Search results for: foodborne pathogens
132 Characterization of Fungal Endophytes in Leaves, Stems and Roots of African Yam Bean (Sphenostylis sternocarpa Hochst ex. A. Rich Harms)
Authors: Iyabode A. Kehinde, Joshua O. Oyekanmi, Jumoke T. Abimbola, Olajumoke E. Ayanda
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African yam bean (AYB), (Sphenostylis stenocarpa) is a leguminous crop that provides nutritionally rich seeds, tubers and leaves for human consumption. AYB potentials as an important food security crop is yet to be realized and thus classified as underutilized crop. Underutilization of the crop has been partly associated with scarce information on the incidence and characterization of fungal endophytes infecting vascular parts of AYB. Accurate and robust detection of these endophytic fungi is essential for diagnosis, modeling, surveillance and protection of germplasm (seed) health. This work aimed at isolating and identifying fungal endophytes associated with leaves, stems and roots of AYB in Ogun State, Nigeria. This study investigated both cultural and molecular properties of endophytic fungi in AYB for its characterization and diversity. Fungal endophytes were isolated and culturally identified. DNA extraction, PCR amplification using ITS primers and analyses of nucleotide sequences of ribosomal DNA fragments were conducted on selected isolates. BLAST analysis was conducted on consensus nucleotide sequences of 28 out of 30 isolates and results showed similar homology with genera of Rhizopus, Cunninghamella, Fusarium, Aspergillus, Penicillium, Alternaria, Diaporthe, Nigrospora, Purpureocillium, Corynespora, Magnaporthe, Macrophomina, Curvularia, Acrocalymma, Talaromyces and Simplicillium. Slight similarity was found with endophytes associated with soybean. Phylogenetic analysis by maximum likelihood method showed high diversity among the general. These organisms have high economic importance in crop improvement. For an instance, Purpureocillium lilacinum showed high potential in control of root rot caused by nematodes in tomatoes. Though some can be pathogens, but many of the fungal endophytes have beneficial attributes to plant in host health, uptake of nutrients, disease suppression, and host immunity.Keywords: molecular characterization, African Yam Bean, fungal endophyte, plant parts
Procedia PDF Downloads 213131 The Effects of Fungicide and Genetics on Fungal Diseases on Wheat in Nebraska With Emphasis on Stem Rust
Authors: Javed Sidiqi, Stephen Baezinger, Stephen Wegulo
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Wheat (Triticum aestivum L.) production continues to be challenged by foliar fungal diseases although significant improvement has been made to manage the diseases through developing resistant varieties and the fungicide use to ensure sufficient wheat is produced to meet the growing population’s need. Significant crop losses have been recorded in the history of grain production and yield losses due to fungal diseases, and the trend continues to threat food security in the world and particularly in the less developed countries. The impact of individual fungal diseases on grain yield has been studied extensively to determine crop losses. However, there is limited research available to find out the combined effects of fungal diseases on grain yield and the ways to effectively manage the diseases. Therefore, the objectives of this research were to study the effect of fungal pathogens on grain yield of pre-released winter wheat genotypes in fungicide treated and untreated plots, and to determine whether S7b gene was present in ‘Gage’ wheat as previously hypothesized. Sixty winter wheat genotypes in fungicide treated and untreated plots were studied across four environments. There was a significant effect of fungicide on grain yield consistently across four environments in three years. Fungicide treated wheat lines demonstrated (4,496 kg/ ha-1) grain yield compared to (3,147 kg/ ha-1) grain yield in untreated wheat lines indicating 43% increased grain yield due to severity of foliar fungal diseases. Furthermore, fungicide application also caused an increase in protein concentration from 153 (g kg-1) to 164 (g kg-1) in treated plots in along with test weight from 73 to 77 (kg hL-1) respectively. Gage wheat variety and ISr7b-Ra were crossed to determine presence of Sr7b in Gage. The F2 and F2:3 segregating families were screened and evaluated for stem rust resistance. The segregation of families fell within 15:1 ratio for two separate resistance genes suggesting that Sr7b segregates independently from an unknown resistance gene in Gage that needs to be characterized for its use in the future wheat breeding program to develop resistant wheat varieties.Keywords: funicide, genetics, foliar diseases, grain
Procedia PDF Downloads 126130 Peculiarities of Microflora of Odontogenic Inflammatory Processes in the Central Kazakhstan Region
Authors: Aliya Tokbergenova, Maida Tusupbekova, Daulet Dzhangaliyev, Alena Lavrinenko
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Background: Odontogenic phlegmons are ranked the first among pyoinflammatory processes in the frequency of hospitalization in maxillofacial surgery in the post-Soviet countries. The main role in etiology is played by obligate anaerobes and aerobes. According to numerous data, the structure of aerobic pathogens is dominated by staphylococci and gram-negative bacteria. Aim: The research aim is to study the microflora of the purulent discharge odontogenic inflammatory processes. Materials and methods: A total of 220 patients have been examined, of which 120 patients aged 25-59 years have been included in the research who did not have comorbidity hospitalized in the maxillofacial hospital in Karaganda (Kazakhstan) from January 2016 to July 2017. The bacteriological research has been carried out on the basis of the multiaccess laboratory of the KSMU, through the Matrix Assisted Laser Desorption/Ionization (MALDI) apparatus. The material sample was pus from the inflammation focus, taken during the operating period. Results: According to the research among 120 patients (100%), 15 patients (12.5%) have had microorganisms not grown. From 105 (87.5%) bacteriological results, it has been revealed the following 1) Streptococcus: 51 (42.5%): Streptococcus beta-haemolytic: 17 (14.2%), Streptococcus pneumoniae: 12 (10%), Streptococcus anginosus: 8 (6.6%), Streptococcus oralis: 8 (6.6%), Streptococcus constellatus: 6 (5.0%); 2) Staphylococci: 27 (22.5%): Staphylococci aureus: 14 (11.7%) and Staphylococci epidermidis: 13 (10.8%); 3) Pseudomonas aeruginosa: 12 (10%); 4) Neisseria: 11 (9.1%): Neisseria mucosa: 5 (4.1%) and Neisseria macacae: 6 (5.0%); 5) Klebsiella pneumoniae: 2 (1.7%); 6) Stenotrophomonas maltophilia: 2 (1.7%). 15 patients (12.5%) experienced complications in the form of 1) The dissemination of the process in 10 patients (8.4%). 2) Osteomyelitis in 3 (2.5%). 3) Mediastinitis in 1 (0.8%). 4) Sinusitis in 1 (0.8%). 15 patients (100%) were carried out repeated bacteriological examination, the following was revealed: 1) Streptococcus: 10 (66.7%): Streptococcus beta-haemolytic: 4 (26.7%), Streptococcus pneumoniae: 2 (13.3%), Streptococcus аnginosus: 2 (13.3%), Streptococcus oralis: 1 (6.7%), Streptococcus constellatus: 1 (6.7%); 2) Staphylococci: 4 (26.7%): Staphylococci aureus: 3 (20%) and Staphylococci epidermidis: 1 (6.7%); 3) Pseudomonas aeruginosa: 1 (6.7%). Conclusions: Thus, according to our research data, streptococci predominate in the odontogenic processes microflora in aerobic flora in the central Kazakhstan region, which refutes the leading role of staphylococci in the development of odontogenic inflammatory processes, thus creating prerequisites for studying new treatment approaches.Keywords: maxillofacial surgery, microflora, odontogenic phlegmons, pyo-inflammatory
Procedia PDF Downloads 192129 Bioinformatic Design of a Non-toxic Modified Adjuvant from the Native A1 Structure of Cholera Toxin with Membrane Synthetic Peptide of Naegleria fowleri
Authors: Frida Carrillo Morales, Maria Maricela Carrasco Yépez, Saúl Rojas Hernández
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Naegleria fowleri is the causative agent of primary amebic meningoencephalitis, this disease is acute and fulminant that affects humans. It has been reported that despite the existence of therapeutic options against this disease, its mortality rate is 97%. Therefore, the need arises to have vaccines that confer protection against this disease and, in addition to developing adjuvants to enhance the immune response. In this regard, in our work group, we obtained a peptide designed from the membrane protein MP2CL5 of Naegleria fowleri called Smp145 that was shown to be immunogenic; however, it would be of great importance to enhance its immunological response, being able to co-administer it with a non-toxic adjuvant. Therefore, the objective of this work was to carry out the bioinformatic design of a peptide of the Naegleria fowleri membrane protein MP2CL5 conjugated with a non-toxic modified adjuvant from the native A1 structure of Cholera Toxin. For which different bioinformatics tools were used to obtain a model with a modification in amino acid 61 of the A1 subunit of the CT (CTA1), to which the Smp145 peptide was added and both molecules were joined with a 13-glycine linker. As for the results obtained, the modification in CTA1 bound to the peptide produces a reduction in the toxicity of the molecule in in silico experiments, likewise, the prediction in the binding of Smp145 to the receptor of B cells suggests that the molecule is directed in specifically to the BCR receptor, decreasing its native enzymatic activity. The stereochemical evaluation showed that the generated model has a high number of adequately predicted residues. In the ERRAT test, the confidence with which it is possible to reject regions that exceed the error values was evaluated, in the generated model, a high score was obtained, which determines that the model has a good structural resolution. Therefore, the design of the conjugated peptide in this work will allow us to proceed with its chemical synthesis and subsequently be able to use it in the mouse meningitis protection model caused by N. fowleri.Keywords: immunology, vaccines, pathogens, infectious disease
