Search results for: bacterial leaf spot

Authors: S.Kauroo, J. Govinden Soulange, D. Marie

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Psiadia species from the Asteraceae are traditionally used in the folk medicine of Mauritius to treat cutaneous and bronchial infections. The present study aimed at validating the phytomedicinal properties of the selected species from the Asteraceae family, namely Psiadia arguta, Psiadia viscosa, Psiadia lithospermifolia, and Distephanus populifolius. Dried hexane, ethyl acetate, and methanol leaf extracts were studied for their antioxidant properties using the DPPH (1, 1-diphenyl-2-picryl-hydrazyl), FRAP (Ferric Reducing Ability of Plasma), and Deoxyribose assays. Antibacterial activity against human pathogenic bacteria namely Escherichia coli (ATCC 27853), Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Klebsiella pneumonia (ATCC27853), Pseudomonas aeruginosa (ATCC 27853), and Bacillus cereus (ATCC 11778) was measured using the broth microdilution assay. Qualitative phytochemical screening using standard methods revealed the presence of coumarins, tannins, leucoanthocyanins, and steroids in all the tested extracts. The measured phenolics level of the selected plant extracts varied from 24.0 to 231.6 mg GAE/g with the maximum level in methanol extracts in all four species. The highest flavonoids and proanthocyanidins content was noted in Psiadia arguta methanolic extracts with 65.7±1.8 mg QE/g and 5.1±0.0 mg CAT/g dry weight (DW) extract, respectively. The maximum free radical scavenging activity was measured in Psiadia arguta methanol and ethyl acetate extracts with IC50 11.3±0.2 and 11.6± 0.2 µg/mL, respectively and followed by Distephanus populifolius methanol extracts with an IC50 of 11.3± 0.8 µg/mL. The maximum ferric reducing antioxidant potential was noted in Psiadia lithospermifolia methanol extracts with a FRAP value of 18.8 ± 0.4 µmol Fe2+/L/g DW. The antioxidant capacity based on DPPH and Deoxyribose values were negatively related to total phenolics, flavonoid and proanthocyanidins content while the ferric reducing antioxidant potential were strongly correlated to total phenolics, flavonoid and proanthocyanidins content. All four species exhibited antimicrobial activity against the tested bacteria (both Gram-negative and Gram-positive). Interestingly, the hexane and ethyl acetate extracts of Psiadia viscosa and Psiadia lithospermifolia were more active than the control antibiotic Chloramphenicol. The Minimum inhibitory concentration (MIC) values for hexane and ethyl acetate extracts of Psiadia viscosa and Psiadia lithospermifolia against the tested bacteria ranged from (62.5 to 500 µg/ml). These findings validate the use of these tested Asteraceae in the traditional medicine of Mauritius and also highlight their pharmaceutical potential as prospective phytomedicines.

Keywords: antibacterial, antioxidant, DPPH, flavonoids, FRAP, Psiadia spp

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Authors: Tsega Asresa, Getahun Tigistu, Melaku Bayih

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Computer vision is a subfield of artificial intelligence that allows computers and systems to extract meaning from digital images and video. It is used in a wide range of fields of study, including self-driving cars, video surveillance, medical diagnosis, manufacturing, law, agriculture, quality control, health care, facial recognition, and military applications. Aromatic medicinal plants are botanical raw materials used in cosmetics, medicines, health foods, essential oils, decoration, cleaning, and other natural health products for therapeutic and Aromatic culinary purposes. These plants and their products not only serve as a valuable source of income for farmers and entrepreneurs but also going to export for valuable foreign currency exchange. In Ethiopia, there is a lack of technologies for the classification and identification of Aromatic medicinal plant parts and disease type cured by aromatic medicinal plants. Farmers, industry personnel, academicians, and pharmacists find it difficult to identify plant parts and disease types cured by plants before ingredient extraction in the laboratory. Manual plant identification is a time-consuming, labor-intensive, and lengthy process. To alleviate these challenges, few studies have been conducted in the area to address these issues. One way to overcome these problems is to develop a deep learning model for efficient identification of Aromatic medicinal plant parts with their corresponding disease type. The objective of the proposed study is to identify the aromatic medicinal plant parts and their disease type classification using computer vision technology. Therefore, this research initiated a model for the classification of aromatic medicinal plant parts and their disease type by exploring computer vision technology. Morphological characteristics are still the most important tools for the identification of plants. Leaves are the most widely used parts of plants besides roots, flowers, fruits, and latex. For this study, the researcher used RGB leaf images with a size of 128x128 x3. In this study, the researchers trained five cutting-edge models: convolutional neural network, Inception V3, Residual Neural Network, Mobile Network, and Visual Geometry Group. Those models were chosen after a comprehensive review of the best-performing models. The 80/20 percentage split is used to evaluate the model, and classification metrics are used to compare models. The pre-trained Inception V3 model outperforms well, with training and validation accuracy of 99.8% and 98.7%, respectively.

Keywords: aromatic medicinal plant, computer vision, convolutional neural network, deep learning, plant classification, residual neural network

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Authors: Satwik Majumder, Dongyun Jung, Jennifer Ronholm, Saji George

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Bovine mastitis is the most common infectious disease in dairy cattle, with major economic implications for the dairy industry worldwide. Continuous monitoring for the emergence of antimicrobial resistance (AMR) among bacterial isolates from dairy farms is vital not only for animal husbandry but also for public health. In this study, the prevalence of AMR in 113 Escherichia coli isolates from cases of bovine clinical mastitis in Canada was investigated. Kirby-Bauer disk diffusion test with 18 antibiotics and microdilution method with three heavy metals (copper, zinc, and silver) was performed to determine the antibiotic and heavy-metal susceptibility. Resistant strains were assessed for efflux and ß-lactamase activities besides assessing biofilm formation and hemolysis. Whole-genome sequences for each of the isolates were examined to detect the presence of genes corresponding to the observed AMR and virulence factors. Phenotypic analysis revealed that 32 isolates were resistant to one or more antibiotics, and 107 showed resistance against at least one heavy metal. Quinolones and silver were the most efficient against the tested isolates. Among the AMR isolates, AcrAB-TolC efflux activity and ß-lactamase enzyme activities were detected in 13 and 14 isolates, respectively. All isolates produced biofilm but with different capacities, and 33 isolates showed α-hemolysin activity. A positive correlation (Pearson r = +0.89) between efflux pump activity and quantity of biofilm was observed. Genes associated with aggregation, adhesion, cyclic di-GMP, quorum sensing were detected in the AMR isolates, corroborating phenotype observations. This investigation showed the prevalence of AMR in E. coli isolates from bovine clinical mastitis. The results also suggest the inadequacy of antimicrobials with a single mode of action to curtail AMR bacteria with multiple mechanisms of resistance and virulence factors. Therefore, it calls for combinatorial therapy for the effective management of AMR infections in dairy farms and combats its potential transmission to the food supply chain through milk and dairy products.

Keywords: antimicrobial resistance, E. coli, bovine mastitis, antibiotics, heavy-metals, efflux pump, ß-lactamase enzyme, biofilm, whole-genome sequencing

Procedia PDF Downloads 216

Authors: Abiodun Amusan, Olugbenga Akinola, Kazeem Akano, María Hernández-Castañeda, Jenna Dick, Akintunde Sowunmi, Geoffrey Hart, Grace Gbotosho

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Introduction: Artemisinin-based Combination Therapy (ACTs) is the cornerstone malaria treatment option in most malaria-endemic countries. Unfortunately, the malaria control effort is constantly being threatened by resistance of Plasmodium falciparum to ACTs. The recent evidence of artemisinin resistance in East Africa and its possibility of spreading to other African regions portends an imminent health catastrophe. This study aimed at evaluating the occurrence, prevalence, and influence of artemisinin-resistance markers on treatment outcomes in Ibadan before and after post-adoption of artemisinin combination therapy (ACTs) in Nigeria in 2005. Method: The study involved day zero dry blood spot (DBS) obtained from malaria patients during retrospective (2000-2005) and prospective (2021) studies. A cohort in the prospective study received oral dihydroartemisinin-piperaquine and underwent a 42-day follow-up to observe treatment outcomes. Genomic DNA was extracted from the DBS samples using a QIAamp blood extraction kit. Fragments of P. falciparum kelch13 (Pfkelch13), P. falciparum coronin (Pfcoronin), P. falciparum multidrug resistance 2 (PfMDR2), and P. falciparum chloroquine resistance transporter (PfCRT) genes were amplified and sequenced on a sanger sequencing platform to identify artemisinin resistance-associated mutations. Mutations were identified by aligning sequenced data with reference sequences obtained from the National Center for Biotechnology Information. Data were analyzed using descriptive statistics and student t-tests. Results: Mean parasite clearance time (PCT) and fever clearance time (FCT) were 2.1 ± 0.6 days (95% CI: 1.97-2.24) and 1.3 ± 0.7 days (95% CI: 1.1-1.6) respectively. Four mutations, K189T [34/53(64.2%)], R255K [2/53(3.8%)], K189N [1/53(1.9%)] and N217H [1/53(1.9%)] were identified within the N-terminal (Coiled-coil containing) domain of Pfkelch13. No artemisinin resistance-associated mutation usually found within the β-propeller domain of the Pfkelch13 gene was found in these analyzed samples. However, K189T and R255K mutations showed a significant correlation with longer parasite clearance time in the patients (P<0.002). The observed Pfkelch13 gene changes did not influence the baseline mean parasitemia (P = 0.44). P76S [17/100 (17%)] and V62M [1/100 (1%)] changes were identified in the Pfcoronin gene fragment without any influence on the parasitological parameters. No change was observed in the PfMDR2 gene, while no artemisinin resistance-associated mutation was found in the PfCRT gene. Furthermore, a sample each in the retrospective study contained the Pfkelch13 K189T and Pfcoronin P76S mutations. Conclusion: The study revealed absence of genetic-based evidence of artemisinin resistance in the study population at the time of study. The high frequency of K189T Pfkelch13 mutation and its correlation with increased parasite clearance time in this study may depict geographical variation of resistance mediators and imminent artemisinin resistance, respectively. The study also revealed an inherent potential of parasites to harbour drug-resistant genotypes before the introduction of ACTs in Nigeria.

