IMViC Test Research Articles

Extended-spectrum beta-laktamase (ESBL) enzyme producing bacteria isolated from rectal swabs of around animals in dairy farms

This study aims to identify bacterial contamination of Klebsiella pneumoniae and Escherichia coli producing Extended Spectrum Beta Lactamase (ESBL) isolated from rectal swabs of local animals on a dairy farm in Deyeng Village, Kediri, East Java. Interviews and observations were conducted with farmers to obtain the required risk factor data (cleanliness of drinking and eating places, drinking water sources, and administration of antibiotics to livestock). The number of samples tested in this study was 34 samples, including 12 chicken samples, 8 goat samples, 5 duck samples, 4 rabbit samples, 3 cat samples and 2 goose samples. Samples were isolated on MCA media + cefotaxime 1 mg/l and isolated again on MCA media without the addition of antibiotics to multiply pure bacterial colonies. The tests carried out to determine the presence of ESBL-producing bacteria are biochemical tests, namely the IMViC test (Indole, Methyl red, Voges-Proskauer, Citrate Utilization), TSIA (Triple Sugar Iron Agar) test, gram staining, and the Double Disk Standard Method test using β-lactam disc antibiotics, namely cefotaxime 30μg, ceftazidime 30μg, and amoxycilline-clavulanate acid 30μg. The results of this research were that 6 samples or 17.6% of ESBL-producing bacteria were identified from poultry groups (chickens, ducks and geese) from dairy farms in Deyeng Village, Kediri Regency, East Java.

Identification of Salmonella sp Bacteria in Processed Balado Shrimp Sold in Food Stalls Around Mulyosari Area

Background & Objective: Food quality is an issue of great concern in developing countries. Because many nutrients are obtained through food. Balado shrimp is a typical Padang dish with a spicy, sweet and savoury taste that is widely admired by all ages from children to adults. The presence of pathogenic bacteria, one of which is Salmonella sp, which contaminates balado shrimp food can cause health problems in the human body until death. The purpose of this study was to determine the presence or absence of Salmonella sp bacteria in processed shrimp balado sold in food stalls in the Mulyosari area. Method: In this study use as many as 30 samples of shrimp balado sold in food stalls Mulyosari area. This sample examination uses a streaking plate procedure and IMViC test. Data analysis using descriptive qualitative. Result: The results of this study showed that there were 3 samples identified Salmonella sp. Of the 30 samples, there was a range of 10% contaminated with Salmonella sp bacteria and 90% were not contaminated with Salmonella sp bacteria. Conclusion: This indicates that positive samples of balado shrimp must be further examined to determine the feasibility of consumption.

ANTIBIOTIC RESISTANCE OF ESBL-PRODUCING E. COLI AND OTHER GRAM-NEGATIVE BACTERIA ISOLATED FROM STREET-VENDED FOODS IN BANGLADESH

The prevalence and impact of antibiotic-resistant pathogens transmitted through food, particularly street-vended foods, is becoming a major public health concern. Although a significant proportion of the urban population in developing countries consumes street-vended foods, the role of these foods in spreading antibiotic resistance has been rarely investigated. In this study, 50 bacterial isolates were obtained from 25 samples representing five categories of street-vended foods: Phuchka, Chatpati, Sausage, Bun, and Salad. The IMVIC test revealed a notably high occurrence of Escherichia coli (n=32) within the collected samples. Three representative isolates were selected for molecular identification using DNA sequencing of 16S rDNA. They were identified as Klebsiella oxytoca, Burkholderia fungorum, and Serratia nematodiphila. The antibiotic susceptibility of the identified isolates (n=35) was investigated using twelve antibiotics following the Kirby-Bauer disk diffusion method. Around 65.63% of the E. coli isolates (n=21) exhibited multidrug resistance. Double Disc Synergy Test (DDST) and Phenotypic Confirmatory Disk Diffusion Test (PCDDT) confirmed ESBL production of Eight multidrug-resistant E. coli isolates (38.09%). The multiple antibiotic resistance (MAR) index showed that 22 E. coli isolates had MAR above 0.2, with resistance mostly against oxacillin, ampicillin, and cefuroxime. The Klebsiella oxytoca isolate showed multidrug resistance viz., ampicillin, oxacillin, cefuroxime, and kanamycin. The Burkholderia fungorum isolate showed no distinct inhibition zone against ampicillin and chloramphenicol. Additionally, the Serratia nematodiphila isolate showed no distinct inhibition zone against three antibiotics, including ampicillin, oxacillin, and cefuroxime. These findings might contribute to the knowledge of emerging antibiotic-resistant foodborne pathogens and raise concerns about the safety of street-vended foods in Bangladesh.