Procedia PDF Downloads 92128 Prevalence of Urinary Tract Infections and Risk Factors among Pregnant Women Attending Ante Natal Clinics in Government Primary Health Care Centres in Akure
Authors: Adepeju Simon-Oke, Olatunji Odeyemi, Mobolanle Oniya
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Urinary tract infection has become the most common bacterial infections in humans, both at the community and hospital settings; it has been reported in all age groups and in both sexes. This study was carried out in order to determine and evaluate the prevalence, current drug susceptibility pattern of the isolated organisms and identify the associated risk factors of UTIs among the pregnant women in Akure, Ondo State, Nigeria. A cross-sectional study was conducted on the urine of pregnant women, and socio-demographic information of the women was collected. A total of 300 clean midstream urine samples were collected, and a general urine microscopic examination and culture were carried out, the Microbact identification system was used to identify gram-negative bacteria. Out of the 300 urine samples cultured, 183(61.0%) yielded significant growth of urinary pathogens while 117(39.0%) yielded either insignificant growth or no growth of any urinary pathogen. Prevalence of UTI was significantly associated with the type of toilet used, symptoms of UTI, and previous history of urinary tract infection (p<0.05). Escherichia coli 58(31.7%) was the dominant pathogen isolated, and the least isolated uropathogens were Citrobacter freudii and Providencia retgerri 2(1.1%) respectively. Gram-negative bacteria showed 77.6%, 67.9%, and 61.2% susceptibility to ciprofloxacin, augmentin, and chloramphenicol, respectively. Resistance against septrin, chloramphenicol, sparfloxacin, amoxicillin, augmentin, gentamycin, pefloxacin, trivid, and streptomycin was observed in the range of 23.1 to 70.1%. Gram-positive uropathogens isolated showed high resistance to amoxicillin (68.4%) and high susceptibility to the remaining nine antibiotics in the range 65.8% to 89.5%. This study justifies that pregnant women are at high risk of UTI. Therefore screening of pregnant women during antenatal clinics should be considered very important to avoid complications. Health education with regular antenatal and personal hygiene is recommended as precautionary measures to UTI.Keywords: pregnant women, prevalence, risk factor, UTIs
Procedia PDF Downloads 147127 Metagenomic Analysis and Pharmacokinetics of Phage Therapy in the Treatment of Bovine Subclinical Mastitis
Authors: Vaibhav D. Bhatt, Anju P. Kunjadia, D. S. Nauriyal, Bhumika J. Joshi, Chaitanya G. Joshi
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Metagenomic analysis of milk samples collected from local cattle breed, kankrej (Bos indicus), Gir (Bos indicus) and Crossbred (Bos indicus X Bos taurus) cattle harbouring subclinical mastitis was carried out by next-generation sequencing (NGS) 454 GS-FLX technology. Around 56 different species including members of Enterobacteriales, Pseudomonadales, Bacillales and Lactobacillales with varying abundance were detected in infected milk. The interesting presence of bacteriophages against Staphylococcus aureus, Escherichia coli, Enterobacter and Yersinia species were observed, especially Enterobacteria and E. coli phages (0∙32%) in Kankrej, Enterobacteria and Staphylococcus phages (1∙05%) in Gir and Staphylococcus phages (2∙32%) in crossbred cattle. NGS findings suggest that phages may be involved in imparting natural resistance of the cattle against pathogens. Further infected milk samples were subjected for bacterial isolation. Fourteen different isolates were identified, and DNA was extracted. Genes (Tet-K, Msr-A, and Mec-A) providing antibiotic resistance to the bacteria were screened by Polymerase Chain Reaction and results were validated with traditional antibiotic assay. Total 3 bacteriophages were isolated from nearby environment of the cattle farm. The efficacy of phages was checked against multi-drug resistant bacteria, identified by PCR. In-vivo study was carried out for phage therapy in mammary glands of female rats “Wister albino”. Mammary glands were infused with MDR isolates for 3 consecutive days. Recovery was observed in infected rats after intramammary infusion of sterile phage suspension. From day 4th onwards, level of C-reactive protein was significant increases up to day 12th . However, significant reduction was observed between days 12th to 18th post treatment. Bacteriophages have significant potential as antibacterial agents and their ability to replicate exponentially within their hosts and their specificity, make them ideal candidates for more sustainable mastitis control.Keywords: bacteriophages, c-reactive protein, mastitis, metagenomic analysis
Procedia PDF Downloads 315126 Impact of Urbanization Growth on Disease Spread and Outbreak Response: Exploring Strategies for Enhancing Resilience
Authors: Raquel Vianna Duarte Cardoso, Eduarda Lobato Faria, José Jorge Boueri
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Rapid urbanization has transformed the global landscape, presenting significant challenges to public health. This article delves into the impact of urbanization on the spread of infectious diseases in cities and identifies crucial strategies to enhance urban community resilience. Massive urbanization over recent decades has created conducive environments for the rapid spread of diseases due to population density, mobility, and unequal living conditions. Urbanization has been observed to increase exposure to pathogens and foster conditions conducive to disease outbreaks, including seasonal flu, vector-borne diseases, and respiratory infections. In order to tackle these issues, a range of cross-disciplinary approaches are suggested. These encompass the enhancement of urban healthcare infrastructure, emphasizing the need for robust investments in hospitals, clinics, and healthcare systems to keep pace with the burgeoning healthcare requirements in urban environments. Moreover, the establishment of disease monitoring and surveillance mechanisms is indispensable, as it allows for the timely detection of outbreaks, enabling swift responses. Additionally, community engagement and education play a pivotal role in advocating for personal hygiene, vaccination, and preventive measures, thus playing a pivotal role in diminishing disease transmission. Lastly, the promotion of sustainable urban planning, which includes the creation of cities with green spaces, access to clean water, and proper sanitation, can significantly mitigate the risks associated with waterborne and vector-borne diseases. The article is based on a review of scientific literature, and it offers a comprehensive insight into the complexities of the relationship between urbanization and health. It places a strong emphasis on the urgent need for integrated approaches to improve urban resilience in the face of health challenges.Keywords: infectious diseases dissemination, public health, urbanization impacts, urban resilience
Procedia PDF Downloads 77125 Cost-Effective and Optimal Control Analysis for Mitigation Strategy to Chocolate Spot Disease of Faba Bean
Authors: Haileyesus Tessema Alemneh, Abiyu Enyew Molla, Oluwole Daniel Makinde
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Introduction: Faba bean is one of the most important grown plants worldwide for humans and animals. Several biotic and abiotic elements have limited the output of faba beans, irrespective of their diverse significance. Many faba bean pathogens have been reported so far, of which the most important yield-limiting disease is chocolate spot disease (Botrytis fabae). The dynamics of disease transmission and decision-making processes for intervention programs for disease control are now better understood through the use of mathematical modeling. Currently, a lot of mathematical modeling researchers are interested in plant disease modeling. Objective: In this paper, a deterministic mathematical model for chocolate spot disease (CSD) on faba bean plant with an optimal control model was developed and analyzed to examine the best strategy for controlling CSD. Methodology: Three control interventions, quarantine (u2), chemical control (u3), and prevention (u1), are employed that would establish the optimal control model. The optimality system, characterization of controls, the adjoint variables, and the Hamiltonian are all generated employing Pontryagin’s maximum principle. A cost-effective approach is chosen from a set of possible integrated strategies using the incremental cost-effectiveness ratio (ICER). The forward-backward sweep iterative approach is used to run numerical simulations. Results: The Hamiltonian, the optimality system, the characterization of the controls, and the adjoint variables were established. The numerical results demonstrate that each integrated strategy can reduce the diseases within the specified period. However, due to limited resources, an integrated strategy of prevention and uprooting was found to be the best cost-effective strategy to combat CSD. Conclusion: Therefore, attention should be given to the integrated cost-effective and environmentally eco-friendly strategy by stakeholders and policymakers to control CSD and disseminate the integrated intervention to the farmers in order to fight the spread of CSD in the Faba bean population and produce the expected yield from the field.Keywords: CSD, optimal control theory, Pontryagin’s maximum principle, numerical simulation, cost-effectiveness analysis
