Search results for: bacterial leaf spot

Authors: Clara Franch de la Cal, Christopher J Morris, Michael McArthur

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Cystic fibrosis (CF) is an autosomal recessive genetic disorder characterized by abnormal transport of chloride and sodium across the lung epithelium, leading to thick and viscous secretions. Within which CF patients suffer from repeated bacterial pulmonary infections, with Pseudomonas aeru-ginosa (PA) eliciting the greatest inflammatory response, causing an irreversible loss of lung func-tion that determines morbidity and mortality. The cell wall of PA is a permeability barrier to many antibacterials and the rise of Mutli-Drug Resistant strains (MDR) is eroding the efficacy of the few remaining clinical options. In addition when PA infection becomes established it forms an antibi-otic-resistant biofilm, embedded in which are slow growing cells that are refractive to drug treat-ment. Making the development of new antibacterials a major challenge. This work describes the development of new type of nanoparticulate oligonucleotide antibacterial capable of tackling PA infections, including MDR strains. It is being developed to both block growth and prevent biofilm formation. These oligonucleotide therapeutics, Transcription Factor Decoys (TFD), act on novel genomic targets by capturing key regulatory proteins to block essential bacterial genes and defeat infection. They have been successfully transfected into a wide range of pathogenic bacteria, both in vitro and in vivo, using a proprietary delivery technology. The surfactant used self-assembles with TFD to form a nanoparticle stable in biological fluids, which protects the TFD from degradation and preferentially transfects prokaryotic membranes. Key challenges are to adapt the nanoparticle so it is active against PA in the context of biofilms and to formulate it for administration by inhalation. This would allow the drug to be delivered to the respiratory tract, thereby achieving drug concentrations sufficient to eradicate the pathogenic organisms at the site of infection.

Keywords: antibacterials, transcriptional factor decoys (TFDs), pseudomonas aeruginosa

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Authors: É. Vincze

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Demand on plant oils has increased to an enormous extent that is due to the change of human nutrition habits on the one hand, while on the other hand to the increase of raw material demand of some industrial sectors, just as to the increase of biofuel production. Besides the determining importance of sunflower in Hungary the production area, just as in part the average yield amount of rapeseed has increased among the produced oil crops. The variety/hybrid palette has changed significantly during the past decade. The available varieties’/hybrids’ palette has been extended to a significant extent. It is agreed that rapeseed production demands professionalism and local experience. Technological elements are successive; high yield amounts cannot be produced without system-based approach. The aim of the present work was to execute the complex study of one of the most critical production technology element of rapeseed production, that was sowing technology. Several sowing technology elements are studied in this research project that are the following: biological basis (the hybrid Arkaso is studied in this regard), sowing time (sowing time treatments were set so that they represent the wide period used in industrial practice: early, optimal and late sowing time) plant density (in this regard reaction of rare, optimal and too dense populations) were modelled. The multifactorial experimental system enables the single and complex evaluation of rapeseed sowing technology elements, just as their modelling using experimental result data. Yield quality and quantity have been determined as well in the present experiment, just as the interactions between these factors. The experiment was set up in four replications at the Látókép Plant Production Research Site of the University of Debrecen. Two different sowing times were sown in the first experimental year (2014), while three in the second (2015). Three different plant densities were set in both years: 200, 350 and 500 thousand plants ha-1. Uniform nutrient supply and a row spacing of 45 cm were applied. Winter wheat was used as pre-crop. Plant physiological measurements were executed in the populations of the Arkaso rapeseed hybrid that were: relative chlorophyll content analysis (SPAD) and leaf area index (LAI) measurement. Relative chlorophyll content (SPAD) and leaf area index (LAI) were monitored in 7 different measurement times.

Keywords: inner quality, plant density, rapeseed, sowing time

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Authors: Priya Tomar, Pallavi Mittal

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As industrialization is inevitable and progress with rapid acceleration, the need for innovative ways to get rid of waste has increased. Recent advancement in bioresource technology paves novel ideas for recycling of factory waste that has been polluting the agro-industry, soil and water bodies. Paper industries in India are in a considerable number, where molasses and impure alcohol are still being used as raw materials for manufacturing of paper. Paper mills based on nonconventional agro residues are being encouraged due to increased demand of paper and acute shortage of forest-based raw materials. The colouring body present in the wastewater from pulp and paper mill is organic in nature and is comprised of wood extractives, tannin, resins, synthetic dyes, lignin and its degradation products formed by the action of chlorine on lignin which imparts an offensive colour to the water. These mills use different chemical process for paper manufacturing due to which lignified chemicals are released into the environment. Therefore, the chemical oxygen demand (COD) of the emanating stream is quite high. This paper presents some new techniques that were developed for the efficiency of bioremediation on paper industry. A short introduction to paper industry and a variety of presently available methods of bioremediation on paper industry and different strategies are also discussed here. For solving the above problem, two bacterial strains (Pseudomonas aeruginosa and Bacillus subtilis) and their consortia (Pseudomonas aeruginosa and Bacillus subtilis) were utilized for the pulp and paper mill effluent. Pseudomonas aeruginosa and Bacillus subtilis named as T–1, T–2, T–3, T–4, T–5, T–6, for the decolourisation of paper industry effluent. The results indicated that a maximum colour reduction is (60.5%) achieved by Pseudomonas aeruginosa and COD reduction is (88.8%) achieved by Bacillus subtilis, maximum pH changes is (4.23) achieved by Pseudomonas aeruginosa, TSS reduction is (2.09 %) achieved by Bacillus subtilis, and TDS reduction is (0.95 %) achieved by Bacillus subtilis. When the wastewater was supplemented with carbon (glucose) and nitrogen (yeast extract) source and data revealed the efficiency of Bacillus subtilis, having more with glucose than Pseudomonas aeruginosa.

Keywords: bioremediation, paper and pulp mill effluent, treated effluent, lignin

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Authors: Karima Oldyerou, B. Meddah, A. Tirtouil

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The food borne infections have a significant impact on public health. Salmonella is the first bacterial cause, especially because of its general availability in the intestinal tract of poultry, pigs and cattle. This bacteria and essential oil of Laurus nobilis subject in this article. In vitro evaluation of the antibacterial activity shows a sensitivity of Salmonella spp. with a MIC of 2.5 mg.ml -1 in vivo after infection of wistar rats and administered orally this essential oil, microbiological results fecal material shows the antibacterial effect of this oil on Salmonella spp.

Keywords: Laurus nobilis, essential oil, salmonella, antibacterial activity, fecal matte

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Authors: Ibrahim M. S. Shnawa, Naeem R. R. Algebory

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The procedure for the assessment of the urinary mucosal cryoglobulin (UMCG) is being reviewed, testified and evaluated. The major features of UMCG are rather similar to that of serum cryoglobulin. Such evident similarities are forming the reality for the existence of the UMCG. There were seven characterizing criteria useable for the identification for UMCG. Upon matching them to the Irish criteria for serum cryoglobulin, some modifications are being proposed to the 16th standards that has been formulated and built as an Irish criterion. The existence of UMCG is being reported for the first time in human chronic infectious bacterial disease.

