API ID Research Articles

Evaluation of the first Candida auris isolates reported from Türkiye in terms of identification by various methods and susceptibility to antifungal drugs

PurposeCandida auris is increasingly being isolated from patients all over the world. It has five clades. In this study, it was aimed to compare the results of biochemical tests obtained using different methods and the antifungal susceptibility profiles of C. auris strains isolated from the first seven cases reported in Türkiye, and evaluate whether this information could be useful as preliminary data in determining the clade of strains in centers that lack the opportunity to apply molecular methods. MethodsIdentification test results obtained using API ID 32 C, API 20 C AUX, VITEK-2 YST, and MALDI-TOF MS; colony color and morphology on Chromagar Candida, CHROMagar Candida Plus media, and cornmeal-Tween 80 agar; susceptibility to antifungals were tested and compared. Antifungal susceptibility test was studied using microdilution method according to the recommendations of EUCAST. Additionally, a pilot study was conducted to investigate the value of CHROMagar Candida Plus. ResultsAll seven strains were identified as Lachancea kluyveri with API ID 32 C, Rhodotorula glutinis; Cryptococcus neoformans with API 20 C AUX, and C. auris with both VITEK-2 YST and MALDI-TOF MS. MIC values for fluconazole were very high (≥64 mg/L) for all seven strains. It was observed that 11 (37.9%) of 29 Candida parapsilosis strains formed colonies with morphology similar to C. auris on CHROMagar Candida Plus medium, leading to false positivity. ConclusionsAlthough there have been many isolations of C. auris in our country in recent years, clade distribution of only a small number of strains is known yet. In this study, when the biochemical properties and antifungal susceptibility profiles of the seven strains were evaluated, it was concluded that they exhibited some characteristics compatible with clade I. It was also observed that strains 1 and 2 may belong to a different clade.

First Report of Pectobacterium aroidearum Causing Soft Rot on Ficus carica in Korea

In July 2021, symptoms of soft rot were observed on the stems of <i>Ficus carica</i> in Yeongam, Jeollanamdo, Korea. To accurately diagnose the cause, infected stem was collected and bacterial strain was isolated. Among these, the pathogenic strain KNUB-08-21 was identified as <i>Pectobacterium aroidearum</i> through 16S rRNA gene sequencing and phylogenetic analysis based on the concatenated sequences of the <i>dnaX</i>, <i>leuS</i>, and <i>recA</i> genes. The affiliation of the isolate with this bacterial species was also confirmed by its biochemical characteristics obtained using API ID 32 GN system. Artificial inoculation confirmed the strain’s pathogenicity in figs, causing significant damage to both stems and fruits. To our knowledge, this is the first report of <i>P. aroidearum</i> causing soft rot disease in <i>F. carica</i> in Korea.

APIMiner: Identifying Web Application APIs Based on Web Page States Similarity Analysis

Modern web applications offer various APIs for data interaction. However, as the number of these APIs increases, so does the potential for security threats. Essentially, more APIs in an application can lead to more detectable vulnerabilities. Thus, it is crucial to identify APIs as comprehensively as possible in web applications. However, this task faces challenges due to the increasing complexity of web development techniques and the abundance of similar web pages. In this paper, we propose APIMiner, a framework for identifying APIs in web applications by dynamically traversing web pages based on web page state similarity analysis. APIMiner first builds a web page model based on the HTML elements of the current web page. APIMiner then uses this model to represent the state of the page. Then, APIMiner evaluates each element’s similarity in the page model and determines the page state similarity based on these similarity values. From the different states of the page, APIMiner extracts the data interaction APIs on the page. We conduct extensive experiments to evaluate APIMiner’s effectiveness. In the similarity analysis, our method surpasses state-of-the-art methods like NDD and mNDD in accurately distinguishing similar pages. We compare APIMiner with state-of-the-art tools (e.g., Enemy of the State, Crawlergo, and Wapiti3) for API identification. APIMiner excels in the number of identified APIs (average 1136) and code coverage (average 28,470). Relative to these tools, on average, APIMiner identifies 7.96 times more APIs and increases code coverage by 142.72%.

