Search results for: colonies

Authors: Muhammad Ameer Nawaz Akram

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The 21st century has been revealed that many individuals around the world are living in urban settlements than in rural zones. The evolution of numerous cities in emerging and newly developed countries is accompanied by the rise of slums. The precise definition of a slum varies countries to countries, but the universal harmony is that slums are dilapidated settlements facing severe poverty and have lacked access to sanitation, water, electricity, good living styles, and land tenure. The slum settlements always vary in unique patterns within and among the countries and cities. The core objective of this study is the spatial identification and classification of slums in Gujrat city Pakistan from very high-resolution GeoEye-1 (0.41m) satellite imagery. Slums were first identified using GPS for sample site identification and ground-truthing; through this process, 425 slums were identified. Then Object-Oriented Analysis (OOA) was applied to classify slums on digital image. Spatial analysis softwares, e.g., ArcGIS 10.3, Erdas Imagine 9.3, and Envi 5.1, were used for processing data and performing the analysis. Results show that OOA provides up to 90% accuracy for the identification of slums. Jalal Cheema and Allah Ho colonies are severely affected by slum settlements. The ratio of criminal activities is also higher here than in other areas. Slums are increasing with the passage of time in urban areas, and they will be like a hazardous problem in coming future. So now, the executive bodies need to make effective policies and move towards the amelioration process of the city.

Keywords: slums, GPS, satellite imagery, object oriented analysis, zonal change detection

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Authors: Aliasghar Golmohammadian, Sona Rostampour Yasouri

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One of the most important intestinal pathogenic strains is E. coli O₁₅₇:H₇. This pathogenic bacterium is transmitted to humans through water and food. E. coli O₁₅₇:H₇ is the main cause of Hemorrhagic colitis (HC), Hemolytic Uremic Syndrome (HUS), Thrombotic Thrombocytopenic Purpura (TTP) and in some cases death. Since E. coli O₁₅₇:H₇ can be transmitted through the consumption of different foods, including vegetables, agricultural products, and fresh dairy products, this study aims to identify and isolate E. coli O₁₅₇:H₇ from wastewater by PCR technique. One hundred twenty samples of water and wastewater were collected by Falcom Sterile from Shahrood and Neka cities. The samples were checked for colony formation after appropriate centrifugation and cultivation in the specific medium of Sorbitol MacConkey Agar (SMAC) and other diagnostic media of E. coli O₁₅₇:H₇. Also, the plates were observed macroscopically and microscopically. Then, the necessary phenotypic tests were performed on the colonies, and finally, after DNA extraction, the PCR technique was performed with specific primers related to rfbE and stx2 genes. The number of 5 samples (6%) out of all the samples examined were determined positive by PCR technique with observing the bands related to the mentioned genes on the agarose gel electrophoresis. PCR is a fast and accurate method to identify the bacteria E. coli O₁₅₇:H₇. Considering that E. coli bacteria is a resistant bacteria and survives in water and food for weeks and months, the PCR technique can provide the possibility of quick detection of contaminated water. Moreover, it helps people in the community control and prevent the transfer of bacteria to healthy and underground water and agricultural and even dairy products.

Keywords: E. coli O₁₅₇:H₇, PCR, water, wastewater

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Authors: Nourmalita Safitri Ningsih, Ridwan, Iqri Puspa Yunanda

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The aim of this study was to utilize the waste from slaughterhouses for chicken feed ingredients is probiotic. Livestock waste produced by livestock activities such as feces, urine, food remains, as well as water from livestock and cage cleaning. The process starts with the isolation of bacteria. Rumen fluid is taken at Slaughterhouse Giwangan, Yogyakarta. Isolation of Lactobacillus ruminus is done by using de Mann Rogosa Sharpe (MRS) medium. In the sample showed a rod-shaped bacteria are streaked onto an agar plates. After it was incubated at 37ºC for 48 hours, after which it is observed. The observation of these lactic acid bacteria it will show a clear zone at about the colony. These bacterial colonies are white, round, small, shiny on the agar plate mikroenkapsul In the manufacturing process carried out by the method of freeze dried using skim milk in addition capsulated material. Then the results of these capsulated bacteria are mixed with feed for livestock. The results from the mixing of capsulated bacteria in feed are to increase the quality of animal feed so as to provide a good effect on livestock. Scanning electron microscope testing we have done show the results of bacteria have been shrouded in skim milk. It can protect the bacteria so it is more durable in use. The observation of the bacteria showed a sheath on Lactobacillus sp. Preservation of bacteria in this way makes the bacteria more durable for use. As well as skim milk can protect bacteria that are resistant to the outside environment. Results of probiotics in chicken feed showed significant weight gain in chickens. Calculation Anova (P <0.005) shows the average chicken given probiotics her weight increased.

Keywords: chicken, probiotics, waste, Lactobacillus sp, bacteria

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Authors: Jack Johnstone

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This research presents postcolonial translation as an approach to eradicate traces of colonialism in former colonies. An example of demonstrating postcolonial translation in the Indonesian context is in Hasan Aspahani's Chairil, a biographical narrative and history book based on the personal life of a well-known Indonesian poet and writer, Chairil Anwar (1922-1949) in Dutch occupied Indonesia. This postcolonial translation approach has been applied in the first five chapters on his early years under Dutch colonization, in an attempt to show a postcolonialised TT. This approach aims to demonstrate the postcolonial refutation of the Dutch colonial language to convey the Indonesian setting to target readers. It is also designed to explicate the summary of the book as well as my attempt to apply postcolonial translation as a strategy to reject the Dutch colonial terms in this book. The data conveys 26 important examples of the ST and TT, in consideration of the chosen three factors of culture, forced-Europeanisation, and cross-genre between a biographical narrative and history under categories of Cultural Bound Objects, Politics and Place. However, the 10 selected examples will be analyzed in the Analysis Chapter, which are discussed at word, sentence, and paragraph level. As well, the translation strategies used, namely retention, substitution and specification on four main examples, on the methods utilized to achieve a postcolonialised translation that attempts to 1) examine the way the alteration of the TT can affect the message portrayed within the ST, 2) show the notion of disagreement between the Dutch colonizers and colonized Indonesians on their views on the way Indonesia should be governed and 3) present a translation that reverses the inequality between the superior colonials and inferior Indigenous Indonesians during the Dutch colonial era.