Procedia PDF Downloads 86124 Characterization of a Broad Range Antimicrobial Substance from Pseudozyma aphidis
Authors: Raviv Harris, Maggie Levy
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Natural product-based pesticides may serve as an alternative to the traditional synthetic pesticides, which have a potentially damaging effect, both to human health and for the environment. Along with plants, microorganisms are a prospective source of such biological pesticides. A unique and active strain of P. aphidis (designated isolate L12, Israel 2004), an epiphytic and non-pathogenic basidiomycete yeast, was isolated in our lab from strawberry leaves. P. aphidis L12 secretions were found to inhibit broad range of plant pathogens. This work demonstrates that metabolites isolated from the biocontrol agent P. aphidis (isolate L12) can inhibit varied fungal and bacterial phytopathogens. Biologically active metabolites were extracted from P. aphidis biomass, using the organic solvent ethyl acetate. The antimicrobial activity of the extract was demonstrated, both in vitro and in planta. Using disk diffusion assays, the following inhibition zones were obtained: 43cm² for Pseudomonas syringae pv. tomato, 28.5cm² for Xanthomonas campestris pv. vesicatoria, 59cm² for Clavibacter michiganensis subsp. michiganensis, 34cm² for Erwinia amylovora and 34cm² for Agrobacterium tumefaciens. Additionally, strong inhibitory activity of the extract against fungi mycelial growth was established, with IC₅₀ values of 606µg ml⁻¹ for Botrytis cinerea, 221µg ml⁻¹ for Pythium spp., 519µg ml⁻¹ for Rhizoctonia solani, 455µg ml⁻¹ for Sclerotinia sclerotiorum, 2270µg ml⁻¹ for Fusarium oxysporum f. sp. lycopersici, and 2038µg ml⁻¹ for Alternaria alternata. The results of the in planta experiments demonstrated a dose-dependent reduction in disease infection. Significant inhibition of B. cinerea lesions on tomato plants was obtained when a spore suspension of this pathogen was treated with extract concentrations higher than 4.2mg ml⁻¹. Concentration of 7mg ml⁻¹ caused a reduction of over 95% in the lesion size of B. cinerea on tomato plants. The strong antimicrobial activity demonstrated both in vitro and in planta against varied phytopathogens, may indicate that the extracted antimicrobial metabolites have potential to serve as natural pesticides in the field.Keywords: antimicrobial, B. cinerea, metabolites, natural pesticides, P. aphidis
Procedia PDF Downloads 231123 Unzipping the Stress Response Genes in Moringa oleifera Lam. through Transcriptomics
Authors: Vivian A. Panes, Raymond John S. Rebong, Miel Q. Diaz
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Moringa oleifera Lam. is known mainly for its high nutritional value and medicinal properties contributing to its popular reputation as a 'miracle plant' in the tropical climates where it usually grows. The main objective of this study is to discover the genes and gene products involved in abiotic stress-induced activity that may impact the M. oleifera Lam. mature seeds as well as their corresponding functions. In this study, RNA-sequencing and de novo transcriptome assembly were performed using two assemblers, Trinity and Oases, which produced 177,417 and 120,818 contigs respectively. These transcripts were then subjected to various bioinformatics tools such as Blast2GO, UniProt, KEGG, and COG for gene annotation and the analysis of relevant metabolic pathways. Furthermore, FPKM analysis was performed to identify gene expression levels. The sequences were filtered according to the 'response to stress' GO term since this study dealt with stress response. Clustered Orthologous Groups (COG) showed that the highest frequencies of stress response gene functions were those of cytoskeleton which make up approximately 14% and 23% of stress-related sequences under Trinity and Oases respectively, recombination, repair and replication at 11% and 14% respectively, carbohydrate transport and metabolism at 23% and 9% respectively and defense mechanisms 16% and 12% respectively. KEGG pathway analysis determined the most abundant stress-response genes in the phenylpropanoid biosynthesis at counts of 187 and 166 pathways for Oases and Trinity respectively, purine metabolism at 123 and 230 pathways, and biosynthesis of antibiotics at 105 and 102. Unique and cumulative GO term counts revealed that majority of the stress response genes belonged to the category of cellular response to stress at cumulative counts of 1,487 to 2,187 for Oases and Trinity respectively, defense response at 754 and 1,255, and response to heat at 213 and 208, response to water deprivation at 229 and 228, and oxidative stress at 508 and 488. Lastly, FPKM was used to determine the levels of expression of each stress response gene. The most upregulated gene encodes for thiamine thiazole synthase chloroplastic-like enzyme which plays a significant role in DNA damage tolerance. Data analysis implies that M. oleifera stress response genes are directed towards the effects of climate change more than other stresses indicating the potential of M. oleifera for cultivation in harsh environments because it is resistant to climate change, pathogens, and foreign invaders.Keywords: stress response, genes, Moringa oleifera, transcriptomics
Procedia PDF Downloads 147122 Rates of Hematophagous Ectoparasite Consumption during Grooming by an Endemic Madagascar Fruit Bat
Authors: Riana V. Ramanantsalama, Aristide Andrianarimisa, Achille P. Raselimanana, Steven M. Goodman
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Few details are available on the consumption of ectoparasites, specifically bat flies (Diptera: Nycteribiidae and Streblidae), by their chiropteran hosts while grooming. Such details could provide information on the dynamics of host-parasite interactions. This study presents data on ectoparasite ingestion rates for an endemic Malagasy fruit bat (Pteropodidae: Rousettus madagascariensis) occupying a cave day roost colony in northern Madagascar. Using quantified behavioral analyses, grooming and associated ingestion rates were measured from infrared videos taken in close proximity to day-roosting bats. The recorded individual bats could be visually identified to age (adult, juvenile) and sex (male, female), allowing analyses of the proportion of time these different classes allocated to consuming ectoparasites via auto-grooming (self) or allo-grooming (intraspecific) per 10 min video recording session. These figures could then be extrapolated to estimates of individual daily consumption rates. Based on video recordings, adults spent significantly more time auto-grooming and allo-grooming than juveniles. The latter group was not observed consuming ectoparasites. Grooming rates and the average number of ectoparasites consumed per day did not differ between adult males and females. The mean extrapolated number consumed on a daily basis for individual adults was 37 ectoparasites. When these figures are overlaid on the estimated number of adult Rousettus occurring at the roost site during the dry season, the projected daily consumption rate was 57,905 ectoparasites. To the best knowledge of the authors of this study, the details presented here represent the first quantified data on bat consumption rates of their ectoparasites, specifically dipterans. These results provide new insights into host-parasite predation dynamics. More research is needed to explore the mechanism zoonotic diseases isolated from bat flies might be transmitted to their bat hosts, specifically those pathogens that can be communicated via an oral route.Keywords: diptera, host-parasite interactions, Madagascar, nycteribiidae, pteropodidae, Rousettus madagascariensis
Procedia PDF Downloads 145121 Unravelling of the TOR Signaling Pathway in Human Fungal Pathogen Cryptococcus neoformans
Authors: Yee-Seul So, Guiseppe Ianiri, Alex Idnurm, Yong-Sun Bahn
