Triple Sugar Iron Agar 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.

Probiotic, bacillus amyloliquefaciens from milk sample of a native cow ‘Kangeyam’ breed in forest fringe village of Coimbatore, Tamil Nadu and value addition thereof

Probiotics include good microorganisms that support the body's ability to operate and stay healthy. These good bacteria have several advantages, including as helping us feel better and warding off bad bacteria when there are too many of them in the stomach. Milk samples from the native indigenous Kangeyam breed cattle reared in the tribal hamlets on the forest fringes of Coimbatore were collected to isolate the beneficial microorganism. The sample is specifically collected from such region as it might have a higher efficiency in comparison with domesticated livestock. The milk samples collected were processed and analyzed for their nutritional value. Milk samples obtained from the native indigenous Kangeyam breed cattle reared in tribal hamlets of forest fringe villages was found to be more efficient in the nutritional aspects as like the difference in the milk from regular domesticated cattle. The quality of the milk was tested and then the probiotics were isolated. The pure culture was obtained in which the basic confirmatory tests such as Gram staining, Motility, IMViC, Catalase, Oxidase, Urease, Gas from glucose, Triple sugar iron test, etc., were done. Culture with more accurate results was made for molecular identification by 16srRNA method, in which the species Bacillus amyloliquefaciens has been confirmed and submitted to NCBI. The probiotic was found to be beneficial. The safety assessment was studied for accepted intake. Then the culture was made frozen, which was further lyophilized to be made into powder. The isolated probiotic with significant health benefits can be considered as a probiotic infused value-added product with commercialization potential.

ESCHERICHIA COLI CONTAMINATION ON IMPORTED FROZEN Buffalo MEAT SOLD AT AIKMEL TRADITIONAL MARKET, AIKMEL DISTRICT, EAST LOMBOK DISTRICT, WEST NUSA TENGGARA PROVINCE

The demand for beef in Indonesia continues to increase. The government is striving to achieve self-sufficiency in beef production, but the high price of beef is driving imports of buffalo meat from India. Buffalo meat has high nutritional value, similar to beef. Since 2015, frozen buffalo meat imports from India have begun to enter Indonesia, particularly in West Nusa Tenggara, as an effort to meet market demand in the modern shopping areas in Mataram City, Lombok. The relatively cheap price of frozen buffalo meat has led to expanded marketing in East Lombok, especially in the traditional market of Aikmel, which is approximately 72 km away from Mataram. The aim of this research is to determine the extent of Escherichia coli contamination and the amount of Escherichia coli contamination per gram of buffalo meat sold in this traditional market. The research samples consist of frozen buffalo meat from India sourced from 10 Indian buffalo meat vendors in the Aikmel market, each weighing 100 grams. The isolation and identification method follows Jawetz et al. (2008), starting with isolation on Eosin Methylene Blue Agar (EMBA), followed by identification with INMVIC (Indole, Methyl Red, Voges Proskauer, Citrate), reinforced by results from TSIA (Triple Sugar Iron Agar) and Urease tests. The research found that E. coli contamination exceeded the specified limit (SNI 7388:2009, 2009) in 9 out of 10 samples taken from 10 Indian buffalo meat vendors in the Aikmel market. The smallest and largest amounts of E. coli contamination in 90% of the contaminated samples were 80x101 and 7x103 colonies/gram, respectively. This condition is crucial for the government to pay attention to, in order to enhance education and supervision of the public regarding the procedures and regulations for the sale of frozen buffalo meat from India, in accordance with the marketing of animal origin products.