Biochemical Properties and Molecular Characterization of Fermented Alcoholic Honey (Muratina) Microbes from Mbeere-Kenya

Muratina is an alcoholic beverage (wine) amongst the communities around The Mt. Kenya, obtained from spontaneous fermentation of honey in a gourd with dried Kigelia africana (Lam.) Benth.fruits. The wine production is still carried out using traditional technology and has never been scaled up. It serves cultural and social value amongst the communities. The objective of this study was to characterize and document the product process and to evaluate the chemical and microbiological quality of Muratina. The production process involves mixing water and honey in a ration of 17L water to 3Kg honey, then allowing it to ferment in a gourd with pre-cured K.africana fruits for 3-5 days at 30°C after which it is ready. Muratina has an alcohol content of 19.66± 0.47 (mL/100ml), pH of 4.06 ± 0.12 and titratable acidity of 7.57± 0.45 (g tartaric acid/100 mL). Microbiological analysis of Muratina showed aerobic mesophiles at 2.1-5.5 x 103 CFU/mL, LAB at 3.2-7.7 x 104 CFU/mL and yeasts at concentration of 5.6 – 7.0 x 103 CFU/mL. Biochemical analysis of LAB isolates revealed various resistances to ox gal, pH and NaCl indicating their potential use as probiotics. All the isolates tested were able to withstand 3% ox-gal, although none were able to grow at pH 1-3. Identification of LAB was carried out using API 50 CHL and the sequencing of the 16s rRNA while those of yeasts was carried out using API ID 32. The isolates were identified as Lactobacillus plantarum, Lactococcus spp and Pediococcus spp. Saccharomyces cereviseae was the major yeast isolated. The high alcohol content of Muratina indicates that it has yeasts that could be of commercial value.

First Report of Pectobacterium brasiliense Causing Bitter Melon Soft Rot Disease in Korea

In the Goesan region, located in Chungcheongbuk-do, Korea, a significant outbreak of soft rot infections was documented in August 2021, affecting fruits of <i>Momordica charantia</i>, commonly known as bitter melon or bitter gourd. The symptoms included a noticeable transition to yellowing in the affected fruits, eventually leading to their collapse. The bacterial strain KNUB-09-21 was isolated from the diseased fruits. Molecular analysis, using the sequences of the 16S rRNA region and three housekeeping genes (<i>dnaX, recA</i>, and <i>leuS</i>), along with the results of compound utilization in the API ID 32 GN system, provide strong evidence for the identification of the isolate KNUB-09-21 as <i>Pectobacterium brasiliense</i>. The pathogenicity of strain KNUB-09-21 on <i>M. charantia</i> was confirmed through a controlled inoculation test. Within two days, inoculated fruits displayed soft rot symptoms closely resembling those observed in naturally affected fruits. This is the first report of soft rot on <i>M. charantia</i> in Korea.

Detection of Listeria Species by Conventional Culture-Dependent and Alternative Rapid Detection Methods in Retail Ready-to-Eat Foods in Turkey.

Foodborne pathogens, like Listeria monocytogenes, continue to inflict substantial financial losses on the food industry. Various methods for detecting Listeria in food have been developed and numerous studies have been conducted to compare the different methods. But, in recent years, new Listeria species have been identified, and currently the genus comprises 26 species. Therefore, it would be a more accurate approach to re-evaluate existing detection methods by considering new species. The present investigation involved the analysis of 42 ready-to-eat (RTE) foods, encompassing a variety of food categories, such as mezes, salads, dairy products, and meat products, with the aim of ascertaining the presence of Listeria. Among the traditional culture-dependent reference methods, the ISO 11290 method was preferred. The process of strain identification was conducted with the API Identification System. Furthermore, to ascertain the existence of L. monocytogenes and Listeria spp., the samples underwent additional analysis employing the VIDAS Immunoassay System, ELISA, and RT-PCR methodologies. Thus, four alternative approaches were employed in this study to compare not only the different methods used to determine Listeria while taking into account the newly identified Listeria species, but also to assess the compliance of retail RTE food items with microbiological criteria pertaining to the genus Listeria. Based on the conducted analyses, L. monocytogenes was conclusively determined to be present in one sample. The presence of Listeria spp. was detected in 30.9% of the samples, specifically in Turkish cig kofte, sliced salami, and salads.