Keywords: artemisinin resistance, plasmodium falciparum, Pfkelch13 mutations, Pfcoronin

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Authors: Himalaya Patir, Bitupon Baruah, Sanjay Gayary, Subhajit Ray

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In recent age significant importance is given for the development of nutritious and health beneficial foods. Fruit juices collected from different fruits when blended that improves not only the physicochemical and nutritional properties but also enhance the sensorial or organoleptic properties. The study was carried out to determine the physico-chemical, nutritional, microbiological analysis and sensory evaluation of mixed fruit juice blend. Juice of orange (Citrus sinensis), apple (Malus domestica), mosambi (Citrus limetta) were blended in the ratio of sample-I (30% apple:30% orange:40% mosambi), sample-II ( 40% apple :30% orange :30% mosambi), sample-III (30% apple :40% orange :30% mosambi) , sample-IV (50% apple :30% orange :20% mosambi), sample-V (30% apple:20% orange:50% mosambi), sample-VI (20% apple :50% orange :30% mosambi) to evaluate all quality characteristics. Their colour characteristics in terms of hue angle, chroma and colour difference (∆E) were evaluated. The physico-chemical parameters analysis carried out were total soluble solids (TSS), total titratable acidity (TTA), pH, acidity (FA), volatile acidity (VA), pH, and vitamin C. There were significant differences (p˂0.05) in the TSS of the samples. However, sample-V (30% apple: 20% orange: 50% mosambi) provides the highest TSS of 9.02gm and significantly differed from other samples (p˂0.05). Sample-IV (50% apple: 30% orange: 20% mosambi) was shown the highest titratable acidity (.59%) in comparison to other samples. The highest value of pH was found as 5.01 for sample-IV (50% apple: 30% orange: 20% mosambi). Sample-VI (20% apple: 50% orange :30% mosambi) blend has the highest hue angle, chroma and colour changes of 72.14,25.29 and 54.48 and vitamin C, i.e. Ascorbic acid (.33g/l) content compared to other samples. The nutritional compositions study showed that, sample- VI (20% apple: 50% orange: 30% mosambi) has the significantly higher carbohydrate (51.67%), protein (.78%) and ash (1.24%) than other samples, while sample-V (30% apple: 20% orange: 50% mosambi) has higher dietary fibre (12.84%) and fat (2.82%) content. Microbiological analysis of all samples in terms of total plate count (TPC) ranges from 44-60 in 101 dilution and 4-5 in 107 dilutions and was found satisfactory. Moreover, other pathogenic bacterial count was found nil. The general acceptability of the mixed fruit juice blend samples were moderately liked by the panellists, and sensorial quality studies showed that sample-V (30% apple: 20% orange: 50% mosambi) contains highest overall acceptability of 8.37 over other samples and can be considered good for consumption.

Keywords: microbiological, nutritional, physico-chemical, sensory properties

Procedia PDF Downloads 178

Authors: Prashant Gupta

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Throughout India, temples have served as cultural centers, commerce hubs, art galleries, educational institutions, and social centers in addition to being places of worship since centuries. Across the country, there are over two million temples, which are crucial economic hubs, attracting devotees and tourists worldwide. In India, we have 53 temples per each 100,000 Indians. As per NSSO survey, the temple economy is worth about $40 billion and 2.32 per cent of GDP based on major temple’s survey, which only includes formal sector. It could be much larger as an actual estimation has not been done yet. In India, 43.1% of total economy represents informal sector. Over 10 billion domestic tourists visit to new destinations every year within India. Even 20 per cent of the 90 million foreign tourists visited Madurai and Mahabalipuram temples which became the most visited tourist spot in 2022. Recently the current central government in power have started revitalizing the ancient Indian civilization by reconstructing and beautifying the major temples of India i.e., Kashi Vishwanath Corridor, Mahakaleshwara Temple, Kedarnath, Ayodhya etc. The reason researcher chose Kashi as a case study because it is known as a Spiritual Capital of India, which is also the abode for the spread of Hinduism, Buddhism, Jainism and Sikkism, which are core Sanatan Dharmic practices. 17,800 Million INR Amount was spend to redevelop Kashi Vishwanath Corridor since 2019. RESEARCH OBJECTIVES 1. To assess historical contribution of temples in socio economic development and revival of Indic Civilization. 2. To examine the role of temples redevelopment for informal sector businesses. 3. To identify the sub-sectors of informal sector businesses 4. To identify products and services of informal businesses for investigation of marketing strategies and business development. PROPOSED METHODS AND PROCEDURES This study will follow a mixed approach, employing both qualitative and quantitative methods of research. To conduct the study, data will be collected from 500 informal business owners through structured questionnaire and interview instruments. The informal business owners will be selected using a systematic random sampling technique. In addition, documents from government offices of the last 10 years of tax collection will be reviewed to substantiate the study. To analyze the study, descriptive and econometric analysis techniques will be employed. EXPECTED CONTRIBUTION OF THE PROPOSED STUDY By studying the contribution of temple re-development on informal business creation and growth, the study will be beneficial to the informal business owners and the government. For the government, scientific and empirical evidence on the contribution of temple re-development for informal business creation and growth to give evidence the study will give based infrastructural development and boosting tax collection. For informal businesses, the study will give them a detailed insight on the nature of their business and the possible future growth potential of their business, and the alternative products and services supplying to their customers in the future. Studying informal businesses will help to identify the key products and services which are majorly profitable and possess potential to multiply and grow through correct product marketing strategies and business development.

Keywords: business development, informal sector businesses, services and products marketing, temple economics

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Authors: Ade O. Oyewole, Sunday O. Okoh, Ruth O. Ishola, Adenike D. Odusote, Chima C. Igwe, Gloria N. Elemo, Anthony I. Okoh

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Azadirachta indica (Neem tree) also referred to as an all-purpose tree is used in a wide range of medical preparations in tropical and subtropical countries for prevention and management of various livestock, crops products and human diseases. In Nigeria however, the potentials of this plant have not been fully exploited thus it causes an environmental nuisance during the fruiting season. With a rise in the demand for herbal personal care products globally extracts from different parts of the neem plant were used as the bio-active ingredients in the formulation of personal care products. In this study, formulated neem soap, body cream, lotion, toothpaste and shampoo are analyzed to determine their antibacterial, antifungal, and toxicity properties. The efficacies of these products for management of infectious diseases, both oral and dermal, were also investigated in vitro. Oil from the neem seeds obtained using a mechanical press and acetone extracts of both the neem bark and leaves obtained by the maceration method were used in the formulation and production of the neem personal care products. The antimicrobial and toxicity properties of these products were investigated by agar diffusion, and haemolytic methods respectively. The five neem products (NPs) exhibited strong antibacterial activities against four multi–drug resistant pathogenic and three none pathogenic bacterial strains (Escherichia coli (180), Listeria ivanovii, Staphylococcus aureus, Enterobacter cloacae, Vibro spp., Streptococcus uberis, Mycobacterium smegmatis), except the neem lotion with insignificant activity against E. coli and S. aureus. The minimum inhibitory concentration (MIC) range was between 0.20-0.40 mg/ mL. The 5 NPs demonstrated moderate activity against three clinical dermatophytes isolates (Tinea corporis, Tinea capitis, and Tinea cruiz) as well as one fungal strain (Candida albican) with the MIC ranging between 0.30 - 0.50 mg/ mL and 0.550 mg/mL respectively. The soap and shampoo were the most active against test bacteria and fungi. The haemolytic analysis results on the 5 NPs indicated none toxicity at 0.50 mg/ mL in sheep red blood cells (SRBC).