PREVALENCE OF ESCHERICHIA COLI, COLIFORM AND NON-COLIFORM BACTERIA IN MUD BUFFALO IN RANGGAGATA AND SERAGE VILLAGES, SOUTHWEST PRAYA SUBDISTRICT, DISTRICT OF CENTRAL LOMBOK, WEST NUSA TENGGARA

Escherichia coli (E. coli) is a common bacterium that inhabits the intestines of humans and animals with hundreds of different strains, both harmful and harmless. One of the diseases that can be caused by E. coli is diarrhea. Diarrhea is a disease in the form of increased frequency of defecation, thinner stool consistency and increased intestinal peristalsis. This study aims to determine the number of E. coli, coliform and non-coliform bacteria in mud buffaloes in Ranggagata and Serage Villages, Southwest Praya Subdistrict, Central Lombok Regency, West Nusa Tenggara with samples used are fresh feces samples, a total of 14 samples. Sample cultivation was carried out on Eosin methyleneblue agar (EMBA) media then samples suspected of E. coli colonies were Gram stained, and biochemical tests on Triple Sugar Iron Agar (TSIA) media, Sulfide Indole Motility (SIM), Methyl red-Voges Proskauer (MR-VP) media, Simmons Citrate and catalase tests. The results showed the number of bacterial populations varied depending on the age of the buffalo with an average number of E. coli bacteria 162.143 ± 196.476, average coliform 272.000± 328.307 and average non-coliform 110.714 ± 201.855. Based on the results of the study it can be concluded that there is an increase in the number of E. coli, coliform and non-coliform as the age of buffalo increases, namely for E. coli is Y = 2.527+0.009x, coliform is Y = 2.525+0.05x and non-coliform bacteria is Y = 3.199+0.007x. Further tests need to be conducted related to other strains or species of E. coli, coliform and non-coliform bacteria in mud buffalo feces.

CONTAMINATION ESCHERICHIA COLI IN BROILER CHICKEN MEAT SOLD AT BADUNG MARKET, DENPASAR, BALI DURING THE TRANSITION SEASON

Escherichia coli is reported as an intensive bacterial contaminant of chicken meat, including broiler meat. Factors that support the occurrence of contamination are the environment, equipment and sellers. This study aims to determine the presence of Escherichia coli bacteria in broiler meat sold in Badung market, Denpasar, Bali during the transitional season. The samples used were fresh broiler meat totaling 15 samples by taking randomly from 15 broiler meat traders. Samples will be taken to the laboratory for isolation and identification of Escherichia coli. The isolation stage uses selective media, namely Eosin Methylen Blue Agar (EMBA), identifying with the gram staining method, and biochemical tests consisting of Triple Sugar Iron Agar (TSIA), Citrate, Indol and Methyl Red tests. The results of the study found 3 (20%) of the 15 samples identified were contaminated by Escherichia coli bacteria. From the results of this study there is still contamination of Escherichia coli bacteria and needs to get more attention from the government in order to get education about the procedures for selling broiler meat according to standards and implement applicable regulations more strictly, because it affects public health.

Detection and evaluation of susceptibility to antibiotics in non-hydrogen sulfide-producing antibiotic-resistant soil microbe: Pseudomonas guariconensis.

Antimicrobial resistance in bacteria is a global threat that can make antibacterial treatments ineffective. One well-known method of antibiotic resistance and a common defensive mechanism in many harmful bacteria is the synthesis of endogenous hydrogen sulfide (H2S) in bacteria. In this study, soil bacteria were screened using the lead acetate agar test and the triple sugar iron test to determine that they were non-endogenous H2S producers. This was further validated by full genome analysis of the identified organism against the gene sequences of H2S-producing genes. Antibacterial resistance of the bacteria was phenotypically analyzed using the Kirby-Bauer disk diffusion method. Then, the effect of exogenous H2S on the antibiotic-resistant bacteria was checked in sodium sulfide, leading to antibiotic re-sensitization.