Keywords: urinary, mucosal, cryoglubulin, standard immunofixation

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Authors: Sumeet Rai, Anuj Tyagi, B. T. Naveen Kumar, Shubhkaramjeet Kaur, Niraj K. Singh

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Since their discovery, indiscriminate use of antibiotics in human, veterinary and aquaculture systems has resulted in global emergence/spread of multidrug-resistant bacterial pathogens. Thus, the need for alternative approaches to control bacterial infections has become utmost important. High selectivity/specificity of bacteriophages (phages) permits the targeting of specific bacteria without affecting the desirable flora. In this study, a lytic phage (Ahp1) specific to Aeromonas hydrophila subsp. hydrophila was isolated from finfish aquaculture pond. The host range of Ahp1 range was tested against 10 isolates of A. hydrophila, 7 isolates of A. veronii, 25 Vibrio cholerae isolates, 4 V. parahaemolyticus isolates and one isolate each of V. harveyi and Salmonella enterica collected previously. Except the host A. hydrophila subsp. hydrophila strain, no lytic activity against any other bacterial was detected. During the adsorption rate and one-step growth curve analysis, 69.7% of phage particles were able to get adsorbed on host cell followed by the release of 93 ± 6 phage progenies per host cell after a latent period of ~30 min. Phage nucleic acid was extracted by column purification methods. After determining the nature of phage nucleic acid as dsDNA, phage genome was subjected to next-generation sequencing by generating paired-end (PE, 2 x 300bp) reads on Illumina MiSeq system. De novo assembly of sequencing reads generated circular phage genome of 42,439 bp with G+C content of 58.95%. During open read frame (ORF) prediction and annotation, 22 ORFs (out of 49 total predicted ORFs) were functionally annotated and rest encoded for hypothetical proteins. Proteins involved in major functions such as phage structure formation and packaging, DNA replication and repair, DNA transcription and host cell lysis were encoded by the phage genome. The complete genome sequence of Ahp1 along with gene annotation was submitted to NCBI GenBank (accession number MF683623). Stability of Ahp1 preparations at storage temperatures of 4 °C, 30 °C, and 40 °C was studied over a period of 9 months. At 40 °C storage, phage counts declined by 4 log units within one month; with a total loss of viability after 2 months. At 30 °C temperature, phage preparation was stable for < 5 months. On the other hand, phage counts decreased by only 2 log units over a period of 9 during storage at 4 °C. As some of the phages have also been reported as glycerol sensitive, the stability of Ahp1 preparations in (0%, 15%, 30% and 45%) glycerol stocks were also studied during storage at -80 °C over a period of 9 months. The phage counts decreased only by 2 log units during storage, and no significant difference in phage counts was observed at different concentrations of glycerol. The Ahp1 phage discovered in our study had a very narrow host range and it may be useful for phage typing applications. Moreover, the endolysin and holin genes in Ahp1 genome could be ideal candidates for recombinant cloning and expression of antimicrobial proteins.

Keywords: Aeromonas hydrophila, endolysin, phage, narrow host range

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Authors: Natesan Balasubramanian, Palpandi Pounpandi, Venkatraman Thamil Priya, Vellasamy Shanmugaiah, Karubbiah Balakrishnan, Mandayam Anandam Thirunarayan

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Streptococcus agalactiae is commonly known as Group B Streptococcus (GBS) and it is the most common cause of life-threatening bacterial infection. GBS first considered as a veterinary pathogen causing mastitis in cattle later becomes a human pathogen for severe neonatal infections. In this present study, a total of 20 new clinical isolates of S. agalactiae were collected from male (6) and female patient (14) with different age group. The isolates were from Urinary tract infection (UTI), blood, pus and eye ulcer. All the 20 S. agalactiae isolates has clear hemolysis properties on blood agar medium and were identified by serogrouping and MALTI-TOF-MS analysis. Antibiotic susceptibility/resistance test was performed for 20 S. agalactiae isolates, further phenotypic resistance pattern was observed for tetracycline, vancomycin, ampicillin and penicillin. Genotypically we found two antibiotic resistance genes such as Betalactem antibiotic resistance gene (Tem) (70%) and tetracycline resistance gene Tet(O) 15% in our isolates. Six virulence factors encoding genes were performed by PCR in twenty GBS isolates, cfb gene (100%), followed by, cylE(90.47%), lmp(85.7%), bca(71.42%), rib (38%) and low frequency in bac gene (4.76%) were determined. Most of the S. agalactiae isolates produced strong biofilm in the polystyrene surface (hydrophobic), and low-level biofilm formation was found in glass tube (hydrophilic) surface. lytR is secreted protein and localized in bacterial cell wall, extra cellular membrane, and cytoplasm. In silico docking studies were performed for lytR protein with four antibiofilm compounds, including a peptide (PR39) with the docking study showed peptide has strong interaction followed by ellagic acid and interaction length is 2.95, 2.97 and 2.95 A°. In ligand EGCGO10 and O11 two atoms intract with lytR (Leu271), with binding bond affinity length is 3.24 and 3.14. The aminoacid Leu 271 is act as an impartant aminoacid, since ellagic acid and EGCG interact with same aminoacid.

Keywords: antibiotics, biofilms, clinical isolates, S. agalactiae, virulence

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Authors: Zhala Ahmad, Zainab Lazim, Haider Hamzah

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Bacterial resistance is a pressing global health issue, with multidrug-resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR) strains to pose a serious threat. In this context, researchers are investigating effective, safe, and affordable metabolites to combat these pathogens. This study focuses on gum essential oil (GEO) extracted from Pistacia atlantica and its activity and the mechanism of action against XDR Gram-negative Acinetobacter baumannii. GEO was extracted by hydrodistillation and analyzed using GC-MS. Eleven A. baumannii isolates were collected from the ward environment of Burn and Plastic Surgery Hospital in Al Sulaymaniyah City, Iraq. They were identified using the VITEK 2 system, 16S rRNA gene, and confirmed with the blaₒₓₐ₋₅₁ gene; A. baumannii ATCC 19606 was used as a reference strain. The isolates were identified as resistant to twelve different antibiotics spanning six distinct antibiotic classes while showing susceptibility to tetracycline and trimethoprim. Over 40 chemical constituents were detected in the gum's essential oils, with α-pinene being the most abundant. GEO was found to inhibit the growth of A. baumannii isolates; the minimum inhibitory concentration (MIC) of GEO was 2.5 µl/ml. GEO induced protein leakage, phosphate, and potassium ion efflux, distorted cell morphology, and cell death in the tested bacteria. GEO exhibited bacterial clearance and anti-adhesion activity using Band-Aids. This study's findings suggest that GEO could be used as a potential alternative treatment for infectious diseases caused by XRD pathogens, shedding further light on the importance of GEO in biomedical applications. Future studies must focus on generating clinically feasible sources of GEO for testing in small animal models before proceeding to human trials, ensuring safe and effective translation from the laboratory to the clinic.

