Most Probable Number Values Research Articles

Potential of Eco-enzyme Averrhoa bilimbi L. Fruit as an Innovation in Tempeh Wastewater Treatment in Plaju District, South Sumatra

Tempeh home industries have been established in residential areas, one of which is in the assisted village of PT Pertamina Internasional RU III Plaju Factory, Palembang, South Sumatra. Eco-enzyme is one of the methods for processing and reducing pollutant organic matter in liquid waste. This study aims to determine the eco-enzyme characteristics of Averrhoa bilimbi L. fruit and exploit its potential to reduce chemical oxygen demand (COD), total suspended solid (TSS), and total dissolved solid (TDS) in tempeh wastewater. Eco-enzymes are characterized by measuring pH, COD, TSS, TDS, and most probable number (MPN). The reduction in COD, TSS and TDS of tempeh wastewater was observed using a simple bioreactor which was treated with the addition of 10% eco-enzyme (three replicates) and without the addition of eco-enzyme (control). The characterization results showed that the eco-enzyme of A. bilimbi L. fruit had a pH of 1.84, COD of 85.33 mg/L, TSS of 440 mg/L, TDS of 15,800 mg/L, protein concentration of 0.459 U/mg, and MPN value of 0.03 CFU/100 mL. The COD value of tempeh liquid waste can be reduced from the initial COD of 256 to 154.67 mg/L. The TSS value of tempeh wastewater decreased by 60% from the initial value of 4,000 mg/L. It takes 18 days to reduce the TSS content below the quality standard. The TDS value of tempeh liquid waste can be reduced by 70% from the initial TDS of 7,333.33 to 1,666.67 mg/L.

Spatiotemporal distribution of thermophilic free-living amoebae in recreational waters: A 5-year survey in Guadeloupe (French West Indies)

Free-living amoebae (FLA) such as Acanthamoeba, Balamuthia mandrillaris, Naegleria fowleri and Sappinia pedata are naturally widespread in freshwater, causing rare but fatal and debilitating infections in humans. Although recent studies have shown an increase in infection rates, there is a paucity of epidemiological studies regarding the presence of these emerging pathogens in water. Herein, we studied the diversity and relative abundance of thermophilic FLA in different recreational baths in a tropical climate for 5 years.From 2018 to 2022, a total of 96 water samples were collected from 7 recreational baths (natural, tiled, regularly cleaned or not, and with temperatures ranging from 27 to 40 °C). DNA was extracted from FLA cultivated at 37 °C to detect thermophilic culturable FLA. Metabarcoding studies were conducted through FLA 18S rRNA gene amplicons sequencing; amplicon sequence variants (ASV) were extracted from each sample and taxonomy assigned against PR2 database using dada2 and phyloseq tools. We also searched for Naegleria sp. and N. fowleri using PCR targeting ITS and NFITS genes (respectively) and we quantified them using an optimized most probable number (MPN) method for FLA. Our results showed that differences in FLA diversity and abundance were observed amongst the 7 baths, but without a clear seasonal distribution. Naegleria, Vermamoeba and Stenamoeba were the most represented genera, while the genera Acanthamoeba and Vahlkampfia were mainly found in 2 baths. The MPN values for Naegleria sp. (NT/l) increased between 2018 and 2022, but the MPN values for N. fowleri (NF/l) seemed to decrease.Globally, our results showed that since we cannot establish a seasonal distribution of FLA, the regular presence of FLA (namely Naegleria and Acanthamoeba) in recreational waters can pose a potential threat in terms of neuroinfections as well as Acanthamoeba keratitis. It is thus imperious to perform the regular control of these baths as a preventive health measure.

MicroMPN: methods and software for high-throughput screening of microbe suppression in mixed populations.

We created a unified set of software and laboratory protocols for screening microbe libraries to assess the suppression of a pathogen in a mixed microbial community. Existing methods of fluorescent labeling were combined with the most probable number (MPN) assay in a microplate format to enumerate the reduction of a pathogenic soil microbe from complex soil matrices. This work provides a fluorescent expression vector available from Addgene, step-by-step laboratory protocols hosted by protocols.io, and MicroMPN, a command-line software for processing plate reader outputs. MicroMPN simplifies MPN estimation from 96- and 384-well microplates. The microplate screening assay is amenable to robotic automation with standard liquid handling robots, further reducing the hands-on processing time. This tool was designed to evaluate synthetic microbial communities for use as microbial inoculates or probiotics. The fluorMPN method is also useful for screening chemical and antimicrobial libraries for pathogen suppression in complex bacterial communities like soil.