First Report of Soft Rot Caused by Pectobacterium brasiliense on Cucumber in Korea

Wilted and water-soaked lesion symptoms were observed on cucumbers in greenhouses located in Daejeon, Chungcheongnam-do, Korea, in June 2021. A bacterial strain, designated KNUB-04-21, was isolated from the cucumbers, which was subsequently identified as <i>Pectobacterium brasiliense</i> through a phylogenetic analysis based on sequences of the 16S rRNA region, <i>dnaX, leuS</i>, and <i>recA</i> genes. The biochemical characteristics of KNUB-04-21 were also similar to those of <i>P. brasiliense</i> through investigation using the API ID 32 GN system. The pathogenicity of KNUB-04-21 was confirmed by inoculating it into healthy cucumber plants. The reisolated strains were also found to be same to the original strain. To our knowledge, this is the first report of <i>P. brasiliense</i> being identified as the causative agent of cucumber soft rot in Korea.

First Report of Melon Soft Rot Disease Caused by Pectobacterium brasiliense in Korea

In May 2021, characteristic soft rot symptoms, including soft, watery, slimy, black rot, wilting, and leaf collapse, were observed on melon plants (<i>Cucumis melo</i>) in Gokseong, Jeollanam-do, Korea. A bacterial strain, designated KNUB-06-21, was isolated from infected plant samples, taxonomically classified, and phylogenetically analyzed using 16S rRNA and housekeeping gene sequencing. Strain KNUB-06-21 was also examined for compound utilization using the API ID 32 GN system and strain KNUB-06-21 was identified as Pectobacterium brasiliense. Subsequent melon stem inoculation studies using strain KNUB-06-21 showed soft rot symptoms similar to field plants. Re-isolated strains shared phenotypic and molecular characteristics with the original <i>P. brasiliense</i> KNUB-06-21 strain. To our knowledge, ours is the first report of <i>P. brasiliense</i> causing melon soft rot disease in Korea.

Incidence of Candida Species Biofilms in Pediatric Cancer Patients Undergoing Chemotherapy Treatment

The oral thrush due to biofilms of Candida species is a common infection for the oral cavity of cancer patients undergoing chemotherapy, leading to oral infections. These biofilms, composed of Candida organisms, pose challenges in terms of diagnosis and treatment due to their increased resistance and ability to persist in the oral environment. This study aimed to determine the frequency of Candida species biofilm colonization in the oral cavity of pediatric cancer patients receiving chemotherapy. By examining the incidence of Candida sp. biofilms, this research provides valuable insights into the oral health challenges specific to children with cancer, emphasizing the need for targeted preventive and management strategies to address these infections effectively. In this study, a total of 100 oral cavity swabs were collected, with 50 swabs obtained from cancer patients undergoing chemotherapy and suspected to have oral candidiasis, and the other 50 swabs collected from non-cancer healthy individuals serving as control. All swabs were cultured on Sabouraud Dextrose agar (SDA) media, followed by microscopic examination of positive samples. The API identification candida system was utilized for the identification of Candida species, along with the implementation of biochemical tests to further study the characteristics of the identified species.The findings demonstrated that out of the 50 cancer patients, approximately 47% (23 patients) exhibited positive yeast growth on SDA agar. Among the 50 non-cancer control cases, approximately 30% (15 cases) showed positive growth on SDA agar. Furthermore, using the API candida system, we confirmed the presence of candida species in 25 cases. Candida albicans was identified as the most prevalent species, followed by Candida parapsilosis, Candida krusei, and Saccharomyces cerevisiae, which were detected less frequently.In conclusion, there were a significant increase in Candida species among cancer patients undergoing chemotherapy treatment in comparison to non-cancer control individuals. This finding highlights the impact of chemotherapy on the prevalence of Candida infections in cancer patients, emphasizing the need for heightened attention and appropriate management strategies in this vulnerable population.

Presence and implication of Candida spp. in patients with peri-implantitis enrolled in a supportive peri-implant therapy program of the Basque Country (Spain). A case-control study.