Keywords: Chairil, Dutch colonialism, Indonesia, postcolonial translation

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Authors: Muhammad Imran Nisar, Fyezah Jehan, Tauseef Akhund, Sadia Shakoor, Kanwal Nayani, Furqan Kabir, Asad Ali, Anita Zaidi

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Pneumococcal Vaccine -10 (PCV 10) was included in the Expanded Program of immunization (EPI) in Sindh, Pakistan in February 2013. This study was carried out immediately before the introduction of PCV 10 to establish baseline pneumococcal carriage and prevalent serotypes in naso-pharynx of children 3-11 months of age in an urban and rural community in Sindh, Pakistan. An additional sample of children aged 12 to 59 months was drawn from the urban community. Nasopharyngeal specimens were collected from a random sample of children. Samples were processed in a central laboratory in Karachi. Pneumococci were cultured on 5% Sheep Blood Agar and serotyping was performed using CDC standardized sequential multiplex PCR assay on bacterial colonies. Serotypes were then categorized into vaccine (PCV-10 and PCV-13) type and non-vaccine types. A total of 670 children were enrolled. Carriage rate for pneumococcus based on culture positivity was 74% and 79.5 % in the infant group in Karachi and Matiari respectively. Carriage rate was 78.2% for children aged 12 to 59 months in Karachi. Proportion of PCV 10 serotypes in infants was 38.8% and 33.5% in Karachi and Matiari respectively. In the older age group in Karachi, the proportion was 30.6%. Most common serotypes were 6A, 6B, 23F, 19A and 18C. This survey establishes vaccine and non-vaccine serotype carriage rate in a vaccine-naïve pediatric population among rural and urban communities in Sindh province. Annually planned surveys in the same communities will inform change in carriage rate after the introduction and uptake of PCV 10 in these communities.

Keywords: Naso-Pharyngeal carriage, Pakistan, PCV10, Pneumococcus

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Authors: Guanghua Yu

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This article has examined the rise of the UK, US, and China at different historical times to explain the argument that it is open access in the economic sphere, as well as institutional building related to the protection of property rights, contract enforcement, financial market, the rule of law, and human resource accumulation that determine economic and human development. Both the UK, after the Glorious Revolution in the seventeenth century, and China, after its adoption of the open door policy at the end of the 1970s, follow such a path of development. The difference between the UK and China in moving toward that path is the different coordination of elites. While the coordination of elites in the UK through parliament played important roles in forcing the government to consider the wider encompassing interest in society after the Glorious Revolution, the coordination of elites in China has mainly been achieved by the Communist Party of China such that the Chinese Government has started to pay greater deal of attention to the wider encompassing interest in the country from 1978. The article has also examined the rise of the US following colonial settlement to independence and institutional building thereafter. The US case is similarly consistent with the argument that open access in the economic sphere and institutional building matter the most to economic development. More decentralized methods of the coordination of elites in the US among colonies (states), the federal governments, and other political groups similarly shaped the path towards open access in the economic sphere and institutional building. As such, open access in the political sphere plays an indirect role in development at best. If that is correct, there are possibilities that different political systems are able to achieve coordination of elites so that governments will turn their attention to development.

Keywords: open access, interconnected institutions, democracy, development

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Authors: Saskia Köbschall

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The idea of ‘being close to nature’ and the ways of achieving this proximity are socially and historically constructed, as are notions of nakedness and nudity. During the colonial period, nudity and clothedness functioned as instruments of racial domination. Nakedness became central to the colonialists’ thinking, to their binary of the ‘civilized’ and those ‘close to nature’, therefore turning the level of perceived unclothedness into a measurement of ‘civilization'. While being ‘one with nature’ continued to be a criterion of backwardness in the colonies, it emerged as a futuristic and avant-garde endeavor in the metropole: In Germany, at the height of its colonial expansion, the Life Reform Movement (Lebensreformbewegung) called for the liberation of the white body from the ‘constraints of civilization’, for its ‘return to nature’ via practices like nudism. Despite this simultaneity, the scholarship of the life reform and the nudist movement in particular does not address the colonial past of the movement or its repercussions in the present. Taking the biography of prominent life reformist Hans Paasche (1881 - 1920) as a starting point, this paper explores the work of imperial legacies in the history and present of the German nudist movement. Paasche started his career as a German colonial officer, participating in the brutal obliteration of the Maji-Maji uprising (1905/06) that claimed the lives of nearly 200.000 people. Once a passionate game hunter, he later became a known nature conservationist; once a self-proclaimed explorer of Africa, he later became one of the most prominent advocates of nudism and vegetarianism. The paper joins conceptual and historical research in order to address the German nudist movement’s colonial past and understand its repercussions in the present.

Keywords: Germany, life reform, colonialism, archives, nudity, nature

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Authors: R. Posia, J. Mistry, K. Kamani

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The health screening of laboratory rodents is essential for ensuring animal health and the validity of biomedical research data. Routine health monitoring is necessary to verify the effectiveness of biosecurity and the specific pathogen free (SPF) status of the colony. The present screening was performed in barrier maintained rat (Rattus norvegicus) colony. Rats were maintained under a controlled environment and strict biosecurity in the facility. The screening was performed on quarterly bases from randomly selected animals from breeding and or maintenance colonies. Selected animals were subject to blood collection under isoflurane anaesthesia. Serum was separated from the collected blood and stored samples at -60 ± 10 °C until further use. A total of 88 samples were collected quarterly bases from animals in a year. In the serological test, enzyme-linked immunosorbent assay (ELISA) was used for screening of serum samples against sialodacryoadenitis virus (SDAV), Sendai virus (SV), and Kilham’s rat virus (KRV). ELISA kits were procured from XpressBio, USA. Test serum samples were run along with positive control, negative control serum in 96 well ELISA plates as per the procedure recommended by the vendor. Test ELISA plate reading was taken in the microplate reader. This screening observed that none of the samples was observed positive for the sialodacryoadenitis virus (SDAV), Sendai virus (SV), and Kilham’s rat virus (KRV), indicating that effectiveness of biosecurity practices followed in the rodent colony. The result of serological screening helps us to declare that our rodent colony is specifically pathogen free for these pathogens.

Keywords: biosecurity, ELISA, specific pathogen free, serological screening, serum

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Authors: Edward Y. W. Chu

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This paper exemplifies how the political and educational considerations have shaped the heavy-handed MOI policy in Hong Kong after its handover to China in 1997. Its result, a significant degeneration of English standard among the non-elite students, will be reported based on a detailed analysis of the public exam statistics available and other empirical studies. The remedial action taken by the Education Bureau out of the economic and educational considerations will be reported with reference to the official documents. The political, economic and educational considerations exemplified in different stages of Mother-tongue MOI policy in Hong Kong are found to be influential in the MOI policy in other Asian countries as well. For example, out of rapid internationalization and marketization, there has been increasing adoption of English as the MOI in post-secondary institutions in China, Japan & South Korea. On the other hand, while colonial languages were firmly made as the MOI in former colonies such as Vietnam and India, they were greatly retrieved upon independence for political and educational reasons. Malaysia also followed the same pattern upon independence but re-introduced partial English MOI policy in late 90s hoping to capitalize favourable globalization benefits. The short-lived policy was abandoned in 2009 because of the perceived political threat of national identity as well as the lack of educational effectiveness. Based on the great majority of Asian countries studied, this paper argues that MOI policy in Asia is much more than an educational issue, and that there is a clear pattern of how decisions of MOI matters are made. Studying the history and development of MOI in Hong Kong and other Asian countries provides a unique angle to view of how Asian countries prepare for the political, economic and educational challenges nowadays.