Keywords: antimicrobial, Azadirachta indica, multi–drug resistant pathogenic bacteria, personal care products

Procedia PDF Downloads 270

Authors: F. Ainlhout, S. Boutaleb, M. C. Diaz-Barradas, M. Zunzunegui

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Argania spinosa is an endemic tree of the southwest of Morocco, occupying 828,000 Ha, distributed mainly between Mediterranean vegetation and the desert. This tree can grow in extremely arid regions in Morocco, where annual rainfall ranges between 100-300 mm where no other tree species can live. It has been designated as a UNESCO Biosphere reserve since 1998. Argania tree is of great importance in human and animal feeding of rural population as well as for oil production, it is considered as a multi-usage tree. Admine forest located in the suburbs of Agadir city, 5 km inland, was selected to conduct this work. The aim of the study was to investigate the temporal variation in root-zone moisture dynamic in response to variation in climatic conditions and vegetation water uptake, using a geophysical technique called Electrical resistivity imaging (ERI). This technique discriminates resistive woody roots, dry and moisture soil. Time-dependent measurements (from April till July) of resistivity sections were performed along the surface transect (94 m Length) at 2 m fixed electrode spacing. Transect included eight Argan trees. The interactions between the tree and soil moisture were estimated by following the tree water status variations accompanying the soil moisture deficit. For that purpose we measured midday leaf water potential and relative water content during each sampling day, and for the eight trees. The first results showed that ERI can be used to accurately quantify the spatiotemporal distribution of root-zone moisture content and woody root. The section obtained shows three different layers: middle conductive one (moistured); a moderately resistive layer corresponding to relatively dry soil (calcareous formation with intercalation of marly strata) on top, this layer is interspersed by very resistant layer corresponding to woody roots. Below the conductive layer, we find the moderately resistive layer. We note that throughout the experiment, there was a continuous decrease in soil moisture at the different layers. With the ERI, we can clearly estimate the depth of the woody roots, which does not exceed 4 meters. In previous work on the same species, analyzing the δ18O in water of xylem and in the range of possible water sources, we argued that rain is the main water source in winter and spring, but not in summer, trees are not exploiting deep water from the aquifer as the popular assessment, instead of this they are using soil water at few meter depth. The results of the present work confirm the idea that the roots of Argania spinosa are not growing very deep.

Keywords: Argania spinosa, electrical resistivity imaging, root system, soil moisture

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Authors: Abejide Dorcas Ropo, , Falusi Olamide Ahmed, Daudu Oladipupo Abdulazeez Yusuf, Salihu Bolaji Zuluquri Neen, Muhammad Liman Muhammad, Gado Aishatu Adamu

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Water stress is a major factor limiting the productivity of crops in the world today. This study evaluated the morpho-agronomic response of twenty-four (24) Nigerian Bambara groundnut landraces to water stress and genetic diversity of some selected accessions using SSR markers. The studies was carried out in the Botanical garden of the Department of Plant Biology, Federal University of Technology, Minna, Niger State, Nigeria in a randomized complete block design using three replicates. Molecular analysis using SSR primers was carried out at the Centre for Bio- Science, International Institute of Tropical Agriculture (IITA) Ibadan, Nigeria in order to characterize ten selected accessions comprising of the seven most drought tolerant and the three most susceptible accessions detected from the morpho-agronomic studies. Results revealed that water stress decreased morpho-agronomic traits such as plant height, leaf area, number of leaves per plant and seed yield etc. A total of 22 alleles were detected by the SSR markers used with a mean number of 4 allelles. Simple Sequence Repeat (SSR) markers MBamCO33, Primer 65 and G358B2-D15 each detected 4 allelles while Primer 3FR and 4FR detected 5 allelles each. The study revealed significantly high polymorphisms in 10 Loci. The mean value of Polymorpic information content was 0.6997 implying the usefulness of the primers used in identifying genetic similarities and differences among the Bambara groundnut genotypes. The SSR analysis revealed a comparable pattern between genetic diversity and drought tolerance of the genotypes. The Unweighted Paired Group Method with Arithmethic Mean (UPGMA) dendrogram showed that at a genetic distance of 0.1, the accessions were grouped into three groups according to their level of tolerance to drought. The two most drought tolerant accessions were grouped together and the 5th and 6th most drought tolerant accession were also grouped together. This suggests that the genotypes grouped together may be genetically close, may possess similar genes or have a common origin. The degree of genetic variants obtained could be useful in bambara groundnut breeding for drought tolerance. The identified drought tolerant bambara groundnut landraces are important genetic resources for drought stress tolerance breeding programme of bambara groundnut. The genotypes are also useful for germplasm conservation and global implications.

Keywords: bambara groundnut, genetic diversity, germplasm, SSR markers, water stress

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Authors: Fatima Bouazza, Khadija Khedid, Lamiae Amallah, Aziz Mouhaddach, Basma Boukour, Jihane Ennadir, Rachida Hassikou

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In order to combine milk and its derived products conservation and flavoring, Moroccans often used aromatic and medicinal plants. These plant extracts are endowed with several nutritive and therapeutic properties. This study constitutes a first national assessment of physico-chemical quality of sheep’s milk from moroccan Sardi breed and the evaluation of the antibacterial effect of three medicinal plants extracts: Aloe barbadensis Miller, Thymus satureioides and Mentha pulegium on flora isolated from this sheep's milk. 100 milk samples were collected in four regions of Morocco. The bacteria isolated were identified by classical and molecular methods (16S rRNA sequencing) and tested, according to the disk method, for their sensitivity to several antibiotics. The physico-chemical analyzes of sheep’s milk concerned the pH, titratable acidity, density, dry extract, freezing point and contents of: fat, proteins, lactose and calcium. The essential oils (EOs) of T. satureioides and M .pulegium were extracted by hydrodistillation and analyzed by GC / MS, while the Aloe vera leaf pulp was analyzed by the methods of Harborne and HPLC. A total number of 125 bacteria have been identified. Significant resistance to chemical antibiotics has been noted in LABs. The average temperature value of milk is around 57.15 °C, the pH is 6.56, the titratable acidity is around 3.4 ° D, the density is 1.035g / cm³ , the total dry extract is around 169.5g / l, the ash (9.8g / l), the freezing point (- 0.556 °C) while the average fat content is 67.85g / l . The samples richest in fat belong to the region of Settat, cradle of the Sardi breed, with a maximum average value of 74.4g / l. The average protein is 56g / l, lactose (39.92g / l), and calcium (1.855g / l). Analysis of the major components of EOs revealed the dominance of borneol in the case of T. satureioides and of pulegone in M. pulegium. Aloe vera gel contains alkaloids, flavonoids, catechic tannins, saponins and 1.60 µg / ml of aloin. The plant extracts have a bactericidal effect on E. coli, Klebsiellaoxytoca and Staphylococci and bacteriostatic effect on LABs of technological interest (Lactobacillus). As a result of this study, it is believed that the consumption of sardi sheep’s milk would be of nutritional benefit. Its richness in fat and proteins predisposes it for biotechnological development in the manufacture of cheese and yogurt. Also, the use of aromatic and medicinal plants, as natural additives would be of great benefit to flavor and maintain its quality.

Keywords: sheep’s milk, lactic flora, antimicrobial power, aloe barbadensis miller, thymus satureioides, mentha pulegium

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Authors: Portia Murphy, Danica Vasic, Anoja W. Gunaratne, Encarnita Sitchon, Teresita Tugonon, Marou Ison, Antoinette Le Busque, Christelle Pagonis, Thomas J. Borody

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Irritable bowel syndrome (IBS) is a chronic gastrointestinal disorder that affects an estimated 11% of the population globally with the most predominant symptoms being abdominal pain, bloating and altered bowel movements. All age groups suffer from IBS although the prevalence of IBS decreases for age groups over 50 years. Women are more likely to suffer from IBS than men. IBS can be categorized into 3 groups based on the type of altered bowel movement: diarrhea-predominant IBS (IBS-D), constipation-predominant IBS (IBS-C) and IBS with mixed bowel habit (IBS-M). The contribution of the gut microbiome to the etiology of IBS is becoming increasingly recognized with rising use of anti-microbial agents. Previous studies on vancomycin and rifaximin used as monotherapy or in combination have been conducted mainly on IBS-C and showed marked improvements in the symptoms. According to our knowledge, no studies reported using these two combinations of antibiotics for IBS-D. Here, we report a consecutive cohort of 18 patients treated with both vancomycin and rifaximin for IBS-D. These patients’ records were reviewed retrospectively. In this cohort, patients ages were between 24-74 years (mean 44 years) and 9 were female. Baseline all patients had diarrhea, 4 with mucus and one with blood. Patients reported other symptoms were abdominal pain (n=11) bloating (n=9), flatulence (n=7), fatigue (n=4) and nausea (n=3). Patients treatments were personalized according to their symptom severity and tolerability and were treated with combination of rifaximin (500 - 3000mg/d) and vancomycin (500mg - 1500mg/d) for an ongoing period. Follow-ups were conducted between 2-32 weeks’ time. Of all patients, 89% patients reported improvement of the symptoms, 1 reported no change and 1 patient’s symptoms got worse. The mechanism of action for both vancomycin and rifaximin involves the inhibition of bacterial cell wall and protein synthesis respectively. The role of these medications in improving the symptoms of this cohort suggests that IBS-D may be microbiome infection driven. In this cohort, similar patient presentations to Clostridium difficile, as well as symptom improvement with the use of rifaximin and particularly vancomycin, suggest that the infectious agent may be an unidentified Clostridium. These preliminary results offer an alternative etiology for IBS-D not previously considered and open the avenue for new research.

