Search results for: Thai isolates
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 946

Search results for: Thai isolates

766 Effects of Microbial Biofertilization on Nodulation, Nitrogen Fixation, and Yield of Lablab purpureus

Authors: Benselama Amel, Ounane S. Mohamed, Bekki Abdelkader

Abstract:

A collection of 20 isolates from fresh Nodules of the legume plant Lablab purpureus was isolated. These isolates have been authenticated by seedling inoculation grown in jars containing sand. The results obtained after two months of culture have revealed that the 20 isolates (100% of the isolates) are able to nodulate their host plants. The results obtained were analyzed statistically by ANOVA using the software statistica and had shown that the effect of the inoculation has significantly improved all the growth parameters (the height of the plant and the dry weight of the aerial parts and roots, and the number of nodules). We have evaluated the tolerance of all strains of the collection to the major stress factors as the salinity, pH and extreme temperature. The osmotolerance reached a concentration up to 1710mm of NaCl. The strains were also able to grow on a wide range of pH, ranging from 4.5 to 9.5, and temperature, between 4°C and 40°C. Also, we tested the effect of the acidity, aluminum and ferric deficit on the Lablab-rhizobia symbiosis. Lablab purpureus has not been affected by the presence of high concentrations of aluminum. On the other hand, iron deficiency has caused a net decrease in the dry biomass of the aerial part. The results of all the phenotypic characters have been treated by the statistical Minitab software, the numerical analysis had shown that these bacterial strains are divided into two distinct groups at a level of similarity of 86 %. The SDS-PAGE was carried out to determine the profile of the total protein of the strains. The coefficients of similarity of polypeptide bands between the isolates and strains reference (Bradyrhizobium, Mesorizobium sp.) confirm that our strain belongs to the groups of rhizobia.

Keywords: SDS-PAGE, rhizobia, symbiosis, phenotypic characterization, Lablab purpureus

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765 Strengthening the Security of the Thai-Myanmar Border Trade of the People in the Mae Sot Customs Checkpoint Area, Tak Province

Authors: Sakapas Saengchai

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A Study on Strengthening the Security of the Thai-Myanmar Border Trade Area of the people in the Mae Sot customs checkpoint area, Tak province, was designed as a qualitative research study. Its objectives were to study the principles of strengthening border trade security and enhancing people's participation. To develop a border trade model that enhances the spatial economy and improves people's quality of life by collecting data using a participant observation method. In-depth interview group chats border checkpoint administrators, Mae Sot customs checkpoint, Tak province, private entrepreneurs, community leaders, and the opening of a community forum to exchange opinions with people in the area. The results of the study found that 1. Security development is to promote crime reduction. Reduce drug trafficking problems Smuggling and human trafficking have been reduced. Including planning and preparation to protect people from terrorism, epidemics, and communicable diseases, including cooperation with Burma on border rules for people and workers, 2. Wealth development is to promote investment. Transport links value chain logistics Cross-border goods and services on the Thai-Myanmar border Both amending regulations and laws to promote fair trade. Emphasis on convenient and fast service as well as promoting the Thai border area to be a tourist attraction that can create prosperity and income for the community in the area By using balanced natural resources, with production and consumption that are environmentally friendly, and emphasizes the participation of the public sector, the private sector, and people from all sectors in the sustainable development of the Thai border.

Keywords: security, border trade, customs, participation, people

Procedia PDF Downloads 149
764 Thai Teenage Prostitution Online

Authors: Somdech Rungsrisawat

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The purposes of this research are to investigate Thai teens’ attitude toward prostitution on the internet, to discover the causes of teenage prostitution and to study the relationship between teenage promiscuity and the causes of teenage prostitution. This study is a mixed research which utilized both qualitative and quantitative approach. The population of this study included teenagers and early adults between 14-21 years old who were studying in high schools, colleges, or universities. A total of 600 respondents was sampled for interviews using a questionnaire, and 48 samples were chosen for an in-depth interview. The findings revealed that the majority of respondents recognized that teenage prostitution on line was real. The reasons for choosing the internet to contact with customers included easy, convenient, safe, and anonymous. Moreover, the internet allowed teen prostitutes to contact customers anywhere and anytime. The correlation showed that promiscuity was related to the trend of teen prostitution. Other factors that contributed to increasing widespread teen prostitution online included their need for quick money to buy luxurious products and to support their extravagant behavior.

Keywords: internet, prostitutes, online, Thai teens

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763 State and Determinant of Caregiver’s Mental Health in Thailand: A Household Level Analysis

Authors: Ruttana Phetsitong, Patama Vapattanawong, Malee Sunpuwan, Marc Voelker

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The majority of care for older people at home in Thai society falls upon caregivers resulting in caregiver’s mental health problem. Beyond individual characteristics, household factors might have a profound effect on the caregiver’s mental health. But reliable data capturing this at the household level have been limited to date. The objectives of the present study were to explore the levels of Thai caregiver’s mental health and to investigate the factors affecting the mental health at household level. Data were obtained from the 2011 National Survey of Thai Older Persons conducted by the National Statistical Office of Thailand. Caregiver’s mental health was measured by using the 15- items-short version of the Thai Mental Health Indicator (TMHI-15) developed by the Department of Mental Health, the Ministry of Public Health. Multivariate logistic regression models were used to explore the impact of potential factors on caregiver’s mental health. The THMI-15 produced an overall average caregiver mental health score of 30.9 out of 45 (SD 5.3). The score can be categorized into good (34.02-45), fair (27.01-34), and poor (0-27). Duration of care for older people, household wealth, and functional dependency of the older people significantly predicted total caregiver’s mental health. Household economic factor was key in predicting better mental health. Compared to those poorest households, the adjusted effect of the fifth quintile household wealth was high (OR=2.34; 95%CI=1.47-3.73). The findings of this study provide a fuller picture to a better understanding of the level and factors that cause the mental health of Thai caregivers. Health care providers and policymakers should consider these factors when designing interventions aimed at alleviating caregiver’s psychological burden when provided care for older people at home.

Keywords: caregiver’s mental health, household, older people, Thailand

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762 Youth Friendly Health Services for Rural Thai Teenagers

Authors: C. Sridawruang

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Young people today has sexual activities differing from those of earlier generations, in that teenagers are likely to have multiple partners, and are frequently in short-term relationships or with partners that are not well known to them. The proportion of teenage mothers in Thailand has increased. Young people were not specifically addressed during the overall very successful HIV-prevention campaigns. Because of this missed opportunity, they are still unaware of the risk of unsafe sexual behavior. Aims: To describe the reproductive health care services in perspectives of rural Thai teenagers Methods: This survey was one part of a mixed method approach taken using survey and focus groups with 439 teenagers aged 12-18 years in 5 villages, Udon Thani, Thailand. The standard questionnaire survey had been used for collecting data. The numeric data was checked and analyzed by using descriptive statistics. Results: Most teenager respondents stated that they do not know where sexual reproductive health services provided for them. Most teenagers felt difficult to access and talk with health staff about sexual related issues. They stated that discussing, or consulting with health providers might not be safe. Teenagers might lose opportunities to access and get advice from health care services. The mean knowledge score of contraception and condom reproductive was 6.34 from a total score 11. Most teenagers especially girls expressed a need for counseling services and reported a need for telephone services. Conclusions: The need of appropriate information focusing on sexual relationships and contraception should be designed to help young people make wise decisions and there should be set health care services for Thai teenagers to make sure that teenagers could access easily. Health care providers need to be trained to improve their knowledge, attitudes and skills in reproductive health care practices for Thai teenagers.