Multiple drug resistant enterobacteria from biofilms in some tap water supplies in a Nigerian university

Tap outlets have been found to contain biofilms, which are a consortium of bacteria, fungi, viruses and protozoa. These microorganisms, especially Gram negative bacteria, could be pathogenic and highly resistant to antibiotics, leading to an increase in the prevalence of diseases worldwide. The university community depends on water from boreholes for consumption and domestic uses; as such, the need to identify the Gram negative bacteria is expedient. The main objective of this research is to determine the potability of the water released from the taps, and ultimately reduce the incidence of biofilm-related water infections. Water samples were collected from five boreholes and five tap outlets were swabbed with a sterile swab stick. Coliforms were estimated from the water samples using the Most Probable Number (MPN) technique. Gram negative bacteria were isolated and identified from the swabbing using standard bacteriological and biochemical tests. The antibiotic resistance patterns of the isolates were also determined, and the genes responsible for the multiple antibiotic resistance were identified using the DNA extraction and Polymerase Chain Reaction (PCR) methods. Bacterial counts of the samples ranged from 6.0×105cfu/mL to 1.4×106cfu/mL. MPN values of the tap water samples ranged from 4 to 1100+cfu/100mL, which exceeded the WHO standards of water quality. A total of 31 bacteria were isolated, of which 8 (25.8%) were Pseudomonas sp, 7 (22.6%) were Proteus sp, 4 (12.9%) were Klebsiella sp, 4 (12.9%) were Escherichia coli, 3 (9.7%) were Enterobacter sp, 3 (9.7%) were Citrobacter sp and 2 (6.5%) were Salmonella sp. All the bacteria showed multiple drug resistance to different antibiotics used, especially the cephalosporins. The gene found to be responsible for the cephalosporin resistance was the TEM - 445. Tap swabbing and water samples were found to contain a high value of coliforms (4 – 1100cfu/100mL), showing heavy faecal pollution, many of which could be pathogenic microorganisms that renders the water unfit for human consumption. The presence of these multidrug resistant microorganisms in the tap outlets could pose a serious threat to public health.

Application of durian rind smoke powder to preserve chicken meatballs at room temperature

Smoke powder is a natural alternative to artificial preservatives for extending the shelf life of food products. This study assessed the use of smoke powder derived from durian rinds as a preservative for chicken meatballs. Durian rinds were pyrolyzed at 300 °C (T1), 340 °C (T2), and 380 °C (T3) to produce liquid smoke. After filtration to separate the remaining carbon, the resulting liquid smoke was purified by distillation at 190 °C and then converted into a smoke powder through the addition of maltodextrin and spray drying. The feasibility of smoke powder as a preservative for chicken meatballs was tested using total volatile base (TVB), total plate count (TPC), pH, and most probable number (MPN) tests for Escherichia coli. The findings demonstrated an upward trend in TVB, TPC, and MPN values over the storage duration. The TVB and TPC tests revealed that meatballs preserved with T3 smoke powder retained acceptable quality upon 64 h of storage. The MPN value of the T3 sample showed that E. coli bacterial contamination could still be tolerated up to 68 h of storage.

Water-Supply Potability Status of Bangladesh University of Health Sciences (BUHS)

Water that is free of disease-producing microorganisms and chemical substances harmful to health is called potable water. Drinking water quality has a microbiological and a physicochemical dimension. In public water supply systems, water should be disinfected. This observational study was conducted through a multiple-tube method. And the water supply channel of Bangladesh University of Health Sciences (BUHS) served as the study's sample. Using an aseptic procedure, 105 ml of water from various sources is collected for each sample on a conical flask. The selected sample was from the four different water supply points at BUHS institution. From sample 1, the MPN (Most Probable Number) index we found the combination of positive sets (0-1-0). The MPN value per 100ml water is (1). From sample 2, the MPN index we found the combination of positive sets (1-1-1). The MPN value per 100ml water is (5). From sample 3, the MPN index we found the combination of positive sets (1-2-1). The MPN value per 100ml water is (7). From sample 4, the MPN index we found the combination of positive sets (0-1-0). The MPN value per 100ml water is (1). In Dhaka city, there is a possibility of contamination of the water sources. Bangladesh University of Health Sciences is a renowned institution with a big number of staff. So, it is important to examine the water condition in this institution. Our work and research reflect a little about it. Gladly and hopefully, we found that there is no pathogenic organism in the water of this institution which is used every day and everybody drinks.