Keywords: antibiotic resistance, Acinetobacter baumannii, essential oils, Pistacia atlantica, alpha-pinene

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Authors: Muhammad Adeel Arshad, Shaukat Ali Bhatti, Faiz-ul Hassan

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Manipulating microbial ecosystem in the rumen is considered as an important strategy to optimize production efficiency in ruminants. In the past, antibiotics and synthetic chemical compounds have been used for the manipulation of rumen fermentation. However, since the non-therapeutic use of antibiotics has been banned, efforts are being focused to search out safe alternative products. In tropics, crop residues and forage grazing are major dietary sources for ruminants. Poor digestibility and utilization of these feedstuffs by animals is a limiting factor to exploit the full potential of ruminants in this area. Hence, there is a need to enhance the utilization of these available feeding resources. One of the potential strategies in this regard is the use of direct-fed microbes. Bacteria and fungi are mostly used as direct-fed microbes to improve animal health and productivity. Commonly used bacterial species include lactic acid-producing and utilizing bacteria (Lactobacillus, Streptococcus, Enterococcus, Bifidobacterium, and Bacillus) and fungal species of yeast are Saccharomyces and Aspergillus. Direct-fed microbes modulate microbial balance in the gastrointestinal tract through the competitive exclusion of pathogenic species and favoring beneficial microbes. Improvement in weight gain and feed efficiency has been observed as a result of feeding direct-fed bacteria. The use of fungi as a direct-fed microbe may prevent excessive production of lactate and harmful oxygen in the rumen leading to better feed digestibility. However, the mechanistic mode of action for bacterial or fungal direct-fed microbes has not been established yet. Various reports have confirmed an increase in dry matter intake, milk yield, and milk contents in response to the administration of direct-fed microbes. However, the application of a direct-fed microbe has shown variable responses mainly attributed to dosages and strains of microbes. Nonetheless, it is concluded that the inclusion of direct-fed microbes may mediate the rumen ecosystem to manage lactic acid production and utilization in both clinical and sub-acute rumen acidosis.

Keywords: microbes, roughages, rumen, feed efficiency, production, fermentation

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Authors: Hamza A. Pantami, Khozizah Shaari, Intan S. Ismail, Chong C. Min

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Introduction: Tilapia is the second-highest harvested freshwater fish species in Malaysia, available in almost all fish farms and markets. Unfortunately, tilapia culture in Malaysia is highly affected by Aeromonas hydrophila and Streptococcus agalactiae, which affect the production rate and consequently pose a direct negative economic impact. Reliance on drugs to control or reduce bacterial infections has been led to contamination of water bodies and development of drug resistance, as well as gave rise to toxicity issues in downstream fish products. Resorting to vaccines have helped curb the problem to a certain extent, but a more effective solution is still required. Using microalgae-based feed to enhance the fish immunity against bacterial infection offers a promising alternative. Objectives: This study aims to evaluate the efficacy of Chlorella vulgaris at lower percentage incorporation in feeds for an immune boost of tilapia in a shorter time. Methods: The study was in two phases. The safety concentration studies at 500 mg/kg-1 and the administration of cultured C. vulgaris biomass via incorporation into fish feed for five different groups in three weeks. Group 1 was the control (0% incorporation), whereas group 2, 3, 4 and 5 received 0.625%, 1.25%, 2.5% and 5% incorporation respectively. The parameters evaluated were the blood profile, serum lysozyme activity (SLA), serum bactericidal activity (SBA), phagocytosis activity (PA), respiratory burst activity (RBA), and lymphoproliferation activity (LPA). The data were analyzed via ANOVA using SPSS (version 16). Further testing was done using Tukey’s test. All tests were performed at the 95% confidence interval (p < 0.05). Results: There were no toxic signs in tilapia fish at 500 mg/kg-1. Treated groups showed significantly better immune parameters compared to the control group (p < 0.05). Conclusions: C. vulgaris crude biomass in a fish meal at a lower incorporation level of 5% can increase specific and non-specific immunity in tilapia fish in a shorter time duration.

Keywords: Chlorella vulgaris, hematology profile, immune boost, lymphoproliferation

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Authors: Isaac Martin, Abigail Lark, Sandra Morales, Eric W. Alton, Jane C. Davies

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Objectives: The cystic fibrosis (CF) lung is a unique microbiological niche, wherein harmful bacteria persist for many years despite antibiotic therapy. Pseudomonas aeruginosa (Pa), the major culprit leading to lung decline and increased mortality, thrives in the lungs of patients with CF due to several factors that have been linked with poor antibiotic performance. Our group is investigating alternative therapies including bacteriophage cocktails with which we have previously demonstrated efficacy against planktonic organisms. In this study, we explored the effects of a 4-phage cocktail on Pa grown in two different conditions, intended to mirror the CF lung: a) alongside standard antibiotic treatment in pre-formed biofilms (structures formed by Pa-secreted exopolysaccharides which provide both physical and cell division barriers to antimicrobials and host defenses and b) in an acidic environment postulated to be present in the CF airway due both to the primary defect in bicarbonate secretion and secondary effects of inflammation. Methods: 16 Pa strains from CF patients at the Royal Brompton Hospital were selected based on sensitivity to a) ceftazidime/ tobramycin and b) the phage cocktail in a conventional plaque assay. To assess efficacy of phage in biofilms, 96 well plates with Pa (5x10⁷ CFU/ ml) were incubated in static conditions, allowing adherent bacterial colonies to form for 24 hr. Ceftazidime and tobramycin (both at 2 × MIC) were added, +/- bacteriophage (4x10⁸ PFU/mL) for a further 24 hr. Cell viability and biomass were estimated using fluorescent resazurin and crystal violet assays, respectively. To evaluate the effect of pH, strains were grown planktonically in shaking 96 well plates at pH 6.0, 6.6, 7.0 and 7.5 with tobramycin or phage, at varying concentrations. Cell viability was quantified by fluorescent resazurin assay. Results: For the biofilm assay, treatment groups were compared with untreated controls and expressed as percent reduction in cell viability and biomass. Addition of the 4-phage cocktail resulted in a 1.3-fold reduction in cell viability and 1.7-fold reduction in biomass (p < 0.001) when compared to standard antibiotic treatment alone. Notably, there was a 50 ± 15% reduction in cell viability and 60 ± 12% reduction in biomass (95% CI) for the 4 biofilms demonstrating the most resistance to antibiotic treatment. 83% of strains tested (n=6) showed decreased bacterial killing by tobramycin at acidic pHs (p < 0.01). However, 25% of strains (n=12) showed improved phage killing at acidic pHs (p < 0.05), with none showing the pattern of reduced efficacy at acidic pH demonstrated by tobramycin. Conclusion: The 4-phage anti-Pa cocktail tested against Pa performs well in pre-formed biofilms and in acidic environments; two conditions intended to mimic the CF lung. To our knowledge, these are the first data looking at the effects of subtle pH changes on phage-mediated bacterial killing in the context of Pa infection. These findings contribute to a growing body of evidence supporting the use of nebulised lytic bacteriophage as a treatment in the context of lung infection.