Keywords: economics, Hong Kong, medium of instruction, politics

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Authors: Kelli G. Thorup, Kristopher A. Blee

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Climate change enhances the occurrence of extreme weather: wildfires, drought, rising summer temperatures, all of which dramatically decline forest growth and increase tree mortality in the mixed-conifer forests of Sierra Nevada, California. However, microbiota living in mutualistic relations with plant rhizospheres have been found to mitigate the effects of suboptimal environmental conditions. The goal of this research is to isolate native beneficial bacteria, plant-growth promoting rhizobacteria (PGPR), that can alleviate heat stress in Pinus ponderosa seedlings. Bacteria were isolated from the rhizosphere of Pinus ponderosa juveniles located in mixed-conifer stand and further characterized for PGP potential based on their ability to produce key growth regulatory phytohormones including auxin, cytokinin, and gibberellic acid. Out of ten soil samples taken, sixteen colonies were isolated and qualitatively confirmed to produce indole-3-acetic acid (auxin) using Salkowski’s reagent. Future testing will be conducted to quantitatively assess phytohormone production in bacterial isolates. Furthermore, bioassays will be performed to determine isolates abilities to increase tolerance in heat-stressed Pinus ponderosa seedlings. Upon completion of this research, a PGPR could be utilized to support the growth and transplantation of conifer seedlings as summer temperatures continue to rise due to the effects of climate change.

Keywords: conifer, heat-stressed, phytohormones, Pinus ponderosa, plant-growth promoting rhizobacteria

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Authors: Munish Garg, Vinni Ahuja

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In the past few years, the high consumption of soft drinks has attracted negative attention world-wide due to its possible adverse effects, leading the health conscious people to find alternative nutraceutical or herbal health drinks. In the present study, a nutraceutical soft drink was developed utilizing some easily available and well known traditional herbs having nutritional potential. The key ingredients were selected as bael, amla, lemon juice, ashwagandha and poppy seeds based on their household routine use in the summer with proven refreshing, cooling and energetic feeling since ages. After several trials made, the final composition of nutraceutical summer soft drink was selected as most suitable combination based on the taste, physicochemical, microbial and organoleptic point of view. The physicochemical analysis of the prepared drink found to contain optimum level of titratable acidity, total soluble solids and pH which were in accordance of the commercial recommendations. There were no bacterial colonies found in the product therefore found within limits. During the nine point’s hedonic scale sensory evaluation, the drink was strongly liked for colour, taste, flavour and texture. The formulation was found to contain flavonoids (80mg/100ml), phenolics (103mg/100ml), vitamin C (250mg/100ml) and has antioxidant potential (75.52%) apart from providing several other essential vitamins, minerals and healthy components. The developed nutraceutical drink provides an economical and feasible option for the consumers with very good taste combined with potential health benefits. The present drink is potentially capable to replace the synthetic soft drinks available in the market.

Keywords: herbal drink, summer drink, nutraceuticals, soft drink

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Authors: Virginia Andrea Garrido Meirelles

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Noah Webster was a lexicographer and a language reformer and is considered the ‘Father of American Scholarship and Education’ because of the exceptional contributions he made as a teacher and grammarian. The goal of this study is to show that the success of his plan can be explained by the fact that it matched the language-in-education policies of his time. To accomplish that goal the present study analyzes the Massachusetts School Laws of 1642, 1647 and 1648 and compares them to the preface of the first edition of The Grammatical Institute of the English Language. The referred laws were three legislative acts enacted in the Massachusetts Colony and replicated almost identically in the other New England colonies. The purpose of those laws was to eradicate pauperism and poverty, on the one side, and to disseminate the idea of right citizenship, on the other. However, until the Declaration of Independence in 1776, all the primers used in the colony were printed in Britain. In 1783, Noah Webster published the first part of his Grammatical Institute of the English Language. In this book, the author states that his goal is to promote the republican principles that guide the civil rights of that time. The material included many texts taken from the Bible to inspire aversion to inadequate behavior and preference for service and good manners. In addition, its goal was to present ‘a new plan of reducing the pronunciation of our language to an easy standard,’ and in that way, create a unified language to abolish ignorance and language corruption. The comparison between the laws and Webster’s Spelling Book shows that the book is the result of the historical and political situation when it was conceived and it satisfied the requirements of the language-in-education policies of the time.

Keywords: American English, language policy, the Massachusetts school laws, webster's spelling book

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Authors: Emmanuel Nikiema

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This paper examines article incorporation in Mauritian Creole, a French Lexifier Creole which exhibits three forms of article incorporation as illustrated in (1-3). While various analyses of article incorporation have been proposed in the literature, fewer studies have explored the motivation of this widespread phenomenon in Mauritian Creole (MC) as opposed to other French Lexifier Creoles spoken in the Caribbean. For example, Mauritian Creole exhibits 4 times more CV incorporation than Haitian Creole, and 40 times more than Reunion Creole. (1) Consonantal type (C): loraz ‘thunder storm’, lete ‘summer’, zwazo ‘bird’, nide ‘idea’. (2) Syllabic type (CV): lapo ‘skin’, liku ‘neck’, ledo ‘back’, leker ‘heart’, diber ‘butter’. (3) Bi-consonantal (CVC): delo ‘water’, dizef ‘egg’, lizye ‘eye’, dilwil ‘oil’. The goal of this study is twofold: 1) uncover the rules governing the three types of article incorporation in MC, and 2) account for its remarkable occurrence in MC as opposed to its quasi-absence in Reunion Creole. We have collected a corpus of over 700 cases and organized it into three categories (C; CV and CVC). For example, there are 471 examples of CV incorporation in MC against 112 in Haitian Creole and only 12 in Reunion Creole. Two questions can be raised: 1) what is the motivation and distribution of the three types of incorporation in MC, and 2) how can one account for the high volume of incorporation in MC as opposed to its quasi-absence in Reunion Creole? We suggest that article incorporation in MC is related to the structure of nouns in Bantou languages. While previous authors have largely used population settlement data in the colonies during the Creole formation period to justify their analyses, we propose an account based on the syntactic structure of Bantou nouns. This analysis will shed light on the contribution of African languages to the formation of MC, and on to why MC has exhibited more article incorporation cases than any other French Lexifier Creole.