Detection of “Non-culturable” <i>Mycobacterium tuberculosis</i> Cells by Culture Methods

The specific features of the reactivation of “non-culturable” (NC) Mycobacterium tuberculosis (MTB) cells for the following propagation in liquid medium were defined, as well as the approach for the quantification of these cells by culture methods was suggested. When NC cells obtained in vitro were inoculated into standard Sauton’s liquid medium, a prolonged (up to 20 days) lag-phase is observed, in which no cell propagation is occurred. During the lag-phase, dormant cells secrete substances of unknown nature that inhibit or completely arrest the reactivation and growth of cultures when initial dormant cells concentration is above 107–108 cells/mL. Addition of meat-peptone broth (3.25 g/L) into a standard Sauton’s medium with a 10-fold reduced concentration of glycerol significantly stimulated the reactivation and propagation of the initially dormant cells inoculated at a concentration above the threshold. This modification of the medium composition made it possible to detect up to 103 times more cells in the MTB population by the method of the Most Probable Number (MPN) of cells. Increased number of the detected dormant cells in the population (up to an average of 2.5 orders of magnitude) was also achieved by adding to the reactivation medium recombinant protein RpfB (5 ng/mL), a protein of the Rpf family – resuscitation promoting factor of dormant bacteria. Perhaps the action of a Rpf enzyme is related to the products of its enzymatic activity, since an increased MPN value in the dormant cells population was also observed when the products of mycobacteria peptidoglycan hydrolysis obtained by the coaction of RpfB and endopeptidase RipA were added. The addition of sonicated peptidoglycan fragments at a concentration up to 1 μg/mL had a similar effect. The obtained results may be used as approaches to optimize liquid media composition and culture conditions aiming to identify in clinical samples the pathogen of tuberculosis remain in “non-culturable” state.

Bacteriological Study of Some Sachet Water Sold in Ogbomoso Metropolis and its Public Health Significance

The introduction of sachet water to consumers was to provide safe, hygienic and affordable instant drinking water to the public. Although this is a laudable idea, current trends seem to suggest that sachet drinking water could be a route of transmission of diseases. The objective of this study was to check the bacteriological safety or otherwise of some sachet water packaged in Ogbomoso metropolis. Using a simple random procedure, fifteen brands of sachet water were bought from vendors at different locations. The samples were analyzed using multiple tubes methods and biochemical assays. Results were recorded as Most Probable Number (MPN) of coliform per 100ml of water. The bacteriological quality of the samples was assessed based on the World Health Organization (WHO) classification system for drinking water. Zero (0%) of the sample was Excellent, two (2) (13.3%) were Satisfactory, one (1) (6.7%) was Suspicious and twelve (12) (80%) were Unsatisfactory using the MPN values recorded. The samples were contaminated with faecal coliform. Escherichia coli was detected in some of the samples which could have caused diarrhea, cramps, nausea and other symptoms in the consumers. The level of contamination could be due to inadequate treatment of water samples by the producers, improper use of filters or post-production contamination. The findings suggest the need to enforce the laws that govern the operation of such production outfits as well as educating consumers on the need to purchase sachet water from manufacturers that have been licensed to produce water and whose product bears the stamp of the National Agency for Food and Drugs Administration and Control (NAFDAC). And producers should ensure proper purification of water and maintain hygienic environment for producing the sachet water.

Dampak Perilaku Masyarakat Membuang Sampah Terhadap Kualitas Air Anak Sungai Kahayan Mendawai 7 Ujung Kota Palangka Raya

Settlements that develop around rivers will result in problems with the disposal of household waste to the riverbanks. There is quite a lot of household waste that enters the river. This study aims to determine the pollution index of the tributaries of the Kahayan Mendawai 7 Palangka Raya River to the fish in the river. The analysis was carried out using water quality analysis based on the MPN (Most Probable Number) Coliform, consisting of three test stages, namely an estimation test using Lactose Broth (KL) medium, a confirmation test using Brilliant Green Lactose Broth (BGLBB) medium, and a certainty test. using Mac Concey Agar (MCA) medium. This type of research is a direct survey method and laboratory analysis. this research was carried out in October 2021. The results of this study were obtained based on the formula and table of MPN values, three series of tubes were obtained with the total MPN Cokiform value in the estimating test of 2,400 cells/100 ml sample, the MPN value confirming test 28 cells/100 ml sample, and the number of Escherichia coli colonies in the certainty test were 69 colonies, so the water quality of the tributary of the Kahayan Mendawai 7 Ujung City of Palangka Raya was in the category of polluted and unfit for consumption.Keywords: Household Waste, Water Quality