Keywords: biofilm, cystic fibrosis, pH, Pseudomonas aeruginosa, lytic bacteriophage

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Authors: Lohendy Munoz-Vargas, Stephen O. Opiyo, Rose Digianantonio, Michele L. Williams, Asela Wijeratne, Gregory Habing

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Non-typhoidal Salmonella enterica is a zoonotic pathogen with critical importance in animal and public health. The persistence of Salmonella on farms affects animal productivity and health, and represents a risk for food safety. The intestinal microbiota plays a fundamental role in the colonization and invasion of this ubiquitous microorganism. To overcome the colonization resistance imparted by the gut microbiome, Salmonella uses invasion strategies and the host inflammatory response to survive, proliferate, and establish infections with diverse clinical manifestations. Cattle serve as reservoirs of Salmonella, and periparturient cows have high prevalence of Salmonella shedding; however, to author`s best knowledge, little is known about the association between the gut microbiome and the onset of Salmonella shedding during the periparturient period. Thus, the objective of this study was to assess the association between changes in bacterial communities and the onset of Salmonella shedding in cattle approaching parturition. In a prospective cohort study, fecal samples from 98 dairy cows originating from four different farms were collected at four time points relative to calving (-3 wks, -1 wk, +1 wk, +3 wks). All 392 samples were cultured for Salmonella. Sequencing of the V4 region of the 16S rRNA gene using the Illumina platform was completed to evaluate the fecal microbiome in a selected sample subset. Analyses of microbial composition, diversity, and structure were performed according to time points, farm, and Salmonella onset status. Individual cow fecal microbiomes, predominated by Bacteroidetes, Firmicutes, Spirochaetes, and Proteobacteria phyla, significantly changed before and after parturition. Microbial communities from different farms were distinguishable based on multivariate analysis. Although there were significant differences in some bacterial taxa between Salmonella positive and negative samples, our results did not identify differences in the fecal microbial diversity or structure for cows with and without the onset of Salmonella shedding. These data suggest that determinants other than the significant changes in the fecal microbiome influence the periparturient onset of Salmonella shedding in dairy cattle.

Keywords: dairy cattle, microbiome, periparturient, Salmonella

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Authors: Hadiatullah, T. S. D. Desak Ketut, A. A. Ayu, A. N. Isna, D. P. Ririn

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Spirogyra sp. is genus of microalgae which has a high carbohydrate content that used as a best medium for bacterial fermentation to produce cellulose. This study objective to determine the effect of pulp banana in the fermented paper production process using Spirogyra sp. and characterizing of the paper product. The method includes the production of bacterial cellulose, assay of the effect fermented paper interpolating with banana pulp using Spirogyra sp., and the assay of paper characteristics include gram-mage paper, water assay absorption, thickness, power assay of tensile resistance, assay of tear resistance, density, and organoleptic assay. Experiments were carried out with completely randomized design with a variation of the concentration of sewage treatment in the fermented paper production interpolating banana pulp using Spirogyra sp. Each parameter data to be analyzed by Anova variance that continued by real difference test with an error rate of 5% using the SPSS. Nata production results indicate that different carbon sources (glucose and sugar) did not show any significant differences from cellulose parameters assay. Significantly different results only indicated for the control treatment. Although not significantly different from the addition of a carbon source, sugar showed higher potency to produce high cellulose. Based on characteristic assay of the fermented paper showed that the paper gram-mage indicated that the control treatment without interpolation of a carbon source and a banana pulp have better result than banana pulp interpolation. Results of control gram-mage is 260 gsm that show optimized by cardboard. While on paper gram-mage produced with the banana pulp interpolation is about 120-200 gsm that show optimized by magazine paper and art paper. Based on the density, weight, water absorption assays, and organoleptic assay of paper showing the highest results in the treatment of pulp banana interpolation with sugar source as carbon is 14.28 g/m2, 0.02 g and 0.041 g/cm2.minutes. The conclusion found that paper with nata material interpolating with sugar and banana pulp has the potential formulation to produce super-quality paper.

Keywords: cellulose, fermentation, grammage, paper, Spirogyra sp.

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Authors: S. Mamoucha, J. Rahul, N. Christodoulakis

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Introduction: Nature has been the source of medicinal agents since the dawn of the human existence on Earth. Currently, millions of people, in the developing world, rely on medicinal plants for primary health care, income generation and lifespan improvement. In Greece, more than 5500 plant taxa are reported while about 250 of them are considered to be of great pharmaceutical importance. Among the plants used for medical purposes, Vitex agnus-castus L. (Verbenaceae) is known since ancient times. It is a small tree or shrub, widely distributed in the Mediterranean basin up to the Central Asia. It is also known as chaste tree or monks pepper. Theophrastus mentioned the shrub several times, as ‘agnos’ in his ‘Enquiry into Plants’. Dioscorides mentioned the use of V. agnus-castus for the stimulation of lactation in nursing mothers and the treatment of several female disorders. The plant has important medicinal properties and a long tradition in folk medicine as an antimicrobial, diuretic, digestive and insecticidal agent. Materials and methods: Leaves were cleaned, detached, fixed, sectioned and investigated with light and Scanning Electron Microscopy (SEM). Histochemical tests were executed as well. Specific histochemical reagents (osmium tetroxide, H2SO4, vanillin/HCl, antimony trichloride, Wagner’ s reagent, Dittmar’ s reagent, potassium bichromate, nitroso reaction, ferric chloride and di methoxy benzaldehyde) were used for the sub cellular localization of secondary metabolites. Results: Light microscopical investigations of the elongated leaves of V. agnus-castus revealed three layers of palisade parenchyma, just below the single layered adaxial epidermis. The spongy parenchyma is rather loose. Adaxial epidermal cells are larger in magnitude, compared to those of the abaxial epidermis. Four different types of capitate, secreting trichomes, were localized among the abaxial epidermal cells. Stomata were observed at the abaxial epidermis as well. SEM revealed the interesting arrangement of trichomes. Histochemical treatment on fresh and plastic embedded tissue sections revealed the nature and the sites of secondary metabolites accumulation (flavonoids, steroids, terpenes). Acknowledgment: This work was supported by IKY - State Scholarship Foundation, Athens, Greece.

Keywords: Vitex agnus-castus, leaf anatomy, histochemical reagents, secondary metabolites

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Authors: Fuad Abdullah Alatawi

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Microbial infections-based diseases are a significant public health issue around the world, mainly when antibiotic-resistant bacterium types evolve. In this research, we explored the anti-bacterial and anti-cancer potency of iron-oxide (Fe₂O₃) nanoparticles prepared from F. macrocarpa fruit extract. The chemical composition of F. macrocarpa fruit extract was used as a reducing and capping agent for nanoparticles’ synthesis was examined by GC-MS/MS analysis. Then, the prepared nanoparticles were confirmed by various biophysical techniques, including X-ray powder diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), UV-Vis Spectroscopy, and Transmission Electron Microscopy (TEM) and Energy Dispersive Spectroscopy (EDAX), and Dynamic Light Scattering (DLS). Also, the antioxidant capacity of fruit extract was determined through 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), Fluorescence Recovery After Photobleaching (FRAP), Superoxide Dismutase (SOD) assays. Furthermore, the cytotoxicity activities of Fe₂O₃ NPs were determined using the (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (MTT) test on MCF-7 cells. In the antibacterial assay, lethal doses of the Fe₂O₃NPs effectively inhibited the growth of gram-negative and gram-positive bacteria. The surface damage, ROS production, and protein leakage are the antibacterial mechanisms of Fe₂O₃NPs. Concerning antioxidant activity, the fruit extracts of F. macrocarpa had strong antioxidant properties, which were confirmed by DPPH, ABTS, FRAP, and SOD assays. In addition, the F. microcarpa-derived iron oxide nanomaterials greatly reduced the cell viability of (MCF-7). The GC-MS/MS analysis revealed the presence of 25 main bioactive compounds in the F. microcarpa extract. Overall, the finding of this research revealed that F. microcarpa-derived Fe₂O₃ nanoparticles could be employed as an alternative therapeutic agent to cure microbial infection and breast cancer in humans.