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Tor1 is a serine/threonine protein kinase that is widely conserved across eukaryotic species. Tor1 was first identified in Saccharomyces cerevisiae as a target of rapamycin (TOR). The TOR pathway has been implicated in regulating cellular responses to nutrients, proliferation, translation, transcription, autophagy, and ribosome biogenesis. Here we identified two homologues of S. cerevisiae Tor proteins, CNAG_06642 (Tor1) and CNAG_05220 (Tlk1, TOR-like kinase 1), in Cryptococcus neoformans causing a life-threatening fungal meningoencephalitis. Both Tor1 and Tlk1 have rapamycin-binding (RB) domains but Tlk1 has truncated RB form. To study the TOR-signaling pathway in the fungal pathogen, we attempt to construct the tor1Δ and tlk1Δ mutants and phenotypically analyze them. Although we failed to construct the tor1Δ mutant, we successfully construct the tlk1Δ mutant. The tlk1Δ mutant does not exhibit any discernable phenotypes, suggesting that Tlk1 is dispensable in C. neoformans. The essentiality of TOR1 is independently confirmed by constructing the TOR1 promoter replacement strain by using a copper transporter 4 (CTR4) promoter and the TOR1/tor1 heterozygous mutant in diploid C. neoformans strain background followed by sporulation analysis. To further analyze the function of Tor1, we construct TOR1 overexpression mutant using a constitutively active histone H3 in C. neoformans. We find that the Tor1 overexpression mutant is resistant to rapamycin but the tlk1Δ mutant does not exhibit any altered resistance to rapamycin, further confirming that Tor1, but not Tlk1, is critical for TOR signaling. Furthermore, we found that Tor1 is involved in response to diverse stresses, including genotoxic stress, oxidative stress, thermo-stress, antifungal drug treatment, and production of melanin. To identify any TOR-related transcription factors, we screened C. neoformans transcription factor library that we constructed in our previous study and identified several potential downstream factors of Tor1, including Atf1, Crg1 and Bzp3. In conclusion, the current study provides insight into the role of the TOR signaling pathway in human fungal pathogens as well as C. neoformans.Keywords: fungal pathogen, serine/threonine kinase, target of rapamycin, transcription factor
Procedia PDF Downloads 221120 A Hedonic Valuation Approach to Valuing Combined Sewer Overflow Reductions
Authors: Matt S. Van Deren, Michael Papenfus
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Seattle is one of the hundreds of cities in the United States that relies on a combined sewer system to collect and convey municipal wastewater. By design, these systems convey all wastewater, including industrial and commercial wastewater, human sewage, and stormwater runoff, through a single network of pipes. Serious problems arise for combined sewer systems during heavy precipitation events when treatment plants and storage facilities are unable to accommodate the influx of wastewater needing treatment, causing the sewer system to overflow into local waterways through sewer outfalls. CSOs (Combined Sewer Overflows) pose a serious threat to human and environmental health. Principal pollutants found in CSO discharge include microbial pathogens, comprising of bacteria, viruses, parasites, oxygen-depleting substances, suspended solids, chemicals or chemical mixtures, and excess nutrients, primarily nitrogen and phosphorus. While concentrations of these pollutants can vary between overflow events, CSOs have the potential to spread disease and waterborne illnesses, contaminate drinking water supplies, disrupt aquatic life, and effect a waterbody’s designated use. This paper estimates the economic impact of CSOs on residential property values. Using residential property sales data from Seattle, Washington, this paper employs a hedonic valuation model that controls for housing and neighborhood characteristics, as well as spatial and temporal effects, to predict a consumer’s willingness to pay for improved water quality near their homes. Initial results indicate that a 100,000-gallon decrease in the average annual overflow discharged from a sewer outfall within 300 meters of a home is associated with a 0.053% increase in the property’s sale price. For the average home in the sample, the price increase is estimated to be $18,860.23. These findings reveal some of the important economic benefits of improving water quality by reducing the frequency and severity of combined sewer overflows.Keywords: benefits, hedonic, Seattle, sewer
Procedia PDF Downloads 177119 Sialic Acid Profile and Sialidase Activity in HIV-Infected Individuals
Authors: Hadiza Abdullahi
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Sialic Acids and sialidases have been implicated in many disease states particularly bacterial and viral infections which are common opportunist infections of HIV disease. Their role in HIV/AIDS is contemplated. A study was carried out to determine Sialic Acid profile and Sialidase Activity in HIV infected and Apparently Healthy individuals, and also determine the relationship between the sialic acid levels and sialidase activity. Blood samples were collected from 200 subjects (150 HIV infected individuals and 50 apparently healthy individuals divided into four groups- HIV ART Naïve, HIV Stable (on ART but have been stable with no clinical episodes), HIV-OI (on ART with opportunistic infections), and Apparently Healthy). Complete Blood Count, Erythrocyte Surface Sialic Acid (ESSA), Free Serum Sialic Acid (FSSA) concentrations and Sialidase activity were determined for all 200 subjects. Analysis of variance (ANOVA) was used to compare the results of the different groups of HIV infected individuals as well as controls. The mean haemoglobin (HGB), Packed Cell Volume (PCV) and Red Blood Cells (RBC) concentrations were significantly lower (P ≤ 0.05) in the HIV groups compared with the apparently healthy groups. Anaemia and neutropaenia were the most common heamatological abnormalities observed in this study with highest prevalence of anaemia found in the ART naive group. The mean FSSA was 0.4±0.4mg/ml. There was a significant difference (p ≤ 0.05) between some groups. The highest levels of FSSA was observed in the HIV ART naïve (0.65±0.5mg/ml). The mean ESSA value for the study population was 0.54±0.35mg/ml with no significant difference (p ≤ 0.05) between groups. The mean sialidase activity values were 0.52±0.1 µmol/min/µl, 0.40±0.1 µmol/min/µl, 0.45±0.1 µmol/min/µl and 0.41±0.1 µmol/min/µl for the HIV ART naïve, HIV stable, HIV/OIs and apparently healthy groups respectively. No significant difference (p ≤ 0.05) was found between groups and also in gender and age. The finding in this study of higher mean sialidase activity and FSSA levels in the ART naïve HIV group compared with other groups indicate that the virus and other opportunistic pathogens may be sialidase producers in vivo which cleave off sialic acids from erythrocytes surface, leading to high levels of FSSA, anaemia and neutropaenia seen in this group. The higher ESSA concentration found in the HIV stable group along with lowest FSSA concentration in the group suggests the presence of sialyltransferases.Keywords: erythrocyte surface sialic acid, free serum sialic acid, HIV, sialidase
Procedia PDF Downloads 212118 Isolation and Identification of Salmonella spp and Salmonella enteritidis, from Distributed Chicken Samples in the Tehran Province using Culture and PCR Techniques
Authors: Seyedeh Banafsheh Bagheri Marzouni, Sona Rostampour Yasouri
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Salmonella is one of the most important common pathogens between humans and animals worldwide. Globally, the prevalence of the disease in humans is due to the consumption of food contaminated with animal-derived Salmonella. These foods include eggs, red meat, chicken, and milk. Contamination of chicken and its products with Salmonella may occur at any stage of the chicken processing chain. Salmonella infection is usually not fatal. However, its occurrence is considered dangerous in some individuals, such as infants, children, the elderly, pregnant women, or individuals with weakened immune systems. If Salmonella infection enters the bloodstream, the possibility of contamination of tissues throughout the body will arise. Therefore, determining the potential risk of Salmonella at various stages is essential from the perspective of consumers and public health. The aim of this study is to isolate and identify Salmonella from chicken samples distributed in the Tehran market using the Gold standard culture method and PCR techniques based on specific genes, invA and ent. During the years 2022-2023, sampling was performed using swabs from the liver and intestinal contents of distributed chickens in the Tehran province, with a total of 120 samples taken under aseptic conditions. The samples were initially enriched in buffered peptone water (BPW) for pre-enrichment overnight. Then, the samples were incubated in selective enrichment media, including TT broth and RVS medium, at temperatures of 37°C and 42°C, respectively, for 18 to 24 hours. Organisms that grew in the liquid medium and produced turbidity were transferred to selective media (XLD and BGA) and incubated overnight at 37°C for isolation. Suspicious Salmonella colonies were selected for DNA extraction, and PCR technique was performed using specific primers that targeted the invA and ent genes in Salmonella. The results indicated that 94 samples were Salmonella using the PCR technique. Of these, 71 samples were positive based on the invA gene, and 23 samples were positive based on the ent gene. Although the culture technique is the Gold standard, PCR is a faster and more accurate method. Rapid detection through PCR can enable the identification of Salmonella contamination in food items and the implementation of necessary measures for disease control and prevention.Keywords: culture, PCR, salmonella spp, salmonella enteritidis
Procedia PDF Downloads 72117 Mechanistic Understanding of the Difference in two Strains Cholerae Causing Pathogens and Predicting Therapeutic Strategies for Cholera Patients Affected with new Strain Vibrio Cholerae El.tor. Using Constrain-based Modelling
Authors: Faiz Khan Mohammad, Saumya Ray Chaudhari, Raghunathan Rengaswamy, Swagatika Sahoo