Keywords: article incorporation, creole languages, description, phonology

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Authors: C. A. Ologunde

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Escherichia coli have been a major microorganisms associated with, and isolated from feacal samples either in adult or children all over the world. Strains of these organisms are resistant to cephalosporins and fluoroquinolone (FQ) antimicrobial agents among hospitalized patients and FQs are the most frequently prescribed antimicrobial class in hospitals, and the level of resistant of E. coli to these antimicrobial agents is a risk factor that should be assessed. Hence, this study was conducted to determine the prevalence and risk factors for colonization with fluoroquinolone (FQ)-resistant E. coli in hospitalized patients in Ado-Ekiti. Rectal swabs were obtained from patients in hospitals in the study area and FQ-resistant E. coli were isolated and identified by means of Nalidixic acid multi-disk and a 1-step screening procedure. Species identification and FQ resistance were confirmed by automated testing (Vitek, bioMerieux, USA). Individual colonies were subjected to pulse-field gel electrophoresis (PAGE) to determine macro-restriction polymorphism after digestion of chromosomal DNA. FQ-resistant E. coli was detected in the stool sample of 37(62%) hospitalized patient. With multivariable analyses, the use of FQ before hospitalization was the only independent risk factor for FQ-resistant E. coli carriage and was consistent for FQ exposures for the 3-12 months of study. Pulsed-field gel electrophoresis of FQ-resistant E. coli identified conal spread of 1(one) strain among 18 patients. Loss (9 patients) or acquisition (10 residents) of FQ-resistant E. coli was documented and was associated with de novo colonization with genetically distinct strains. It was concluded that FQ-resistant E. coli carriage was associated with clonal spread. The differential effects of individual fluoroquinolone on antimicrobial drug resistance are an important area for future study, as hospitals manipulate their formularies with regard to use of individual fluoroquinolone, often for economic reasons.

Keywords: E. coli, fluoroquinolone, risk factors, feacal carriage, hospitalized patients, Ado-Ekiti

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Authors: Uzma Imtiaz

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Ecological feminism adheres to the basic philosophy that patriarchy is the wellspring of natural and gender domination. It explores the relationship between women and nature in a patriarchal society. Eco-feminism argues that women and nature have an intrinsic association and exploitation of women is the exploitation of nature itself. It further views the world as a holistic institution that offers equal opportunities for men and women. Eco-feminism rejects male domination in a patriarchal society where men and women do not get equal rights to survival. Furthermore, it investigates modern capitalist practices that exert unjust male dominance over nature and women. Cultural eco-feminist theorists argue that industrialization and modern science are male-centered and exhibit male chauvinistic views in attempts to control females’ ability to reproduce. This research intends to analyze an eco-feminist novel by Laila Halaby from the eco-feminism theoretical framework of Maria Mies and Vandana Shiva. The feminist dystopian novel throws light on the double-faced processes of capitalism and housewifization that destroy the autonomy of women over their bodies and life. Moreover, this study aims to highlight the unjust capitalistic processes and policies that turn other countries and women into colonies to exploit them by white men in the name of progress and civilization. The novel brings the patriarchal ways of dominance over women into question. This research paper concludes that women and men should get equal opportunities to survive in society, and women should have given rights over their bodies to decide their future. The research is qualitative in nature, so the method of close reading is selected to analyze the hypodermic effect of patriarchy in society. This study is valuable in highlighting the exploitative ways of men to subjugate women and nature and helps to give awareness to women against gender exploitation in society.

Keywords: housewifization, exploitation, capitalist patriarchy, female victimization

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Authors: Ali Aydin, Mert Sudagidan, Aysen Coban, Alparslan Kadir Devrim

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Pseudomonas spp. (specifically, P. fluorescens and P. fragi) are considered the principal spoilage microorganisms of refrigerated poultry meats. The higher the level psychrophilic spoilage Pseudomonas spp. on carcasses at the end of processing lead to decrease the shelf life of the refrigerated product. The aim of the study was the identification of psychrophilic Pseudomonas spp. having proteolytic and lipolytic activities from poultry meats by 16S rRNA and rpoB gene sequencing, investigation of protease and lipase related genes and determination of proteolytic activity of Pseudomonas spp. In the of isolation procedure, collected chicken meat samples from local markets and slaughterhouses were homogenized and the lysates were incubated on Standard method agar and Skim Milk agar for selection of proteolytic bacteria and tributyrin agar for selection of lipolytic bacteria at +4 °C for 7 days. After detection of proteolytic and lipolytic colonies, the isolates were firstly analyzed by biochemical tests such as Gram staining, catalase and oxidase tests. DNA gene sequencing analysis and comparison with GenBank revealed that 126 strong enzyme Pseudomonas spp. were identified as predominantly P. fluorescens (n=55), P. fragi (n=42), Pseudomonas spp. (n=24), P. cedrina (n=2), P. poae (n=1), P. koreensis (n=1), and P. gessardi (n=1). Additionally, protease related aprX gene was screened in the strains and it was detected in 69/126 strains, whereas, lipase related lipA gene was found in 9 Pseudomonas strains. Protease activity was determined using commercially available protease assay kit and 5 strains showed high protease activity. The results showed that psychrophilic Pseudomonas strains were present in chicken meat samples and they can produce important levels of proteases and lipases for food spoilage to decrease food quality and safety.

Keywords: Pseudomonas, chicken meat, protease, lipase

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Authors: Rekaya A. Shabbir, Marco Mingarelli, Glenn Flux, Ananya Choudhury, Tim A. D. Smith

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Introduction: Heterogeneity of dose distributions across a tumour is problematic for targeted radiotherapy. Gold nanoparticles (AuNPs) enhance dose-distributions of targeted radionuclides. The aim of this study is to demonstrate if tumour dose-distribution of targeted AuNPs radiolabelled with either of two radioisotopes (¹⁷⁷Lu and ⁹⁰Y) in breast cancer cells produced homogeneous dose distributions. Moreover, in vitro and in vivo studies were conducted to study the importance of receptor level on cytotoxicity of EGFR-targeted AuNPs in breast and colorectal cancer cells. Methods: AuNPs were functionalised with DOTA and OPPS-PEG-SVA to optimise labelling with radionuclide tracers and targeting with Erbitux. Radionuclides were chelated with DOTA, and the uptake of the radiolabelled AuNPs and targeted activity in vitro in both cell lines measured using liquid scintillation counting. Cells with medium (HCT8) and high (MDA-MB-468) EGFR expression were incubated with targeted ¹⁷⁷Lu-AuNPs for 4h, then washed and allowed to form colonies. Nude mice bearing tumours were used to study the biodistribution by injecting ¹⁷⁷Lu-AuNPs or ⁹⁰Y-AuNPs via the tail vein. Heterogeneity of dose-distribution in tumours was determined using autoradiography. Results: Colony formation (% control) was 81 ± 4.7% (HCT8) and 32 ± 9% (MDA-MB-468). High uptake was observed in the liver and spleen, indicating hepatobiliary excretion. Imaging showed heterogeneity in dose-distributions for both radionuclides across the tumours. Conclusion: The cytotoxic effect of EGFR-targeted AuNPs is greater in cells with higher EGFR expression. Dose-distributions for individual radiolabelled nanoparticles were heterogeneous across tumours. Further strategies are required to improve the uniformity of dose distribution prior to clinical trials.

