. Four (4) monomorfic fragments were found both in the group of fish tolerant and susceptible to low salinities. Primer UBC-122 generated 6 (85,7 %) and 9 (90,0 %) polymorfic fragments in the fish tolerant and susceptible to low salinities, respectively.

Keywords: genetic diversity, epinephelus fuscoguttatus, V. parahaemolyticus, PCR-RAPD, low extreme salinity

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Authors: Karla Sanchez-Ortiz, Yunuen Tapia-Torres, John Larsen, Felipe Garcia-Oliva

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Due to food demand production, the use of synthetic nitrogenous fertilizer has increased in agricultural soils to replace the N losses. Nevertheless, the intensive use of synthetic nitrogenous fertilizer in conventional agriculture negatively affects the soil and therefore the environment, so alternatives such as organic agriculture have been proposed for being more environmentally friendly. However, further research in soil is needed to see how agricultural management affects the dynamics of C and N. The objective of this research was to evaluate the C and N dynamics in the soil with three different agricultural management: an agricultural plot with intensive inorganic fertilization, a plot with semi-organic management and an agricultural plot with recent abandonment (2 years). For each plot, the soil C and N dynamics and the enzymatic activity of NAG and β-Glucosidase were characterized. Total C and N concentration of the plant biomass of each site was measured as well. Dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) was higher in abandoned plot, as well as this plot had higher total carbon (TC) and total nitrogen (TN), besides microbial N and microbial C. While the enzymatic activity of NAG and β-Glucosidase was greater in the agricultural plot with inorganic fertilization, as well as nitrate (NO₃) was higher in fertilized plot, in comparison with the other two plots. The aboveground biomass (AB) of maize in the plot with inorganic fertilization presented higher TC and TN concentrations than the maize AB growing in the semiorganic plot, but the C:N ratio was highest in the grass AB in the abandoned plot. The C:N ration in the maize grain was greater in the semi-organic agricultural plot. These results show that the plot under intensive agricultural management favors the loss of soil organic matter and N, degrading the dynamics of soil organic compounds, promoting its fertility depletion.

Keywords: mineralization, nitrogen cycle, soil degradation, soil nutrients

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Authors: Mina Ganji Morad, Maziar Azadisoleimanieh, Sina Ganji Morad

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A green roof is formed by green plants alive and has many positive impacts in the regional climatic, as well as indoor. Green roof system to prevent solar radiation plays a role in the cooling space. The cooling is done by reducing thermal fluctuations on the exterior of the roof and by increasing the roof heat capacity which cause to keep the space under the roof cool in the summer and heating rate increases during the winter. A roof garden is one of the recommended ways to reduce energy consumption in large cities. Despite the scale of the city green roofs have effective functions, such as beautiful view of city and decontaminating the urban landscape and reduce mental stress, and in an exchange of energy and heat from outside to inside spaces. This article is based on a review of 20 articles and 10 books and valid survey results on the positive effects of green roofs to prevent energy waste in the building. According to these publications, three of the conventional roof, green roof typical and green roof with certain administrative details (layers of glass) and the use of resistant plants and shrubs have been analyzed and compared their heat transfer. The results of these studies showed that one of the best green roof systems for mountainous climate is tree and shrub system that in addition to being resistant to climate change in mountainous regions, will benefit from the other advantages of green roof. Due to the severity of climate change in mountainous areas it is essential to prevent the waste of buildings heating and cooling energy. Proper climate design can greatly help to reduce energy.

Keywords: green roof, heat transfer, reducing energy consumption, mountainous areas, sustainable architecture

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Authors: Tanmaya Pradhan, K. Midhun, M. Joy Thomas

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Increasing the shelf life and improving the quality are important objectives for the success of packaged liquid food industry. One of the methods by which this can be achieved is by deactivating the micro-organisms present in the liquid food through pasteurization. Pasteurization is done by heating, but some serious disadvantages such as the reduction in food quality, flavour, taste, colour, etc. were observed because of heat treatment, which leads to the development of newer methods instead of pasteurization such as treatment using UV radiation, high pressure, nuclear irradiation, pulsed electric field, etc. In recent years the use of the pulsed electric field (PEF) for inactivation of the microbial content in the food is gaining popularity. PEF uses a very high electric field for a short time for the inactivation of microorganisms, for which we require a high voltage pulsed power source. Pulsed power sources used for PEF treatments are usually in the range of 5kV to 50kV. Different pulse shapes are used, such as exponentially decaying and square wave pulses. Exponentially decaying pulses are generated by high power switches with only turn-on capacity and, therefore, discharge the total energy stored in the capacitor bank. These pulses have a sudden onset and, therefore, a high rate of rising but have a very slow decay, which yields extra heat, which is ineffective in microbial inactivation. Square pulses can be produced by an incomplete discharge of a capacitor with the help of a switch having both on/off control or by using a pulse forming network. In this work, a pulsed power-based system is designed with the help of high voltage capacitors and solid-state switches (IGBT) for the inactivation of pathogenic micro-organism in liquid food such as fruit juices. The high voltage generator is based on the Marx generator topology, which can produce variable amplitude, frequency, and pulse width according to the requirements. Liquid food is treated in a chamber where pulsed electric field is produced between stainless steel electrodes using the pulsed output voltage of the supply. Preliminary bacterial inactivation tests were performed by subjecting orange juice inoculated with Escherichia Coli bacteria. With the help of the developed pulsed power source and the chamber, the inoculated orange has been PEF treated. The voltage was varied to get a peak electric field up to 15kV/cm. For a total treatment time of 200µs, a 30% reduction in the bacterial count has been observed. The detailed results and analysis will be presented in the final paper.