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Cholera caused by pathogenic gut bacteria Vibrio Cholerae (VC), is a major health problem in developing countries. Different strains of VC exhibit variable responses subject to different extracellular medium (Nag et al, Infect Immun, 2018). In this study, we present a new approach to model the variable VC responses in mono- and co-cultures, subject to continuously changing growth medium, which is otherwise difficult via simple FBA model. Nine VC strain and seven E. coli (EC) models were assembled and considered. A continuously changing medium is modelled using a new iterative-based controlled medium technique (ITC). The medium is appropriately prefixed with the VC model secretome. As the flux through the bacteria biomass increases secretes certain by-products. These products shall add-on to the medium, either deviating the nutrient potential or block certain metabolic components of the model, effectively forming a controlled feed-back loop. Different VC models were setup as monoculture of VC in glucose enriched medium, and in co-culture with VC strains and EC. Constrained to glucose enriched medium, (i) VC_Classical model resulted in higher flux through acidic secretome suggesting a pH change of the medium, leading to lowering of its biomass. This is in consonance with the literature reports. (ii) When compared for neutral secretome, flux through acetoin exchange was higher in VC_El tor than the classical models, suggesting El tor requires an acidic partner to lower its biomass. (iii) Seven of nine VC models predicted 3-methyl-2-Oxovaleric acid, mysirtic acid, folic acid, and acetate significantly affect corresponding biomass reactions. (iv) V. parhemolyticus and vulnificus were found to be phenotypically similar to VC Classical strain, across the nine VC strains. The work addresses the advantage of the ITC over regular flux balance analysis for modelling varying growth medium. Future expansion to co-cultures, potentiates the identification of novel interacting partners as effective cholera therapeutics.Keywords: cholera, vibrio cholera El. tor, vibrio cholera classical, acetate
Procedia PDF Downloads 162116 Profiling the Volatile Metabolome in Pear Leaves with Different Resistance to the Pear Psylla Cacopsylla bidens (Sulc) and Characterization of Phenolic Acid Decarboxylase
Authors: Mwafaq Ibdah, Mossab, Yahyaa, Dor Rachmany, Yoram Gerchman, Doron Holland, Liora Shaltiel-Harpaz
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Pear Psylla is the most important pest of pear in all pear-growing regions, in Asian, European, and the USA. Pear psylla damages pears in several ways: high-density populations of these insects can cause premature leaf and fruit drop, diminish plant growth, and reduce fruit size. In addition, their honeydew promotes sooty mold on leaves and russeting on fruit. Pear psyllas are also considered vectors of pear pathogens such as Candidatus Phytoplasma pyri causing pear decline that can lead to loss of crop and tree vigor, and sometimes loss of trees. Psylla control is a major obstacle to efficient integrated pest management. Recently we have identified two naturally resistance pear accessions (Py.760-261 and Py.701-202) in the Newe Ya’ar live collection. GC-MS volatile metabolic profiling identified several volatile compounds common in these accessions but lacking, or much less common, in a sensitive accession, the commercial Spadona variety. Among these volatiles were styrene and its derivatives. When the resistant accessions were used as inter-stock, the volatile compounds appear in commercial Spadona scion leaves, and it showed reduced susceptibility to pear psylla. Laboratory experiments and applications of some of these volatile compounds were very effective against psylla eggs, nymphs, and adults. The genes and enzymes involved in the specific reactions that lead to the biosynthesis of styrene in plant are unknown. We have identified a phenolic acid decarboxylase that catalyzes the formation of p-hydroxystyrene, which occurs as a styrene analog in resistant pear genotypes. The His-tagged and affinity chromatography purified E. coli-expressed pear PyPAD1 protein could decarboxylate p-coumaric acid and ferulic acid to p-hydroxystyrene and 3-methoxy-4-hydroxystyrene. In addition, PyPAD1 had the highest activity toward p-coumaric acid. Expression analysis of the PyPAD gene revealed that its expressed as expected, i.e., high when styrene levels and psylla resistance were high.Keywords: pear Psylla, volatile, GC-MS, resistance
Procedia PDF Downloads 146115 Removal of Heavy Metals from Municipal Wastewater Using Constructed Rhizofiltration System
Authors: Christine A. Odinga, G. Sanjay, M. Mathew, S. Gupta, F. M. Swalaha, F. A. O. Otieno, F. Bux
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Wastewater discharged from municipal treatment plants contain an amalgamation of trace metals. The presence of metal pollutants in wastewater poses a huge challenge to the choice and applications of the preferred treatment method. Conventional treatment methods are inefficient in the removal of trace metals due to their design approach. This study evaluated the treatment performance of a constructed rhizofiltration system in the removal of heavy metals from municipal wastewater. The study was conducted at an eThekwni municipal wastewater treatment plant in Kingsburgh - Durban in the province of KwaZulu-Natal. The construction details of the pilot-scale rhizofiltration unit included three different layers of substrate consisting of medium stones, coarse gravel and fine sand. The system had one section planted with Phragmites australis L. and Kyllinga nemoralis L. while the other section was unplanted and acted as the control. Influent, effluent and sediment from the system were sampled and assessed for the presence of and removal of selected trace heavy metals using standard methods. Efficiency of metals removal was established by gauging the transfer of metals into leaves, roots and stem of the plants by calculations based on standard statistical packages. The Langmuir model was used to assess the heavy metal adsorption mechanisms of the plants. Heavy metals were accumulated in the entire rhizofiltration system at varying percentages of 96.69% on planted and 48.98% on control side for cadmium. Chromium was 81% and 24%, Copper was 23.4% and 1.1%, Nickel was 72% and 46.5, Lead was 63% and 31%, while Zinc was 76% and 84% on the on the water and sediment of the planted and control sides of the rhizofilter respectively. The decrease in metal adsorption efficiencies on the planted side followed the pattern of Cd>Cr>Zn>Ni>Pb>Cu and Ni>Cd>Pb>Cr>Cu>Zn on the control side. Confirmatory analysis using Electron Scanning Microscopy revealed that higher amounts of metals was deposited in the root system with values ranging from 0.015mg/kg (Cr), 0.250 (Cu), 0.030 (Pb) for P. australis, and 0.055mg/kg (Cr), 0.470mg/kg (Cu) and 0.210mg/kg,(Pb) for K. nemoralis respectively. The system was found to be efficient in removing and reducing metals from wastewater and further research is necessary to establish the immediate mechanisms that the plants display in order to achieve these reductions.Keywords: wastewater treatment, Phragmites australis L., Kyllinga nemoralis L., heavy metals, pathogens, rhizofiltration
Procedia PDF Downloads 262114 Fastidious Enteric Pathogens in HIV
Authors: S. Pathak, R. Lazarus
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A 25-year-old male HIV patient (CD4 cells 20/µL and HIV viral load 14200000 copies/ml) with a past medical history of duodenal ulcer, pneumocystis carinii pneumonia, oesophageal candidiasis presented with fever and a seizure to hospital. The only recent travel had been a religious pilgrimage from Singapore to Malaysia 5 days prior; during the trip he sustained skin abrasions. The patient had recently started highly active antiretroviral therapy 2 months prior. Clinical examination was unremarkable other than a temperature of 38.8°C and perianal warts. Laboratory tests showed a leukocyte count 12.5x109 cells/L, haemoglobin 9.4 g/dL, normal biochemistry and a C-reactive protein 121 mg/L. CT head and MRI head were unremarkable and cerebrospinal fluid analysis performed after a delay (due to technical difficulties) of 11 days was unremarkable. Blood cultures (three sets) taken on admission showed Gram-negative rods in the anaerobic bottles only at the end of incubation with culture result confirmed by molecular sequencing showing Helicobacter cinaedi. The patient was treated empirically with ceftriaxone for seven days and this was converted to oral co-amoxiclav for a further seven days after the blood cultures became positive. A Transthoracic echocardiogram was unremarkable. The patient made a full recovery. Helicobacter cinaedi is a gram-negative anaerobic fastidious organism affecting patients with comorbidity. Infection may manifest as cellulitius, colitis or as in this case as bloodstream infection – the latter is often attributed to faeco-oral infection. Laboratory identification requires prolonged culture. Therapeutic options may be limited by resistance to macrolides and fluoroquinolones. The likely pathogen inoculation routes in the case described include gastrointestinal translocation due to proctitis at the site of perianal warts, or breach of the skin via abrasions occurring during the pilgrimage. Such organisms are increasing in prevalence as our patient population ages and patients have multiple comorbidities including HIV. It may be necessary in patients with unexplained fever to prolong incubation of sterile sites including blood in order to identify this unusual fastidious organism.Keywords: fastidious, Helicobacter cinaedi, HIV, immunocompromised