Keywords: cancer cells, dose distributions, radionuclide therapy, targeted gold nanoparticles

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Authors: Sheila Shahidi, Hootan Rezaee, Abosaeed Rashidi, Mahmood Ghoranneviss

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Plasma treatment has an explosive increase in interest and use in industrial applications as for example in medical, biomedical, automobile, electronics, semiconductor and textile industry. A lot of intensive basic research has been performed in the last decade in the field of textiles along with technical textiles. Textile manufacturers and end-users alike have been searching for ways to improve the surface properties of natural and man-made fibers. Specifically, there is a need to improve adhesion and wettability. Functional groups may be introduced onto the fiber surface by using gas plasma treatments, improving fiber surface properties without affecting the fiber’s bulk properties. In this research work, ZnO nanoparticles (ZnO-NPs) were insitue synthesized by sonochemical method at room temperature on both untreated and plasma pretreated cotton woven fabric. Oxygen and nitrogen plasmas were used for pre-functionalization of cotton fabric. And the effect of oxygen and nitrogen pre-functionalization on adhesion properties between ZnO nanoparticles and cotton surface were studied. The results show that nanoparticles with average sizes of 20-100 nm with different morphologies have been created on the surface of samples. Synthesis of ZnO-NPs was varied in the morphological transformation by changes in zinc acetate dehydrate concentration. Characterizations were carried out using Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD), Inductive coupled plasma (ICP) and Spectrophotometery. The antibacterial activities of the fabrics were assessed semi-quantitatively by the colonies count method. The results show that the finished fabric demonstrated significant antibacterial activity against S. aureus in antibacterial test. The wash fastness of both untreated and plasma pretreated samples after 30 times of washing was investigated. The results showed that the parameters of plasma reactor plays very important role for improving the antibacterial durability.

Keywords: antibacterial activity, cotton, fabric, nanoparticles, plasma

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Authors: Abu Salim Mustafa

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Polymerase chain reaction (PCR) assays targeting genomic DNA segments have been established for the detection of Helicobacter pylori in clinical specimens. However, the data on comparative evaluations of various targets in detection of H. pylori are limited. Furthermore, the frequencies of vacA (s1 and s2) and cagA genotypes, which are suggested to be involved in the pathogenesis of H. pylori in other parts of the world, are not well studied in Kuwait. The aim of this study was to evaluate PCR assays for the detection and genotyping of H. pylori by targeting the amplification of DNA targets from four genomic segments. The genomic DNA were isolated from 72 clinical isolates of H. pylori and tested in PCR with four pairs of oligonucleotides primers, i.e. ECH-U/ECH-L, ET-5U/ET-5L, CagAF/CagAR and Vac1F/Vac1XR, which were expected to amplify targets of various sizes (471 bp, 230 bp, 183 bp and 176/203 bp, respectively) from the genomic DNA of H. pylori. The PCR-amplified DNA were analyzed by agarose gel electrophoresis. PCR products of expected size were obtained with all primer pairs by using genomic DNA isolated from H. pylori. DNA dilution experiments showed that the most sensitive PCR target was 471 bp DNA amplified by the primers ECH-U/ECH-L, followed by the targets of Vac1F/Vac1XR (176 bp/203 DNA), CagAF/CagAR (183 bp DNA) and ET-5U/ET-5L (230 bp DNA). However, when tested with undiluted genomic DNA isolated from single colonies of all isolates, the Vac1F/Vac1XR target provided the maximum positive results (71/72 (99% positives)), followed by ECH-U/ECH-L (69/72 (93% positives)), ET-5U/ET-5L (51/72 (71% positives)) and CagAF/CagAR (26/72 (46% positives)). The results of genotyping experiments showed that vacA s1 (46% positive) and vacA s2 (54% positive) genotypes were almost equally associated with VaCA+/CagA- isolates (P > 0.05), but with VacA+/CagA+ isolates, S1 genotype (92% positive) was more frequently detected than S2 genotype (8% positive) (P< 0.0001). In conclusion, among the primer pairs tested, Vac1F/Vac1XR provided the best results for detection of H. pylori. The genotyping experiments showed that vacA s1 and vacA s2 genotypes were almost equally associated with vaCA⁺/cagA^- isolates, but vacA s1 genotype had a significantly increased association with vacA⁺/cagA⁺ isolates.

Keywords: H. pylori, PCR, detection, genotyping

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Authors: Vongsathorn Ngampuak, Yutachai Chookaew, Wipawee Dejtisakdi

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Dunaliella salina has great potential as a system for generating commercially valuable products, including beta-carotene, pharmaceuticals, and biofuels. Our goal is to improve this potential by enhancing growth rate and other properties of D. salina under optimal growth conditions. We used ethyl methane sulfonate (EMS) to generate random mutants in D. salina KU11, a strain classified in Thailand. In a preliminary experiment, we first treated D. salina cells with 0%, 0.8%, 1.0%, 1.2%, 1.44% and 1.66% EMS to generate a killing curve. After that, we randomly picked 30 candidates from approximately 300 isolated survivor colonies from the 1.44% EMS treatment (which permitted 30% survival) as an initial test of the mutant screen. Among the 30 survivor lines, we found that 2 strains (mutant #17 and #24) had significantly improved growth rates and cell number accumulation at stationary phase approximately up to 1.8 and 1.45 fold, respectively, 2 strains (mutant #6 and #23) had significantly decreased growth rates and cell number accumulation at stationary phase approximately down to 1.4 and 1.35 fold, respectively, while 26 of 30 lines had similar growth rates compared with the wild type control. We also analyzed cell size for each strain and found there was no significant difference comparing all mutants with the wild type. In addition, mutant #24 had shown an increase of biomass accumulation approximately 1.65 fold compared with the wild type strain on day 5 that was entering early stationary phase. From these preliminary results, it could be feasible to identify D. salina mutants with significant improved growth rate, cell accumulation and biomass production compared to the wild type for the further study; this makes it possible to improve this microorganism as a platform for biotechnology application.