Keywords: Escherichia coli bacteria, high voltage generator, microbial inactivation, pulsed electric field, pulsed forming line, solid-state switch

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Authors: Ajuna B. Henry

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In recent decades, climate change has demanded biological pesticides; more Bt strains are being discovered worldwide, some containing novel insecticidal genes while others have been modified through molecular approaches for increased yield, toxicity, and wider host target. In this study, B. thuringiensis strain AH-2 (Bt-2) was isolated from the soil and tested for insecticidal activity against Aphis gossypii (Hemiptera: Aphididae) and Lepidoptera insect pests: fall webworm (Hyphantria cunea) and diamondback moth (Plutella xylostella). A commercial strain B. thuringiensis subsp. kurstaki (Btk), and a chemical pesticide, imidacloprid (for Hemiptera) and chlorantraniliprole (for Lepidoptera), were used as positive control and the same media (without bacterial inoculum) as a negative control. For aphidicidal activity, Bt-2 caused a mortality rate of 70.2%, 78.1% or 88.4% in third instar nymphs of A. gossypii (3N) at 10%, 25% or 50% culture concentrations, respectively. Moreover, Bt-2 was effectively produced in cost-effective (PB) supplemented with either glucose (PBG) or sucrose (PBS) and maintained high aphicidal efficacy with 3N mortality rates of 85.9%, 82.9% or 82.2% in TSB, PBG or PBS media, respectively at 50% culture concentration. Bt-2 also suppressed adult fecundity by 98.3% compared to only 65.8% suppression by Btk at similar concentrations but was slightly lower than chemical treatment, which caused 100% suppression. Partial purification of 60 – 80% (NH4)2SO4 fraction of Bt-2 aphicidal proteins purified on anion exchange (DEAE-FF) column revealed a 105 kDa aphicidal protein with LC50 = 55.0 ng/µℓ. For Lepidoptera pests, chemical pesticide, Bt-2, and Btk cultures, mortality of 86.7%, 60%, and 60% in 3rd instar larvae of P. xylostella, and 96.7%, 80.0%, and 93.3% in 6th instar larvae of H. cunea, after 72h of exposure. When the entomopathogenic strains were cultured in a cost-effective PBG or PBS, the insecticidal activity in all strains was not significantly different compared to the use of a commercial medium (TSB). Bt-2 caused a mortality rate of 60.0%, 63.3%, and 50.0% against P. xylostella larvae and 76.7%, 83.3%, and 73.3% against H. cunea when grown in TSB, PBG, and PBS media, respectively. Bt-2 (grown in cost-effective PBG medium) caused a dose-dependent toxicity of 26.7%, 40.0%, and 63.3% against P. xylostella and 46.7%, 53.3%, and 76.7% against H. cunea at 10%, 25% and 50% culture concentration, respectively. The partially purified Bt-2 insecticidal proteins fractions F1, F2, F3, and F4 (extracted at different ratios of organic solvent) caused low toxicity (50.0%, 40.0%, 36.7%, and 30.0%) against P. xylostella and relatively high toxicity (56.7%, 76.7%, 66.7%, and 63.3%) against H. cunea at 100 µg/g of artificial diets. SDS-PAGE analysis revealed that a128kDa protein is associated with toxicity of Bt-2. Our result demonstrates a medium and strong larvicidal activity of Bt-2 against P. xylostella and H. cunea, respectively. Moreover, Bt-2 could be potentially produced using a cost-effective PBG medium which makes it an effective alternative biocontrol strategy to reduce chemical pesticide application.

Keywords: biocontrol, insect pests, larvae/nymph mortality, cost-effective media, aphis gossypii, plutella xylostella, hyphantria cunea, bacillus thuringiensi

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