Procedia PDF Downloads 378113 Ceratocystis manginecans Causal Agent of a Destructive Mangoes in Pakistan
Authors: Asma Rashid, Shazia Iram, Iftikhar Ahmad
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Mango sudden death is an emerging problem in Pakistan. As its prevalence is observed in almost all mango growing areas and severity varied from 2-5% in Punjab and 5-10% in Sindh. Symptoms on affected trees include bark splitting, discoloration of the vascular tissue, wilting, gummosis and at the end rapid death. Total of n= 45 isolates were isolated from different mango growing areas of Punjab and Sindh. Pathogenicity of these fungal isolates was tested through artificial inoculation method on different hosts (potato tubers, detached mango leaves, detached mango twigs and mango plants) under controlled conditions and all were proved pathogenic with varying degree of aggressiveness in reference to control. The findings of the present study proved that out of these four methods, potato tubers inoculation method was the most ideal as this fix the inoculums on the target site. Increased fungal growth and spore numbers may be due to soft tissues of potato tubers from which Ceratocystis isolates can easily pass. Lesion area on potato tubers was in the range of 7.09-0.14 cm2 followed by detached mango twigs which were ranged from 0.48-0.09 cm2). All pathological results were proved highly significant at P<0.05 through ANOVA but isolate to isolate showed non-significant behaviour but they have the positive effect on lesion area. Re-isolation of respective fungi was achieved with 100 percent success which results in the verification of Koch’s postulates. DNA of fungal pathogens was successfully extracted through phenol chloroform method. Amplification was done through ITS, b-tubulin gene, and Transcription Elongation Factor (EF1-a) gene primers and the amplified amplicons were sequenced and compared from NCBI which showed 99-100 % similarity with Ceratocystis manginecans. Fungus Ceratocystis manginecans formed one of strongly supported sub-clades through phylogenetic tree. Results obtained through this work would be supportive in establishment of relation of isolates with their region and will give information about pathogenicity level of isolates that would be useful to develop the management policies to reduce the afflictions in orchards caused by mango sudden death.Keywords: artificial inoculation, mango, Ceratocystis manginecans, phylogenetic, screening
Procedia PDF Downloads 248112 Isolation, Identification and Screening of Pectinase Producing Fungi Isolated from Apple (Malus Domestica)
Authors: Shameel Pervez, Saad Aziz Durrani, Ibatsam Khokhar
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Pectinase is an enzyme that breaks down pectin, a compound responsible for structural integrity of the plant. Pectin is difficult to break down mechanically and the cost is very high, that is why many industries including food industries use pectinase enzyme produced by microbes for pectin breakdown. Apple (Malus domestica) is an important fruit in terms of market value. Every year, millions of apples are wasted due to post-harvest rot caused by fungi. Fungi are natural decomposers of our ecosystem and are infamous for post-harvest rot of apple fruit but at the same time they are prized for their high production of valuable extracellular enzymes such as pectinase. In this study, fungi belonging to different genus were isolated from rotten apples. Rotten samples of apple were picked from different markets of Lahore. After surface sterilization, the rotten parts were cut into small pieces and placed onto MEA media plates for three days. Afterwards, distinct colonies were picked and purified by sub-culturing. The isolates were identified to genus level through the study of basic colony morphology and microscopic features. The isolates were then subjected to screening for pectinase activity on MS media to compare pectinase production and were then subsequently tested for pathogenic activity through wound suspension method to evaluate the pathogenic activity of isolates in comparison with their pectinolytic activity. A total of twelve fungal strains were isolates from rotten apples. They were belonging to genus Penicillium, Alternaria, Paecilomyces and Rhizopus. Upon screening for pectinolytic activity, isolates Pen 1, Pen 4, and Rz showed high pectinolytic activity and were further subjected to DNA isolation and partial sequencing for species identification. The results of partial sequencing were combined with in-depth study of morphological features revealing Pen 1 as Penicillium janthinellum, Pen 4 as Penicillium griseofulvum, and Rz as Rhizopus microsporus. Pathogenic activity of all twelve isolates was evaluated. Penicillium spp. were highly pathogenic and destructive and same was the case with Paecilomyces sp. and Rhizopus sp. However, Alternaria spp. were found to be more consistent in their pathogenic activity, on all types of apples.Keywords: apple, pectinase, fungal pathogens, penicillium, rhizopus
Procedia PDF Downloads 63111 Investigating the Use of Seaweed Extracts as Biopesticides
Authors: Emma O’ Keeffe, Helen Hughes, Peter McLoughlin, Shiau Pin Tan, Nick McCarthy
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Biosecurity is emerging as one of the most important issues facing the agricultural and forestry community. This is as a result of increased invasion from new pests and diseases with the main protocol for dealing with these species being the use of synthetic pesticides. However, these chemicals have been shown to exhibit negative effects on the environment. Seaweeds represent a vast untapped resource of bio-molecules with a broad range of biological activities including pesticidal. This project investigated both the antifungal and antibacterial activity of seaweed species against two problematic root rot fungi, Armillaria mellea and Heterobasidion annosum and ten quarantine bacterial plant pathogens including Xanthomonas arboricola, Xanthomonas fragariae, and Erwinia amylovora. Four seaweed species were harvested from the South-East coast of Ireland including brown, red and green varieties. The powdered seaweeds were extracted using four different solvents by liquid extraction. The poisoned food technique was employed to establish the antifungal efficacy, and the standard disc diffusion assay was used to assess the antibacterial properties of the seaweed extracts. It was found that extracts of the green seaweed exhibited antifungal activity against H. annosum, with approximately 50% inhibition compared to the negative control. The protectant activities of the active extracts were evaluated on disks of Picea sitchensis, a plant species sensitive to infection from H. annosum and compared to the standard chemical control product urea. The crude extracts exhibited very similar activity to the 10% and 20% w/v concentrations of urea, demonstrating the ability of seaweed extracts to compete with commercially available products. Antibacterial activity was exhibited by a number of seaweed extracts with the red seaweed illustrating the strongest activity, with a zone of inhibition of 15.83 ± 0.41 mm exhibited against X. arboricola whilst the positive control (10 μg/disk of chloramphenicol) had a zone of 26.5 ± 0.71 mm. These results highlight the potential application of seaweed extracts in the forestry and agricultural industries for use as biopesticides. Further work is now required to identify the bioactive molecules that are responsible for this antifungal and antibacterial activity in the seaweed extracts, including toxicity studies to ensure the extracts are non-toxic to plants and humans.Keywords: antibacterial, antifungal, biopesticides, seaweeds
Procedia PDF Downloads 172110 Study of Growth Behavior of Some Bacterial Fish Pathogens to Combined Selected Herbal Essential Oil
Authors: Ashkan Zargar, Ali Taheri Mirghaed, Zein Talal Barakat, Alireza Khosravi, Hamed Paknejad
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With the increase of bacterial resistance to the chemical antibiotics, replacing it with ecofriendly herbal materials and with no adverse effects in the host body is very important. Therefore, in this study, the effect of combined essential oil (Thymus vulgaris-Origanum magorana and Ziziphora clinopodioides) on the growth behavior of Yersinia ruckeri, Aeromonas hydrophila and Lactococcus garvieae was evaluated. The compositions of the herbal essential oils used in this study were determined by gas chromatography-mass spectrometry (GC-MS) while, the investigating of antimicrobial effects was conducted by the agar-disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and bacterial growth curves determination relied on optical density (OD) at 630 nm. The main compounds were thymol (40.60 %) and limonene (15.98 %) for Thymus vulgaris while carvacrol (57.86 %) and thymol (13.54 %) were the major compounds in Origanum magorana. As regards Ziziphora clinopodiodes, α-pinene (22.6 %) and carvacrol (21.1 %) represented the major constituents. Concerning Yersinia ruckeri, disc-diffusion results showed that t.O.z (50 % Origanum majorana) combined essential oil was presented the best inhibition zone (30.66 mm) but it was exhibited no significant differences with other tested commercial antibiotics except oxytetracycline (P <0/05). The inhibitory activity and the bactericidal effect of the t.O.z, unveiled by the MIC= 0.2 μL /mL and MBC= 1.6 μL /mL values, were clearly the best between all combined oils. The growth behaviour of Yersinia ruckeri was affected by this combined essential oil and changes in temperature and pH conditions affected herbal oil performance. As regard Aeromonas hydrophila, its results were so similar to Yersinia ruckeri results and t.O.z (50 % Origanum majorana) was the best between all combined oils (inhibition zone= 26 mm, MIC= 0.4 μL /mL and MBC= 3.2 μL /mL, combined essential oil was affected bacterial growth behavior). Also for Lactococcus garvieae, t.O.z (50 % Origanum majorana) was the best between all combined oils having the best inhibition zone= 20.66 mm, MIC= 0.8 μL /mL and MBC= 1.6 μL /mL and best effect on inhibiting bacterial growth. Combined herbal essential oils have a good and noticeable effect on the growth behavior of pathogenic bacteria in the laboratory, and by continuing research in the host, they may be a suitable alternative to control, prevent and treat diseases caused by these bacteria.Keywords: bacterial pathogen, herbal medicine, growth behavior, fish