Keywords: youth friendly health services, rural, Thai, teenagers

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761 Prevalence and Antibiotic Susceptibility of Bacterial Isolates from Mastitis Milk of Cow and Buffalo in Udaipur, India

Authors: Hardik Goswami, Gayatri Swarnakar

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-Mastitis disease has been known as one of the most costly diseases of dairy cattle and observed as an inflammatory disease of cow and buffalo udder. Mastitis badly affected animal health, quality of milk and economics of milk production along with cause’s great economic loss. Bacteria have been representing the most common etiological agents of mastitis. The antibiotic sensitivity test was important to attain accurate treatment of mastitis. The aim of present research work was to explore prevalence and antibiotic susceptibility pattern of bacterial isolates recovered from cow and buffalo clinical mastitis milk sample. During the period of April 2010 to April 2014, total 1487 clinical mastitis milk samples of cow and buffalo were tested to check the prevalence of mastitis causing bacterial isolates. Milk samples were collected aseptically from the udder at the time of morning milking. The most prevalent bacterial isolates were Staphylococcus aureus (24.34%) followed by coliform bacteria (15.87%), coagulase negative Staphylococcus aureus (13.85%), non-coliform bacteria (13.05%), mixed infection (12.51%), Streptococcus spp. (10.96%). Out of 1487, 140 (9.42%) mastitis milk samples showed no growth on culture media. Identification of bacteria made on the basis of Standard Microbial features and procedures. Antibiotic susceptibility of bacterial isolates was investigated by Kirby-Bauer disk diffusion method. In vitro Antibiotic susceptibility test of bacterial isolates revealed higher sensitivity to Gentamicin (74.6%), Ciprofloxacin (62.1%) and Amikacin (59.4%). The lower susceptibility was shown to Amoxicillin (21.6%), Erythromycin (26.4%) and Ceftizoxime (29.9%). Antibiotic sensitivity pattern revealed Gentamicin are the possible effective antibiotic against the major prevalent mastitis pathogens. Present research work would be helpful in increase production, quality and quantity of milk, increase annual income of dairy owners and improve health of cow and buffaloes.

Keywords: antibiotic, buffalo, cow, mastitis, prevalence

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760 Efficiency Analysis of Trader in Thailand and Laos Border Trade: Case Study of Textile and Garment Products

Authors: Varutorn Tulnawat, Padcharee Phasuk

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This paper investigates the issue of China’s dumping on border trade between Thailand and Laos. From the pass mostly, the border trade goods are traditional textile and garment mainly served locals and tourists which majority of traders is of small and medium size. In the present day the competition is fierce, the volume of trade has expanded far beyond its original intent. The major competitors in Thai-Laos border trade are China, Vietnam and also South Korea. This research measures and compares the efficiency and ability to survive the onslaught of Thai and Laos firm along Thailand (Nong Kai province) and Laos (Vientiane) border. Two attack strategies are observed, price cutting and incense such as full facilitation for big volume order. Data Envelopment Analysis (DEA) is applied to data surveyed from 90 Thai and Laos entrepreneurs. The expected results are the proportion of efficiency and inefficiency firms. Points of inefficiency and suggested improvement are also discussed.

Keywords: border trade, dea, textile, garment

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759 Metadiscourse in Chinese and Thai Request Emails: Analysis and Pedagogical Application

Authors: Chia-Ling Hsieh, Kankanit Potikit

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Metadiscourse refers to linguistic resources employed by writers to organize text and interact with readers. While metadiscourse has received considerable attention within the field of discourse analysis, few studies have explored the use of metadiscourse in email, one of the most popular forms of computer-mediated communication. Furthermore, the diversity of cross-linguistic research required to uncover the influence of cultural factors on metadiscourse use is lacking. The present study compares metadiscourse markers employed in Chinese and Thai-language request emails with the purpose of discovering cross-cultural similarities and differences that are meaningful and applicable to foreign language teaching. The analysis is based on a corpus of 200 request emails: 100 composed in Chinese and 100 in Thai, with half of the emails from each language data set addressed to professors and the other half addressed to classmates. Adopting Hyland’s model as an analytical framework, two primary categories of metadiscourse are identified. Textual metadiscourse helps to create text coherence, while interpersonal metadiscourse functions to convey authorial stance. Results of the study make clear that both Chinese and Thai-language emails use significantly more interpersonal markers than textual markers, indicating that email, as a unique communicative medium, is characterized by high degrees of concision and interactivity. Users of both languages further deploy similar patterns in writing emails to recipients of different social statuses. Compared with emails addressed to classmates, emails addressed to professors are notably longer and include more transition and engagement markers. Nevertheless, cultural factors do play a role. Emails composed in Thai, for example, include more textual markers than those in Chinese, as Thai favors formal expressions and detailed explanations, while in contrast, emails composed in Chinese employ more interpersonal markers than those in Thai, since Chinese tends to emphasize recipient involvement and attitudinal warmth. These findings thereby demonstrate the combined effects of email as a communicative medium, social status, and cultural values on metadiscourse usage. The study concludes by applying these findings to pedagogical suggestions for teaching email writing to Chinese and Thai language learners based on similarities and differences in metadiscourse strategy between the two languages.

Keywords: discourse analysis, email, metadiscourse, writing instruction

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758 Isolation and Characterization of Indigenous Rhizosphere Bacteria Producing Gibberellin Acid from Local Soybeans in Three Different Areas of South Sulawesi

Authors: Asmiaty Sahur, Ambo Ala, Baharuddin Patanjengi, Elkawakib Syam'un

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This study aimed to isolate and characterize the indigenous Rhizosphere bacteria producing Gibberellin Acid as plant growth isolated from local soybean of three different areas in South Sulawesi, Indonesia. Several soil samples of soybean plants were collected from the Rhizosphere of local soybeans in three different areas of South Sulawesi such as Soppeng, Bone and Takalar. There were 56 isolates of bacteria were isolated and grouped into gram-positive bacteria and gram negative bacteria .There are 35 isolates produce a thick slime or slimy when cultured on media Natrium Broth and the remaining of those produced spores. The results showed that of potential bacterial isolated produced Gibberellin Acid in high concentration. The best isolate of Rhizosphere bacteria for the production of Gibberellin Acid is with concentration 2%. There are 4 isolates that had higher concentration are AKB 19 (4.67 mg/ml) followed by RKS 17 (3.80 mg/ml), RKS 25 (3.70 mg / ml) and RKS 24 (3.29 mg/ml) respectively.