Keywords: ficus microcarpa, iron oxide, antibacterial activity, cytotoxicity

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Authors: A. F. C. Nassar, D. K. Tessler, L. Okuda, C. Del Fava, D. P. Chiebao, A. H. C. N. Romaldini, A. P. Alvim, M. J. Sanchez-Vazquez, M. S. Rosa, J. C. Pompei, R. Harakava, M. C. S. Araujo, G. H. F. Marques, E. M. Pituco

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Glanders is a zoonotic disease of Equidae caused by the bacterium Burkholderia mallei presented in acute or chronic clinical forms with inflammatory nodules in the respiratory tract, lymphangitis and caseous lymph nodes. There is not a treatment with veterinary drugs to this life-threatening disease; thus, its occurrence must be notified to official animal health services and any infected animal must be eliminated. This study aims to detect B. mallei from horses euthanized in outbreaks of glanders in Brazil, providing a better understanding of the bacterial characteristics and determine a proper protocol for isolation. The work was carried out with the collaboration of the Ministry of Agriculture and the Sao Paulo State Animal Health Department, while its procedures were approved by the Committee of Ethics in Animal Experimentation from the Instituto Biologico (CETEA n°156/2017). To the present time, 16 horses from farms with outbreaks of glanders detected by complement fixation test (CFT) serology method were analyzed. During the necropsy, samples of possibly affected organs (lymph nodes, lungs, heart, liver, spleen, kidneys and trachea) were collected for bacterial isolation, molecular tests and pathology. Isolation was performed using two enriched mediums, a potato infusion agar with 5% sheep blood, 4% glycerol and antibiotics (penicilin100U/ mL), and another with the same ingredients except the antibiotic. A PCR protocol was modified for this study using primers design to identify a region of the Flip gen of B. mallei. Thru isolation, 12.5% (2/16) animals were confirmed positive using only the enriched medium with antibiotic and confirmed by PCR: from mediastinal and submandibular lymph nodes and lungs in one animal and from mediastinal lymph node in the other. The detection of the bacterium using PCR showed positivity of 100% (16/16) horses from 144 samples of organs. Pathology macroscopic lesions observed were catarrhal nasal discharge, fetlock ulcers, emaciation, lymphangitis in limbs, suppurative lymphangitis, lymph node enlargement, star shaped liver, and spleen scars, adherence of the renal capsule, pulmonary hemorrhage, and miliary nodules. Microscopic lesions were suppurative bronchopneumonia with microabscesses and Langhans giant cells in lungs; lymph nodes with abscesses and intense lymphoid reaction; hemosiderosis and abscesses in spleen. Positive samples on PCR will be sequenced later and analyzed comparing with previous records in the literature. A throughout description of the recent acute cases of glanders occurring in Brazil and characterization of the bacterium related will contribute to advances in the knowledge of the pathogenicity, clinical symptoms, and epidemiology of this zoonotic disease. Acknowledgment: This project is sponsored by FAPESP.

Keywords: equines, bacterial isolation, zoonosis, PCR, pathology

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Authors: M. Honarpardaz, Z. Zhang, J. Derkx, A. Trangärd, J. Larsson

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Pilot stages are one of the most common positioners and regulators in industry. In this paper, we present two novel concepts for pilot stages with low power consumption to regulate a pneumatic device. Pilot 1, first concept, is designed based on a conventional frame core electro-magnetic actuator and a leaf spring to control the air flow and pilot 2 has an axisymmetric actuator and spring made of non-oriented electrical steel. Concepts are simulated in a system modeling tool to study their dynamic behavior. Both concepts are prototyped and tested. Experimental results are comprehensively analyzed and compared. The most promising concept that consumes less than 8 mW is highlighted and presented.

Keywords: electro-magnetic actuator, multidisciplinary system, low power consumption, pilot stage

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Authors: Pulcherie Matsodoum Nguemte, Adrien Wanko Ngnien, Guy Valerie Djumyom Wafo, Ives Magloire Kengne Noumsi, Pierre Francois Djocgoue

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The identification of plant species with the capacity to grow on hydrocarbon-polluted soils is an essential step for phytoremediation. In view of developing phytoremediation in Cameroon, floristic surveys have been conducted in 4 cities (Douala, Yaounde, Limbe, and Kribi). In each city, 13 hydrocarbon-polluted, as well as unpolluted sites (control), have been investigated using quadrat method. 106 species belonging to 76 genera and 30 families have been identified on hydrocarbon-polluted sites, unlike the control sites where floristic diversity was much higher (166 species contained in 125 genera and 50 families). Poaceae, Cyperaceae, Asteraceae and Amaranthaceae have higher taxonomic richness on polluted sites (16, 15,10 and 8 taxa, respectively). Shannon diversity index of the hydrocarbon-polluted sites (1.6 to 2.7 bits/ind.) were significantly lower than the control sites (2.7 to 3.2 bits/ind.). Based on a relative frequency > 10% and abundance > 7%, this study highlights more than ten plants predisposed to be effective in the cleaning-up attempts of soils contaminated by hydrocarbons. Based on the floristic indicators, 6 species (Eleusine indica (L.) Gaertn., Cynodon dactylon (L.) Pers., Alternanthera sessilis (L.) R. Br. ex DC †, Commelinpa benghalensis L., Cleome ciliata Schum. & Thonn. and Asystasia gangetica (L.) T. Anderson) were selected for a study to determine their capacity to remediate a soil contaminated with fuel oil (82.5 ml/ kg of soil). The experiments lasting 150 days takes into account three modalities - Tn: uncontaminated soils planted (6) To contaminated soils unplanted (3) and Tp: contaminated soil planted (18) – randomized arranged. 3 on 6 species (Eleusine indica, Cynodon dactylon, and Alternanthera sessilis) survived the climatic and soil conditions. E. indica presents a significantly higher growth rate for density and leaf area while C. dactylon had a significantly higher growth rate for stem size and leaf numbers. A. sessilis showed stunted growth and development throughout the experimental period. The species Eleusine indica (L.) Gaertn. and Cynodon dactylon (L.) Pers. can be qualified as polluo-tolerant plant species; polluo-tolerance being the ability of a species to survive and develop in the midst subject to extreme physical and chemical disturbances.