Antimicrobial susceptibility patterns of urinary tract infections causing bacterial isolates and associated risk factors among HIV patients in Tigray, Northern Ethiopia

BackgroundUrinary tract infections, a prevalent global infectious disease, are clinical issues not well studied in HIV-positive individuals. UTIs have become a global drug resistance issue, but the prevalence and antibiotic susceptibility patterns of UTI-causing bacteria among HIV patients in Tigray, Ethiopia, are poorly understood. This study aims to identify the prevalence of UTI-causing bacteria, their antibiotic susceptibility patterns, and associated risk factors in HIV patients attending ART clinics at Mekelle General Hospital and Ayder Comprehensive Specialized Hospital in Tigray, Northern Ethiopia.MethodClean-catch midstream urine samples (10–15 mL) were collected from HIV patients who are attending ART clinics at Mekelle General Hospital and Ayder Comprehensive Specialized Hospital. Samples were analyzed based on standard microbiological protocols using cysteine-lactose electrolyte deficient (CLED) agar. Pure colonies of bacterial isolates were obtained by sub-culturing into Mac-Conkey, Manitol Salt agar and blood agar plates. The bacterial isolates were then identified using macroscopic, microscopic, biochemical, and Gram staining methods. Gram-negative bacteria were identified using biochemical tests like triple sugar iron agar, Simon’s citrate agar, lysine iron agar, urea, motility test, and indol test, whereas Gram-positive isolates were identified using catalase and coagulase tests. The Kirby-Bauer disk diffusion technique was used to analyze the antimicrobial susceptibility pattern of bacterial isolates. Data was analyzed using SPSS version 25.0.ResultsAmong the 224 patients, 28 (12.5%) of them had been infected by UTIs-causing bacteria. E. coli was the dominant bacterium (16 (57%)) followed by K. pneumoniae (4 (14%)), and S. aureus (3 (11%)). Of the total bacterial isolates, 22 (78.6%) of them developed multi-drug resistance. All Gram-positive (100%) and 75% of Gram-negative bacterial isolates were found to be resistant to two or more drugs. Patients with a history of UTIs, and with CD4 count < 200 cells/ mm3, were more likely to have significant bacteriuria. Compared to male patients, female patients were more affected by the UTIs-causing bacteria. More than 93% of the UTIs-causing bacterial isolates were susceptible to nitrofurantoin, ceftriaxone, ciprofloxacin, and gentamycin; whereas they are highly resistant to ampicillin (96%), cotrimoxazole (82%) and tetracycline (71%).ConclusionsMost of the bacterial isolates were highly resistant to ampicillin, cotrimoxazole, and tetracycline. Female patients were more affected by the UTIs causing bacteria. The highest prevalence (12.5%) of UTIs in HIV patients needs special attention for better management and monitoring. Previous UTI history and immune suppression are predictors of UTIs, highlighting the need for intervention measures involving molecular studies to identify resistant bacteria genes and promote patient immune reconstitution.

Molecular detection of hlyF gene on multidrug resistance of avian pathogenic Escherichia coli isolated from ducks on wet markets of Surabaya, Indonesia

Abstract. Kendek IA, Putri MFR, Wibisono FJ, Effendi MH, Tyasningsih W, Ugbo EN, Agumah NB. 2024. Molecular detection of hlyF gene on multidrug resistance of avian pathogenic Escherichia coli isolated from ducks on wet markets of Surabaya, Indonesia. Biodiversitas 25: 1246-1252. Avian Pathogenic Escherichia coli (APEC) is a strain of pathogen that can cause colibacillosis in poultry, including ducks worldwide. This disease can be influenced by various virulence genes, one of which is the hlyF gene which can cause systemic disorders and potentially become zoonotic in poultry. This study aimed to detect the hlyF virulence gene from the multidrug resistance (MDR) properties of Escherichia coli originating from 158 duck cloacal swab samples from seven live markets in Surabaya. Samples were isolated and identified using Eosin Methylene Blue Agar (EMBA) and MacConkey Agar (MCA) media, Gram staining, and then continued with biochemical tests of Triple Sugar Iron Agar (TSIA), Simmons Citrate Agar (SCA), Sulfide Indole Motility (SIM), Methyl red (MR) and Voges-Proskauer (VP). Samples were tested for MDR properties and continued with confirmation of the hlyF virulence gene using PCR. Based on the results of this study, it showed that 85% (134/158) were positive for Escherichia coli. Escherichia coli bacteria experienced resistance to the antibiotic erythromycin 96% (129/134), ciprofloxacin 16% (22/134), antibiotics gentamicin 15% (20/134), aztreonam 4.4% (6/134) and chloramphenicol 3% (5/58). The MDR test results in this study were 15% (20/134), while the PCR test results for the hlyF gene were 60% (12/20). Therefore, Escherichia coli was found to have the hlyF virulence gene, an MDR in Escherichia coli bacteria at the Surabaya live market. Furthermore, APEC strains with the hlyF gene, an MDR, potentially affect public health.