Keywords: rhizosphere, bacteria, gibberellin acid, soybeans

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757 Competitiveness of Animation Industry: The Case of Thailand

Authors: T. Niracharapa

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The research studied and examined the competitiveness of the animation industry in Thailand. Data were collected based on articles, related reports and websites, news, research, and interviews of key persons from both public and private sectors. The diamond model was used to analyze the study. The major factor driving the Thai animation industry forward includes a quality workforce, their creativity and strong associations. However, discontinuity in government support, infrastructure, marketing, IP creation and financial constraints were factors keeping the Thai animation industry less competitive in the global market.

Keywords: animation, competitiveness, government, Thailand, market

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756 Diversity and Distribution of Cytochrome P450 2C9 Genes Related with Medical Cannabis in Thai Patients

Authors: Tanakrit Doltanakarn

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Introduction: These days, cannabis is being accepted in many countries due to the fact that cannabis could be use in medical. The medical cannabis is used to treat and reduce the pain many diseases. For example, neuropathic pain, Parkinson, autism disorders, cancer pain reduce the adverse effect of chemotherapy, diabetes, and migraine. Active ingredients in cannabis that modulate patients' perceptions of their conditions include Δ9‐tetrahydrocannabinol (THC), cannabidiol (CBD), flavonoids, and terpenes. However, there is an adverse effect of cannabis, cardiovascular effects, psychosis, schizophrenia, mood disorder, and cognitive alternation. These effects are from the THC and CBD ingredients in the cannabis. The metabolize processes of delta-9 THC to 11-OH-delta 9 -THC (inactive form), THC were cause of adverse effects. Interestingly, the distributions of CYP2C9 gene (CYP2C9*2 and CYP2C9*3, poor metabolizer) that might affect incidences of adverse effects in patients who treated with medical cannabis. Objective: The aim of this study we want to investigate the association between genetic polymorphism of CYP2C9 frequency and Thai patients who treated with medical cannabis. Materials and Methods:We recruited sixty-five unrelated Thai patients from the College of Pharmacy, Rangsit University. DNA were extracted using Genomic DNA Mini Kit. Genotyping of CYP2C9*2 (430C>T, rs1799853) and CYP2C9*3 (1075A>C, rs1057910) were genotyped by the TaqMan Real-time PCR assay. Results: Among these 31 medicals cannabis-induced ADRs patients, they were diagnosed with 22 (33.85%) tachycardia and 3 (4.62%) arrhythmia. There were 34 (52.31%) medical cannabis-tolerant controls who were included in this study.40 (61.53%) Thai patients were female, and 25 (38.46%) were male, with median age of 57 (range 27 – 87) years. In this study, we found none of the medical cannabis-induced ADRs carried CYP2C9*2 variant along with medical cannabis-tolerant control group. CYP2C9*3 variant (intermediate metabolizer, IM) was found just only one of thirty-one (3.23%) in the medical cannabis-induced ADRs and two of thirty-fourth (5.88%) in the tolerant controls. Conclusions: Thus, the distribution of CYP2C9 alleles offer a comprehensive view of pharmacogenomics marker in Thai population that could be used as a reference for worldwide to investigate the pharmacogenomics application.

Keywords: medical cannabis, adverse effect, CYP2C9, thai patients

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755 Error Analysis in English Essays Writing of Thai Students with Different English Language Experiences

Authors: Sirirat Choophan Atthaphonphiphat

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The objective of the study is to analyze errors in English essay writing of Thai (Suratthani Rajabhat University)’s students with different English language experiences. 16 subjects were divided into 2 groups depending on their English language experience. The data were collected from English essay writing about 'My daily life'. The finding shows that 275 tokens of errors were found from 240 English sentences. The errors were categorized into 4 types based on frequency counts: grammatical errors, mechanical errors, lexical errors, and structural errors, respectively. The findings support all of the researcher’s hypothesizes, i.e. 1) the students with low English language experience made more errors than those with high English language experience; 2) all errors in English essay writing of Suratthani Rajabhat University’s students, the interlingual errors are more than the intralingual ones; 3) systemic and structural differences between English (target language) and Thai (mother-tongue language) lead to the errors in English essays writing of Suratthani Rajabhat University’s students.

Keywords: applied linguistics, error analysis, interference, language transfer

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754 Analysis of Population and Growth Rate Methanotof Bateria as Reducers Methane Gases Emission in Rice Field

Authors: Maimuna Nontji

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The life cycle of rice plant has three phases of growth; they are the vegetative, reproductive and maturation phase. They greatly affect the life of dynamics metanotrof bacterial as reducer methane emissions in the rice field, both of population and on the rate of growth. The aim of this study was to analyze the population and growth rate of methanotrof isolates which has been isolated in previous studies. Isolates were taken at all the life cycle of rice plant. Population of analysis was conducted by standard plate count method and growth rate was analysed by logarithmic calculation. The results showed that each isolate varied in population and growth rate. The highest population was obtained in the isolates Gowa Methanotrof Reproductive (GMR 8) about 7.06 x 10 11 cfu / ml on 3 days of incubation and the lowest population was obtained in the Gowa Methanotrof Maturation (GMP 5) about 0.27 x 10 11 cfu / ml on 7 day of incubation. Some isolate were demonstrated in long growth rate about 5 days of incubation and another are 3 days.

Keywords: emission, methanotrof, methane, population

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753 The Design Inspired by Phra Maha Chedi of King Rama I-IV at Wat Phra Chetuphon Vimolmangklaram Rajwaramahaviharn

Authors: Taechit Cheuypoung

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The research will focus on creating pattern designs that are inspired by the pagodas, Phra Maha Chedi of King Rama I-IV, that are located in the temple, Wat Phra Chetuphon Vimolmangklararm Rajwaramahaviharn. Different aspects of the temple were studied, including the history, architecture, significance of the temple, and techniques used to decorate the pagodas, Phra Maha Chedi of King Rama I-IV. Moreover, composition of arts and the form of pattern designs which all led to the outcome of four Thai application pattern. The four patterns combine Thai traditional design with international scheme, however, maintaining the distinctiveness of the glaze mosaic tiles of each Phra Maha Chedi. The patterns consist of rounded and notched petal flowers, leaves and vine, and various square shapes, and original colors which are updated for modernity. These elements are then grouped and combined with new techniques, resulting in pattern designs with modern aspects and simultaneously reflecting the charm and the aesthetic of Thai craftsmanship which are eternally embedded in the designs.