Identification of Escherichia coli Multidrug Resistance in Cattle in Abattoirs

Escherichia coli is a typical flora of an animals' digestive tract. Based on these details, this study was done on the detection and identification of multidrug-resistant E. coli in cattle in Surabaya abattoirs. Each cattle rectum swab sample was streaked into EMB agar media and followed by Gram staining. The IMViC test was used to confirm the presence of E. coli. Based on morphological culture features, Gram staining, and biochemical testing, the sample examination results revealed that 41 samples (41%) of the 100 cattle rectal swab samples that were isolated were confirmed to be positive for E. coli. The profile of antibiotic resistance from the results of the E. coli resistance test to antibiotics showed that out of a total of 41 isolates of E. coli, 12 isolates (29.27%) were detected as resistant to 1 class of antibiotics tested, while 2 isolates (4.88%) were resistant to 2 classes of antibiotics, and 3 isolates (7.32%) were confirmed to be multidrug resistance (MDR).

Screening of Bacillus sp. (OQ654027) Mediated Seed Bio-priming Enhance Plant-Growth-Promotion for Sustainable Crop Production of Groundnut and Chickpea

Plant growth-promoting bacteria are valuable microbes that enhance the plant growth, development, and yield. Several seed priming methods are able to improvise seed germination, seedling potency, to overwhelmed abiotic stress and biotic stress management which gets prime attention. Generally, plant-growth promoting bacteria are utilized for biopriming technique that is able to survive under stress environmental conditions. The aims of the present investigation were to screening ofKC9bacterial isolate as PGPB in groundnut and chickpea and study the effects ofKC9 bacterial isolation seed, seedling germination and growth during the development of both seeds. In this study KC9 has been isolated from the soil sample of chickpea crop field at Saurashtra region. Preliminary screening was conducted based on their identification via morphological, biochemical i.e., catalase, starch hydrolysis, sugar utilization, motility, IMViC test, gram’s nature and by 16S rRNA gene nucleotide sequences and identified as Bacillus sp. that has been submitted to the NCBI for Accession number. Further examine the production of indole acetic acid (IAA), Ammonia (NH3), HCN, Siderophore and Phosphate solubilization. KC9 (Bacillus sp.) was recognized as most efficient plant-growth promoting bacteria. For the pot experiment seed biopriming method is employing to coat the seeds of chickpea and groundnut with Bacillus inoculant consider as treated and seeds sown with double distilled water as control. Bacillus coated seed biopriming also minimize the seed germination time period that takes only 3-4 days to proper seed radicle development. By the statistical approach of paired t-test of pot experiment on chickpea and groundnut seeds and seedlings were treated with Bacillus strain exhibited significantly higher levels at of seed germination, seedling growth via root length, shoot length and number of leaves in compared to non-treated control plants. In this present investigation , impact of seedling germination on the inoculated condition with KC9 of groundnut and chickpea, determining remarkable changes in total chlorophyll content (11.81 µg/g and 10.40 µg/g), relative water content(RWC) obtained 86% and 83%., total free amino acids (92.14±0.03µg/ml whereas 37.5±0.02µg/ml), total phenolic acid content (76.28±0.03mg GAE/100g and chickpea is 52.83±0.08 mg GAE/100g) and total flavonoid content increases with the treatment of KC9 (Bacillus sp.) 626±0.05QE/100g is recorded in groundnut seedlings and chickpea is 0.085±0.01QE/100g respectively.