Keywords: Cameroon, cleaning-up, floristic surveys, phytoremediation

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Authors: Arojojoye Oluwatosin Adetola, Nwaechefu Olajumoke Olufunlayo, Ademola Adetokunbo Oyagbemi, Jeremiah Moyinoluwalogo Afolabi, Asaolu Racheal Oluwabukola

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The effects of man’s activities on the environment include depletion of natural resources alongside pollution of water bodies. Petroleum exploration in the Niger Delta region of Nigeria has compromised the aquatic environment with grave consequences on the entire ecosystem. In this study, we assessed the environmental safety of Araromi River, located in an oil-producing area in Ondo State, in the Niger Delta region of Nigeria by determining the levels of heavy metals (copper, cadmium, chromium, nickel, lead) and some biomarkers of oxidative stress (malondialdehyde, glutathione-S-transferase, glutathione peroxidase, catalase, superoxide dismutase, myeloperoxidase and reduced glutathione) in Clarias gariepinus (350-400g) from the river using standard methods. Clarias gariepinus from a clean fish farm in the same geographical location as the reference site (Ilesannmi fishery) was used as a control. Water samples from both sites were also analysed for some physicochemical parameters, heavy metals, and bacterial contamination. Our findings show a significant increase in malondialdehyde level (index of lipid peroxidation) as well as alterations in antioxidant status in the organs of Clarias gariepinus from Araromi River compared with control. A significant increase in bacterial contaminants, heavy metal pollutants, and particulate matter deposits were also observed in the water sample from Araromi River compared with control. In conclusion, high levels of indicators of environmental pollution observed in the water sample from Araromi River coupled with induction of oxidative stress in Clarias gariepinus from the river show that Araromi River is polluted; therefore, consumption of fishes and other aquatic organisms from the river may be unsafe for the people in that community.

Keywords: Araromi River, Clarias gariepinus, environmental pollution, heavy metals, oxidative stress

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Authors: Nadia Obaed, Lexi Frankel, Amalia Ardeljan, Denis Nigel, Anniki Witter, Omar Rashid

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Breast cancer is the most common cancer among women, with a lifetime risk of one in eight of all women in the United States. Although breast cancer is prevalent throughout the world, the uneven distribution in incidence and mortality rates is shaped by the variation in population structure, environment, genetics and known lifestyle risk factors. Furthermore, the bacterial profile in healthy and cancerous breast tissue differs with a higher relative abundance of bacteria capable of causing DNA damage in breast cancer patients. Previous bacterial infections may change the composition of the microbiome and partially account for the environmental factors promoting breast cancer. One study found that higher amounts of Staphylococcus, Bacillus, and Enterobacteriaceae, of which Escherichia coli (E. coli) is a part, were present in breast tumor tissue. Based on E. coli’s ability to damage DNA, it is hypothesized that there is an increased risk of breast cancer associated with previous E. coli infection. Therefore, the purpose of this study was to evaluate the correlation between E. coli infection and the incidence of breast cancer. Holy Cross Health, Fort Lauderdale, provided access to the Health Insurance Portability and Accountability (HIPAA) compliant national database for the purpose of academic research. International Classification of Disease 9th and 10th Codes (ICD-9, ICD-10) was then used to conduct a retrospective analysis using data from January 2010 to December 2019. All breast cancer diagnoses and all patients infected versus not infected with E. coli that underwent typical E. coli treatment were investigated. The obtained data were matched for age, Charlson Comorbidity Score (CCI score), and antibiotic treatment. Standard statistical methods were applied to determine statistical significance and an odds ratio was used to estimate the relative risk. A total of 81286 patients were identified and analyzed from the initial query and then reduced to 31894 antibiotic-specific treated patients in both the infected and control group, respectively. The incidence of breast cancer was 2.51% and present in 2043 patients in the E. coli group compared to 5.996% and present in 4874 patients in the control group. The incidence of breast cancer was 3.84% and present in 1223 patients in the treated E. coli group compared to 6.38% and present in 2034 patients in the treated control group. The decreased incidence of breast cancer in the E. coli and treated E. coli groups was statistically significant with a p-value of 2.2x10-16 and 2.264x10-16, respectively. The odds ratio in the E. coli and treated E. coli groups was 0.784 and 0.787 with a 95% confidence interval, respectively (0.756-0.813; 0.743-0.833). The current study shows a statistically significant decrease in breast cancer incidence in association with previous Escherichia coli infection. Researching the relationship between single bacterial species is important as only up to 10% of breast cancer risk is attributable to genetics, while the contribution of environmental factors including previous infections potentially accounts for a majority of the preventable risk. Further evaluation is recommended to assess the potential and mechanism of E. coli in decreasing the risk of breast cancer.

Keywords: breast cancer, escherichia coli, incidence, infection, microbiome, risk

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Authors: Suchithra V., Shreedhanya, Kavya Menon, Vidya Niranjan

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Skin metagenomics has a wide range of applications with direct relevance to the health of the organism. It gives us insight to the diverse community of microorganisms (the microbiome) harbored on the skin. In the recent years, it has become increasingly apparent that the interaction between skin microbiome and the human body plays a prominent role in immune system development, cancer development, disease pathology, and many other biological implications. Next Generation Sequencing has led to faster and better understanding of environmental organisms and their mutual interactions. This project is studying the human skin microbiome of different individuals having varied skin conditions. Bacterial 16S rRNA data of skin microbiome is downloaded from SRA toolkit provided by NCBI to perform metagenomics analysis. Twelve samples are selected with two controls, and 3 different categories, i.e., sex (male/female), skin type (moist/intermittently moist/sebaceous) and occlusion (occluded/intermittently occluded/exposed). Quality of the data is increased using Cutadapt, and its analysis is done using FastQC. USearch, a tool used to analyze an NGS data, provides a suitable platform to obtain taxonomy classification and abundance of bacteria from the metagenome data. The statistical tool used for analyzing the USearch result is METAGENassist. The results revealed that the top three abundant organisms found were: Prevotella, Corynebacterium, and Anaerococcus. Prevotella is known to be an infectious bacterium found on wound, tooth cavity, etc. Corynebacterium and Anaerococcus are opportunist bacteria responsible for skin odor. This result infers that Prevotella thrives easily in sebaceous skin conditions. Therefore it is better to undergo intermittently occluded treatment such as applying ointments, creams, etc. to treat wound for sebaceous skin type. Exposing the wound should be avoided as it leads to an increase in Prevotella abundance. Moist skin type individuals can opt for occluded or intermittently occluded treatment as they have shown to decrease the abundance of bacteria during treatment.

Keywords: bacterial 16S rRNA , next generation sequencing, skin metagenomics, skin microbiome, taxonomy

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Authors: David Sarpong, Waleed Khursheed, Ernest Danquah, Erin Murphy

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Shigella is a pathogenic bacterium responsible for shigellosis, a severe diarrheal disease that claims the lives of immunocompromised individuals worldwide. To develop therapeutics against this disease, an understanding of the molecular mechanisms underlying the pathogen’s physiology is crucial. Small non-coding RNAs (sRNAs) have emerged as important regulators of bacterial physiology, including as components of toxin-antitoxin systems. In this study, we investigated the role of RyfA in S. flexneri physiology and virulence. RyfA, originally identified as an sRNA in Escherichia coli, is conserved within the Enterobacteriaceae family, including Shigella. Whereas two copies of ryfA are present in S. dysenteriae, all other Shigella species contain only one copy of the gene. Additionally, we identified a putative open reading frame within the RyfA transcript, suggesting that it may be a dual-functioning gene encoding a small protein in addition to its sRNA function. To study ryfA in vitro, we cloned the gene into an inducible plasmid and observed the effect on bacterial growth. Here, we report that RyfA production inhibits the growth of S. flexneri, and this inhibition is dependent on the contained open reading frame. In-silico analyses have revealed the presence of two divergently transcribed sRNAs, RyfB1 and RyfB2, which share nucleotide complementarity with RyfA and thus are predicted to function as anti-toxins. Our data demonstrate that RyfB2 has a stronger antitoxin effect than RyfB1. This regulatory pattern suggests a novel form of a toxin-antitoxin system in which the activity of a single toxin is inhibited to varying degrees by two sRNA antitoxins. Studies are ongoing to investigate the regulatory mechanism(s) of the antitoxin genes, as well as the downstream targets and mechanism of growth inhibition by the RyfA toxin. This study offers distinct insights into the regulatory mechanisms underlying Shigella physiology and may inform the development of new anti-Shigella therapeutics.