Molecular detection of iron gene on multidrug resistant avian fecal Escherichia coli isolated from broiler on traditional markets, Surabaya, Indonesia

Abstract. Putri MFR, Kendek IA, Wibisono FJ, Effendi MH, Rahardjo D, Tyasningsih W, Ugbo EN. 2023. Molecular detection of iron gene on multidrug resistant avian fecal Escherichia coli isolated from broiler on traditional markets, Surabaya, Indonesia. Biodiversitas 24: 6454-6460. Avian Fecal Escherichia coli (AFEC) is the cause of colibacillosis, which often infects the poultry industry throughout the world. The virulence gene influences the iroN factor, which causes systemic infections in poultry. This research aimed to determine Multidrug Resistance (MDR) in AFEC obtained from traditional markets in Surabaya. There were 96 cloacal swab samples from broiler chickens. The samples were isolated and identified using culture media in the form of Eosin Methylene Blue Agar (EMBA), gram stain, Triple Sugar Iron Agar (TSIA), Sulfide Indole Motility (SIM), Simmons Citrate Agar (SCA), Voges Proskauer (MR-VP) indole and methyl test; detection of multidrug resistance using Mueller-Hinton Agar (MHA) media. The results of the isolation and identification of Escherichia coli bacteria in this study showed that 60.4% of the isolates were positive for Escherichia coli. Escherichia coli is resistant to tetracycline antibiotics by 56%, ciprofloxacin by 55%, antibiotics aztreonam 29%, kanamycin 20%, and chloramphenicol 18%. The multidrug resistance test result on Escherichia coli was 25.8%. The PCR test results for the iroN gene were 40%. Therefore, there are MDR and iroN genes in avian fecal Escherichia coli in Surabaya traditional markets; APEC with iroN gene poses the potential to affect public health.

Isolation and Identification of Gram Negative Bacteria in The Lumen Fluid of Colorectal Cancer Patients

Colorectal cancer is one of the major health problems in Indonesia and the third most common cause of death worldwide. Colorectal cancer is a cancer of the colon and the rectum. There is considerable research evidence demonstrating the important role of complex gut microbiota communities in developing intestinal tumorigenesis. This study aims to isolate and identify gram-negative bacteria found in the colonic lumen fluid of colorectal cancer patients using the gram-staining method and biochemical test. From microscopic identification results, macroscopic gram staining, motility test, Simon citrates agar (SCA), and triple sugar iron agar (TSIA) tests against H1 and H2 isolates described gram-negative stem bacteria included in the Enterobacteriaceae family belonging to the species Shigella sp.

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.