Isolation and Identification of Bacteria Present on Spectacles

The human eyes have a special anatomy that makes them vulnerable to infectious agents, including bacteria, as well as direct environmental exposure. The eyelids, conjunctiva, and cornea are the main parts of the eyes that are most frequently infected. The numerous microorganisms have been linked to eye infections caused by microbial contamination and colonization of eyeglasses. A study reported the presence of bacterial contaminants on spectacles used by students at Shri Shivaji Science and Arts College Chikhli, in July 2023 to August 2023, Such as Pseudomonas sp., Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus sp. Total 40 samples are collect from different students such as 14 samples are collected from 11th Students, 12 samples are collected from 12th Students, 08 samples are collected from B.Sc. Students and 06 Samples collected from M.Sc. Students were taken by using sterile cotton swabs that had been moistened with sterile distilled water. After that it was inoculated on the Blood agar and MacConkey agar by streak plate method and incubated for 24 hours at 37°C. After incubation for 24 hours at 37°C, for confirmed the growing microorganism by using different tests such as Gram Staining, Hanging-Drop method, IMViC test, Carbohydrate fermentation, Enzyme production (Catalase Test and Oxidase Test), and Triple Sugar Iron Agar test are performed as per required for confirmation. All samples shows positive results, when we examined results, it shows different bacteria are present such as Bacillus sp. (55%), Pseudomonas sp. (42.50%), Staphylococcus aureus (22.50%), and Staphylococcus epidermidis (12.50%). The results shows that the there was contamination by pathogenic bacteria on spectacles used by the students of Shri Shivaji Science and Arts College Chikhli. It means spectacles provide suitable condition for survive of pathogenic bacteria then its leads to the cause of eyes infection.

Detection of extended-spectrum β-lactamase-producing Escherichia coli genes isolated from cat rectal swabs at Surabaya Veterinary Hospital, Indonesia.

Escherichia coli causes a bacterial illness that frequently affects cats. Diseases caused by E. coli are treated using antibiotics. Because of their proximity to humans, cats possess an extremely high risk of contracting antibiotic resistance genes when their owners touch cat feces containing E. coli that harbor resistance genes. This study was conducted to identify multidrug-resistant E. coli and extended-spectrum β-lactamase (ESBL)-producing genes from cat rectal swabs collected at Surabaya City Veterinary Hospital to determine antibiotic sensitivity. Samples of cat rectal swabs were cultured in Brilliant Green Bile Lactose Broth medium and then streaked on eosin methylene blue agar medium for bacterial isolation, whereas Gram-staining and IMViC tests were conducted to confirm the identification results. The Kirby-Bauer diffusion test was used to determine antibiotic sensitivity, and the double-disk synergy test was used to determine ESBL-producing bacteria. Molecular detection of the genes TEM and CTX-M was performed using a polymerase chain reaction. Based on morphological culture, Gram-staining, and biochemical testing, the results of sample inspection showed that of the 100 cat rectal swab samples isolated, 71 (71%) were positive for E. coli. Furthermore, 23 E. coli isolates (32.39%) demonstrated the highest resistance to ampicillin. Four isolates were confirmed to be multidurg-resistant and ESBL-producing strains. Molecular examination revealed that three E. coli isolates harbored TEM and CTX-M. In conclusion, pet owners must be educated on the use of antibiotics to improve their knowledge about the risks of antibiotic resistance.