Keywords: Dunaliella salina, ethyl methyl sulfonate, growth rate, biomass

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Authors: Andreev V. A., Kovalenko T. N., Privolnev V. V., Chernavin A. V., Knyazeva E. R.

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Aim: To evaluate the antagonistic activity of Streptococcus salivarius K12 (SsK12) against ENT and oral cavity infection pathogens (S. pneumoniae, S. pyogenes, S. aureus), gram-negative bacteria (E. coli, P. aeruginosa) and C. albicans. Materials and methods: The probiotic strain SsK12 was isolated from the dietary supplement containing at least 1 × 109 CFU per tablet. The tablet was dissolved in the enrichment broth. The resulting suspension was seeded on 5% blood agar and incubated at 35°C in 4-6% CO2 for 48 hours. The raised culture was identified as Streptococcus salivarius with MALDI-TOF mass spectrometry method. The evaluation of SsK12 antagonistic activity was carried out using a perpendicular streak technique. The daily SsK12 culture was inoculated as heavy streaks with a loop at one side of Petri dish with the Muller-Hinton agar (MHA) and incubated for 24 hours at 350 C in anaerobic conditions. It was supposed that bacteriocins would diffuse over the whole area of the agar media. On the next day S. pneumoniae, S. pyogenes, S. aureus, E. coli, P. aeruginosa and C. albicans clinical isolates were streaked at the clear side of MHA Petri dish. MHA Petri dish inoculated with SsK12 (one part) and with the respective clinical isolates (another part) streaked perpendicularly on the same day was used as the control. Results: There was no growth of S. pyogenes on the Petri dish with SsK12 daily culture; the growth of a few colonies of S. pneumonia was noted. The growth of S. aureus, E. coli, P. aeruginosa and C. albicans was noted along the inoculated streak. On the control Petri dish with simultaneous inoculating of the SsK12 strain and the test cultures, the growth of all the testes isolates was noted. Conclusions: (1) SsK12 possesses perfect antagonistic activity against S. pyogenes and good activity against S. pneumoniae. (2) There was no antagonistic activity of SsK12 against S. aureus, E. coli, P. aeruginosa and C. albicans. (3) SsK12 antagonistic properties make it possible to use this probiotic strain for prophylaxis of recurrent ENT infections.

Keywords: probiotics, SsK12, streptococcus salivarius K12, antagonistic activity

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Authors: Çiğdem Sezer, Aksem Aksoy, Leyla Vatansever

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This study has been done for the purpose of evaluation of public health and identifying of enterotoxigenic Staphyloccocus aureus in kitchen of catering. In the kitchen of catering, samples have been taken by swabs from surface of equipments which are in the salad section, meat section and bakery section. Samples have been investigated with classical cultural methods in terms of Staphyloccocus aureus. Therefore, as a 10x10 cm area was identified (salad, cutting and chopping surfaces, knives, meat grinder, meat chopping surface) samples have been taken with sterile swabs with helping FTS from this area. In total, 50 samples were obtained. In aseptic conditions, Baird-Parker agar (with egg yolk tellurite) surface was seeded with swabs. After 24-48 hours of incubation at 37°C, the black colonies with 1-1.5 mm diameter and which are surrounded by a zone indicating lecithinase activity were identified as S. aureus after applying Gram staining, catalase, coagulase, glucose and mannitol fermentation and termonuclease tests. Genotypic characterization (Staphylococcus genus and S.aureus species spesific) of isolates was performed by PCR. The ELISA test was applied to the isolates for the identification of staphylococcal enterotoxins (SET) A, B, C, D, E in bacterial cultures. Measurements were taken at 450 nm in an ELISA reader using an Ridascreen-Total set ELISA test kit (r-biopharm R4105-Enterotoxin A, B, C, D, E). The results were calculated according to the manufacturer’s instructions. A total of 50 samples of 97 S. aureus was isolated. This number has been identified as 60 with PCR analysis. According to ELISA test, only 1 of 60 isolates were found to be enterotoxigenic. Enterotoxigenic strains were identified from the surface of salad chopping and cutting. In the kitchen of catering, S. aureus identification indicates a significant source of contamination. Especially, in raw consumed salad preparation phase of contamination is very important. This food can be a potential source of food-borne poisoning their terms, and they pose a significant risk to consumers have been identified.

Keywords: Staphylococcus aureus, enterotoxin, catering, kitchen, health

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Authors: Ana Paola Echavarria, Mariuxi Medina, Haydelba D'Armas, Carmita Jaramillo, Diana San Martin

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The use of medicinal plants is one of the oldest and most extended medical therapies that goes back to prehistoric times, and nowadays, they are also used in the preparation of phytopharmaceuticals with options to cure diseases. The test for the determination of fungi was carried out in the Pharmacy Pilot Plant (treatment of the leaves of the plant species) and the Microbiology Laboratory (determination of fungi of the plant species, using growth medium called Sabouraud agar plus the vegetal sample), of the Academic Unit of Chemical Sciences and Health, of the Universidad Tecnica de Machala. Subsequently, colony counting was performed, both macroscopic, which is determined in the growth medium of the seeding, and microscopic, to identify the germinative forms using blue lactophenol. The procedure was repeated in duplicate to replicate the results data. The determination of the total fungal content of the following plant species was evaluated: Cymbopogon citratus (lemon verbena), Melissa officinalis (lemon balm), Taraxacum officinale (dandelion), Artemisia absinthium (absinthe), Piper carpunya (guaviduca), Moringa oleifera (moringa), Coriandrum sativum (coriander), Momordica charantia (achochilla), Borago officinalis (borage), Aloysia citriodora (cedron), Ambrosia artemisifolia (altamisa) and Ageratum conyzoides (mastrante). The results obtained showed that all the samples of the twelve plant species studied developed filamentous fungi, with great variability of them, within the permissible limits and contemplated by the Ecuadorian Institute of Normalization (INEN), being suitable as raw material for its use in the preparation of nutraceuticals and medicinal products or phytodrugs; with the exception of A. conyzoides (mastranto) which is the only species that exceeds the regulation in the average of dilutions.