Keywords: sRNA, shigella, toxin-antitoxin, Type 1 toxin antitoxin

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Authors: J. Adeppa, S. Samanta, O. K. Raina

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Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.

Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR

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Authors: Shih-Kuei Lin

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The characterization of the arbitrage-free dynamics of interest rates is developed in this study under the presence of Markov jump risks, when the term structure of the interest rates is modeled through simple forward rates. We consider Markov jump risks by allowing randomness in jump sizes, independence between jump sizes and jump times. The Markov jump diffusion model is used to capture empirical phenomena and to accurately describe interest jump risks in a financial market. We derive the arbitrage-free model of simple forward rates under the spot measure. Moreover, the analytical pricing formulas for a cap and a floor are derived under the forward measure when the jump size follows a lognormal distribution. In our empirical analysis, we find that the LIBOR market model with Markov jump risk better accounts for changes from/to different states and different rates.

Keywords: arbitrage-free, cap and floor, Markov jump diffusion model, simple forward rate model, volatility smile, EM algorithm

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Authors: Dagmara A. Niedziela, Mark P. Murphy, Orla M. Keane, Finola C. Leonard

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Staphylococcus aureus is the most frequent cause of clinical and subclinical bovine mastitis in Ireland. Mastitis caused by S. aureus is often chronic and tends to recur after antibiotic treatment. This may be due to several virulence factors, including attributes that enable the bacterium to internalize into bovine mammary epithelial cells, where it may evade antibiotic treatment, or evade the host immune response. Four bovine-adapted lineages (CC71, CC97, CC151 and ST136) were identified among a collection of Irish S. aureus mastitis isolates. Genotypic variation of mastitis-causing strains may contribute to different presentations of the disease, including differences in milk somatic cell count (SCC), the main method of mastitis detection. The objective of this study was to investigate the influence of bacterial strain and lineage on host immune response, by employing cell culture methods in vitro as well as an in vivo infection model. Twelve bovine adapted S. aureus strains were examined for internalization into bovine mammary epithelial cells (bMEC) and their ability to induce an immune response from bMEC (using qPCR and ELISA). In vitro studies found differences in a variety of virulence traits between the lineages. Strains from lineages CC97 and CC71 internalized more efficiently into bovine mammary epithelial cells (bMEC) than CC151 and ST136. CC97 strains also induced immune genes in bMEC more strongly than strains from the other 3 lineages. One strain each of CC151 and CC97 that differed in their ability to cause an immune response in bMEC were selected on the basis of the above in vitro experiments. Fourteen first-lactation Holstein-Friesian cows were purchased from 2 farms on the basis of low SCC (less than 50 000 cells/ml) and infection free status. Seven cows were infected with 1.73 x 102 c.f.u. of the CC97 strain (Group 1) and another seven with 5.83 x 102 c.f.u. of the CC151 strain (Group 2). The contralateral quarter of each cow was inoculated with PBS (vehicle). Clinical signs of infection (temperature, milk and udder appearance, milk yield) were monitored for 30 days. Blood and milk samples were taken to determine bacterial counts in milk, SCC, white blood cell populations and cytokines. Differences in disease presentation in vivo between groups were observed, with two animals from Group 2 developing clinical mastitis and requiring antibiotic treatment, while one animal from Group 1 did not develop an infection for the duration of the study. Fever (temperature > 39.5⁰C) was observed in 3 animals from Group 2 and in none from Group 1. Significant differences in SCC and bacterial load between groups were observed in the initial stages of infection (week 1). Data is also being collected on cytokines and chemokines secreted during the course of infection. The results of this study suggest that a strain from lineage CC151 may cause more severe clinical mastitis, while a strain from lineage CC97 may cause mild, subclinical mastitis. Diversity between strains of S. aureus may therefore influence the clinical presentation of mastitis, which in turn may influence disease detection and treatment needs.

Keywords: Bovine mastitis, host immune response, host-pathogen interactions, Staphylococcus aureus

Procedia PDF Downloads 157

Authors: Guna Raj Dhungana, Roshan Nepal, Apshara Parajuli, , Archana Maharjan, Shyam K. Mishra, Pramod Aryal, Rajani Malla

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Introduction: Klebsiella pneumoniae is a well-known opportunistic human pathogen, primarily causing healthcare-associated infections. The global emergence of carbapenemase-producing K. pneumoniaeis a major public health burden, which is often extensively multidrug resistant.Thus, because of the difficulty to treat these ‘superbug’ and menace and some term as ‘apocalypse’ of post antibiotics era, an alternative approach to controlling this pathogen is prudent and one of the approaches is phage mediated control and/or treatment. Objective: In this study, we aimed to isolate novel bacteriophage against carbapenemase-producing K. pneumoniaeand characterize for potential use inphage therapy. Material and Methods: Twenty lytic phages were isolated from river water using double layer agar assay and purified. Biological features, physiochemical characters, burst size, host specificity and activity spectrum of phages were determined. One most potent phage: Phage TU_Kle10O was selected and characterized by electron microscopy. Whole genome sequences of the phage were analyzed for presence/absence of virulent factors, and other lysin genes. Results: Novel phage TU_Kle10O showed multiple host range within own genus and did not induce any BIM up to 5th generation of host’s life cycle. Electron microscopy confirmed that the phage was tailed and belonged to Caudovirales family. Next generation sequencing revealed its genome to be 166.2 Kb. bioinformatical analysis further confirmed that the phage genome ‘did not’ contain any ‘bacterial genes’ within phage genome, which ruled out the concern for transfer of virulent genes. Specific 'lysin’ enzyme was identified phages which could be used as 'antibiotics'. Conclusion: Extensively multidrug resistant bacteria like carbapenemase-producing K. pneumoniaecould be treated efficiently by phages.Absence of ‘virulent’ genes of bacterial origin and presence of lysin proteins within phage genome makes phages an excellent candidate for therapeutics.