UJI CEMARAN MIKROBA Coliform DAN IDENTIFIKASI Escherichia coli PADA SEDIAAN JAMU QUSTHUL HINDI YANG BEREDAR DI PASARAN

The need to maintain endurance during the Covid-19 pandemic has increased the intensity of people consuming herbal medicine. The Qusthul Hindi herbal medicine comes from the qust plant. This plant has the potential to treat various human ailments, including respiratory, digestive, and metabolic disorders. Making herbal medicine that is still very simple allows herbal medicine to be contaminated by microbes. This study aims to identify the presence of Coliform microbial contamination and E. coli bacteria in Qusthul Hindi herbal preparations circulating in the market. The five samples of Qusthul Hindi herbal medicine used in this study were obtained from several shops in Indonesian e-commerce. The Coliform microbial contamination test was conducted using the Most Probable Number (MPN) method. The MPN test consists of 2 stages: a presumptive test using Lactose Broth media and a confirmed test using Escherichia coli Broth media. MPN calculation is based on the number of positive test tubes. Positive test tube observations can be seen by observing the emergence of turbidity or the formation of gas contained in the Durham tube. The MPN value is calculated from the combination of positive tubes in the MPN table. The identification of Escherichia coli was tested using Eosin Methylene Blue Agar (EMBA) selective media, and then gram staining was performed. From the results of the estimator test, confirmatory test, selective medium test to gram staining, it showed that five Qusthul Hindi herbs tested were positively contaminated by Coliform microbes and all samples showed positive results for Escherichia coli contamination.

Antibiotic resistant bacteria isolated from Agulu lake in Anambra State of Nigeria West Africa

This research investigated the antibiotic resistant bacteria isolated from Agulu Lake in Anambra state of Nigeria. Water experiment was carried out on four (4) water samples aseptically collected from different points in the Agulu Lake. The samples were analyzed for physicochemical features, Most Probable Number (MPN), Gram stain, microscopy, motility, biochemical traits and antibiotic susceptibility tests using standard microbiological instrumentation. The results were analyzed through inferential statistics. The bacterial isolates were characterized based on their Gram stain, microscopy, motility and biochemical properties. The degree of susceptibility of the isolates against conventional antibiotics was determined using antibiotics’ impregnated discs assay. The physicochemical characteristics revealed the following average results: acidic pH of 6.0, temperature of 34 0C and turbid. The highest MPN value was recorded by water sample collected from site D while the lowest MPN value was found at site B of the stream. The results of Gram stain, microscopy, motility and biochemical characterization inferred the presence of Pseudomonas species and Escherichia coli in the water samples. The study confirmed that Agulu Lake in Anambra state of Nigeria contains bacteria that are resistant to most conventional antibiotics. So, there is a call for massive information dissemination to all for caution towards health promotion.

Detection of Escherichia albertii in Retail Oysters

Escherichia albertii is an emerging foodborne pathogen. Owing to its distribution in river water, it is important to determine the presence of E. albertii in aquaculture-related foods. In this study, we investigated the distribution of E. albertii in retail oyster samples. A total of 427 raw oyster samples (385 Pacific oysters and 42 Japanese rock oysters) were enriched in modified Escherichia coli broth (mEC) or mEC supplemented with novobiocin (NmEC) at 42°C. The cultures were used for E. albertii-specific nested PCR assay, as well as for E. albertii isolation using deoxycholate hydrogen sulfide lactose agar (DHL), DHL supplemented with rhamnose and xylose, and MacConkey agar supplemented with rhamnose and xylose. The population of E. albertii in nested PCR-positive samples was determined using the most-probable-number (MPN) method. E. albertii isolates were subjected to biochemical and genetic characterization. E. albertii was detected in 5 (1.6%) of 315 Pacific oyster samples (one piece each), 2 (2.9%) of 70 Pacific oyster samples (25 g each), and 2 (4.8%) of 42 Japanese rock oyster samples procured from four geographically distinct regions. A total of 64 E. albertii strains were isolated from eight of the nine nested PCR assay-positive oyster samples, and the MPN value was under the detection limit (<3 MPN/10 g). A specific season or month for detecting E. albertii was not observed in this study, suggesting that the pathogen is present in seawater. All the E. albertii isolates, except one, were positive for the virulence factor eae, indicating that these isolates have the potential to infect humans.