Keywords: clostridium deficile, diarrhea predominant Irritable Bowel Syndrome, microbiome, vancomycin/rifaximin combination

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Authors: Swati Tomar, Sunil Kumar Gupta

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Anammox is a promising and cost effective alternative to conventional treatment systems that facilitates direct oxidation of ammonium nitrogen under anaerobic conditions with nitrite as an electron acceptor without addition of any external carbon sources. The present study investigates the process kinetics of laboratory scale anammox hybrid reactor (AHR) which combines the dual advantages of attached and suspended growth. The performance & behaviour of AHR was studied under varying hydraulic retention time (HRTs) and nitrogen loading rate (NLRs). The experimental unit consisted of 4 numbers of 5L capacity anammox hybrid reactor inoculated with mixed seed culture containing anoxic and activated sludge. Pseudo steady state (PSS) ammonium and nitrite removal efficiencies of 90.6% and 95.6%, respectively, were achieved during acclimation phase. After establishment of PSS, the performance of AHR was monitored at seven different HRTs of 3.0, 2.5, 2.0, 1.5, 1.0, 0.5 and 0.25 d with increasing NLR from 0.4 to 4.8 kg N/m3d. The results showed that with increase in NLR and decrease in HRT (3.0 to 0.25 d), AHR registered appreciable decline in nitrogen removal efficiency from 92.9% to 67.4 %, respectively. The HRT of 2.0 d was considered optimal to achieve substantial nitrogen removal of 89%, because on further decrease in HRT below 1.5 days, remarkable decline in the values of nitrogen removal efficiency were observed. Analysis of data indicated that attached growth system contributes an additional 15.4 % ammonium removal and reduced the sludge washout rate (additional 29% reduction). This enhanced performance may be attributed to 25% increase in sludge retention time due to the attached growth media. Three kinetic models, namely, first order, Monod and Modified Stover-Kincannon model were applied to assess the substrate removal kinetics of nitrogen removal in AHR. Validation of the models were carried out by comparing experimental set of data with the predicted values obtained from the respective models. For substrate removal kinetics, model validation revealed that Modified Stover-Kincannon is most precise (R2=0.943) and can be suitably applied to predict the kinetics of nitrogen removal in AHR. Lawrence and McCarty model described the kinetics of bacterial growth. The predicted value of yield coefficient and decay constant were in line with the experimentally observed values.

Keywords: anammox, kinetics, modelling, nitrogen removal, sludge wash out rate, AHR

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Authors: Michelle Themalil, Celia Bueno, Rulla Al- Araji

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Objective and Study: There are no established guidelines for antibiotic prophylaxis in children with central venous catheters (CVCs) on home parenteral nutrition (HPN), leading to varying practices across UK Centres. We hypothesize that children with intestinal failure are at increased risk for bacteraemia due to altered anatomy, dysmotility, inflammation, biofilm formation in long-term CVCs, and the use of central lines during procedures. Given the bacteraemia rates of up to 8% in upper and 25% in lower endoscopy for adults without central lines, we argue that prophylactic antibiotics are reasonable, given the increased risks faced by this high-risk group of children. Methods: We conducted a five-year review of patients with central venous access receiving home parenteral nutrition (HPN) who underwent endoscopies with antibiotic prophylaxis at our center (tertiary). We documented and analyzed post-procedure infections and their associated risk factors. Results: A total of 15 patients on HPN underwent 29 endoscopic procedures, including 4 upper, 9 combined upper and lower, and 16 combined upper, lower, and ileoscopy. Confirmed infection rates remained at 0% up to 28 days post-procedure. The agreed-upon prophylaxis regimen was implemented, with ciprofloxacin and metronidazole administered as the primary antibiotics. Notably, only 51.7% of patients received a peripheral cannula despite recommendations to avoid central line use during anesthesia, and 20.6% had small intestinal bacterial overgrowth. Conclusions: This study is the first to investigate post-endoscopy infection rates in pediatric patients on HPN. Despite a small sample size, we observed a 0% infection rate, significantly lower than reported rates in adults. These findings suggest that further research is warranted to explore the implications of antibiotic prophylaxis in this unique patient cohort and to establish guidelines that may enhance patient safety during endoscopic procedures.

Keywords: post endosopy infections, central venous access, home parenteral nutrition, intestinal failure

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Authors: Otuwose E. Agyeno, Adeniyi A. Jayeola, Bashir A. Ajala

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P. esculentus is indigenous to Nigeria yet no wild relation has been encountered or reported. This has made it difficult to establish proper lineages between the varieties and landraces under cultivation. The present work is the first to determine the apormophy of 135 morphoanatomical characters in organs of 46 accessions drawn from 23 populations of this species based on dicta. The character states were coded in accession x character-state matrices and only 83 were informative and utilised for neighbour joining clustering based on euclidean values, and heuristic search in parsimony analysis using PAST ver. 3.15 software. Compatibility and evolutionary trends between accessions were then explored from values and diagrams produced. The low consistency indices (CI) recorded support monophyly and low homoplasy in this taxon. Agglomerative schedules based on character type and source data sets divided the accessions into mainly 3 clades, each of complexes of accessions. Solenostemon rotundifolius (Poir) J.K Morton was the outgroup (OG) used, and it occurred within the largest clades except when the characters were combined in a data set. The OG showed better compatibility with accessions of populations of landrace Isci, and varieties Riyum and Long’at. Otherwise, its aerial parts are more consistent with those of accessions of variety Bebot. The highly polytomous clades produced due to anatomical data set may be an indication of how stable such characters are in this species. Strict consensus trees with more than 60 nodes outputted showed that the basal nodes were strongly supported by 3 to 17 characters across the data sets, suggesting that populations of this species are more alike. The OG was clearly the first diverging lineage and closely related to accessions of landrace Gwe and variety Bebot morphologically, but different from them anatomically. It was also distantly related to landrace Fina and variety Long’at in terms of root, stem and leaf structural attributes. There were at least 5 other clades with each comprising of complexes of accessions from different localities and terrains within the study area. Spherical stem in cross section, size of vascular bundles at the stem corners as well as the alternate and whorl phyllotaxy are attributes which may have facilitated each other’s evolution in all accessions of the landrace Gwe, and they may be innovative since such states are not characteristic of the larger Lamiaceae, and Plectranthus L’Her in particular. In conclusion, this study has provided valuable information about infraspecific diversity in this taxon. It supports recognition of the varietal statuses accorded to populations of P. esculentus, as well as the hypothesis that the wild gene might have been distributed on the Jos Plateau. However, molecular characterisation of accessions of populations of this species would resolve this problem better.

Keywords: clustering, lineage, morphoanatomical characters, Nigeria, phylogenetics, Plectranthus esculentus, population

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Authors: Simona Dostalova, Dita Munzova, Ana Maria Jimenez Jimenez, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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The boom of nanomedicine in recent years has led to the development of numerous new nanomaterials that can be used as nanocarriers in the drug delivery. These nanocarriers can either be synthetic or natural-based. The disadvantage of many synthetic nanocarriers is their toxicity in patient’s body. Protein cages that can naturally be found in human body do not exhibit such disadvantage. However, the release of cargo from some protein cages in target cells can be problematic. As a special type of protein cages can serve the capsid of many viruses, including phage. Phages infect bacterial cells; therefore they are not harmful to human cells. The targeting of phage particles to cancer cells can be solved by producing of empty phage capsids during which the targeting moieties (e.g. peptides) can be cloned into genes of phage capsid to decorate its surface. Moreover, the produced capsids do not contain viral nucleic acid and are therefore not infectious to beneficial bacteria in the patient’s body. The protein cage composed of viral capsid is larger than other frequently used apoferritin cage but its size is still small enough to benefit from passive targeting by Enhanced Permeability and Retention effect. In this work, bacteriophage λ was used, both whole and its empty capsid for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin, etoposide and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2 % maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000 g and 4 °C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 µg/ml). Free molecules of drugs were removed by dialysis. The encapsulation was verified using spectrophotometric and electrochemical methods. The amount of encapsulated drug linearly increased with the amount of applied drug (determination coefficient R2=0.8013). 76% of applied drug was encapsulated in phage λ particles (concentration of 10 µg/ml), even with the highest applied concentration of drugs, 200 µg/ml. Only 1% of encapsulated drug was detected in phage DNA. Similar results were obtained with encapsulation in phage empty capsid. Therefore, it can be concluded that the encapsulation of drugs into phage particles is efficient and mostly occurs by interaction of drugs with protein capsid.