COLISEPTICEMIA INFECTION IN A 20 DAYS OLD BROILER CHICKEN IN TIMUHUN VILLAGE, KLUNGKUNG

Colisepticemia is one of poultry infectious disease caused by Avian pathogenic Escherichia coli. The aim of this study was identifying the causative of an infected broiler chickens aged 20 days. The research was conducted at a close house farm in Timuhun Village, Banjarangkan, Klungkung Regency, Bali with a population of 16,000 broiler chickens. The number of sick chickens was 163 and 55 died. The chicken samples were brought to the Veterinary Pathology Laboratory, Udayana Faculty of Veterinary Medicine for necropsy. Organ samples that underwent changes were taken to the Veterinary Bacteriology and Parasitology Laboratory. In bacteriological examination, organ samples were isolated using nutrient agar and eosyne methylene blue agar (EMBA) media. Furthermore, the results of bacterial isolation were identified using triple sugar iron agar (TSIA) media, Simmon Citrate Agar (SCA), Sulfid Indole Motility (SIM), Methyl Red-Voges Proskauer (MR-VP) and testing sugars (glucose and lactose). In the parasitology laboratory, fecal samples were examined using native, sedimentation and flotation methods. The results of the epidemiological examination of the disease, 1% morbidity, 0.3% mortality and 33.7% case fatality rate (CFR). Post-mortem examination found typical anatomical pathology in the form of polyserotysis and fibrin accumulation in the pericardium, perihepatic, pleural and mesentery. The results of the fecal examination did not find any agents and/or worm eggs. Bacterial isolation on EMBA media grew metallic green round colonies with black dots. Gram stain shows short rods and is colored red. Based on the results of pathology, parasitology and bacteriology examinations, the etiology of this cases caused by Avian Pathogenic Escherichia coli (APEC). Elevating sanitation was needed to reduce the cases.

A study of Isolation of Salmonella species from blood culture &amp; it’s Antimicrobial Resistant Pattern.

Background: The main aim of this study was to monitor the antimicrobial resistant pattern of Salmonella isolates to select proper antibiotic &amp; prevent drug resistance in Salmonella species. Material and Methods: Total 250 blood culture bottles were collected from patients clinically suspected enteric fever and loaded to BACTEC 9050. Out of them 200 signalled positive samples were inoculated on chocolate and Mac Conkey agar plates and incubated overnight at 370 C in the incubator and identified by colony characteristics, procedures like Gram staining, wet preparation for motility and bio-chemical reactions like oxidase test, catalase test, triple sugar iron agar, Citrate test, using Salmonella antisera like Poly O, O-9, and H-d. The clinical isolates were subjected to antibiotic sensitivity test on Mueller-Hinton agar, using modified Kirby Bauer disc diffusion method as per Clinical Laboratory Standard Institute (CLSI) guidelines. Results: Total 61 Salmonella were isolated. 49 were Salmonella typhi and 12 were Salmonella paratyphi. Overall rate of resistance of 49 S. typhi isolates was 2 % to ampicillin, 4 % to Azithromycin, 4 % to fluoroquinolones and 2 % to co-trimoxazole. S. paratyphi isolates were 100% sensitive to ampicillin, co-trimoxazole, chloramphenicol and 3rd generation cephalosporins. Conclusion: Enteric fever is one such infection which poses challenges in antimicrobial resistance. Continuous surveillance is important to track bacterial resistance and to treat infections in a cost-effective manner.

The Potential of Tapak Dara (Catharanthus roseus) Leaves Endophytic Bacteria BETD5 as Antioxidant and Anticancer Against T47D Breast Cancer Cells

This study aimed to measure antioxidant activity, to characterize biochemical properties and to measure cytotoxicity activity in BETD5 isolates. The method used in this research was experimental method. Parameters observed were antioxidant activity and cytotoxicity activity consisting of IC50 (Inhibition Concentration) and biochemical properties. Isolate used was extract of endophytic bacteria of Tapak Dara (BETD5). antioxidant activity was conducted using various sample extract concentration, namely 5ppm, 10ppm, 25ppm, 50ppm and 100ppm. The biochemical tests carried out included Triple Sugar Iron Agar (TSIA) test, Simmons Citrate (SC) test, motility test, indole test, catalase test and Methyl Red (MR) test. Cytotoxicity test on cell line T47D was performed by MTT assay using concentration (%) of 3.125; 6.25; 12.5; 25 and 50. The results showed that antioxidant activity of BETD5 was strong because IC50 value obtained was 78.485 ppm. Based on biochemical tests, BETD5 was suspected belong to the genus Staphylococcus. Cytotoxicity activity of BETD5 obtained an IC50 value of 15.58 ppm. This indicates that endophytic bacterial extract of Tapak Dara (BETD5) had moderate cytotoxicity against the T47D breast cancer cell line. GCMS test results showed that the endophytic bacterial isolate BETD5 had the highest secondary metabolite content, namely cis-Ocimene (24.31%).