Detection of microbial contamination in chicken meat from local markets in Surabaya, East Java, Indonesia.

Background and Aim:Chicken meat can be contaminated by microorganisms anywhere in the supply chain, from farm to market, and these microorganisms can be transmitted to humans through direct contact, contact with the environment, and food consumption. The microbial contamination has a serious impact on public health. This study aimed to analyze the microbial contamination of chicken meat sampled from local markets in Surabaya, East Java, Indonesia.Materials and Methods:A total of 60 samples of fresh chicken meat obtained from 10 traditional markets (six samples per market) were examined for the presence of bacteria. Staphylococcus aureus, Salmonella spp., and Escherichia coli were identified using Gram staining, culturing, and biochemical tests. The most probable number (MPN) method was used to identify E. coli.Results:Most chicken meat samples were positive for S. aureus (58.3%), Salmonella spp. (48.3%), and E. coli (40%). The samples were considered positive for E. coli if the MPN value was higher than 1×101 CFU/g.Conclusion:High microbial contamination was found in all the chicken meat sampled from local markets in Surabaya. Such contamination can lead to foodborne diseases so, proper hygiene and sanitation standards should be followed from slaughterhouses to the end-users.

The use of alternative water sources as a means of adaptation to water shortages in Nsimeyong, Yaounde city: a quality assessment

Nsimeyong (Damas), one of the residential areas of Yaounde, Capital city of Cameroon experiences water shortages making residents rely on other sources of water including streams, springs, boreholes and wells. Water samples were aseptically collected from the processing plants and from five sites of Nsimeyong area. This study was aimed at assessing suspected heavy metals and coliforms of water collected from fifteen alternative water supply sources in this area. Heavy metals (Pb and Cd) were determined using an Atomic Absorption Spectrophotometer. The International Organization for Standardization (ISO) 9308-2:1990b last reviewed and confirmed in 2017 was used to determine the Most Probable Number (MPN) of total coliforms by the five-tube MPN table. The presence of presumptive E. coli was confirmed on Sugar, Indole, and Motility (SIM) media at 44⁰C and by the addition of Kovac's reagent to tryptone water. Water samples showed higher concentrations of Pb beyond the maximum permissible level (0.1mg/l). With the exception of processed tap water at home, all other alternative sources of water used in Damas registered Cd above the World Health Organization recommendation (0.03mg/l). Unlike Cd (p = 0.0001), the difference among Pb content of water samples at the collection sites was not statistically significant (p = 0.1416). More than 66% of water samples (of which closed to 39% were open wells) had MPN value for total coliform beyond the permissible level (<10) in drinking water. Moreover, E.coli and Klebsiella spp were detected in 16.7% and 11.1% of water samples from alternative sources, respectively. Additional household treatment such as filtration, to remove heavy metals and boiling before use of such water is necessary to achieve the standard bacteriological quality for human consumption thereby reducing risk of exposure to waterborne diseases. This study constitutes the baseline information to guide the policy makers in the development and implementation of the water safety plan in Nsimeyong area.

Prevalence and Characteristics of Salmonella spp. from a Pig Farm in Shanghai, China.