Keywords: colonies, fungi, medicinal plants, microbiological quality, Sabouraud agar

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Authors: John Onolame Unuofin, Uchechukuw U. Nwodo, Anthony I. Okoh

Abstract:

Laccase is constantly gaining status as important biocatalyst in biotechnology. The illimitable potential of its industrial applications and the corresponding aggressive need for phenomenal volumes of extracellularly secreted laccases have called for its interminable production from sources which are able to meet this demand within a relatively short period of time, preferably bacteria. In response to this call, this study was designed to source for laccase-producing bacteria from different environmental matrices. Three sampling environments were chosen such as wastewater treatment plants, University of Fort Hare vicinity and the Hogback woodland, all within the Eastern Cape, South Africa. Samples such as effluents, sediments, leaf litters, degrading wood and rock scrapings were selectively enriched with some model aromatic compounds and were further screened qualitatively and quantitatively on five phenolic substrates ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), Guaiacol, 1-Naphthol, Potassium Ferric Cyanide and Syringaldazine). Basis for selection was their ability to elicit a colour change on at least three of the above mentioned agar based assay substrates. The choice isolates were further identified based on 16S rRNA molecular identification techniques. 33 isolates were screened out of the 40 representative distinct colonies during the qualitative plate screens, while quantitative screens selected out 11 bacterial isolates. They were, based on molecular identification, desginated as members of the genera Pseudomonas, Stenotrophomonas and Citrobacter of the gammaproteobacteria and Bordetalla and Achromobacter of the betaproteobacteria respectively. We therefore conclude based on our outcomes that we may have isolated efficient laccase-producing bacteria, which might be of beneficial significance in catalysis and biotechnology.

Keywords: beta proteobacteria, catalysis, gammaproteobacteria, laccase

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

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Most conventional chemotherapeutic agents, which are used for the treatment of cancers, not only eradicate cancer cells but also affect normal hematopoietic Stem cells (HSCs) that lead to severe pancytopenia during treatment. Therefore, a need exists for distinct approaches to treat cancer without or with minimum effect on normal HSCs. Parthenolide (PTL), a herbal product occurring naturally in the plant Feverfew, is a potential chemotherapeutic agent for the treatment of many cancers, such as acute myeloid leukemia (AML) and chronic lymphocytic leukemia (CLL). In this study we investigated the effect of different PTL concentrations on the viability of normal HSCs and also on the ability of these cells to form colonies after they have been treated with PTL in vitro. Methods: Normal 24 bone marrow and cord blood samples were included in this study after obtaining informed consent. The mononuclear cells were isolated using density gradient separation. Cells were cultured with different PTL concentrations for 24 hours. Post-culture cell viability was assessed using 7ADD in a flow cytometry-based test. In addition, a colony-forming unit assay (CFU) was carried out to assess the effect of PTL on HSCs. the expression of NFҝB was also assessed using a PE-labeled anti-pNFκBP65 antibody. Results: This study showed that there was no statistically significant difference in the percentage of cell death between untreated and PTL-treated cells with 5 μM PTL (p = 0.7), 10 μM PTL (p = 0.4) and 25 μM (p = 0.09) respectively. However, at higher doses, PTL caused a significant increase in the percentage of cell death. These results were significant when compared to untreated control (p < 0.001). The response of cord blood cells (n=4), on the other hand, was slightly different from that for bone marrow cells in that the percentage of cell death was significant at 100 μM PTL. Therefore, cord blood cells seemed more resistant than bone marrow cells. Conclusion: At concentrations ≤25 μM, PTL has a minimum or no effect on HSCs in vitro. Cord blood HSCs are more resistant to PTL compared to bone marrow HSCs.

Keywords: stem cell, parthenolide, ALL, NFKB

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Authors: Haruka Ito, Toru Maruyama, Michihiro Ito, Chuya Shinzato, Hiroyuki Fujimura, Yoshikatsu Nakano, Shoichiro Suda, Sachiyo Aburatani, Haruko Takeyama

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Clarifying symbiotic relationships is one of the most important theme for understanding the marine eco-system. Coral reef has been regarded as an important environmental resource. Coral holobiont composed by coral, symbiotic microalgae zooxanthellae, and bacteria have complexed relationship. Zooxanthellae mainly supply organic matter to the host corals through their photosynthetic activity. The symbiotic relationship is indispensable for corals but may easily collapses due to the rise of seawater temperature. However, the molecular mechanism how seawater temperature influences their relationships still remain unclear. In this study, the transcriptomic analysis has applied to elucidate the coral-zooxanthellae relationships under high seawater temperature stress. To observe reactions of corals and zooxanthellae against the rise of seawater temperature, meta-gene expression in coral have been analyzed. The branches from six different colonies of a stony coral, Acropora tenuis, were sampled at nine times by 2016 at two locations, Ishikawabaru and South of Sesoko Island, Okinawa, Japan. The mRNAs extracted from the branches including zooxanthellae were sequenced by illumina HiSeq. Gene Set Enrichment Analysis (GSEA) based on hyper geometric distribution was performed. The seawater temperature at 2016 summer was unusually high, which was caused by El Niño event, and the number of zooxanthellae in coral was decreased in August. GSEA derived the several specific genes expressed in A. tenuis under heat stress conditions. The upregulated genes under heat stress highly related with infection immunity. The downregulated genes significantly contained cell cycle related genes. Thu, it is considered that heat stress cause disorder in cell metabolism of A. tenuis, resulting in serious influence to coral holobiont.

Keywords: coral, symbiosis, thermal stress response, transcriptome analysis

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Authors: Etleva Hamzaraj, Eva Kica, Anila Paparisto, Pranvera Lazo

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Worldwide demographic developments of recent decades have been associated with negative environmental consequences. For this reason, there is a growing interest in assessing the state of natural ecosystems or assessing human impact on them. In this respect, this study aims to evaluate the change in water quality of the Mat River for a period of about ten years to highlight human impact. In one year, period of study, several biological and environmental parameters are determined to evaluate river water quality, and the data collected are compared with those of a similar study in 2007. Samples are collected every month in five stations evenly distributed along the river. Total coliform bacteria, the number of heterotrophic bacteria in water, and benthic macroinvertebrates are used as biological parameters of water quality. The most probable number index is used for evaluation of total coliform bacteria in water, while the number of heterotrophic bacteria is determined by counting colonies on plates with Plate Count Agar, cultivated with 0.1 ml sample after series dilutions. Benthic macroinvertebrates are analyzed by the number of individuals per taxa, the value of biotic index, EPT Richness Index value and tolerance value. Environmental parameters like pH, temperature, and electrical conductivity are measured onsite. As expected, the bacterial load was higher near urban areas, and the pollution increased with the course of the river. The maximum concentration of fecal coliforms was 1100 MPN/100 ml in summer and near the most urbanized area of the river. The data collected during this study show that after about ten years, there is a change in water quality of Mat River. According to a similar study carried out in 2007, the water of Mat River was of ‘excellent’ quality. But, according to this study, the water was classified as of ‘excellent’ quality only in one sampling site, near river source, while in all other stations was of ‘good’ quality. This result is based on biological and environmental parameters measured. The human impact on the quality of water of Mat River is more than evident.