MICRIBIOLOGICAL ASSESSMENT OF DRINKING WATER AND PERVASIVENESS OF WATER BORN DISEASES IN DUNGARPUR, RAJASTHAN

INTRODUCTION: This study assessed E. coli counts in drinking water from different sources and illustrates the high prevalence of gastrointestinal diseases in Dungarpur districts of southern Rajasthan, India. METHODOLOGY: The microbiological parameters studied were pathogen detection, total coliform count and total viable count. Microbiological analysis of water samples were performed as described in Standard Methods for the Examination of Water and Wastewater (APHA - AWWA, 2005 and Chhabra, 2008). Total coliform count performed by the Most Probable Number (MPN) method, which is commonly used as an indicator of potability of water. Presence of the bacteria from Enterobacteriacae family was confirmed using IMViC test. The data of Indoor and Outdoor patients for such disease were collected.RESULT AND DISCUSSION: Seasonal fluctuation in coliform counts was found to be prominent and varied from over 200 MPN/ 100 ml in winter to more than 1600 (over the measurable limit) in rainy season in three water bodies. Escherichia coli counts ranged from 17 to 500 MPN/100 ml for all the sampled sites. E. coli and Total coliform were highest in rainy season. Results of IMViC test indicates that in rainy season except for Margia Dam all the water bodies were laden with all five categories of enteric bacteria namely E. coli, Shigella, Citrobacter and Klebsiella/ Enterobacter. The contamination is significantly higher during the wet season compared to dry season, due to WBD patients does exhibit a trend that in late summer and rainy month’s incidence of the disease symptoms intensifies. CONCLUSION: Dungarpur district was analyzed and same pattern of seasonal variation in microbial counts of water samples and similar hospital data trend was observed. So, somehow the microbial contamination of surface water and run-off reaches the ground-water table. Therefore, hospital data confirms that the source of drinking water plays a strong, positive and significant association with waterborne diseases.

Identification of Escherichia coli and Salmonella on Fishery Product from Juanda Airport, East Java, Indonesia

Fishery products are foodstuffs that are prone to pathogenic microbial contamination which can cause food poisoning and disease in consumers. Therefore, fishery products need attention related to food safety. Pathogenic bacteria that often contaminate fishery products include E. coli and Salmonella which can cause disease in humans. This study aims to examine E. coli and Salmonella bacteria that contaminate fishery products that will be exported from Juanda Airport, East Java. The research method used was microscopic examination using conventional methods consisting of isolation and biochemical tests (for E. coli bacteria); prediction test, affirmation test and IMVIC test (for Salmonella bacteria). The results showed that fishery products consisting of Barramundi Skin On, Frozen Vannamei, Frozen Black Tiger Shrimp, Breaded Shrimp, Classic Frog Thighs, Skip Jack, Frozen Octopus, Frozen Mackarel, Frozen Sardine, and Frozen Squid were contaminated by bacteria that are not E. coli and Salmonella groups.

Mung beans (Phaseolus radiates L.): Utilization as components of the growth medium Bacillus subtilis

Bacillus subtilis is a bacterial contaminant widely discovered in laboratories and used in educational needs. Synthetic media are relatively expensive, which makes it difficult for microbiology laboratories to meet large-scale bacterial growth media demands. This has encouraged researchers to find alternative media with cheaper and easily available prices, namely Mung Beans, which are known to contain protein nutrients that are good for bacterial growth, which is as much as 24%. The purpose of this study was to determine the potential of mung beans’ alternative media as a medium for bacterial growth. This research is a descriptive study and the sample used is 1 tube of a pure isolate of Bacillus subtilis. The variable of this research is mung beans as an alternative medium for the growth of Bacillus subtilis. The results were obtained by observed macroscopically and microscopically, followed by a carbohydrate fermentation test and an IMViC test. Then showed that the colonies growing on the alternative medium of mung beans were in accordance with the characteristics of the colonies growing on Nutrient Agar medium (control), which indicated that these characteristics were Bacillus subtilis. Based on the research that has been done, it can be concluded that there is a qualitative growth of Bacillus subtilis in mung beans alternative media, which means that mung beans alternative media has the potential as a growth medium for Bacillus subtilis.