The peri-implant sulcus is a good niche for infectious colonization such as Candida spp. In this study, the level of Candida spp. fungal colonization is analyzed in patients with peri-implantitis under supportive peri-implant therapy, as well as its correlation with the main clinicopathological data. A case-control study was carried out on 161 patients treated with dental implants, 80 with PI and 81 without PI, which corresponded to 91 women and 70 men, whose mean age was 60.90 years. A specific protocol was completed for the clinical and implant data. Microbiological samples were taken by oral rinse and with paper tips from the peri-implant sulcus. For the quantitative and qualitative analysis Candida Chromogenic Agar/CONDA plates were incubated for 72 h at 36 + 1°C. Fungal growth was considered active when having more than 50 CFU. Specific Candida spp. cultures were later confirmed by API ID 32C and PCR. Fungal growth was achieved in 28% of oral rinse and 6.75% of peri-implant fluid samples. No significant differences were recognized between study groups. Most of the cultures (>65%) showed more than 50 CFU. The most frequent species were Candida albicans and Candida parapsilosis. There was no association between different PI risk factors and fungal data. The presence of Candida spp. in the oral cavity of patients with dental implants was related to total edentulism and the use of implant-fixed complete prosthesis implant-retained removable prosthesis. These results suggest that there is no link between PI and presence of Candida in patients with dental implants undergoing regular supportive periodontal therapy.

Selection of mixed starters for the preparation of traditional Moroccan bread

The main objective of this work was the selection of mixed starters with a combination of Lactic Acid Bacteria (LAB) and yeast strains for traditional bread production in Morocco. For this, a total of 21 LAB strains and 36 yeast strains were isolated from different traditional sourdough. Dough fermentation were assessed by monitoring physicochemical parameters including, titratable acidity, decrease of pH and lactic acid, ethanol and CO2 production. A total of six yeasts and four LAB were selected for their technological performances. Morphological, physiological and biochemical identification performed using API identification kits confirmed that these strains belonged to Saccharomyes cerevisiae, Candida humilis and Saccharomyces exiguus species, and Lactobacillus plantarum and Lactobacillus casei species for yeasts and LAB respectively. The yeast S3-L2 and the LAB ODBL5 strains exhibited the best performances among the selected ones; S3-L2 yeast strain were able to produce ΔV=23mL of CO2 and showed the highest values of ethanol and biomass production (2.87 g/L and 1.25 10^9 UFC/mL, respectively). Whereas OD-BL5 LAB strains produced 13.9 g/L of lactate in dough. These findings lead to consider these two strains very good candidates for the formulation of an effective mixed starter for bread preparation. Subsequently, sensorial analysis results showed that bread prepared using mixed starter No. 24 composed of the two selected species exhibited better exterior appearance, golden and crispy crust, large volume and honeycomb crumb, compared to the control.

Skin Bacteriome and its Resistance to Antibiotics in Free Range Pigs

The cutaneous microbiome and also its resistance to antibiotics is exposed to change, depending on different habitat factors. This research investigates the composition of cutaneous microflora and its antibiotic resistance in pigs raised on free range farms (mix breed swine, from low input small farms). Swabs were collected from the skin surface and subjected to classical microbiological methods (simple broth and nutrient agar cultivation, colony isolation and biochemical API identification). The antibacterial resistance to gentamicin, streptomycin, oxitetracycline, tylosin, amoxacillin-clavulanic acid, marbofloxacin, tulatromycin, cefotaxime and doxycycline was estimated by Kirby Bauer method and multiple antibiotic resistance (MAR) index was calculated. Strains from Staphylococcus (sciuri and warnerii), Shigella spp., Kytococcus sedentarius, Salmonella spp. and Citrobacter freundii genera and species were identified in the collected samples. The most resistant was a S. warnerii strain, but the MAR index was high (0.33) in 50% of the strains. The most efficient antibiotic was cefotaxime and the least efficient was oxitetracycline. The results indicated the presence of antibiotic resistant ubiquitous and pathogenic strains in the investigated pigs which need caution, since they could express pathogenicity under appropriate conditions which low input farming system could provide.