Isolasi dan Identifikasi Bakteri Clostridium Botulinum pada Minyak Jelantah

The aims of this study were to determine whether Clostridium botulinum bacteria were present in used cooking oil, and to look at the characteristics of Clostridium botulinum bacteria in used cooking oil. In this study, isolation of bacteria from used cooking oil using blood agar media, identification of bacteria using gram staining and biochemical tests such as gelatin test, triple sugar iron agar (TSIA) test, simon citrate test, and motility test were carried out. The results of the study obtained 2 out of 3 samples containing Clostridium botulinum bacteria. These bacteria are more common in used cooking oil, which has a more blackish brown color, lots of crumbs left over from food, and temperatures around 30-55℃. In conclusion, there is Clostridium botulinum bacteria found in used cooking oil. Clostridium botulinum colonies are grey, alpha hemolytic, circular to irregular in shape with scalloped edges or rhizoids, and flat to raised surfaces.&#x0D; Keywords: Clostridium botulinum, Isolation, Identification, Cooking Oil

Isolasi dan Uji Aktivitas Selulolitik Bakteri Asal Limbah Bagas

Bagasse is one of the by-products of granulated sugar industries or processed beverages that become waste in the environment. The bagasse will be overgrown by cellulolytic bacteria that produce cellulase enzymes. This study aims to isolate, characterize, and measure bacterial cellulase activity qualitatively and quantitatively. The research phase included isolation and characterization of bacteria, qualitative cellulase activity test, preparing standard curves and bacterial growth curves, crude extract cellulase production, constructing standard glucose curves, quantitative enzyme activity tests, and data analysis. Six isolates with macroscopic characteristics varied in color, elevation, margins, and colony shape; three bacillus isolates, one short bacillus isolate, and two cocci isolates were obtained. All isolates showed positive test results of amylum, triple sugar iron agar, and catalase; five citrate-positive isolates, and three motile-positive isolates. From the cellulase qualitative test, two bacterial isolates had the highest cellulolytic index, namely AT1 (1.79) and BAW3 (1.72) on 0.5% CMC media. The quantitative cellulase test results of the two selected isolates were 0.01176 U/mL for AT1 and 0.01170 U/mL for BAW3, both of which are still classified as low degradation capabilities.&#x0D; &#x0D; Keywords: cellulase activity, cellulolytic index, Sugarcane waste

Microbiota from Intestinal Gut of Dawkinsia filamentosa (Valenciennes,1844) as Indicator of Environmental Condition