Analysis of Escherichia coli Microbial Contamination and Total Coliform Bacteria in Refill Drinking Water in Pondok Cabe Ilir Village, South Tangerang City

Water is an essential nutrient for human health. It is important to maintain adequate drinking water intake to prevent dehydration, which can cause hypothermia, dizziness, constipation, and kidney stones. Currently, water-filling stations are an alternative source of drinking water because of limited access to clean water at affordable prices. The purpose of this study was to determine bacterial contamination at a drinking water station in Pondok Cabe Ilir, South Tangerang, Banten, based on Permenkes RI No. 492/Menkes/Per/IV/2010. The study was conducted using the Most Probable Number (MPN) method and the IMViC test, Triple Sugar Iron test, H2S production test, and motility test to identify Escherichia coli. The results showed that one out of five refilled drinking water samples contained Coliform bacteria above the threshold according to Minister of Health Regulation No. 492/Menkes/Per/IV/2010.

Complementation of Biochemical and Physiological Assays with Functional PGPR Based Assays to Screen Potential Isolates

The work deals with the biochemical characterization of rhizospheric isolates including Indole, Methyl Red, Voges Proskauer, Citrate test (IMViC test) based quantitative biochemical assay, PGPR properties like nitrogen assimilation (in microbes), phosphate solubilization etc. A special aspect of finding the correlation between the biochemical tests and PGPR properties were also studied. Methyl red test and tryptophanase assay (by indole test) were found to be positively and negatively correlated with phosphate solubilization. Vogues Proskeur and citrate utilization tests were negatively correlated with the phosphate solubilization. In future, these biochemical tests can be used as determining factors to identify phosphate solubilizing bacteria. Moreover, a group of bacteria was identified by the scatter plot analysis which shows low acid production with high phosphate solubilization. Lastly, we have given a new approach of screening rhizospheric bacteria based on motility on nitrogen deficient media.

Seasonal Fluctuation of Escherichia coli in Nainital Lake, Uttarakhand State, India

The season has a significant impact on the presence of microorganisms in water sources. Various contaminants and bacteria are transported into water sources by heavy rains. Because of the poor microbiological condition of such water bodies, regular monitoring is required. Routine monitoring, on the other hand, considers the microbial quality of the water column.The objective of this research was to investigate the microbiological quality of Nainital Lake in Uttarakhand, using Escherichia coli as a faecal indicator organism, and examine how seasonal fluctuations (dry and rainy seasons) affected its abundance. During 2018 and 2019, samples were taken from Nainital Lake sampling sites during the summer (dry) and rainy (wet) seasons and presence of E. coli was examined using various media, IMViC and catalase tests. E. coli was discovered at all of the locations and its concentration was influenced by seasonal variations. In both years, the cfu/ml count during the rainy season was observed higher in comparison to count during the summer. The transmission of this bacteria into the water column as a result of events such as floods or human activities might harm the health of those who consume untreated lake water for recreation and other domestic reasons. As a result, community education for health care practitioners and the general public about the proper use of lake water is essential. In order to preserve human health and other aquatic life, regular water quality monitoring is also required.

Bioremediation of textile dye effluents using indigenous earthworm Lampito mauritii (Kinberg)

The present study is intended to assess and monitor the Bioremediation of textile dye effluent using Lampito mauritii earthworm. The study focus on the bioremediation of surface water of SIPCOT dye industry Cuddalore district over a period of one year and has revealed variations among the different parameters at different study sites. For the analysis of surface water three sampling sites were selected for the study. In the area, from SIPCOT dye industry to Cuddalore district the parameters concentration was decreased and it is useful for drinking purpose. Heavy metals such as copper and zinc are essential elements for metabolic activities however they show toxic effects when exposed to higher levels whereas lead, mercury, cadmium and are not essential for metabolism which they exhibit toxic properties. The heavy metals of surface water in the study area are in the order of Zn > Cu > Pb > Cd > Hg (mg/L) respectively. Mercury, cadmium were noticed in least level in the non-industrial area whereas industrial area concentration of metals were slightly increased than the prescribed limits. The examination of water samples for bacteriological parameter includes MPN index, total heterotrophic bacterial colony count, morphological and biochemical characteristics. MPN is a suitable and widely used method to determine the microbial quality of water. Due to heavy rain the microbial load was increased during monsoon season. Based on morphology and biochemical characteristics, identify the species of the organism namely Escherichia coli, Vibrio cholerae, Salmonella typhi, Klebsiella pneumoniae, Enterobacter aerogenes and Pseudomonas aeruginosa. Based on motility, shape, size and structure, the organism was identified and based on IMVIC tests of biochemical reaction with production of acid and gas the organism was identified.