Keywords: Chedi, Pagoda, pattern, Wat

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752 Thai Teachers' Growth Mindset as Related to Thai Students' Achievements

Authors: Chintida Vichitsophaphan, Piyapat Chitpirom, Chaichana Nimnuan, Teerakiat Jareonsettasin

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The purpose of this research is to study the influence of a growth mindset, as defined by Prof. Dweck, in Thai teachers and Thai students’ achievements and success. The participants of the study were teachers of schools in Office of the Basic Education Commission in Bangkok. The teachers were recruited from high achievement schools and low achievement schools (based on average National Standard Test Score). Participants were divided into two groups: 85 teachers in 3 high achievement schools and 213 teachers in 6 low achievement schools. They were asked to complete the Carol Dweck’s Implicit Theories Scale – (Adults) 8 items. Data were analyzed including the use of mean, standard deviation and t-test to test hypothesis. The finding of this study revealed that teachers who were in the high achievement schools have higher scores in Carol Dweck’s Implicit Theories Scale (x ̅= 35.5, SE = .58) than teachers who were in the low achievement schools (x ̅= 33.9, SE = .35) at .05 level. The difference is statistically significant (t (296) = 2.44, p = .015) with the effect size of 0.31. In conclusion, teachers’ growth mindset from high achievement schools have higher scores than teachers’ growth mindset from low achievement schools, and this is statistically significant. From the study, it can be concluded that growth mindset development for teachers has a tendency to increase students’ achievements. For these reasons, it is necessary to implement such training and development in our education system in larger scale, and even nationwide policies.

Keywords: fixed mindset, growth mindset, students’ achievement, teachers’ growth mindset

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751 The Relationship of Service Marketing Mix and Intention to Repurchase of Thai Dessert

Authors: Siri-Orn Champatong

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This research aimed to study the relationship between attitudes toward marketing mix and customers’ repurchase intention of Thai dessert in case of Ekachai Salee Suphan’s shop in Thailand. This study employed by survey and quantitative research and the questionnaire was used to collect the data from 385 sampled of customers who visited at Ekachai Salee Suphan's shop in Thailand. The descriptive statistics and Pearson’s correlation coefficient analysis was used to analyze data. The research found that the customers’ perception on the overall and individual aspects of attitudes toward marketing mix include products, prices, promotions, and physical characteristics were at very good level, but distribution channels, staffs, and management process were at good level. Furthermore, the hypothesis tests found that attitudes toward overall marketing mix had relationship with individual aspects of intention that were the repurchase intention, willingness to recommend others to repurchase, and possibility to repurchase with statistical significance level of 0.01.

Keywords: intention, repurchase, service marketing mix, Thai dessert

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750 Genotypic Characterization of Gram-Positive Bacteria Isolated on Ornamental Animals Feed

Authors: C. Miranda, R. Soares, S. Cunha, L. Ferreira, G. Igrejas, P. Poeta

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Different animal species, including ornamental animals, are reported as potential reservoirs of antibiotic resistance genes. Consequently, these resistances can be disseminated in the environment and transferred to humans. Moreover, multidrug-resistant bacteria reduce the efficacy of antibiotics, as the case of vancomycin-resistant enterococci. Enterococcus faecalis and E. faecium are described as the main nosocomial pathogens. In this line, the aim of this study was to characterize resistance and virulence genes of enterococci species isolated from samples of food supplied to ornamental animals during 2020. The 29 enterococci isolates (10 E. faecalis and 19 E. faecium) were tested for the presence of the resistance genes for the following antibiotics: erythromicyn (ermA, ermB and ermC), tetracycline (tetL, tetM, tetK and tetO), quinupristin/dalfopristin (vatD and vatE), gentamicin (aac(6’)-aph(2’’)-Ia), chloramphenicol (catA), streptomycin (ant(6)-Ia) and vancomycin (vanA and vanB). The same isolates were also tested for 10 virulence factors genes (esp, ace, gelE, agg, fsr, cpd, cylA, cylB, cylM and cylLL). The resistance and virulence genes were performed by PCR, using specific primers and conditions. Negative and positive controls were used in all PCR assays. The most prevalent resistance genes detected in both enterococci species were ermB (n=15, 52%), ermC (n=7, 24%), tetK (n=8, 28%) and vatE (n=4, 14%). Resistance genes for vancomycin were found in ten (34%) E. faecalis and ten (34%) E. faecium isolates. Only E. faecium isolates showed the presence of ermA (n=2, 7%), tetL (n=13, 45%) and ant(6)-Ia gene (n=4, 14%). A total of nine (31%) enterococci isolates were classified as multidrug-resistant bacteria (3 E. faecalis and 6 E. faecium). In three E. faecalis and one E. faecium were not detected resistance genes. The virulence genes detected in both species were agg (n=6, 21%) and cylLL (n=11, 38%). In general, each isolate showed only one of these virulence genes. Five E. faecalis and eleven E. faecium isolates were negative for all analyzed virulence genes. These preliminary results showed the presence of multidrug-resistant enterococci in food supplied to ornamental animals, in particular vancomycin-resistant enterococci. This genotypic characterization reinforces the relevance to public health in the control of antibiotic-resistant bacteria.

Keywords: antibiotic resistance, enterococci, feed, ornamental animals

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749 Lack of Association between IL-10 Promoter Gene Polymorphisms and Tuberculosis Susceptibility in Thai Population

Authors: Manaphol Kulpraneet, Anirut Limtrakul, Surangrat Srisurapanon, Piyatida Tangteerawatana

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Tuberculosis (TB) remains a global health care disease world-wide. Control of the global TB epidemic has been impaired by the lack of an effective vaccine, by the emergence of drug resistant forms of Mycobacterium tuberculosis and by lack of sensitive and rapid diagnostics. Cytokines play a major role in defense against M. tuberculosis infection. Polymorphisms in the genes encoding various cytokines have been associated with tuberculosis susceptibility. Polymorphisms of the regulatory cytokine gene, the interleukin (IL)-10 is associated with the risk of tuberculosis (TB) in different populations. However, IL-10 gene polymorphism and susceptibility to TB in Thai is still unknown. The purpose of this study was to evaluate whether the common IL-10 promoter gene polymorphisms are associated with TB in Thai population. Forty eight patients with newly diagnosed pulmonary tuberculosis were studied. DNA samples were extracted from leukocytes and used to investigate -1087A/G, -819C/T, -252C/A (rs1800896, rs1800871, rs1800872) in IL-10 gene using restriction fragment length polymorphism (PCR-RFLP) methods. In this study, the genotype and allele frequencies of IL-10-1087A/G, -819C/T, -252C/A polymorphism did not significantly different between TB patients and healthy controls ((genotype: p=0.38, p=0.92, p=1; allele: p=0.57, p=0.77, p=0.89, respectively). The lack of association between common IL-10 promoter polymorphisms and TB susceptibility in this study may provide clue for better understanding of IL-10-1087A/G, -819C/T, -252C/A polymorphism and TB susceptibility in Thai population, which might facilitate the rationale design of vaccines. However, further studies in large scales population are required for confirmation.