Keywords: cytostatics, drug delivery, nanocarriers, phage capsid

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Authors: Ane Escobar, Paula Angelomé, Mihaela Delcea, Marek Grzelczak, Sergio Enrique Moya

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The antibacterial capacity of bone-anchoring implants can be improved by the use of antibiotics that can be delivered to the media after the surgery. Mesoporous films have shown great potential in drug delivery for orthopedic applications, since pore size and thickness can be tuned to produce different surface area and free volume inside the material. This work shows the synthesis of mesoporous titania films (MTF) by sol-gel chemistry and evaporation-induced self-assembly (EISA) on top of glass substrates. Pores with a diameter of 12nm were observed by Transmission Electron Microscopy (TEM). A film thickness of 100 nm was measured by Scanning Electron Microscopy (SEM). Gentamicin was used to study the antibiotic delivery from the film by means of High-performance liquid chromatography (HPLC). The Staphilococcus aureus strand was used to evaluate the effectiveness of the penicillin loaded films toward inhibiting bacterial colonization. MC3T3-E1 pre-osteoblast cell proliferation experiments proved that MTFs have a good biocompatibility and are a suitable surface for MC3T3-E1 cell proliferation. Moreover, images taken by Confocal Fluorescence Microscopy using labeled vinculin, showed good adhesion of the MC3T3-E1 cells to the MTFs, as well as complex actin filaments arrangement. In order to improve cell proliferation Bone Morphogenetic Protein-2 (BMP-2) was adsorbed on top of the mesoporous film. The deposition of the protein was proved by measurements in the contact angle, showing an increment in the hydrophobicity while the protein concentration is higher. By measuring the dehydrogenase activity in MC3T3-E1 cells cultured in dually functionalized mesoporous titatina films with gentamicin and BMP-2 is possible to find an improvement in cell proliferation. For this purpose, the absorption of a yellow-color formazan dye, product of a water-soluble salt (WST-8) reduction by the dehydrogenases, is measured. In summary, this study proves that by means of the surface modification of MTFs with proteins and loading of gentamicin is possible to achieve an antibacterial effect and a cell growth improvement.

Keywords: antibacterial, biocompatibility, bone morphogenetic protein-2, cell proliferation, gentamicin, implants, mesoporous titania films, osteoblasts

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Authors: Yu-Wen Lin, Chin-Sheng Tang, Wan-Yi Chen

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Various biological specimens, drugs, and chemicals exist in the hospital. The medical staffs and hypersensitive inpatients expose might expose to multiple hazards while they work or stay in the hospital. Therefore, the indoor air quality (IAQ) of the hospital should be paid more attention. Respiratory care wards (RCW) are responsible for caring the patients who cannot spontaneously breathe without the ventilators. The patients in RCW are easy to be infected. Compared to the bacteria concentrations of other hospital units, RCW came with higher values in other studies. This research monitored the IAQ of the RCW and checked the compliances of the indoor air quality standards of Taiwan Indoor Air Quality Act. Meanwhile, the influential factors of IAQ and the impacts of ventilator modules, with humidifier or with filter, were investigated. The IAQ of two five-bed wards and one nurse station of a RCW in a regional hospital were monitored. The monitoring was proceeded for 16 hours or 24 hours during the sampling days with a sampling frequency of 20 minutes per hour. The monitoring was performed for two days in a row and the AIQ of the RCW were measured for eight days in total. The concentrations of carbon dioxide (CO₂), carbon monoxide (CO), particulate matter (PM), nitrogen oxide (NOₓ), total volatile organic compounds (TVOCs), relative humidity (RH) and temperature were measured by direct reading instruments. The bioaerosol samples were taken hourly. The hourly air change rate (ACH) was calculated by measuring the air ventilation volume. Human activities were recorded during the sampling period. The linear mixed model (LMM) was applied to illustrate the impact factors of IAQ. The concentrations of CO, CO₂, PM, bacterial and fungi exceeded the Taiwan IAQ standards. The major factors affecting the concentrations of CO, PM₁ and PM₂.₅ were location and the number of inpatients. The significant factors to alter the CO₂ and TVOC concentrations were location and the numbers of in-and-out staff and inpatients. The number of in-and-out staff and the level of activity affected the PM₁₀ concentrations statistically. The level of activity and the numbers of in-and-out staff and inpatients are the significant factors in changing the bacteria and fungi concentrations. Different models of the patients’ ventilators did not affect the IAQ significantly. The results of LMM can be utilized to predict the pollutant concentrations under various environmental conditions. The results of this study would be a valuable reference for air quality management of RCW.

Keywords: respiratory care ward, indoor air quality, linear mixed model, bioaerosol

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Authors: Kichan Lee, Kwang-Ho Choi, Mi-Hye Hwang, Young Min Son, Bang-Hun Hyun, Byeong Yeal Jung

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Recently, worldwide food safety authorities have been strengthening food hygiene in order to curb foodborne illness outbreaks. The hygiene status of Korean slaughterhouses has been monitored annually by Animal and Plant Quarantine Agency and provincial governments through foodborne pathogens investigation using slaughtered pig and cattle meats. This study presented the prevalence of food-borne pathogens from 2014 to 2016 in Korean slaughterhouses. Sampling, microbiological examinations, and analysis of results were performed in accordance with ‘Processing Standards and Ingredient Specifications for Livestock Products’. In total, swab samples from 337 pig carcasses (100 samples in 2014, 135 samples in 2015, 102 samples in 2016) and 319 cattle carcasses (100 samples in 2014, 119 samples in 2015, 100 samples in 2016) from twenty slaughterhouses were examined for Listeria monocytogenes, Campylobacter jejuni, Campylobacter coli, Salmonella spp., Staphylococcus aureus, Clostridium perfringens, Yersinia enterocolitica, Escherichia coli O157:H7 and non-O157 enterohemorrhagic E. coli (EHEC, serotypes O26, O45, O103, O104, O111, O121, O128 and O145) as foodborne pathogens. The samples were analyzed using cultural and PCR-based methods. Foodborne pathogens were isolated in 78 (23.1%) out of 337 pig samples. In 2014, S. aureus (n=17) was predominant, followed by Y. enterocolitica (n=7), C. perfringens (n=2) and L. monocytogenes (n=2). In 2015, C. coli (n=14) was the most prevalent, followed by L. monocytogenes (n=4), S. aureus (n=3), and C. perfringens (n=2). In 2016, S. aureus (n=16) was the most prevalent, followed by C. coli (n=13), L. monocytogenes (n=2) and C. perfringens (n=1). In case of cattle carcasses, foodborne bacteria were detected in 41 (12.9%) out of 319 samples. In 2014, S. aureus (n=16) was the most prevalent, followed by Y. enterocolitica (n=3), C. perfringens (n=3) and L. monocytogenes (n=2). In 2015, L. monocytogenes was isolated from 4 samples, S. aureus from three, C. perfringens, Y. enterocolitica and Salmonella spp. from one, respectively. In 2016, L. monocytogenes (n=6) was the most prevalent, followed by C. perfringens (n=3) C. jejuni (n=1), respectively. It was found that 10 carcass samples (4 cattle and 6 pigs) were contaminated with two bacterial pathogen tested. Interestingly, foodborne pathogens were more detected from pig carcasses than cattle carcasses. Although S. aureus was predominantly detected in this study, other foodborne pathogens were also isolated in slaughtered meats. Results of this study alerted the risk of foodborne pathogen infection for humans from slaughtered meats. Therefore, the authors insisted that it was important to enhance hygiene level of slaughterhouses according to Hazard Analysis and Critical Control Point.