PSEUDOMONAS FLUORESCENS IN SHEEP MILK GREEK YOGHURT FROM VLASINA – A BIOCHEMICAL CHARACTERIZATION

Pseudomonas fluorescens is an aerobic, rod-shaped, non-sporulating gram-negative bacteria, mostlyfound in soil, decaying organic matter and feces. This species contaminates the milk mainly through animal feeddust, silage, utensils and polluted water. In addition to casein digestion, this psychotropic and lipolytic speciesgenerates butyric and caproic acids by fermentation of milk fat, which release a strong unpleasant odor and give arancid and bitter taste to dairy products. In this study we tested 48 hours old sheep's sour milk, produced in ahousehold in Vlasina, southeastern Serbia, using traditional method. Standard bacteriological protocols were usedfor isolation and identification Pseudomonas fluorescens strains. Biochemical identification was performed usingthe commercial API 32GN E system, and 60473057073 profile was obtained (Pseudomonas fluorescens, %Id=99.5and T=0.74). Positive biochemical tests were: N-acetyl-glukosamine (NAG), D-Ribose (RIB), Sodium malonate(MNT), Sodium acetate (ACE), Lactic acid (LAT), L-Alanine (ALA), D-Mannitol (MAN), D-Glucose (GLU), LArabinose(ARA), Capric acid (CAP), Valeric acid (VALT), Trisodium citrate (CIT), L-Histidine (HIS), Potasium2-ketogluconate (2KG), 3-Hydroxybutyric acid (3OBU), 4-Hydroxybenzoic acid (pOBE), L-Serine (SER), LProline(PRO) and Oxidase (OX). Negative biochemical tests were: L-Rhamnose (RHA), Inositol (INO), DSaccharose(SAC), D-Maltose (MAL), Itaconic acid (ITA), Suberic acid (SUB), Salicin (SAL), D-Melibose (MEL),L-Fucose (FUC), D-Sorbitol (SOR), Propionic acid (PROP), Potasium 5-ketogluconate (5KG), Glycogen (GLYG)and 3-hydroxybenzoic acid (mOBE). Based on the example of the strain Pseudomonas fluorescens, the API ID 32GN system proved to be precise and very efficient in the identification of this lipolytic type. Given thatpasteurization and cooling processes do not entirely inhibit the enzym activity and growth of this psychotrophicbacteria, informing individual milk and milk product manufacturers in the Vlasina region about good manufacturingpractices would limit contamination and bacteriological deterioration.

P466 Isolation of Cryptococcus neoformans and other yeast from pigeon droppings in Khartoum state, Sudan

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMObjectivesThe aim of this study was to investigate the presence of Cryptococcus neoformans and other yeast species in pigeon droppings in Khartoum state, Sudan.MethodsSample collection A total of 120 samples were collected from pigeon droppings from Khartoum state. Pigeon droppings were collected using sterile wooden spatulas and placed directly in clean plastic bags.Processing of sampleApproximately 2 g of pigeon droppings were added to 10 ml of sterile saline. The samples were allowed to stand for 20 min with frequent vortex.Isolation and Identification of yeast isolatesA loopful of supernatant fluid from prepared samples was taken and streaked onto Sabouraud's dextrose agar media supplemented with 0.05 mg/ml chloramphenicol incubated at 37°C for 1-2 days. All yeast isolates were identified by direct microscopic examination using the lacto-phenol-cotton-blue stain India ink stain and Gram's stain. Further confirmatory tests were conducted using Corn Meal Agar (CMA) with tween 80, germ tube production test using horse serum, and urease test. Confirmation of identification was done using API 20C AUX and API ID 32C AUX (bioMérieux®, Madrid, Spain).Results Cryptococcus neoformans is the most common species isolated from pigeon droppings as shown in Table 1. Different Candida species have been isolated Figure 1.Isolated yeastNumber Cryptococcus neoformans 42 Candida spp34 Cryptococcus albidus 5 Stephanoscus ciferrii 4 Rhodotorula glutinis 4 Rhodotorula mucilaginosa 3 Geotrichum capitatum 2 Zygosaccharomyces spp1ConclusionThe present study concluded that there is a potential role of pigeons as a reservoir for C. neoformans and other zoonotic yeasts in the environment that can affect humans and animals.

Screening For Potential Exopolysaccharide Producers From Lactobacillus spp Isolated From Locally Fermented Milk (Nono)