Keywords: IL-10, cytokines, single nucleotide polymorphism (SNP), tuberculosis

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748 Phylogenetic Diversity and Antibiotic Resistance in Sediments of Aegean Sea

Authors: Ilknur Tuncer, Nihayet Bizsel

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The studies in bacterial diversity and antimicrobial resistance in coastal areas are important to understand the variability in the community structures and metabolic activities. In the present study, antimicrobial susceptibility and phylogenetic analysis of bacteria isolated from stations with different depths and influenced by terrestrial and marine fluxes in eastern Aegean Sea were illustrated. 51% of the isolates were found as resistant and 14% showed high MAR index indicating the high-risk sources of contamination in the environment. The resistance and the intermediate levels and high MAR index of the study area were 38–60%, 11–38% and 0–40%, respectively. According to 16S rRNA gene analysis, it was found that the isolates belonged to two phyla Firmicutes and Gammaproteobacteria with the genera Bacillus, Halomonas, Oceanobacillus, Photobacterium, Pseudoalteromonas, Psychrobacter, and Vibrio. 47% of Bacillus strains which were dominant among all isolates were resistant. In addition to phylogenetically diverse bacteria, the variability in resistance, intermediate and high MAR index levels of the study area indicated the effect of geographical differences.

Keywords: bacterial diversity, multiple antibiotic resistance, 16S rRNA genes, Aegean Sea

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747 Mechanical and Physical Properties of Various Types of Dental Floss

Authors: Supanitayanon Lalita, Dechkunakorn Surachai, Anuwongnukroh Niwat, Srikhirin Toemsak, Roongrujimek Pitchaya, Tua-Ngam Peerapong

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Objective: To compare maximum load, percentage of elongation, physical characteristics of 4 types of dental floss: (1) Thai Silk Floss (silk, waxed), (2) Oral B® Essential Floss (nylon, waxed), (3) Experimental Floss Xu (nylon, unwaxed), (4) Experimental Floss Xw (nylon, waxed). Materials & method: Four types of floss were tested (n=30) with a Universal Testing Machine (Instron®). Each sample (30 cm long, 5 cm segment) was fixed, and pulled apart with load cell of 100 N and a test speed of 100 mm/min. Physical characteristics were investigated by digital microscope under 2.5×10 magnification, and scanning electron microscope under 1×100 and 5×100 magnification. The size of the filaments was measured in micron (μm) and the fineness were measured in Denier. Statistical analysis: For mechanical properties, the maximum load and the percentage of elongation were presented as mean ± SD. The distribution of the data was calculated by the Kolmogorov-Smirnov test. One-way ANOVA and multiple comparison (Tukey HSD) were used to analyze the differences among the groups with the level of a statistical difference at p < 0.05. Results: The maximum load of Floss Xu, Floss Xw, Oral B and Thai Silk were 47.39, 46.46, 25.38, and 23.70 N, respectively. The percentage of elongation of Oral B, Floss Xw, Floss Xu and Thai Silk were 72.43, 44.62, 31.25, and 16.44%, respectively. All 4 types of dental floss showed statistically differences in both the maximum load and percentage of elongation at p < 0.05, except for maximum load between Floss Xw and Floss Xu that showed no statistically significant difference. Physical characteristics of Thai silk revealed the most disintegrated, the smallest, and the least fine filaments. Conclusion: Floss Xu had the highest maximum load. Oral B had the highest percentage of elongation. Wax coating on Floss X increased the elongation but had no significant effect on the maximum load. The physical characteristics of Thai Silk resulted in the lowest mechanical properties values.

Keywords: dental floss, maximum load, mechanical property, percentage of elongation, physical property

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746 The Effects of Negative Electronic Word-of-Mouth and Webcare on Thai Online Consumer Behavior

Authors: Pongsatorn Tantrabundit, Lersak Phothong, Ong-art Chanprasitchai

Abstract:

Due to the emergence of the Internet, it has extended the traditional Word-of-Mouth (WOM) to a new form called “Electronic Word-of-Mouth (eWOM).” Unlike traditional WOM, eWOM is able to present information in various ways by applying different components. Each eWOM component generates different effects on online consumer behavior. This research investigates the effects of Webcare (responding message) from product/ service providers on negative eWOM by applying two types of products (search and experience). The proposed conceptual model was developed based on the combination of the stages in consumer decision-making process, theory of reasoned action (TRA), theory of planned behavior (TPB), the technology acceptance model (TAM), the information integration theory and the elaboration likelihood model. The methodology techniques used in this study included multivariate analysis of variance (MANOVA) and multiple regression analysis. The results suggest that Webcare does slightly increase Thai online consumer’s perceptions on perceived eWOM trustworthiness, information diagnosticity and quality. For negative eWOM, we also found that perceived eWOM Trustworthiness, perceived eWOM diagnosticity and quality have a positive relationship with eWOM influence whereas perceived valence has a negative relationship with eWOM influence in Thai online consumers.

Keywords: consumer behavior, electronic word-of-mouth, online review, online word-of-mouth, Thai online consumer, webcare

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745 Antibacterial Potentials of the Leaf Extracts of Siam Weed (Chromolaena odorata) on Wound Isolates

Authors: M. E. Abalaka, O. A. Falusi, M. Galadima, D. Damisa

Abstract:

The antimicrobial activity of aqueous, ethanolic and methanolic extracts of Chromolaena odorata (Siam weed) was evaluated against four wound isolates: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae at the concentrations of 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml respectively. S. aureus and E. coli showed high susceptibility to the various extracts than the other test isolates. The aqueous extract showed activity against Staphylococcus aureus with a mean diameter of zone of inhibition of 16 ± 3.00 at concentration of 200mg/ml and as low as 8 ± 0.00 at concentration of 25mg/ml; E. coli showed susceptibility with a mean diameter of zone of inhibition of 18 ± 2.00 and 10 ± 0.00 at a concentration of 200mg/ml and 25mg/ml respectively. Pseudomonas aeruginosa and Klebsiella pneumoniae were resistant to the aqueous extract. Methanol extract showed activity against Staphylococcus aureus with a mean diameter of zone of inhibition at 28 ± 4.00 and 12 ± 2.30 at a concentration of 200mg/ml and 25mg/ml respectively; while E. coli was susceptible with mean diameter of zone of inhibition of 18 ± 2.00 and as low as 12 ± 0.00 at a concentration of 200mg/ml and 50mg/ml respectively, Pseudomonas aeruginosa showed considerable susceptibility with mean diameter of zone of inhibition of 13 ± 1.00 and 12 ± 0.00 at a concentration of 200mg/ml and 100mg/ml respectively. The ethanol extract showed activity against S. aureus with a mean diameter zone of inhibition of 15 ± 2.00 and 9 ± 0.00 at a concentration of 200mg/ml and 25mg/ml respectively: E. coli showed susceptibility with a mean diameter zone of inhibition of 20 ± 4.00 and 13 ± 2.00 at a concentration of 200mg/ml and 25mg/ml respectively. Pseudomonas aeruginosa showed considerable susceptibility with a mean diameter zone of inhibition of 13 ± 1.00 and 9 ± 0.00 at a concentration of 200mg/ml and 100mg/ml respectively. The results above indicate the efficacy and potency of the crude extracts of Chromolaena odorata leaf on the tested wound isolates.