Salmonella spp. is a major foodborne pathogen that is distributed among most pork production chains worldwide. This study aimed to investigate the dynamic changes in Salmonella spp. along the pig breeding process monthly from April 2018 to March 2019 in a pig farm in Shanghai, China, and identify the potential critical control points during the production. In total, 239 Salmonella spp. isolates were obtained from 1389 samples, in which Salmonella were detected from 26.3% (222/843) of fecal samples, 7.1% (17/240) of feed samples, and 0.0% (0/306) of both water and insect samples. Seven different serotypes were identified, with the predominant serotype being Salmonella Derby (21.8%), followed by Salmonella Typhimurium (18.8%), Salmonella Rissen (16.3%), Salmonella Mbandaka (12.6%), and Salmonella 1,4,[5],12:i:- (11.8%). Most probable number (MPN) analysis revealed that the load of Salmonella spp. gradually increased along the pig production chain, while the highest number of Salmonella spp. isolates was at the fattening stage (MPN value, 11-15 MPN/g). The pulsed-field gel electrophoresis showed that both Salmonella Typhimurium and Salmonella Derby isolates were grouped to six clusters. The antimicrobial resistance analyzed demonstrated that 80.0% of the isolates were of multidrug resistance and resistant to sulfamethoxazole (84.5%), lincomycin (89.4%), ampicillin (96.9%), oxytetracycline (93.8%), and tetracycline (95.1%). We further evaluated the Salmonella spp. Resistance to quaternary ammonium compounds (QACs) showed an increasing trend along with the testing period indicating that the use of QACs could induce the resistance of Salmonella spp. to QACs. Our study confirmed the dynamic changes in Salmonella spp. over time and space in this pig farm and identified feed and the fattening house as the key points for the prevention and control of Salmonella spp. contamination.

Analysis of wastes height reduction, macrobiology and microbiology colony (fecal coli) on dry leaves composting process

The scattered leaf wastes can be turned into high nutrition organic fertilizer by utilizing vermicomposting methods. The wastes will be consumed by earthworms reducing the mass of the waste and the excrement can be used as vermicompost. One of the important things regarding the potential of earthworms in vermicomposting is the consumption rate or the decrease in the waste pile heights. Also, the number of microorganism colonies can be determined using the MPN (Most Probable Number) method based on the parameters of fecal coli bacteria in SNI 19-7030-2004. The vermicompost produced in this study comes from banyan leaves (Ficus benjamina) and bamboo leaves (Bambusa bambos). It takes 56 days for the vermicompost to ripen. The decrease in pile heights was measured every three days using a ruler and resulted in an average reduction of 2.18 cm/day for Ficus benjamina leaves and 1.0 cm/day for Bambusa bamboos leaves. Samples of microorganisms were taken on the 28th and 56th day and the MPN value was calculated. The MPN value obtained on the 28th day was 290 MPN/g and on the 56th day was 460 MPN/g, indicating that the colony had reached the predetermined quality standard.

Detection of Escherichia coli Contamination using Most Probable Number (MPN) methods in Chicken Meats in Market of Surabaya

Foodborne diseases are diseases transmitted through contaminated food and are one of the main public health problems. The bacteria that usually contaminates chicken meat is Escherichia coli. E. coli contamination in chicken meat can affect consumer health. Chicken meat processing and market conditions affect the level of bacteria contamination. This study aims to measure the contamination of E. coli bacteria in chicken meat in Surabaya's markets. The method which used to count and estimate the number of E. coli bacteria found in chicken meat in this study was Most Probable Number (MPN). The results showed 20 samples (33.3%) from 60 samples gave negative results. This indicated that only 20 samples of chicken meat were safe for consumption because the MPN value is &lt;1 × 101 CFU/g while the remaining 40 samples (66.7%) were not safe for consumption because the MPN value is&gt; 1 × 101 CFU/g. The standard of SNI 3924: 2009 concerning the minimum requirement for the amount of E. coli contamination in chicken meat is 1×101 CFU/g so that the contamination of E. coli which found in chicken meat in Surabaya was high.

Evaluating Methods for Detecting Escherichia albertii in Chicken Meat

Escherichia albertii is an emerging foodborne pathogen. The source of the E. albertii infection in most foodborne outbreaks is unknown because E. albertii is difficult to isolate from suspected food or water. E. albertii has a broad host range among birds and can be isolated from chicken meat. In this study, PCR assay, enrichment, and isolation conditions for detecting E. albertii in chicken meat were evaluated. The growth of 47 E. albertii strains isolated in Japan between 1994 and 2018 and a type strain was evaluated in modified EC broth (mEC) and mEC supplemented with novobiocin (NmEC) and on media containing carbohydrates. The enzyme used for the nested PCR, the enrichment conditions, the most-probable-number (MPN) method, and agar media were also evaluated with chicken meat. To distinguish E. albertii from presumptive non-E. albertii bacteria, desoxycholate hydrogen sulfide lactose agar (DHL), MacConkey agar (MAC), and these agars supplemented with rhamnose and xylose (RX-DHL and RX-MAC, respectively) were used. All E. albertii strains grew in mEC and NmEC at both 36 and 42°C and did not utilize rhamnose, sucrose, or xylose. Both the first and nested PCRs with TaKaRa Ex Taq, which was 10 to 100 times more active than the other enzymes, produced positive results in enrichment culture of 25 g of chicken meat inoculated with >20 CFU of E. albertii and incubated in mEC and NmEC at 42°C for 22 ± 2 h. Thus, the first PCR was sensitive enough to detect E. albertii in chicken meat. The MPN values in mEC and NmEC were 0.5- and 2.3-fold higher than the original inoculated bacterial levels, respectively. E. albertii in chicken meat was more efficiently isolated with enrichment in NmEC (70.1 to 100%) and plating onto RX-DHL (85.4%) and RX-MAC (100%) compared with enrichment in mEC (53.5 to 83.3%) and plating onto DHL (70.1%) and MAC (92.4%). Thus, optimized conditions for the surveillance of E. albertii contamination in food and investigations of E. albertii outbreaks, including the infectious dose, were clarified.