Keywords: carcass, cattle, foodborne, Korea, pathogen, pig

Procedia PDF Downloads 344

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

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

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

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Authors: Wei Ji

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This study was conducted in the Kansas City metropolitan area of the United States, which has experienced significant urban sprawling in recent decades. The remote sensing of land cover changes in this area spanned over four decades from 1972 through 2010. The project was implemented in two stages: the first stage focused on detection of long-term trends of urban land cover change, while the second one examined how to detect the coupled effects of human impact and climate change on urban landscapes. For the first-stage study, six Landsat images were used with a time interval of about five years for the period from 1972 through 2001. Four major land cover types, built-up land, forestland, non-forest vegetation land, and surface water, were mapped using supervised image classification techniques. The study found that over the three decades the built-up lands in the study area were more than doubled, which was mainly at the expense of non-forest vegetation lands. Surprisingly and interestingly, the area also saw a significant gain in surface water coverage. This observation raised questions: How have human activities and precipitation variation jointly impacted surface water cover during recent decades? How can we detect such coupled impacts through remote sensing analysis? These questions led to the second stage of the study, in which we designed and developed approaches to detecting fine-scale surface waters and analyzing coupled effects of human impact and precipitation variation on the waters. To effectively detect urban landscape changes that might be jointly shaped by precipitation variation, our study proposed “urban wetscapes” (loosely-defined urban wetlands) as a new indicator for remote sensing detection. The study examined whether urban wetscape dynamics was a sensitive indicator of the coupled effects of the two driving forces. To better detect this indicator, a rule-based classification algorithm was developed to identify fine-scale, hidden wetlands that could not be appropriately detected based on their spectral differentiability by a traditional image classification. Three SPOT images for years 1992, 2008, and 2010, respectively were classified with this technique to generate the four types of land cover as described above. The spatial analyses of remotely-sensed wetscape changes were implemented at the scales of metropolitan, watershed, and sub-watershed, as well as based on the size of surface water bodies in order to accurately reveal urban wetscape change trends in relation to the driving forces. The study identified that urban wetscape dynamics varied in trend and magnitude from the metropolitan, watersheds, to sub-watersheds in response to human impacts at different scales. The study also found that increased precipitation in the region in the past decades swelled larger wetlands in particular while generally smaller wetlands decreased mainly due to human development activities. These results confirm that wetscape dynamics can effectively reveal the coupled effects of human impact and climate change on urban landscapes. As such, remote sensing of this indicator provides new insights into the relationships between urban land cover changes and driving forces.

Keywords: urban land cover, human impact, climate change, rule-based classification, across-scale analysis

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Authors: G. A. Gevorgyan, A. S. Mamyan, L. G. Stepanyan, L. R. Hambaryan

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Bacterio- and phytoplankton growth rates in 'Yerevanyan lich' reservoir and the Hrazdan river in Yerevan city, Armenia were investigated in April and June-August, 2015. Phytoplankton sampling and analysis were performed by the standard methods accepted in hydrobiological studies. The quantitative analysis of aerobic, coliform and E. coli bacteria is done by the 'RIDA COUNT' medium sheets (coated with ready-to-use culture medium). The investigations showed that the insufficient management of household discharges in Yerevan city caused the organic and fecal pollution of the Hrazdan river in this area which in turn resulted in an increase in bacterial count and increased sanitary and pathogenic risks to the environment and human health. During the investigation in April, the representatives of diatom algae prevailed quantitatively in the coastal area of 'Yerevanyan lich' reservoir, nevertheless, a significant change in the phytoplankton community in June occurred: due to green algae bloom in the reservoir, the quantitative parameters of phytoplankton increased significantly. This was probably conditioned by a seasonal increase in the water temperature in the conditions of the sufficient concentration of nutrients. However, a succession in phytoplankton groups during July-August occurred, and a dominant group (according to quantitative parameters) in the phytoplankton community was changed as follows: green algae-diatom algae-blue-green algae. Rapid increase in the quantitative parameters of diatom and blue-green algae in the reservoir may have been conditioned by increased organic matter level resulted from green algae bloom. Algal bloom in 'Yerevanyan lich' reservoir caused changes in phytoplankton community and an increase in bacterioplankton count not only in the reservoir but also in the Hrazdan river sites located in the downstream from the reservoir. Thus, the insufficient management of urban discharges and aquatic ecosystems in Yerevan city led to unfavorable changes in water quality and microbial and phytoplankton communities in “Yerevanyan lich” reservoir and the Hrazdan river which in turn caused increased sanitary and pathogenic risks to the environment and human health.

Keywords: algal bloom, bacterioplankton, phytoplankton, Hrazdan river, Yerevanyan lich reservoir

Procedia PDF Downloads 275

Authors: Temesgen Sidamo, Prakruti S. Rao, Eleni Akllilu, Workineh Shibeshi, Yumi Park, Yong-Soon Cho, Jae-Gook Shin, Scott K. Heysell, Stellah G. Mpagama, Ephrem Engidawork

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The fluoroquinolones (FQs) are used off-label for the treatment of multidrug-resistant tuberculosis (MDR-TB), and for evaluation in shortening the duration of drug-susceptible TB in recently prioritized regimens. Within the class, levofloxacin (LFX) and moxifloxacin (MXF) play a substantial role in ensuring success in treatment outcomes. However, sub-therapeutic plasma concentrations of either LFX or MXF may drive unfavorable treatment outcomes. To the best of our knowledge, the pharmacokinetics of LFX and MXF in Ethiopian patients with MDR-TB have not yet been investigated. Therefore, the aim of this study was to develop a population pharmacokinetic (PopPK) model of levofloxacin (LFX) and moxifloxacin (MXF) and assess the percent probability of target attainment (PTA) as defined by the ratio of the area under the plasma concentration-time curve over 24-h (AUC0-24) and the in vitro minimum inhibitory concentration (MIC) (AUC0-24/MIC) in Ethiopian MDR-TB patients. Steady-state plasma was collected from 39 MDR-TB patients enrolled in the programmatic treatment course and the drug concentrations were determined using optimized liquid chromatography-tandem mass spectrometry. In addition, the in vitro MIC of the patients' pretreatment clinical isolates was determined. PopPK and simulations were run at various doses, and PK parameters were estimated. The effect of covariates on the PK parameters and the PTA for maximum mycobacterial kill and resistance prevention was also investigated. LFX and MXF both fit in a one-compartment model with adjustments. The apparent volume of distribution (V) and clearance (CL) of LFX were influenced by serum creatinine (Scr), whereas the absorption constant (Ka) and V of MXF were influenced by Scr and BMI, respectively. The PTA for LFX maximal mycobacterial kill at the critical MIC of 0.5 mg/L was 29%, 62%, and 95% with the simulated 750 mg, 1000 mg, and 1500 mg doses, respectively, whereas the PTA for resistance prevention at 1500 mg was only 4.8%, with none of the lower doses achieving this target. At the critical MIC of 0.25 mg/L, there was no difference in the PTA (94.4%) for maximum bacterial kill among the simulated doses of MXF (600 mg, 800 mg, and 1000 mg), but the PTA for resistance prevention improved proportionately with dose. Standard LFX and MXF doses may not provide adequate drug exposure. LFX PopPK is more predictable for maximum mycobacterial kill, whereas MXF's resistance prevention target increases with dose. Scr and BMI are likely to be important covariates in dose optimization or therapeutic drug monitoring (TDM) studies in Ethiopian patients.

Keywords: population PK, PTA, moxifloxacin, levofloxacin, MDR-TB patients, ethiopia

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Authors: Ibrahim Siddig Hamid, Ikram Mohamed Eltayeb

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Trachyspermum ammi belongs to the family Apiaceae, is used traditionally for the treatment of gastrointestinal ailments, lack of appetite and bronchial problems as well used as antiseptic, antimicrobial, antipyretic, febrifugal and in the treatment of typhoid fever. Peganum harmala belongs to the family Zygophyllaceae it has been reported to have an antibacterial activity and used to treat depression and recurring fevers. It also used to kill algae, bacteria, intestinal parasites and molds. In Sudan, the combination of two plants are traditionally used for the treatment of bacillary dysentery. Bacillary dysentery is caused by one or more types of Shigella species bacteria mainly Shigella dysenteri and shigella flexneri. Bacillary dysentery is mainly found in hot countries like Sudan with poor hygiene and sanitation. Bacillary dysentery causes sudden onset of high fever and chills, abdominal pain, cramps and bloating, urgency to pass stool, weight loss, and dehydration and if left untreated it can lead to serious complications including delirium, convulsions and coma. A serious infection like this can be fatal within 24 hours. The objective of this study is to investigate the in vitro susceptibility of Sh. flexneri and Sh. dysenteriae to the T. ammi and P. harmala. T. ammi and P. harmala were extracted by 96% ethanol using Soxhlet apparatus. The antimicrobial activity of the extracts was investigated according to the disc diffusion method. The discs were prepared by soaking sterilized filter paper discs in 20 microliter of serially diluted solutions of each plant extract with the concentrations (100, 50, 25, 12.5, 6.25mg/dl) then placing them on Muller Hinton Agar plates that were inoculated with bacterial suspension separately, the plates were incubated for 24 hours at 37c and the minimum inhibitory concentration of the extract which was the least concentration of the extract to inhibit fungal growth was determined. The results showed the high antimicrobial activity of T. ammi extract with an average diameter zone ranging from 18-20 mm and its minimum inhibitory concentration was found to be 25 mg/ml against the two shigella species. P. harmala extract was found to have slight antibacterial effect against the two bacteria. This result justified the Sudanese traditional use of Trachyspermum ammi plant for the treatment of bacillary dysentery.