Keywords: antibacterial, Chromolaena odorata, leaf extracts, test isolates

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744 Antimicrobial Resistance Patterns of Campylobacter from Pig and Cattle Carcasses in Poland

Authors: Renata Szewczyk, Beata Lachtara, Kinga Wieczorek, Jacek Osek

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Campylobacter is recognized as the main cause of bacterial gastrointestinal infections in Europe. A main source of the pathogen is poultry and poultry meat; however, other animals like pigs and cattle can also be reservoirs of the bacteria. Human Campylobacter infections are often self-limiting but in some cases, macrolide and fluoroquinolones have to be used. The aim of this study was to determine antimicrobial resistance patterns (AMR) of Campylobacter isolated from pig and cattle carcasses. Between July 2009 and December 2015, 735 swabs from pig (n = 457) and cattle (n = 278) carcasses were collected at Polish slaughterhouses. All samples were tested for the presence of Campylobacter by ISO 10272-1 and confirmed to species level using PCR. The antimicrobial susceptibility of Campylobacter isolates was determined by a microbroth dilution method with six antimicrobials: gentamicin (GEN), streptomycin (STR), erythromycin (ERY), nalidixic acid (NAL), ciprofloxacin (CIP) and tetracycline (TET). It was found that 167 of 735 samples (22.7%) were contaminated with Campylobacter. The vast majority of them were of pig origin (134; 80.2%), whereas for cattle carcasses Campylobacter was less prevalent (33; 19.8%). Among positive samples C. coli was predominant species (123; 73.7%) and it was isolated mainly from pig carcasses. The remaining isolates were identified as C. jejuni (44; 26.3%). Antimicrobial susceptibility indicated that 22 out of 167 Campylobacter (13.2%) were sensitive to all antimicrobials used. Fourteen of them were C. jejuni (63.6%; pig, n = 6; cattle, n = 8) and 8 was C. coli (36.4%; pig, n = 4; cattle, n = 4). Most of the Campylobacter isolates (145; 86.8%) were resistant to one or more antimicrobials (C. coli, n = 115; C. jejuni, n = 30). Comparing the AMR for Campylobacter species it was found that the most common pattern for C. jejuni was CIP-NAL-TET (9; 30.0%), whereas CIP-NAL-STR-TET was predominant among C. coli (47; 40.9%). Multiresistance, defined as resistance to three or more classes of antimicrobials, was found in 57 C. coli strains, mostly obtained from pig (52 isolates). On the other hand, only one C. jejuni strain, isolated from cattle, showed multiresistance with pattern CIP-NAL-STR-TET. Moreover, CIP-NAL-STR-TET was characteristic for most of multiresistant C. coli isolates (47; 82.5%). For the remaining C. coli the resistance patterns were CIP-ERY-NAL-TET (7 strains; 12.3%) and for one strain of each patterns: ERY-STR-TET, CIP-STR-TET, CIP-NAL-GEN-STR-TET. According to the present findings resistance to erythromycin was observed only in 11 C. coli (pig, n = 10; cattle, n = 1). In conclusion, the results of this study showed that pig carcasses may be a serious public health concern because of contamination with C. coli that might features multiresistance to antimicrobials.

Keywords: antimicrobial resistance, Campylobacter, carcasses, multi resistance

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743 Clinical and Molecular Characterization of Mycoplasmosis in Sheep in Egypt

Authors: Walid Mousa, Mohamed Nayel, Ahmed Zaghawa, Akram Salama, Ahmed El-Sify, Hesham Rashad, Dina El-Shafey

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Mycoplasmosis in small ruminants constitutes a serious contagious problem in smallholders causing severe economic losses worldwide. This study was conducted to determine the clinical, Minimum Inhibitory Concentration (MIC) and molecular characterization of Mycoplasma species associated in sheep breeding herds in Menoufiya governorate, Egypt. Out of the examination of 400 sheep, 104 (26%) showed respiratory manifestations, nasal discharges, cough and conjunctivitis with systemic body reaction. Meanwhile, out of these examined sheep, only 56 (14%) were positive for mycoplasma isolation onto PPLO(Pleuropneumonia-like organisms) specific medium. The MIC for evaluating the efficacy of sensitivity of Mycoplasma isolates against different antibiotics groups revealed that both the Linospectin and Tylosin with 2ug, 0.25ug/ml concentration were the most effective antibiotics for Mycoplasma isolates. The application of PCR was the rapid, specific and sensitive molecular approach for detection of M. ovipneumoniae, and M. arginine at 390 and 326 bp, respectively, in all tested isolates. In conclusion, the diagnosis of Mycoplsamosis in sheep is important to achieve effective control measures and minimizing the disease dissemination among sheep herds.

Keywords: MIC, mycoplasmosis, PCR, sheep

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742 Symbiotic Functioning, Photosynthetic Induction and Characterisation of Rhizobia Associated with Groundnut, Jack Bean and Soybean from Eswatini

Authors: Zanele D. Ngwenya, Mustapha Mohammed, Felix D. Dakora

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Legumes are a major source of biological nitrogen, and therefore play a crucial role in maintaining soil productivity in smallholder agriculture in southern Africa. Through their ability to fix atmospheric nitrogen in root nodules, legumes are a better option for sustainable nitrogen supply in cropping systems than chemical fertilisers. For decades, farmers have been highly receptive to the use of rhizobial inoculants as a source of nitrogen due mainly to the availability of elite rhizobial strains at a much lower compared to chemical fertilisers. To improve the efficiency of the legume-rhizobia symbiosis in African soils would require the use of highly effective rhizobia capable of nodulating a wide range of host plants. This study assessed the morphogenetic diversity, photosynthetic functioning and relative symbiotic effectiveness (RSE) of groundnut, jack bean and soybean microsymbionts in Eswatini soils as a first step to identifying superior isolates for inoculant production. According to the manufacturer's instructions, rhizobial isolates were cultured in yeast-mannitol (YM) broth until the late log phase and the bacterial genomic DNA was extracted using GenElute bacterial genomic DNA kit. The extracted DNA was subjected to enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and a dendrogram constructed from the band patterns to assess rhizobial diversity. To assess the N2-fixing efficiency of the authenticated rhizobia, photosynthetic rates (A), stomatal conductance (gs), and transpiration rates (E) were measured at flowering for plants inoculated with the test isolates. The plants were then harvested for nodulation assessment and measurement of plant growth as shoot biomass. The results of ERIC-PCR fingerprinting revealed the presence of high genetic diversity among the microsymbionts nodulating each of the three test legumes, with many of them showing less than 70% ERIC-PCR relatedness. The dendrogram generated from ERIC-PCR profiles grouped the groundnut isolates into 5 major clusters, while the jack bean and soybean isolates were grouped into 6 and 7 major clusters, respectively. Furthermore, the isolates also elicited variable nodule number per plant, nodule dry matter, shoot biomass and photosynthetic rates in their respective host plants under glasshouse conditions. Of the groundnut isolates tested, 38% recorded high relative symbiotic effectiveness (RSE >80), while 55% of the jack bean isolates and 93% of the soybean isolates recorded high RSE (>80) compared to the commercial Bradyrhizobium strains. About 13%, 27% and 83% of the top N₂-fixing groundnut, jack bean and soybean isolates, respectively, elicited much higher relative symbiotic efficiency (RSE) than the commercial strain, suggesting their potential for use in inoculant production after field testing. There was a tendency for both low and high N₂-fixing isolates to group together in the dendrogram from ERIC-PCR profiles, which suggests that RSE can differ significantly among closely related microsymbionts.