Keywords: bacteriophage, Klebsiella pneumoniae, MDR, phage therapy, carbapenemase,

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Authors: Haftay Abraha Tadesse, Dawit Gebreegziabiher Hagos, Atsebaha Gebrekidan Kahsay, Mahumd Abdulkader

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Background: Salmonella species and Escherichia coli (E. coli) are important foodborne pathogens affecting humans and animals. They are among the most important causes of infection that are associated with the consumption of contaminated food. This study was aimed to determine the prevalence, antimicrobial susceptibility patterns and associated risk factors for Salmonella species and E. coli in raw meat from butchery houses of Mekelle, Northern Ethiopia. Method: A cross-sectional study was conducted from January to December 2019. Socio-demographic data and risk factors were collected using a predesigned questionnaire. Meat samples were collected aseptically from the butchery houses and transported using icebox to Mekelle University, College of Veterinary Sciences for the isolation and identification of Salmonella species and E. coli. Antimicrobial susceptibility patterns were determined using Kirby disc diffusion method. Data obtained were cleaned and entered into Statistical Package for the Social Sciences version 22 and logistic regression models with odds ratio were calculated. P-value < 0.05 was considered as statistically significant. Results: A total of 153 out of 384 (39.8%) of the meat specimens were found to be contaminated. The contamination of Salmonella species and E. coli were 15.6% (n=60) and 20.8%) (n=80), respectively. Mixed contamination (Salmonella species and E. coli) was observed in 13 (3.4 %) of the analyzed. Poor washing hands regularly (AOR = 8.37; 95% CI: 2.75-25.50) and not using gloves during meat handling (AOR=11. 28; 95% CI:(4.69 27.10) were associated with overall bacterial contamination. About 100% of the tested isolates were sensitive to ciprofloxacin, gentamicin, Co trimoxazole , sulphamethoxazole, ceftriaxone, and trimethoprim and ciprofloxacin, gentamicin, and norfloxacine of E. coli and Salmonella species, respectively, while the resistance of amoxyclav_amoxicillin and erythromycin were both isolated bacteria species. The overall multidrug resistance pattern for Salmonella and E. coli were 51.4% (n=19) and 31.8% (14), respectively. Conclusion: Of the 153 (153/384) contaminated raw meat, 60 (15.6%) and 80 (20.8%) were contaminated by Salmonella species and E. coli, respectively. Poor handwashing practice and not using glove during meat handling showed a significant association with bacterial contamination. Multidrug-resistant showed in Salmonella species, and E. coli were 19 (51.4%) and 14 (31.8%), respectively.

Keywords: antimicrobial susceptibility test, butchery houses, E. coli, raw meat, salmonella species

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Authors: Manuela Amorim, Cláudia S. Marques, Maria Conceição Calhau, Hélder J. Pinheiro, Maria Manuela Pintado

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Recently it has been recognized peptides from different food sources with biological activities. However, no relevant study has proven the potential of brewer yeast peptides in the modulation of gut microbiota. The importance of human intestinal microbiota in maintaining host health is well known. Probiotics, prebiotics and the combination of these two components, can contribute to support an adequate balance of the bacterial population in the human large intestine. The survival of many bacterial species inhabiting the large bowel depends essentially on the substrates made available to them, most of which come directly from the diet. Some of these substrates can be selectively considered as prebiotics, which are food ingredients that can stimulate beneficial bacteria such as Lactobacilli or Bifidobacteria growth in the colon. Moreover, conventional food can be used as vehicle to intake bioactive compounds that provide those health benefits and increase people well-being. In this way, the main objective of this work was to study the potential prebiotic activity of brewer yeast peptide extract (BYP) obtained via hydrolysis of yeast proteins by cardosins present in Cynara cardunculus extract for possible use as a functional ingredient. To evaluate the effect of BYP on the modulation of gut microbiota in diet-induced obesity model, Wistar rats were fed either with a standard or a high-fat diet. Quantified via 16S ribosomal RNA (rRNA) expression by quantitative PCR (qPCR), genera of beneficial bacteria (Lactobacillus spp. and Bifidobacterium spp.) and three main phyla (Firmicutes, Bacteroidetes and Actinobacteria) were assessed. Results showed relative abundance of Lactobacillus spp., Bifidobacterium spp. and Bacteroidetes was significantly increased (P < 0.05) by BYP. Consequently, the potential health-promoting effects of WPE through modulation of gut microbiota were demonstrated in vivo. Altogether, these findings highlight the possible intervention of BYP as gut microbiota enhancer, promoting healthy life style, and the incorporation in new food products, leads them bringing associated benefits endorsing a new trend in the improvement of new value-added food products.

Keywords: functional ingredients, gut microbiota, prebiotics, brewer yeast peptide extract

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Authors: Diana M. Torres, Anngie K. Hernandez, Andrea Villareal, Magda R. Gomez, Sadao Kobayashi

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Colombian Garlic crops exhibited mild mosaic, yellow stripes, and deformation. This group of symptoms suggested a viral infection. Several viruses belonging to the genera Potyvirus, Carlavirus and Allexivirus are known to infect garlic and lower their yield worldwide, but in Colombia, there are no studies of viral infections in this crop, only leek yellow stripe virus (LYSV) has been reported to our best knowledge. In Colombia, there are no management strategies for viral diseases in garlic because of the lack of information about viral infections on this crop, which is reflected in (i) high prevalence of viral related symptoms in garlic fields and (ii) high dispersal rate. For these reasons, the purpose of the present study was to evaluate the viral status of garlic in Colombia, which can represent a major threat on garlic yield and quality for this country 55 symptomatic leaf samples were collected for virus detection by RT-PCR and mechanical inoculation. Total RNA isolated from infected samples were subjected to RT-PCR with primers 1-OYDV-G/2-OYDV-G for Onion yellow dwarf virus (OYDV) (expected size 774pb), 1LYSV/2LYSV for LYSV (expected size 1000pb), SLV 7044/SLV 8004 for Shallot latent virus (SLV) (expected size 960pb), GCL-N30/GCL-C40 for Garlic common latent virus (GCLV) (expected size 481pb) and EF1F/EF1R for internal control (expected size 358pb). GCLV, SLV, and LYSV were detected in infected samples; in 95.6% of the analyzed samples was detected at least one of the viruses. GCLV and SLV were detected in single infection with low prevalence (9.3% and 7.4%, respectively). Garlic generally becomes coinfected with several types of viruses. Four viral complexes were identified: three double infection (64% of analyzed samples) and one triple infection (15%). The most frequent viral complex was SLV + GCLV infecting 48.1% of the samples. The other double complexes identified had a prevalence of 7% (GCLV + LYSV and SLV + LYSV) and 5.6% of the samples were free from these viruses. Mechanical transmission experiments were set up using leaf tissues of collected samples from infected fields, different test plants were assessed to know the host range, but it was restricted to C. quinoa, confirming the presence of detected viruses which have limited host range and were detected in C. quinoa by RT-PCR. The results of molecular and biological tests confirm the presence of SLV, LYSV, and GCLV; this is the first report of SLV and LYSV in garlic plants in Colombia, which can represent a serious threat for this crop in this country.

Keywords: SLV, GCLV, LYSV, leek yellow stripe virus, Allium sativum

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Authors: Deepali Lal, Sudha Summerwar, Jyoutsna Pandey

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The resistance of pesticide by the pest is an important matter of concern.The pesticide of plant origin having nontoxic biodegradable and environmentally friendly qualities. The frequent spraying of toxic chemicals is developing resistance to the pesticide. Leaf powder of the plants like Argimone mexicana and Calotropis procera is prepared, Different doses of these plant extracts are given to the Fourth in star stages of Helicoverpa armigera through feeding methods, to find their efficacy the experimental findings will be put under analysis using various parameters. The effect on paritrophic membrane is also studied.

Keywords: distillation plant, acetone, alcohol, pipette, castor leaves, grams pods, larvae of helicoverpa armigera, plant extract, vails, jars, cotton

Procedia PDF Downloads 319