Keywords: harmala, peganum, shigella, trachyspermum

Procedia PDF Downloads 243

Authors: Ghasem Yazdanpanah, Mona Kakavand, Hassan Niknejad

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Objectives: An important issue in tissue engineering (TE) is hemocompatibility. The current engineered vessels are seriously at risk of thrombus formation and stenosis. Amnion (AM) is the innermost layer of fetal membranes that consists of epithelial and mesenchymal sides. It has the advantages of low immunogenicity, anti-inflammatory and anti-bacterial properties as well as good mechanical properties. We recently introduced the amnion as a natural biomaterial for tissue engineering. In this study, we have evaluated hemocompatibility of amnion as potential biomaterial for tissue engineering. Materials and Methods: Amnions were derived from placentas of elective caesarean deliveries which were in the gestational ages 36 to 38 weeks. Extracted amnions were washed by cold PBS to remove blood remnants. Blood samples were obtained from healthy adult volunteers who had not previously taken anti-coagulants. The blood samples were maintained in sterile tubes containing sodium citrate. Plasma or platelet rich plasma (PRP) were collected by blood sample centrifuging at 600 g for 10 min. Hemocompatibility of the AM samples (n=7) were evaluated by measuring of activated partial thromboplastin time (aPTT), prothrombin time (PT), hemolysis, and platelet aggregation tests. P-selectin was also assessed by ELISA. Both epithelial and mesenchymal sides of amnion were evaluated. Glass slide and expanded polytetrafluoroethylene (ePTFE) samples were defined as control. Results: In comparison with glass as control (13.3 ± 0.7 s), prothrombin time was increased significantly while each side of amnion was in contact with plasma (p<0.05). There was no significant difference in PT between epithelial and mesenchymal surfaces (17.4 ± 0.7 s vs. 15.8 ± 0.7 s, respectively). However, aPPT was not significantly changed after incubation of plasma with amnion epithelial and mesenchymal surfaces or glass (28.61 ± 1.39 s, 31.4 ± 2.66 s, glass, 30.76 ± 2.53 s, respectively, p>0.05). Amnion surfaces, ePTFE and glass samples have less hemolysis induction than water considerably (p<0.001), in which no differences were detected. Platelet aggregation measurements showed that platelets were less stimulated by the amnion epithelial and mesenchymal sides, in comparison with ePTFE and glass. In addition, reduction in amount of p-selectin, as platelet activation factor, after incubation of samples with PRP indicated that amnion has less stimulatory effects on platelets than ePTFE and glass. Conclusion: Amnion as a natural biomaterial has the potential to be used in tissue engineering. Our results suggest that amnion has appropriate hemocompatibility to be employed as a vascular substitute.

Keywords: amnion, hemocompatibility, tissue engineering, biomaterial

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Authors: A. Díaz-Roa, P. I. Silva Junior, F. J. Bello

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Sarconesiopsis magellanica (Diptera: Calliphoridae) is a medically important necrophagous fly which is used for establishing the post-mortem interval. Dipterous maggots release diverse proteins and peptides contained in larval excretion and secretion (ES) products playing a key role in digestion. The most important mechanism for combating infection using larval therapy depends on larval ES. These larvae are protected against infection by a diverse spectrum of antimicrobial peptides (AMPs), one already known like lucifensin. Special interest in these peptides has also been aroused regarding understanding their role in wound healing since they degrade necrotic tissue and kill different bacteria during larval therapy. The action of larvae on wounds occurs through 3 mechanisms of action: removal of necrotic tissue, stimulation of granulation tissue, and antibacterial action of larval ES. Some components of the ES include calcium, urea, allantoin ammonium bicarbonate and reducing the viability of Gram positive and Gram negative bacteria. The Lucilia sericata fly larvae have been the most used, however, we need to evaluate new species that could potentially be similar or more effective than fly above. This study was thus aimed at identifying and characterizing S. magellanica AMPs contained in ES products for the first time and compared them with the common fly used L. sericata. These products were obtained from third-instar larvae taken from a previously established colony. For the first analysis, ES fractions were separate by Sep-Pak C18 disposable columns (first step). The material obtained was fractionated by RP-HPLC by using Júpiter C18 semi-preparative column. The products were then lyophilized and their antimicrobial activity was characterized by incubation with different bacterial strains. The first chromatographic analysis of ES from L. sericata gives 6 fractions with antimicrobial activity against Gram-positive bacteria Micrococus luteus, and 3 fractions with activity against Gram-negative bacteria Pseudomonae aeruginosa while the one from S. magellanica gaves 1 fraction against M. luteus and 4 against P. aeruginosa. Maybe one of these fractions could correspond to the peptide already known from L. sericata. These results show the first work for supporting further experiments aimed at validating S. magellanica use in larval therapy. We still need to search if we find some new molecules, by making mass spectrometry and ‘de novo sequencing’. Further studies are necessary to identify and characterize them to better understand their functioning.

Keywords: antimicrobial peptides, larval therapy, Lucilia sericata, Sarconesiopsis magellanica

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Authors: Omaima A. Sharaf, Tarek A. Moussa, Said M. Badr El-Din, H. Moawad

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Background: Polycyclic aromatic hydrocarbons (PAHs) pose great environmental and human health concerns for their widespread occurrence, persistence, and carcinogenic properties. PAHs releases due to anthropogenic activities to the wider environment have led to higher concentrations of these contaminants than would be expected from natural processes alone. This may result in a wide range of environmental problems that can accumulate in agricultural ecosystems, which threatened to become a negative impact on sustainable agricultural development. Thus, this study aimed to evaluate the physico-chemical, and microbial properties of the compost leachate (CL) to assess its role as nutrient and microbial source (biostimulation/bioaugmentation) for developing a cost-effective bioremediation technology for PAHs contaminated sites. Material and Methods: PAHs-degrading bacteria were isolated from CL that was collected from a composting site located in central Scotland, UK. Isolation was carried out by enrichment using phenanthrene (PHR), pyrene (PYR) and benzo(a)pyrene (BaP) as the sole source of carbon and energy. The isolates were characterized using a variety of phenotypic and molecular properties. Six different isolates were identified based on the difference in morphological and biochemical tests. The efficiency of these isolates in PAHs utilization was assessed. Further analysis was performed to define taxonomical status and phylogenic relation between the most potent PAHs-utilizing bacterial strains and other standard strains, using molecular approach by partial 16S rDNA gene sequence analysis. Results indicated that the 16S rDNA sequence analysis confirmed the results of biochemical identification, as both of biochemical and molecular identification of the isolates assigned them to Bacillus licheniformis, Pseudomonas aeruginosa, Alcaligenes faecalis, Serratia marcescens, Enterobacter cloacae and Providenicia which were identified as the prominent PAHs-utilizers isolated from CL. Conclusion: This study indicates that the CL samples contain a diverse population of PAHs-degrading bacteria and the use of CL may have a potential for bioremediation of PAHs contaminated sites.

Keywords: polycyclic aromatic hydrocarbons, physico-chemical analyses, compost leachate, microbial and biochemical analyses, phylogenic relations, 16S rDNA sequence analysis

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Authors: A. Alyaaria, b, J. P. Wignerona, A. Ducharneb, Y. Kerrc, P. de Rosnayd, R. de Jeue, A. Govinda, A. Al Bitarc, C. Albergeld, J. Sabaterd, C. Moisya, P. Richaumec, A. Mialonc

Abstract:

Global Level-3 surface soil moisture (SSM) maps from the passive microwave soil moisture and Ocean Salinity satellite (SMOSL3) have been released. To further improve the Level-3 retrieval algorithm, evaluation of the accuracy of the spatio-temporal variability of the SMOS Level 3 products (referred to here as SMOSL3) is necessary. In this study, a comparative analysis of SMOSL3 with a SSM product derived from the observations of the Advanced Microwave Scanning Radiometer (AMSR-E) computed by implementing the Land Parameter Retrieval Model (LPRM) algorithm, referred to here as AMSRM, is presented. The comparison of both products (SMSL3 and AMSRM) were made against SSM products produced by a numerical weather prediction system (SM-DAS-2) at ECMWF (European Centre for Medium-Range Weather Forecasts) for the 03/2010-09/2011 period at global scale. The latter product was considered here a 'reference' product for the inter-comparison of the SMOSL3 and AMSRM products. Three statistical criteria were used for the evaluation, the correlation coefficient (R), the root-mean-squared difference (RMSD), and the bias. Global maps of these criteria were computed, taking into account vegetation information in terms of biome types and Leaf Area Index (LAI). We found that both the SMOSL3 and AMSRM products captured well the spatio-temporal variability of the SM-DAS-2 SSM products in most of the biomes. In general, the AMSRM products overestimated (i.e., wet bias) while the SMOSL3 products underestimated (i.e., dry bias) SSM in comparison to the SM-DAS-2 SSM products. In term of correlation values, the SMOSL3 products were found to better capture the SSM temporal dynamics in highly vegetated biomes ('Tropical humid', 'Temperate Humid', etc.) while best results for AMSRM were obtained over arid and semi-arid biomes ('Desert temperate', 'Desert tropical', etc.). When removing the seasonal cycles in the SSM time variations to compute anomaly values, better correlation with the SM-DAS-2 SSM anomalies were obtained with SMOSL3 than with AMSRM, in most of the biomes with the exception of desert regions. Eventually, we showed that the accuracy of the remotely sensed SSM products is strongly related to LAI. Both the SMOSL3 and AMSRM (slightly better) SSM products correlate well with the SM-DAS2 products over regions with sparse vegetation for values of LAI < 1 (these regions represent almost 50% of the pixels considered in this global study). In regions where LAI>1, SMOSL3 outperformed AMSRM with respect to SM-DAS-2: SMOSL3 had almost consistent performances up to LAI = 6, whereas AMSRM performance deteriorated rapidly with increasing values of LAI.