Procedia PDF Downloads 71109 Molecular Farming: Plants Producing Vaccine and Diagnostic Reagent
Authors: Katerina H. Takova, Ivan N. Minkov, Gergana G. Zahmanova
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Molecular farming is the production of recombinant proteins in plants with the aim to use the protein as a purified product, crude extract or directly in the planta. Plants gain more attention as expression systems compared to other ones due to the cost effective production of pharmaceutically important proteins, appropriate post-translational modifications, assembly of complex proteins, absence of human pathogens to name a few. In addition, transient expression in plant leaves enables production of recombinant proteins within few weeks. Hepatitis E virus (HEV) is a causative agent of acute hepatitis. HEV causes epidemics in developing countries and is primarily transmitted through the fecal-oral route. Presently, all efforts for development of Hepatitis E vaccine are focused on the Open Read Frame 2 (ORF2) capsid protein as it contains epitopes that can induce neutralizing antibodies. For our purpose, we used the CMPV-based vector-pEAQ-HT for transient expression of HEV ORF2 in Nicotiana benthamina. Different molecular analysis (Western blot and ELISA) showed that HEV ORF2 capsid protein was expressed in plant tissue in high-yield up to 1g/kg of fresh leaf tissue. Electron microscopy showed that the capsid protein spontaneously assembled in low abundance virus-like particles (VLPs), which are highly immunogenic structures and suitable for vaccine development. The expressed protein was recognized by both human and swine HEV positive sera and can be used as a diagnostic reagent for the detection of HEV infection. Production of HEV capsid protein in plants is a promising technology for further HEV vaccine investigations. Here, we reported for a rapid high-yield transient expression of a recombinant protein in plants suitable for vaccine production as well as a diagnostic reagent. Acknowledgments -The authors’ research on HEV is supported with grants from the Project PlantaSYST under the Widening Program, H2020 as well as under the UK Biotechnological and Biological Sciences Research Council (BBSRC) Institute Strategic Programme Grant ‘Understanding and Exploiting Plant and Microbial Secondary Metabolism’ (BB/J004596/1). The authors want to thank Prof. George Lomonossoff (JIC, Norwich, UK) for his contribution.Keywords: hepatitis E virus, plant molecular farming, transient expression, vaccines
Procedia PDF Downloads 151108 Observation on Microbiological Profile of Type2 Diabetic Foot Ulcer and Its Antimicrobial Sensitivity Pattern in a Tertiary Care Hospital in Eastern India
Authors: Pampita Chakraborty, Sukumar Mukherjee
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Diabetes Mellitus (DM) is commonly encountered metabolic disorder in clinical practice. An estimated 25 percent of DM patients develop foot problems. Foot ulceration and infection are one of the major causes of morbidity, hospitalization or even amputation. Objective: To isolate and identify bacterial pathogens in Diabetic Foot Ulcer (DFU) and to observe its antimicrobial sensitivity pattern. Methodology: A prospective study was conducted for a period of 9 months at the Department of Microbiology, GD Hospital & Diabetes Institute, Kolkata. 75 DFU patients were recruited in the study. Specimens for microbiological studies obtained from ulcer base were examined as gram stained smear and was cultured aerobically on Nutrient agar, Blood agar and MacConkey agar plates. Antimicrobial sensitivity test was performed by disc diffusion techniques according to CLSI guidelines. Result: In this study out of 75cases, 73% (55/75) were male and 27% (20/75) were females with mean (SD) age of 51.11(±10) years. Out of 75 pus cultures, 63(84%) showed growth of microorganism making total of 81 bacterial isolates with 71.42% of monomicrobial infection and 28.57% of polymicrobial infection. Out of 81 isolates 53(65.43%) were gram negative and 21(25.92%) were gram positive. E.coli was relatively common isolate 21(26%) followed by Staphylococcus aureus 15(18.5%), Klebsiella pneumonia 14(17.28%), Pseudomonas aeruginosa 12 (14.81%), Proteus spp. 3 (3.70%), and Enterococcus faecalis 6 (7.40%). 75% of Gram-negative microorganism were extended Beta-lactamase enzyme (ESBL) producer and around 20 % of Klebsiella and Proteus spp. were carbapenemase enzyme producer. Among Gram positive, around 50% of S.aureus was MRSA, sensitive only to Vancomycin, Teicoplanin & Linezolid. Conclusion: More prevalence of monomicrobial gram-negative bacteria than gram-positive bacteria in DFU was observed. This study emphasizes that Beta-Lactam group of antibiotics should not be the empirical treatment of choice for Gram-negative isolates; instead alternatives like Carbapenems, Amikacin could be a better option. On the other hand, Vancomycin and Linezolid are preferred for most of the infection with gram-positive aerobes. Continuous surveillance of resistant bacteria is required for empiric therapy.Keywords: antibiotic resistant, antimicrobial susceptibility, diabetic foot ulcer, surveillance
Procedia PDF Downloads 369107 Integrating One Health Approach with National Policies to Improve Health Security post-COVID-19 in Vietnam
Authors: Yasser Sanad, Thu Trang Dao
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Introduction: Implementing the One Health (OH) approach requires an integrated, interdisciplinary, and cross-sectoral methodology. OH is a key tool for developing and implementing programs and projects and includes developing ambitious policies that consider the common needs and benefits of human, animal, plant, and ecosystem health. OH helps humanity readjust its path to environmentally friendly and impartial sustainability. As co-leader of the Global Health Security Agenda’s Zoonotic Disease Action Package, Vietnam pioneered a strong OH approach to effectively address early waves of the COVID-19 outbreak in-country. Context and Aim: The repeated surges in COVID-19 in Vietnam challenged the capabilities of the national system and disclosed the gaps in multi-sectoral coordination and resilience. To address this, FHI 360 advocated for the standardization of the OH platform by government actors to increase the resiliency of the system during and post COVID-19. Methods: FHI 360 coordinated technical resources to develop and implement evidence-based OH policies, promoting high-level policy dialogue between the Ministries of Health, Agriculture, and the Environment, and policy research to inform developed policies and frameworks. Through discussions, an OH-building Partnership (OHP) was formed, linking climate change, the environment, and human and animal health. Findings: The OHP Framework created a favorable policy environment within and between sectors, as well as between governments and international health security partners. It also promoted strategic dialogue, resource mobilization, policy advocacy, and integration of international systems with National Steering Committees to ensure accountability and emphasize national ownership. Innovative contribution to policy, practice and/or research: OHP was an effective evidence-based research-to-policy platform linking to the National One Health Strategic Plan (2021-2025). Collectively they serve as a national framework for the implementation and monitoring of OH activities. Through the adoption of policies and plans, the risk of zoonotic pathogens, environmental agent spillover, and antimicrobial resistance can be minimized through strengthening multi-sectoral OH collaboration for health security.Keywords: one health, national policies, health security, COVID-19, Vietnam
Procedia PDF Downloads 105106 Performance Evaluation of On-Site Sewage Treatment System (Johkasou)
Authors: Aashutosh Garg, Ankur Rajpal, A. A. Kazmi