Keywords: water quality, coliform bacteria, MPN index, benthic macroinvertebrates, biotic index

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Authors: Adrizal, Irsan Ryanto, Sri Juwita, Adika Sugara, Tino Bapirco

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The objective of the research was to study the influence of the inclusion of the cabbage waste into a buffalo rations made of sugarcane waste on the feed formula and characteristic of complete feed silage. Research carried out a two-stage i.e. the feed formulation and experiment of making complete feed silage. Feed formulation is done by linear programming. Data input is the price of feed stuffs and their nutrient contents as well as requirements for rations, while the output is the use of each feed stuff and the price of complete feed. The experiment of complete feed silage was done by a completely random design 4 x 4. The treatments were 4 inclusion levels of the cabbage waste i.e. 0%,(T1) 5%(T2), 10%(T3) and 15% (T4), with 4 replications. The result of feed formulation for T1 was cabbage (0%), sugarcane top (17.9%), bagasse (33.3%), Molasses (5.0%), cabagge (0%), Thitonia sp (10.0%), rice brand (2.7%), palm kernel cake (20.0%), corn meal (9.1%), bond meal (1.5%) and salt (0.5%). The formula of T2 was cabagge (5%), sugarcane top (1.7%), bagasse (45.2%), Molasses (5.0%), , Thitonia sp (10.0%), rice brand (3.6%), palm kernel cake (20.0%), corn meal (7.5%), bond meal (1.5%) and salt (0.5%). The formula of T3 was cabbage (10%), sugarcane top (0%), bagasse (45.3%), Molasses (5.0%), Thitonia sp (10.0%), rice brand (3.8%), palm kernel cake (20.0%), corn meal (3.9%), bond meal (1.5%) and salt(0.5%). The formula of T4 was cabagge (15.0%), sugarcane top (0%), bagasse (44.1%), Molasses (5.0%), Thitonia sp (10.0%), rice brand (3.9%), palm kernel cake (20.0%), corn meal (0%), bond meal (1.5%) and salt (0.5%). An increase in the level of inclusion of the cabbage waste can decrease the cost of rations. The cost of rations (IDR/kg on DM basis) were 1442, 1367, 1333, and 1300 respectively. The rations formula were not significantly (P > 0.05) influent the on fungal colonies, smell, texture and color of the complete ration silage, but the pH increased significantly (P < 0.05). It concluded that inclusion of cabbage waste can minimize the cost of buffalo ration, without decreasing the silage quality of complete feed.

Keywords: buffalo, cabbage, complete feed, sillage characteristic, sugarcane waste

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Authors: Rawia M. Khalil, Ahmed A. Abd El Rahman, Mahfouz A. Kassem, Mohamed S. El Ridi, Mona M. Abou Samra, Ghada E. A. Awad, Soheir S. Mansy

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Solid lipid nanoparticles (SLNs) have gained great attention for the topical treatment of skin associated fungal infection as they facilitate the skin penetration of loaded drugs. Our work deals with the preparation of nystatin loaded solid lipid nanoparticles (NystSLNs) using the hot homogenization and ultrasonication method. The prepared NystSLNs were characterized in terms of entrapment efficiency, particle size, zeta potential, transmission electron microscopy, differential scanning calorimetry, rheological behavior and in vitro drug release. A stability study for 6 months was performed. A microbiological study was conducted in male rats infected with Candida albicans, by counting the colonies and examining the histopathological changes induced on the skin of infected rats. The results showed that SLNs dispersions are spherical in shape with particle size ranging from 83.26±11.33 to 955.04±1.09 nm. The entrapment efficiencies are ranging from 19.73±1.21 to 72.46±0.66% with zeta potential ranging from -18.9 to -38.8 mV and shear-thinning rheological Behavior. The stability studies done for 6 months showed that nystatin (Nyst) is a good candidate for topical SLN formulations. A least number of colony forming unit/ ml (cfu/ml) was recorded for the selected NystSLN compared to the drug solution and the commercial Nystatin® cream present in the market. It can be fulfilled from this work that SLNs provide a good skin targeting effect and may represent promising carrier for topical delivery of Nyst offering the sustained release and maintaining the localized effect, resulting in an effective treatment of cutaneous fungal infection.

Keywords: candida infections, hot homogenization, nystatin, solid lipid nanoparticles, stability, topical delivery

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Authors: Leila Rashki Ghaleno, Mohamad Amin Hajari, Leila Montazeri, Abdolhossein Shahverdi, Mojtaba Rezazadeh Valojerdi

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Background: Spermatogonia stem cells (SSCs) exhibit pluripotency, enabling them to undergo differentiation into many cell lineages, including neurons, glia, endothelial cells, and hepatocytes when cultured in vitro. Although the specific mechanisms are not yet fully understood, it has been observed that biopolymer agents, such as hyaluronic acid (HA) and alginate (Alg), have the potential to induce transdifferentiation of SSCs. The current work aimed to examine the process of in vitro spermatogenesis and the conversion of mouse testicular cells into hepatocytes and erythrocyte-like cells utilizing the HA-Alg hydrogel. Method: After being extracted from the testes of a 5-day postpartum mouse (5 DPP), the testicular cells were separated into two enzymatic stages and then put into a composite hydrogel containing 0.5% HA and 1% alginate. On days 14 and 28 of culture, the colonies' growth, the cells' viability, and their histology were assessed. Result: Despite observing significant cell proliferation on day 14 and the development of circular-shaped organoids on day 28, it was noted that the organoids generated in the HA-Alg medium tended to maintain their circular morphology on day 28. Notably, the testicular cells underwent transdifferentiation into cell types resembling erythrocytes and hepatocytes. The hepatocyte-like cells exhibited the presence of glycogen and lipid deposits, indicating their hepatocyte-like characteristics. Interestingly, immunostaining analysis revealed the secretion of albumin and the presence of VEGFR on day 14. However, on day 28, albumin expression was not detected, while the expression of Sox9 (a marker for hepatocytes), Vegf, CD34, and C-kit (markers for erythrocytes) showed increased levels in the gene expression evaluation. Conclusion: The present findings indicated that HA-Alg could be a potent and effective agent for the transdifferentiation of testis cells into erythrocyte and hepatocyte-like cells, as recent studies have confirmed the transformation of SSCs into hepatocyte cells during in vitro culture.

Keywords: 3D culture, mouse testicular cell, hyaluronic acid, liver organoids

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