Keywords: remote sensing, microwave, soil moisture, AMSR-E, SMOS

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Authors: Miaomiao Zhang, Sabrina Beckmann, Haluk Ertan, Rocky Chau, Mike Manefield

Abstract:

Phenazines are a large class of nitrogen-containing aromatic heterocyclic compounds, which are almost exclusively produced by bacteria from diverse genera including Pseudomonas and Streptomyces. Phenazine-1-carboxylic acid (PCA) as one of 'core' phenazines are converted from chorismic acid before modified to other phenazine derivatives in different cells. Phenazines have attracted enormous interests because of their multiple roles on biocontrol, bacterial interaction, biofilm formation and fitness of their producers. However, in spite of ecological importance, degradation as a part of phenazines’ fate only have extremely limited attention now. Here, to isolate PCA-degrading bacteria, 200 mg L-1 PCA was supplied as sole carbon, nitrogen and energy source in minimal mineral medium. Quantitative PCR and Reverse-transcript PCR were employed to study abundance and activity of functional gene MFORT 16269 in PCA degradation, respectively. Intermediates and products of PCA degradation were identified with LC-MS/MS. After enrichment and isolation, a PCA-degrading strain was selected from soil and was designated as Rhodanobacter sp. PCA2 based on full 16S rRNA sequencing. As determined by HPLC, strain PCA2 consumed 200 mg L-1 (836 µM) PCA at a rate of 17.4 µM h-1, accompanying with significant cells yield from 1.92 × 105 to 3.11 × 106 cells per mL. Strain PCA2 was capable of degrading other phenazines as well, including phenazine (4.27 µM h-1), pyocyanin (2.72 µM h-1), neutral red (1.30 µM h-1) and 1-hydroxyphenazine (0.55 µM h-1). Moreover, during the incubation, transcript copies of MFORT 16269 gene increased significantly from 2.13 × 106 to 8.82 × 107 copies mL-1, which was 2.77 times faster than that of the corresponding gene copy number (2.20 × 106 to 3.32 × 107 copies mL-1), indicating that MFORT 16269 gene was activated and played roles on PCA degradation. As analyzed by LC-MS/MS, decarboxylation from the ring structure was determined as the first step of PCA degradation, followed by cleavage of nitrogen-containing ring by dioxygenase which catalyzed phenazine to nitrosobenzene. Subsequently, phenylhydroxylamine was detected after incubation for two days and was then transferred to aniline and catechol. Additionally, genomic and proteomic analyses were also carried out for strain PCA2. Overall, the findings presented here showed that a newly isolated strain Rhodanobacter sp. PCA2 was capable of degrading phenazines through decarboxylation and cleavage of nitrogen-containing ring, during which MFORT 16269 gene was activated and played important roles.

Keywords: decarboxylation, MFORT16269 gene, phenazine-1-carboxylic acid degradation, Rhodanobacter sp. PCA2

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Authors: O. Antypenko, I. Vasilieva, S. Kovalenko

Abstract:

Investigations for new effective and less toxic antimicrobials agents are always up-to-date. The tetrazole derivatives are quite interesting objects as for synthesis as well as for pharmacological screening. Thus, some derivatives of tetrazole demonstrated antimicrobial activity, namely 5-phenyl-tetrazolo[1,5-c]quinazoline was effective one against Staphylococcus aureus and Esherichia faecalis (MIC = 250 mg/L). Besides, investigation of the 9-bromo(chloro)-5-morpholin(piperidine)-4-yl-tetrazolo[1,5-c]quinazoline’s antimicrobial activity against Esherichia coli and Enterococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus revealed that sensitivity of Gram-positive bacteria to the compounds was higher than that of Gram-negative bacteria. So, our previously synthesized, 31 derivatives of 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas were decided to test for their in vitro antibacterial activity against Gram-positive bacteria (Staphylococcus aureus ATCC 25923, Enterobacter aerogenes, Enterococcus faecalis ATCC 29212), Gram-negative bacteria (Pseudomonas aeruginosa ATCC 9027, Escherichia coli ATCC 25922, Klebsiella pneumoniae 68) and antifungal properties against Candida albicans ATCC 885653. Agar-diffusion method was used for determination of the preliminary activity compared to well-known reference antimicrobials. All the compounds were dissolved in DMSO at a concentration of 100 μg/disk, using inhibition zone diameter (IZD, mm) as a measure for the antimicrobial activity. The most active turned to be 3 structures, that inhibited several bacterial strains: 1-ethyl-3-(5-fluoro-2-(1H-tetrazol-5-yl)phenyl)urea (1), 1-(4-bromo-2-(1H-tetrazol-5-yl)-phenyl)-3-(4-(trifluoromethyl)phenyl)urea (2) and 1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (3). IZM (mm) was 40 (Escherichia coli), 25 (Klebsiella pneumonia) for compound 1; 12 (Pseudomonas aeruginosa), 15 (Staphylococcus aureus), 10 (Enterococcus faecalis) for compound 2; 25 (Staphylococcus aureus), 15 (Enterococcus faecalis) for compound 3. The most sensitive to the activity of the substances were Gram-negative bacteria Pseudomonas aeruginosa. While none of compound effected on Candida albicans. Speaking about, reference drugs: Amikacin (30 µg/disk) showed 27 and Ceftazide (30 µg/disk) 25 against Pseudomonas aeruginosa. That is, unfortunately, higher than studied 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas. Obtained results will be used for further purposeful optimization of the leading compounds in the more effective antimicrobials because of the ever-mounting problem of microorganism’s resistance.

Keywords: antimicrobial, antifungal, compounds, 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas

Procedia PDF Downloads 358

Authors: Maria L. G. Lourenco, Keylla H. N. P. Pereira, Viviane Y. Hibaru, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

Abstract:

Neonatal sepsis is a systemic response of acute infection by bacteria that may lead to high mortality in a litter. This study aims to report a case of sepsis by alpha-hemolytic Streptococcus and Mannheimia haemolytica in neonate dogs. A pregnant, mixed-breed bitch at approximately the 60th day of pregnancy was admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, and subjected to a c-section due to uterine atony and fetuses no heartbeats on the ultrasound examination. The mother presented leukopenia of 1.6 thousand leukocytes, and there was no other information regarding previous clinical history. Among the offspring, four were stillborn, and five were born alive. On clinical examination, neonates weighed between 312 and 384 grams. Reflexes were present, and the newborn's body temperature was between 89.9 ºF and 96.4 ºF. Neonates also presented clinical signs of neonatal infection: omphalitis, abdomen, and extremities with cyanotic color, hematuria, and diarrhea (meconium). Complementary tests revealed leukopenia. The presence of alpha hemolytic streptococcus and Mannheimia haemolytica was revealed in the bacterial culture. The bacteria were sensitive to cephalosporins and penicillin on the antibiogram. Treatment for sepsis was instituted with the drug ceftriaxone, at a dose of 50 mg per kilogram, administered intravenous (jugular vein). Subsequently administered subcutaneous, every 12 hours, for seven days. Heated fluid therapy was performed, with Ringer lactate, at a dose of 4 ml per 100 grams of weight, intravenous. Heating measures were instituted. Blood plasma was also administered, at a dose of 2 mL per 100 grams of weight, administered subcutaneous, as a source of passive immunity. A maternal milk substitute was instituted, and lactation was discontinued since the mother was unable to nurse due to the infection. The mother was neutered during the c-section and treated with ceftriaxone (50 mg/kg). After seven days, the newborns presented normal clinical signs and no alterations in the hemogram. Early diagnosis and intervention were essential for the survival of these patients.

Keywords: neonatal infection, puppies, bacteria, newborn

Procedia PDF Downloads 121