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The efficiency of an on-site wastewater treatment system named Johkasou was evaluated based on its pollutant removal efficiency over 10 months. This system was installed at IIT Roorkee and had a capacity of treating 7 m3/d of sewage water, sufficient for a group of 30-50 people. This system was fed with actual wastewater through an equalization tank to eliminate the fluctuations throughout the day. Methanol and ammonium chloride was added into this equalization tank to increase the Chemical Oxygen Demand (COD) and ammonia content of the influent. The outlet from Johkasou is sent to a tertiary unit consisting of a Pressure Sand Filter and an Activated Carbon Filter for further treatment. Samples were collected on alternate days from Monday to Friday and the following parameters were evaluated: Chemical Oxygen Demand (COD), Biochemical Oxygen Demand (BOD), Total Suspended Solids (TSS), and Total Nitrogen (TN). The Average removal efficiency for Chemical Oxygen Demand (COD), Biochemical Oxygen Demand (BOD), Total Suspended Solids (TSS), and Total Nitrogen (TN) was observed as 89.6, 97.7, 96, and 80% respectively. The cost of treating the wastewater comes out to be Rs 23/m3 which includes electricity, cleaning and maintenance, chemical, and desludging costs. Tests for the coliforms were also performed and it was observed that the removal efficiency for total and fecal coliforms was 100%. The sludge generation rate is approximately 20% of the BOD removal and it needed to be removed twice a year. It also showed a very good response against the hydraulic shock load. We performed vacation stress analysis on the system to evaluate the performance of the system when there is no influent for 8 consecutive days. From the result of stress analysis, we concluded that system needs a recovery time of about 48 hours to stabilize. After about 2 days, the system returns again to original conditions and all the parameters in the effluent become within the limits of National Green Tribunal (NGT) standards. We also performed another stress analysis to save the electricity in which we turned the main aeration blower off for 2 to 12 hrs a day and the results showed that we can turn the blower off for about 4-6 hrs a day and this will help in reducing the electricity costs by about 25%. It was concluded that the Johkasou system can remove a sufficient amount of all the physiochemical parameters tested to satisfy the prescribed limit set as per Indian Standard.Keywords: on-site treatment, domestic wastewater, Johkasou, nutrient removal, pathogens removal
Procedia PDF Downloads 113105 Effects of Caprine Arthritis-Encephalitis Virus (CAEV) Infection on the Expression of Cathelicidin Genes in Goat Blood Leukocytes
Authors: Daria Reczynska, Justyna Jarczak, Michal Czopowicz, Danuta Sloniewska, Karina Horbanczuk, Wieslaw Jarmuz, Jaroslaw Kaba, Emilia Bagnicka
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Since people, animals and plants are constantly exposed to pathogens they have developed very complex systems of defense. Among ca. 1000 antimicrobial peptides from different families so far identified, approximately 30 belonging to cathelicidin family can be found in mammals. Cathelicidins probably constitute the first line of defense because they can act at a physiological salt concentration which is present in healthy tissues. Moreover, the low salt concentration which is present in infected tissues inhibits their activity. In goat bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), myeloid antimicrobial peptide 28 (MAP28), myeloid antimicrobial peptide 34 (MAP34 A and B), goat bactenecin3.4 (ChBac3.4) were identified. Caprine arthritis-encephalitis (CAE) caused by small ruminant lentivirus (SRLV) is economic problem. The main CAE symptoms are weight loss, arthritis, pneumonia and mastitis (significant elevation of the somatic cell count and deterioration of some technological parameters). The study was conducted on 24 dairy goats. The animals were divided into two groups: experimental (SRLV-infected) and control (non-infected). The blood samples were collected five times: on the 1st, 7th, 30th, 90th and 150thday of lactation. The levels of transcripts of BAC7.5, BAC5, MAP28 and MAP34 genes in blood leucocytes were measured using qPCR method. There were no differences in mRNA levels of studied genes between stages of lactation. The differences were observed in expressions of BAC5, MAP28 and MAP34 genes with lower levels in the experimental group. There was no difference in BAC7.5 expression between groups. The decreased levels of transcripts of cathelicidin genes in blood leucocytes of SRLV-infected goats may indicate the disturbances of homeostasis in organisms. It can be concluded that SRLV infection seems to inhibit expression of cathelicidin genes. The study was financed by a grant from the National Scientific Center No. UMO-2013/09/B/NZ/03514.Keywords: goat, CAEV, cathelicidins, blood leukocytes, gene expression
Procedia PDF Downloads 283104 Tick Infestation and its Implications on Health and Welfare of Cattle under Pastoral System in Nigeria
Authors: Alabi Olufemi, Adeyanju Taiwo, Oloruntoba Oluwasegun, Adeleye Bobola, Alabi Oyekemi
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The pastoral system is a predominant form of cattle production in Nigeria, characterized by extensive grazing on communal lands. However, this system is challenged by various factors, including tick infestation, which significantly affects cattle health and welfare hence this investigation which aims to provide an in-depth understanding of tick infestation in the context of Nigerian pastoral systems, emphasizing its impact on cattle health and welfare. The country harbors a diverse array of tick species that affect cattle. These ticks belong to different genera, including Rhipicephalus, Amblyomma, and Hyalomma, among others. Each species has unique characteristics, life cycles, and host preferences, contributing to the complexity of tick infestation dynamics in pastoral settings. Tick infestation has numerous detrimental effects on cattle health. The direct effects include blood loss, anemia, skin damage due to feeding, and the transmission of pathogens that cause diseases such as anaplasmosis, babesiosis, and theileriosis. Indirectly, tick infestation can lead to reduced productivity, weight loss, and increased susceptibility to other diseases.The welfare of cattle in Nigerian pastoral systems is significantly impacted by tick infestation. Infested cattle often exhibit signs of distress, including restlessness, reduced grazing activity, and altered behavior. Furthermore, the discomfort caused by tick bites can lead to chronic stress, compromising the overall welfare of the animals. Effective tick control is crucial for mitigating the impact of infestation on cattle health and welfare. Strategies such as acaricide application, pasture management, genetic selection for tick resistance cattle, and vaccination against tick-borne diseases are commonly used. Tick infestation presents a significant challenge to cattle production under the pastoral system in Nigeria. It not only impacts cattle health but also compromises their welfare. Addressing the issue of tick infestation requires a multifaceted approach that integrates effective control strategies with sustainable management practices. Further research is needed to develop tailored interventions that account for the unique characteristics of Nigerian pastoral systems, ultimately ensuring the well-being and productivity of cattle in these settings.Keywords: tick infestation, pastoral system, welfare, cattle
Procedia PDF Downloads 55103 Antioxidant Activity and Microbiological Quality of Functional Bread Enriched with Morus Alba Leaf Extract during Storage
Authors: Joanna Kobus-Cisowska, Daria Szymanowska, Piotr Szulc, Oskar Szczepaniak, Marcin Dziedzinski, Szymon Byczkiewicz
Abstract:
A wide range of food products is offered on the market. However, increasing consumer awareness of the impact of food on health causes a growing interest in enriched products. Cereal products are an important element of the daily diet of man. In the literature, no data was found on the impact of Morus alba preparations on the content of active ingredients and properties of wholemeal bread. Mulberry leaves (Morus alba L) are a rich source of bioactive compounds with multidirectional antioxidant activity, which means that they can be a component of new foods that prevent disease or support therapy and improve the patient's health. The aim of the study was to assess the impact of the addition of white mulberry leaf extract on the antioxidant activity of bread. It has been shown that bread can be a carrier of biologically active substances from mulberry leaves, because the addition of mulberry at a sensory acceptable level and meeting microbiological requirements significantly influenced the increase in the content of bioactive ingredients and the antioxidant activity of bread. The addition of mulberry leaf water extract to bread increased the level of flavonols and phenolic acids, in particular protocatechic, chlorogenic gallic and caffeic acid and isoquercetin and rutine, and also increased the antioxidant potential, which were microbiological stable during 5 days storage. It has been shown also that the addition of Morus alba preparations has a statistically significant effect on anti-radical activity. In addition, there were no differences in activity in DPPH · and ABTS · + tests between post-storage samples. This means that the compounds responsible for the anti-radical activity present in the bread were not inactivated during storage. It was found that the tested bread was characterized by high microbiological purity, which is indicated by the obtained results of analyzes performed for the titers of indicator microorganisms and the absence of pathogens. In the tested products from the moment of production throughout the entire storage period, no undesirable microflora was found, which proves their safety and guarantees microbiological stability during the storage period.Keywords: antioxidants, bread, extract, quality
Procedia PDF Downloads 174