Molecular characterization of Escherichia coli isolated from milk samples with regard to virulence factors and antibiotic resistance

Background and Aim:Raw milk is considered an essential source of nutrition during all stages of human life because it offers a valuable supply of protein and minerals. Importantly, milk is considered a good media for the growth and contamination of many pathogenic bacteria, especially food-borne pathogens such as Escherichia coli. Thus, the objective of this study was to characterize E. coli and detect its virulence factors and antibiotic resistance from raw milk samples.Materials and Methods:Raw milk samples (n=100) were collected from different localities in Qena, Egypt, and investigated for the presence of E. coli using different biochemical tests, IMViC tests, serotyping to detect somatic antigen type, and molecularly by polymerase chain reaction (PCR) tests. The presence of different virulence and antimicrobial genes (hly, eae, stx1, stx2, blaTEM, tetA(A), and tetB genes) in E. coli isolates was evaluated using PCR.Results:The results demonstrated that 10 out of 100 milk samples were contaminated with E. coli. Depending on serology, the isolates were classified as O114 (one isolate), O27 (two isolates), O111 (one isolate), O125 (two isolates), and untypeable (five isolates) E. coli. The sequencing of partially amplified 16S rRNA of the untypeable isolates resulted in one isolate, which was initially misidentified as untypeable E. coli but later proved as Enterobacter hormaechei. Moreover, antibacterial susceptibility analysis revealed that nearly all isolates were resistant to more than 3 families of antibiotics, particularly to b-lactams, clindamycin, and rifampin. PCR results demonstrated that all E. coli isolates showed an accurate amplicon for the blaTEM and tetA(A) genes, four isolates harbored eae gene, other four harbored tetB gene, and only one isolate exhibited a positive stx2 gene.Conclusion:Our study explored vital methods for identifying E. coli as a harmful pathogen of raw milk using 16S rRNA sequencing, phylogenetic analysis, and detection of virulence factors and antibiotic-resistant genes.

Detection of Class 1 Integron Encoding Gene in Multidrug Resistance (MDR) Citrobacter freundii Isolated from Healthy Broiler Chicken

This study was aimed to find out that broiler chicken farms have problems with antibiotic resistance Citrobacter freundii and determined the prevalence and class 1 integron encoding gene. Multidrug resistance Citrobacter freundii was collected from broiler chicken among one hundred and sixty cloacal swab samples from 32 farms in Blitar for 3 months. The method of bacterial inoculation used MacConkey agar and biochemical test was conducted by IMViC and TSIA test. Citrobacter freundii for antibiotic sensitivity pattern was tested by disk diffusion, and the multidrug resistance encoding gene was tested by PCR. This study exposed 160 samples, and 13.75% (22/160) samples were positive of Citrobacter freundii. The antibiotic sensitivity pattern showed high resistances against ampicillin and erythromycin (77.27%), tetracycline (59.09%), trimetropim-sulfamethoxazole (50%), and streptomycin (22.72%). Isolates that were detected as multidrug resistance were continued with PCR testing to prove the existence of a class 1 integron encoding gene. Multidrug resistance Citrobacter freundii isolated from broiler chicken farms in Blitar were 81.82% (18/22), and were indicated that five were positive Class 1 Integron encoding gene. The results of this study showed that the prevalence and distribution of multidrug resistance Citrobacter freundii were high, so it can cause the spread of antimicrobial resistance to public health. Class 1 integron encoding gene was found 22.72% from multidrug resistance Citrobacter freundii by PCR. It was concluded that broiler chicken farms need assessment management to reduce and avoid multidrug resistance bacteria in animals and human. Therefore, the use of appropriate antibiotics is a good step to reduce the incidence of MDR in poultry.