Antibiotics resistant Escherichia coli isolated from aquatic ecosystems in Palembang, South Sumatra, Indonesia

Abstract. Verawaty M, Apriyani N, Tarigan LR, Apriyan ET, Laurenta WC, Apriani N, Muharani. 2020. Antibiotics resistant Escherichia coli isolated from aquatic ecosystems in Palembang, South Sumatra, Indonesia. Biodiversitas 21: 86-97. Antibiotics-resistant bacteria (ARB) are one of the emerging water contaminants currently gaining serious global concern due to their adverse risk to ecosystems, wildlife, and public health. This study determined antibiotics resistant Escherichia coli isolated from aquatic ecosystems in Palembang. Most probable number (MPN) was used for bacterial estimation and the Kirby-Bauer method was used for susceptibility test against antimicrobial agents. The results indicated that 82% of E. coli isolates from 28 sampling sites were resistant to ampicillin, 57% to tobramycin, and 71% to tetracycline. The isolates showed intermediate profile to kanamycin (50%), 57% to cotrimoxazole, 50% to cefixime, and 54% to gentamycin. These isolates still showed sensitivity towards ciprofloxacin (86%) and chloramphenicol (61%). Total coliform (TC) numbers ranged from 0 to &gt;1600 MPN/100 mL. Sampling sites with high MPN values of ≥1600 MPN/100 mL were Sekanak watersheds (SW1, SW8, SW11, SW12, SW13, and SW14) followed by SW2 and SW9 with a value of 1600 MPN/100 mL, while samples from cattle and fish farms (CW) varied from 0 to 170.000 MPN/100 mL. TC of samples collected from retention ponds (RP) ranged from 0 (RPJSC) to 1.600.000 MPN/100 mL (RPSH3). The Escherichia coli (EC) counts varied from 1.7 x 103 (RPSH2) to ≥1.6 × 104 MPN/100 mL (SW11, SW12, SW13, and SW14). Several samples (SW3, RPPI, RPTS, RPSB, RPIBA, and RPOPI) have no E. coli. The results indicated some of the sampling locations that exceeds the quality standard of water have been regulated by the Governor of South Sumatra and the Indonesian Government.

Escherichia coli O157:H7 in Drin Prevalence of Escherichia coli O157:H7 in Drink from Different Food Premises in Kota Samarahan Sarawak

Contamination of drinks with E. coli O157:H7 served in food premises such as restaurants can cause haemorrhagic colitis and haemolytic uremic syndrome to humans. The presence or absence of faecal pathogen was demonstrated using coliform group as indicator microorganisms. Therefore, this study was conducted to detect the presence of E. coli O157:H7 in drinking water from food restaurant premise in Kota Samarahan and Kuching to ensure safe and potable drinking water is served to the consumer. A total of thirty (n=30) drink samples including six types of each of the samples are cold plain water, iced tea, iced milo, syrup and iced milk tea. Most Probable Number (MPN) procedure was used in this study to enumerate the MPN values of coliform bacteria in each drink collected. A total of 53.33% (16/30) of the drink samples showed positive E. coli detection. Then, the PCR assay showed 6.25% (one out of 16 isolates) samples were positive and carried stx1 gene produced by E. coli O157:H7 in iced milo sample types. This study showed the drinks collected from food premises was contaminated with faecal contamination, which was not safe to drink by the consumer. Therefore, preventive actions should be taken to prevent foodborne illness outbreak in future