Keywords: genetic diversity, relative symbiotic effectiveness, inoculant, N₂-fixing

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741 Pathogenic Escherichia Coli Strains and Their Antibiotic Susceptibility Profiles in Cases of Child Diarrhea at Addis Ababa University, College of Health Sciences, Tikur Anbessa Specialized Hospital, Addis Ababa, Ethiopia

Authors: Benyam Zenebe, Tesfaye Sisay, Gurja Belay, Workabeba Abebe

Abstract:

Background: The prevalence and antibiogram of pathogenic E. coli strains, which cause diarrhea vary from region to region, and even within countries in the same geographical area. In Ethiopia, diagnostic approaches to E. coli induced diarrhea in children less than five years of age are not standardized. The aim of this study was to determine the involvement of pathogenic E. coli strains in child diarrhea and determine the antibiograms of the isolates in children less than 5 years of age with diarrhea at Addis Ababa University College of Health Sciences TikurAnbessa Specialized Hospital, Addis Ababa, Ethiopia. Methods: A purposive study that included 98 diarrheic children less than five years of age was conducted at Addis Ababa University College of Health Sciences, TikurAnbessa Specialized Hospital, Addis Ababa, Ethiopia to detect pathogenic E. coli biotypes. Stool culture was used to identify presumptive E. coliisolates. Presumptive isolates were confirmed by biochemical tests, and antimicrobial susceptibility tests were performed on confirmed E. coli isolates by the disk diffusion method. DNA was extracted from confirmed isolates by a heating method and subjected to Polymerase Chain Reaction or the presence of virulence genes. Amplified PCR products were analyzed by agarose gel electrophoresis. Data were collected on child demographics and clinical conditions using administered questionnaires. The prevalence of E. coli strains from the total diarrheic children, and the prevalence of pathogenic strains from total E. coli isolates along with their susceptibility profiles; the distribution of pathogenic E.coli biotypes among different age groups and between the sexes were determined by using descriptive statistics. Result: Out of 98 stool specimens collected from diarrheic children less than 5 years of age, 75 presumptive E. coli isolates were identified by culture; further confirmation by biochemical tests showed that only 56 of the isolates were E. coli; 29 of the isolates were found in male children and 27 of them in female children. Out of the 58 isolates of E. coli, 25 pathotypes belonging to different classes of pathogenic strains: STEC, EPEC, EHEC, EAEC were detected by using the PCR technique. Pathogenic E. coli exhibited high rates of antibiotic resistance to many of the antibiotics tested. Moreover, they exhibited multiple drug resistance. Conclusion: This study found that the isolation rate of E. coli and the involvement of antibiotic-resistant pathogenic E. coli in diarrheic children is prominent, and hence focus should be given on the diagnosis and antimicrobial sensitivity testing of pathogenic E. coli at Addis Ababa University College of Health Sciences TikurAnbessa Specialized Hospital. Among antibiotics tested, Cefotitan could be a drug of choice to treat E. coli.

Keywords: antibiotic susceptibility profile, children, diarrhea, E. coli, pathogenic

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740 Mutations in rpoB, katG and inhA Genes: The Association with Resistance to Rifampicin and Isoniazid in Egyptian Mycobacterium tuberculosis Clinical Isolates

Authors: Ayman K. El Essawy, Amal M. Hosny, Hala M. Abu Shady

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The rapid detection of TB and drug resistance, both optimizes treatment and improves outcomes. In the current study, respiratory specimens were collected from 155 patients. Conventional susceptibility testing and MIC determination were performed for rifampicin (RIF) and isoniazid (INH). Genotype MTBDRplus assay, which is a molecular genetic assay based on the DNA-STRIP technology and specific gene sequencing with primers for rpoB, KatG, and mab-inhA genes were used to detect mutations associated with resistance to rifampicin and isoniazid. In comparison to other categories, most of rifampicin resistant (61.5%) and isoniazid resistant isolates (47.1%) were from patients relapsed in treatment. The genotypic profile (using Genotype MTBDRplus assay) of multi-drug resistant (MDR) isolates showed missing of katG wild type 1 (WT1) band and appearance of mutation band katG MUT2. For isoniazid mono-resistant isolates, 80% showed katG MUT1, 20% showed katG MUT1, and inhA MUT1, 20% showed only inhA MUT1. Accordingly, 100% of isoniazid resistant strains were detected by this assay. Out of 17 resistant strains, 16 had mutation bands for katG distinguished high resistance to isoniazid. The assay could clearly detect rifampicin resistance among 66.7% of MDR isolates that showed mutation band rpoB MUT3 while 33.3% of them were considered as unknown. One mono-resistant rifampicin isolate did not show rifampicin mutation bands by Genotype MTBDRplus assay, but it showed an unexpected mutation in Codon 531 of rpoB by DNA sequence analysis. Rifampicin resistance in this strain could be associated with a mutation in codon 531 of rpoB (based on molecular sequencing), and Genotype MTBDRplus assay could not detect the associated mutation. If the results of Genotype MTBDRplus assay and sequencing were combined, this strain shows hetero-resistance pattern. Gene sequencing of eight selected isolates, previously tested by Genotype MTBDRplus assay, could detect resistance mutations mainly in codon 315 (katG gene), position -15 in inhA promotes gene for isoniazid resistance and codon 531 (rpoB gene) for rifampicin resistance. Genotyping techniques allow distinguishing between recurrent cases of reinfection or reactivation and supports epidemiological studies.