Aims: To isolate and identify the gut associated micro-organisms present in the gastrointestinal tract of freshwater fish Dawkinsia filamentosa. &#x0D; Place and Duration of Study: Sampling site: The fish samples were collected from the river Gadana, located at the foot hills of Western Ghats, Alwarkurichi, Tenkasi district, Tamil Nadu, India. Place: Sri Paramakalyani Centre for Environmental Sciences, Manonmaniam Sundaranar University, Azhwarkurichi, Tenkasi, Tamilnadu, India (Latitude 8o20’N; Longitutde 77o10’E). Duration: The research was done during December 2021 and April 2022.&#x0D; Methodology: The fish samples were anaesthetized and surface sterilized followed by aseptic dissectionby using sterile scalpel and needle. The gut contents were removed and homogenized well and isolation of Gut micro biome was done by means of serial dilution. The identification of gastrointestinal bacteria was done by performing various biochemical tests such as Gram staining, Motility, Indole test, Methyl red test, Voges Proskauer test, Citrate utilization test, Triple sugar Iron test, Catalase test, Oxidase test, Urease test, Nitrate test, Starch hydrolysis and Carbohydrate Fermentation Test. The fungi present in gastrointestinal tract is identified with lacto phenol cotton blue staining followed by microscopic observations.&#x0D; Results: The overall bacterial load of gastrointestinal material ranged from 47 x10-7 to 219 x10-5 cfu/g. A total of six bacterial strains and four fungi strains were isolated. Several biochemical tests were used to identify the bacterial isolates. The six bacterial isolates were tentatively identified as Pseudomonas sp., Aeromonas sp., Staphylococcus sp., Bacillus sp., Enterobacter sp., and Vibrio sp. and the four fungi species isolated from the gutwere Aspergillus flavus, Aspergillus niger, Beauveria basiana, and Penicillium sp.&#x0D; Conclusion: The current research revealed that, Dawkinsia filamentosa has a variety of microbiota in its gut. Microbial species from freshwater environments may be able to augment fish feed with these bacteria in commercial aquaculture operations. Additionally, these findings have aided in the development of possible remedies, enhanced knowledge of host-microbe interactions in other vertebrates, and enhanced aquaculture practices. The benefits of identifying gut microbiota in fishes is, it helps to study about the host nutrient catabolism and immune defence mechanism against the disease-causing pathogens.

Molecular identification of thermophilic bacteria with antimicrobial activity isolated from hot springs in North Sumatra, Indonesia

Abstract. Ginting CN, Piska F, Harmileni, Fachrial E. 2023. Molecular identification of thermophilic bacteria with antimicrobial activity isolated from hot springs in North Sumatra, Indonesia. Biodiversitas 24: 752-758. Microorganisms such as thermophilic bacteria are promising sources of antimicrobial compounds due to their abundance, which has yet to be widely investigated. Therefore, this study aims to identify thermophilic bacteria isolated from natural hot springs, Sidebuk Debuk, in North Sumatra, Indonesia. The bacteria were isolated from sediments and hot springs water. Isolated 10 bacteria were characterized and identified based on their morphological and biochemical properties, which included gelatinase activity, motility, citrate utilization, triple sugar iron agar test, catalase test, cell shape, color, and Gram staining. Antimicrobial activity was tested using the disc diffusion method against Escherichia coli and Staphylococcus aureus. The most potential isolates were identified molecularly by amplification of 16S rRNA genes using universal primers. The sequences result were trimmed and assembled using BioEdit and submitted to BLAST to determine the homology of the bacteria. The isolate ITU9 was identified as Bacillus megaterium and exhibited the best antimicrobial activity with an inhibition zone of 9.5 mm against S. aureus during 72 h of incubation. This research showed that the thermophilic B. megaterium strain ITU9 was successfully isolated from Sidebuk Debuk hot springs and had stable antimicrobial activity for 72 h against E. coli and S. aureus.

Identification and Pathogenicity of Escherichia coli from Cloacal Swabs

Colibacillosisis is an infectious disease in poultry caused by pathogenic Escherichia coli (E. coli) as primary or secondary agents. This study were aim to identify and test the pathogenicity ofE. coli isolated from chicken cloacal swabs. The samples used in this study were E. coli isolated from chicken cloacal swabs as many as 42 samples. The first stage in this research was subculturing E.coli in Eosin Methylene Blue Agar (EMBA) medium, then identified by Gram stain and biochemistry with Triple Sugar Iron Agar (TSIA), Sulfide Indole Motility (SIM), Simmons Citrate Agar (SCA) and Methyl Red Voges-Proskauer (MRVP). Pathogenicity of E. coli was tested by culturing in Blood Agar to see the results of hemolysin production and on Sorbitol-MacConkey Agar (SMAC) to see the ability of Sorbitol fermentation. The results obtained were 12 isolates was pathogenic E. coli looked from the results of the pathogenicity test in the Blood Agar and SMAC Agar.