PEMERIKSAAN CEMARAN Escherichia coli, Shigella sp DAN Salmonella sp PADA SUSU SAPI PERAH YANG DIPEROLEH DARI PETERNAKAN ASAM KUMBANG KECAMATAN MEDAN SELAYANG

Milk is a drink that is mandatory for consumption by the public, especially for children, therefore, milk must be free from contamination with Escherichia coli, Shigella sp and Salmonella sp bacteria, through the tools used are not clean, the environment is dirty, the hands of workers, and others. This study aims to determine the presence or absence of Escherichia coli, Shigella sp, and Salmonella sp bacteria in Dairy Cow Milk obtained from the Asam Kumbang Farm, Medan Selayang District. The research sample was ten samples of milk from dairy cows in one farm with different cows which were treated with two aseptic and non-aseptic treatments. The research method used is descriptive research with MPN (Most Probable Number) method to count coliform bacteria followed by an examination of LB, BGLB, EMBA media, gram staining, biochemical tests, IMVIC tests, and user testing methods on samples of Shigella sp and Salmonella bacteria sp followed by an examination of NB, SCB, SSA media, gram staining, biochemical tests, and IMVIC. The results showed that one aseptic sample A and 3 non-aseptic samples A, B, C had Escherichia coli bacteria in dairy cows' milk. The quality of milk must comply with the requirements of SNI 3141.1:2011 Fresh Milk Quality and the Maximum Limit of Microbial Contamination in SNI 7388-2009 Fresh Milk.

Plasmid-Mediated Antibiotic Resistant Escherichia coli in Sarawak Rivers and Aquaculture Farms, Northwest of Borneo.

Background: The emergence of plasmid-mediated antibiotic resistance in Escherichia coli in water resources could pose a serious threat to public health. The study aims to investigate the dispersion of plasmid-mediated antibiotic-resistant E. coli from six rivers in Sarawak and two aquaculture farms in Borneo. Methods: A total of 74 water samples were collected for the determination of their bacteria colony count. An IMViC test identified 31 E. coli isolates and tested their susceptibility against twelve clinically important antibiotics. The extraction of plasmid DNA was done using alkali lysis SDS procedures. Characteristics, including plasmid copy number, molecular weight size, resistance rate and multiple antibiotic resistance (MAR), were assessed. Results: Our findings revealed that bacterial counts in rivers and aquaculture farms ranged from log 2.00 to 3.68 CFU/mL and log 1.70 to 5.48 cfu/mL, respectively. Resistance to piperacillin (100%) was observed in all E. coli; resistance to amoxicillin (100%) and ampicillin (100%) was observed in E. coli found in aquaculture farms; resistance to streptomycin (93%) was observed in E. coli found in rivers. All E. coli were resistant to ≥2 antibiotics and formed 26 MAR profiles, ranging from an index of 0.17 to 0.83, indicating that there are high risks of contamination. Some (48.4%) of the E. coli were detected with plasmids (1.2 to >10 kb), whereas 51.6% of the E. coli did not harbor any plasmids. The plasmid copy numbers reported were one plasmid (n = 7), two plasmids (n = 4), ≥ two plasmids (4). E. coli isolated from the Muara Tuang River showed the highest-molecular-weight plasmids. A statistical analysis revealed that there is no significant correlation (r = 0.21, p = 0.253) between the number of plasmids and the MAR index of the tested isolates. Conclusion: The distribution of MAR in E. coli from rivers is higher compared to the aquaculture environment. Our study suggests that MAR in isolates could be chromosome-mediated. Our results suggest that riverbed sediments could serve as reservoirs for MAR bacteria, including pathogens, under different climatic conditions, and their analysis could provide information for public health concerns.