Keywords: M. tuberculosis, rpoB, KatG, inhA, genotype MTBDRplus

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739 Drug Sensitivity Pattern of Organisms Causing Chronic Suppurative Otitis Media

Authors: Fatma M. Benrabha

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The aim of the study was to determine the type and pattern of antibiotic susceptibility of the pathogenic microorganisms causing chronic suppurative otitis media (CSOM), which could lead to better therapeutic decisions and consequently avoidance of appearance of resistance to specific antibiotics. Most frequently isolated agents were Pseudomonas aeruginosa 28.5%; followed by Staphylococcus aureus 18.2%; proteus mirabilis 13.9%; Providencia stuartti 6.7%; Bacteroides melaninogenicus, Aspergillus sp., candida sp., 4.2% each; and other microorganisms were represented in 3-0.2%. Drug sensitivities pattern of Pseudomonas aeruginosa showed that ciprofloxacin was active against the majority of isolates (93.9%) followed by ceftazidime 86.2%, amikacin 76.2% and gentamicin 40.8%. However, Staphylococcus aureus isolates were resistant to penicillin 72.7%, erythromycin 28.6%, cephalothin 18.2%, cloxacillin 8.3% and ciprofloxacin was active against 96.2% of isolates. The resistance pattern of proteus mirabilis were 55.6% to ampicillin, 47.1% to carbencillin, 29.4% to cephalothin, 14.3% to gentamicin and 4.8% to amikacin while 100% were sensitive to ciprofloxacin. We conclude that ciprofloxacin is the best drug of choice in treatment of CSOM caused by the common microorganisms.

Keywords: otitis media, chronic suppurative otitis media (CSOM), microorganism, drug sensitivity

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738 Genotyping and Phylogeny of Phaeomoniella Genus Associated with Grapevine Trunk Diseases in Algeria

Authors: A. Berraf-Tebbal, Z. Bouznad, , A.J.L. Phillips

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Phaeomoniella is a fungus genus in the mitosporic ascomycota which includes Phaeomoniella chlamydospora specie associated with two declining diseases on grapevine (Vitis vinifera) namely Petri disease and esca. Recent studies have shown that several Phaeomoniella species also cause disease on many other woody crops, such as forest trees and woody ornamentals. Two new species, Phaeomoniella zymoides and Phaeomoniella pinifoliorum H.B. Lee, J.Y. Park, R.C. Summerbell et H.S. Jung, were isolated from the needle surface of Pinus densiflora Sieb. et Zucc. in Korea. The identification of species in Phaeomoniella genus can be a difficult task if based solely on morphological and cultural characters. In this respect, the application of molecular methods, particularly PCR-based techniques, may provide an important contribution. MSP-PCR (microsatellite primed-PCR) fingerprinting has proven useful in the molecular typing of fungal strains. The high discriminatory potential of this method is particularly useful when dealing with closely related or cryptic species. In the present study, the application of PCR fingerprinting was performed using the micro satellite primer M13 for the purpose of species identification and strain typing of 84 Phaeomoniella -like isolates collected from grapevines with typical symptoms of dieback. The bands produced by MSP-PCR profiles divided the strains into 3 clusters and 5 singletons with a reproducibility level of 80%. Representative isolates from each group and, when possible, isolates from Eutypa dieback and esca symptoms were selected for sequencing of the ITS region. The ITS sequences for the 16 isolates selected from the MSP-PCR profiles were combined and aligned with sequences of 18 isolates retrieved from GenBank, representing a selection of all known Phaeomoniella species. DNA sequences were compared with those available in GenBank using Neighbor-joining (NJ) and Maximum-parsimony (MP) analyses. The phylogenetic trees of the ITS region revealed that the Phaeomoniella isolates clustered with Phaeomoniella chlamydospora reference sequences with a bootstrap support of 100 %. The complexity of the pathosystems vine-trunk diseases shows clearly the need to identify unambiguously the fungal component in order to allow a better understanding of the etiology of these diseases and justify the establishment of control strategies against these fungal agents.

Keywords: Genotyping, MSP-PCR, ITS, phylogeny, trunk diseases

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737 Phylogenetic Analysis of Georgian Populations of Potato Cyst Nematodes Globodera Rostochiensis

Authors: Dali Gaganidze, Ekaterine Abashidze

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Potato is one of the main agricultural crops in Georgia. Georgia produces early and late potato varieties in almost all regions. In traditional potato growing regions (Svaneti, Samckhet javaheti and Tsalka), the yield is higher than 30-35 t/ha. Among the plant pests that limit potato production and quality, the potato cyst nematodes (PCN) are harmful around the world. Yield losses caused by PCN are estimated up to 30%. Rout surveys conducted in two geographically distinct regions of Georgia producing potatoes - Samtskhe - Javakheti and Svaneti revealed potato cyst nematode Globodera rostochiensi. The aim of the study was the Phylogenetic analyses of Globodera rostochiensi revealed in Georgia by the amplification and sequencing of 28S gen in the D3 region and intergenic ITS1-15.8S-ITS2 region. Identification of all the samples from the two Globodera populations (Samtskhe - Javakheti and Svaneti), i.e., G. rostochiensis (20 isolates) were confirmed by conventional multiplex PCR with ITS 5 universal and PITSp4, PITSr3 specific primers of the cyst nematodes’ (G. pallida, G. rostochiensis). The size of PCR fragment 434 bp confirms that PCN samples from two populations, Samtskhe- Javakheti and Svaneti, belong to G. rostochiensi . The ITS1–5.8S-ITS2 regions were amplified using prime pairs: rDNA1 ( 5’ -TTGATTACGTCCCTGCCCTTT-3’ and rDNA2( 5’ TTTCACTCGCCGTTACTAAGG-3’), D3 expansion regions were amplified using primer pairs: D3A (5’ GACCCCTCTTGAAACACGGA-3’) and D3B (5’-TCGGAAGGAACCAGCTACTA-3’. PCR products of each region were cleaned up and sequenced using an ABI 3500xL Genetic Analyzer. Obtained sequencing results were analyzed by computer program BLASTN (https://blast.ncbi.nlm.nih.gov/Blast.cg). Phylogenetic analyses to resolve the relationships between the isolates were conducted in MEGA7 using both distance- and character-based methods. Based on analysis of G.rostochiensis isolate`s D3 expansion regions are grouped in three major clades (A, B and C) on the phylogenetic tree. Clade A is divided into three subclades; clade C is divided into two subclades. Isolates from the Samtckhet-javakheti population are in subclade 1 of clade A and isolates in subclade 1 of clade C. Isolates) from Svaneti populations are in subclade 2 of clade A and in clad B. In Clade C, subclade two is presented by three isolates from Svaneti and by one isolate (GL17) from Samckhet-Javakheti. . Based on analysis of G.rostochiensis isolate`s ITS1–5.8S-ITS2 regions are grouped in two main clades, the first contained 20 Georgian isolates of Globodera rostochiensis from Svaneti . The second clade contained 15 isolates of Globodera rostochiensis from Samckhet javakheti. Our investigation showed of high genetic variation of D3 and ITS1–5.8S-ITS2 region of rDNA of the isolates of G. rostochiensis from different geographic origins (Svameti, Samckhet-Javakheti) of Georgia. Acknowledgement: The research has been supported by the Shota Rustaveli National Scientific Foundation of Georgia : Project # FR17_235

Keywords: globodera rostochiensi, PCR, phylogenetic tree, sequencing

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