Asymptomatic nasopharyngeal bacterial carriage, multi-drug resistance pattern and associated factors among primary school children at Debre Berhan town, North Shewa, Ethiopia

BackgroundNasopharyngeal carriage of bacteria is the main source for transmission of pathogens across individuals and horizontal spread of organisms in the community. It is an important risk factor for the acquisition of community-acquired respiratory tract infection. It is the major public health problem among children. The asymptomatic carriage of nasopharyngeal bacteria is different globally, particularly in Africa, carriage is higher in children and decreases with increasing age, 63.2% in children less than 5 years, 42.6% in children 5–15 years, and 28.0% in adults older than 15 years.ObjectiveThe aims of this study was to determine asymptomatic nasopharyngeal bacterial carriage, multi-drug resistance pattern and associated factors among primary school children at Debre Berhan town, North Shewa, Ethiopia.MethodsInstitutional based cross-sectional study was conducted at Debre Berhan town primary schools from February 1 to April 30, 2021. Primarily, the schools were stratified into two strata, public and private primary schools. From a total of sixteen government and fourteen private primary schools, five government and five private schools were selected by using a simple random sampling technique. Socio-demographic variables and potential risk factors were assessed using a structured questionnaire. A total of 384 nasopharyngeal swab samples were collected using sterile swabs aseptically; and inoculated on Blood agar, Chocolate agar, MacConkey agar, and Mannitol salt agar. The colony was characterized to isolate bacteria, and bacterial identification was performed by Gram reaction, hemolysis patterns, colonial characteristics and pigmentation, catalase test, coagulase test, mannitol fermentation test, oxidase test, fermentation of carbohydrates, H2S production, motility, formation of indole, triple sugar iron agar (TSI), citrate utilization, lysine decarboxylase or methyl red vogues proskur utilization, urea hydrolysis and satellitism tests. Antimicrobial sensitivity tests were performed by using modified Kirby-Bauer disk diffusion method. Data were entered into statistical package Epi data 4.0.0.6 and transferred to and analyzed using SPSS software version-23. P value of < 0.05 with Odds ratio (OR) and 95% confidence interval (CIs) was considered as statistically significant.ResultsThe overall prevalence of nasopharyngeal carriage of bacterial isolate was 35.7% (95% CI 30.7–40.7%). The predominant isolates were Staphylococcus aureus 54.5% followed by coagulase-negative Staphylococcus 35.8%, and Streptococcus pyogens 4.5%. Most bacterial isolates were susceptible to chloramphenicol, ciprofloxacin, gentamycin, nitrofurantoin, azithromycin, ciprofloxacin; and the overall multidrug resistance pattern of isolated bacteria was 62.03% out of 137 bacterial isolates. Numbers of rooms ≤ 2 per house [AOR = 5.88, 95%CI 1.26–27.57], having history of hospitalization [AOR = 4.08, 95%CI 1.45–11.53], passive smoking [AOR = 4.87, 95%CI 1.49–15.97], family size of > 5 members [AOR = 2.17, 95%CI 1.24–3.81], and number of students in the classroom [AOR = 2.35,95%CI 1.37–4.02] were statistically significant associated risk factors for nasopharyngeal bacteria carriage.ConclusionAsymptomatic nasopharyngeal bacteria carriage in children is alarming for community-acquired infection. The overall multidrug resistance was very high. The risk of the carriage was increased with having a history of passive smoking, being in large family size and number of students per class. Longitudinal follow-up studies would be helpful for better understanding the infection risk in bacterial pathogen carriers.