Incidence of Multidrug-Resistant Escherichia coli Harbouring blaTEM and tetA Genes Isolated from Seafoods in Lagos Nigeria.

The presence of multidrug-resistant Escherichia coli of fecal origin in seafood is a serious concern. Seafood containing MDR E. coli can serve as a medium for the transfer of resistant bacteria to consumers. The aim of the present study is to isolate and identify multidrug-resistant E. coli and associated resistant genes from selected seafood (catfish, crabs and tilapia fish) purchased from wholesalers and retailers at sea landing areas in Lagos state, Nigeria. A total of two hundred and thirty-eight (238) samples consisting of catfish (52), tilapia fish (78) and crab (108) were collected and investigated for the presence of E. coli from the period of June 2018-April 2019. Colonies that showed metallic sheen were considered presumptive E. coli isolates, and positive isolates were chosen for further confirmed by biochemical methods using IMViC tests, Oxidase test, triple sugar iron agar test and sugar fermentation test. Antimicrobial susceptibility of the isolates to eight classes of antibiotics was determined by disc diffusion methods while amplification of suspected antibiotic resistance genes were done by the polymerase chain reaction (PCR) using specific primers. A total of 105 (44.1%) E. coli were isolated from selected samples by standard microbiological procedures. The grand total of 59 (56.2%) isolates showed multiple antibiotic-resistant patterns. The overall result showed high-level resistance to tetracycline 101/105 (96.1%) and trimethoprim 90/105 (85.7%), cefotaxime 67/105 (42.9%) while the highest susceptibility of 101/105 (96.2%) was recorded for amikacin followed by gentamicin 84/105 (80%), meropenem 75/105 (71.4%), ceftazidime (69.5). The presence of tetA and blaTEM was prevalent among the isolates. Our results indicate that seafood may be a reservoir of β-lactam and tetracycline-resistance determinants.

Molecular identification of blaCTX-M and blaTEM genes encoding extended-spectrum ß-lactamase (ESBL) producing Escherichia coli isolated from raw cow’s milk in East Java, Indonesia

Abstract. Ansharieta R, Ramandinianto SC, Effendi MH, Plumeriastuti H. 2021. Molecular identification of blaCTX-M and blaTEM genes encoding extended-spectrum ß-lactamase (ESBL) producing Escherichia coli isolated from raw cow’s milk in East Java, Indonesia. Biodiversitas 22: 1600-1605. The emergence of extended-spectrum ?-lactamase (ESBL) producing bacteria and its increasing level has become public health issue. The presence of these bacteria in food of animal origin is quite alarming. The objective of this study was to detect and characterize Escherichia coli producing ESBL encoding genes, isolated from 200 raw cow milk samples in East Java, Indonesia. The results of this study showed that 70.5% of isolates were confirmed as E. coli, based on the morphological growth of colonies on the EMB Agar and biochemical IMViC tests. In this study, the double-disc synergy test (DDST) method was used to confirm the ESBL, and previously sorted out presumptively by using Aztreonam antibiotic disc. The antibiotics used were amoxicillin-clavulanate, ceftazidime, and cefotaxime for DDST. In addition, ESBL confirmation with Multiplex PCR method for blaCTX-M and blaTEM genes were done. The presence of ESBL-producing by E. coli isolated from raw cow’s milk in East Java were 2.12% (3/141). The PCR results showed that the double blaCTX-M and blaTEM gene harbored by 2 ESBL isolates and one blaTEM gene as many as 1 ESBL isolate. Thus, the findings of our study indicate that milk can be a good reservoir of bacteria carrying blaCTX-M and blaTEM ESBL resistance genes with the potential to affect human health.