Exopolysaccharides (EPS) are exogenous bacterial sugar polymers with many applications in dairy, pharmaceutical and cosmetic industries, using it as thickeners, stabilizers and gelling agents. The study aimed to screen for potential exopolysaccharide producers from Lactobacillus spp. isolated from locally fermented milk (nono). Twenty-five nono Samples were collected from Wambai market, Kano. Lactic acid bacteria were isolated using de Man Ragosa and Sharpe Agar. Isolates were identified by API 50 CHL kit and web, and screened for EPS production in which the EPS was extracted and quantified using the phenol-sulphuric method. Next, the influence of carbon source (Glucose, Sucrose and Lactose) and concentrations on EPS were evaluated on some of the isolated strains. The functional groups of the EPS were confirmed using FTIR. The isolated Lactobacillus spp. were all Gram positive, catalase and oxidase negative, API identification yielded; Lactobacillus acidophilus 1, Lb. brevis 1, Lb. fermentum, Lb. paracasei ssp paracasei, Lb. acidophilus 3. Ten isolates yielded EPS in the range of 248.33mg/l - 07.83mg/l. The FTIR analysis of extracted EPS produced peaks around 3,300–881cm-1. Hence the study has brought to light the presence of potential EPS producing LAB in nono, which could be further exploited to harness their potential.

Antifungal activity and genomic analysis of Bacillus velezensis X49

Proteolytic enzymes and lipopeptides contain broad-spectrum antimicrobial activities, which have great potential for research and development. A microbial strain X49 obtained from protease screening plate showed antifungal activities against six fungi. Biochemical analysis, 16S rRNA sequencing, API identification system, and electron microscope analysis were carried out to identify the bacterium. Azocasein method was used to analyze the protease activity. Lipopeptides were extracted for antifungal analysis. The result indicated that strain X49 grew in the range of 10-50 ℃ and pH 4.0-9.0. Moreover, it survived in 10% NaCl, showing good halotolerance. Strain X49 was identified as Bacillus velezensis. Genomic analysis of B. velezensis X49 revealed eleven genes encoding serine protease. The ID 1_894 belonging to S8 subtilisin family was 99% similar to the serine protease with known antifungal ability. On the other hand, thirty genes encoding non-ribosomal peptide synthetase involved in the lipopeptide biosynthesis, including surfactin, iturin, fengycin, bacitracin, and gramicidin, were identified. Part of the extracellular proteolytic activity remained under high temperature. After co-fermentation of B. velezensis X49 with Zingiber officinale Rosc., the antifungal activity of the lipopeptide extract from the co-fermentation was greatly improved. In conclusion, B. velezensis X49 showed clear inhibitory effect on both plant and human pathogens. The active substances co-fermented with Chinese herbs and microbes can be utilized for further drug development.

Isolation, Identification and Characterization of Potential Probiotics from Fermented Food Products

Potential probiotics were isolated, identified and characterised from ogi, fufu, nunu, palmwine and fermented tigernut milk. Pour plate method was used for the isolation of lactic acid bacteria (LAB) on De Man Rogosa Sharpe (MRS) media. Three (3) pure colonies were distinctly obtained from each of the fermented food sources with microbial mean counts ranging from 8.13 - 8.25 Log10 Cfu/ml. Isolated strains were identified and characterized using morphological, API-50 CHL (Bio-merieux, France) and Polymerase chain reaction (PCR) analysis. Ten (10) of the isolated microorganisms were identified as Lactobacilli, two (2) Micrococci and a Lactococcus. The Lactobacilli were catalase negative and oxidase negative rod-shaped bacteria. The identification accuracy of the isolates based on similarities from the computer-aided API and PCR GenBank databases ranged from 49.00 - 99.90% and 79.00 - 99.00% respectively. The amplification pattern of the 16S regions of the sequenced isolates showed DNA fragments with 500 - 1000 base pairs. The LAB strains identified are Lactobacillus fermentum NBRC 15885, Leuconostoc mesenteroides LM, Lactobacillus plantarum CIP 10315.1, Lactobacillus plantarum NBRC 15891, Lactobacillus parabuchneri LMG 11457, Lactobacillus pentosus 124-3 and Lactobacillus brevis ATCC 14869. These strains had high correlation in both the API and PCR identification techniques that was used in this study. Potential probiotic lactic acid bacteria can be isolated and identified from ogi, fufu, nunu, paimwine and fermented tigernut milk.

Phenotypic and genotypic characterisation of Lactobacillus and yeast isolates from a traditional New Zealand Māori potato starter culture

Parāroa Rēwena is a traditional Māori sourdough produced by fermentation using a potato starter culture. The microbial composition of the starter culture is not well characterised, despite the long history of this product. The morphological, physiological, biochemical and genetic tests were conducted to characterise 26 lactic acid bacteria (LAB) and 15 yeast isolates from a Parāroa Rēwena potato starter culture. The results of sugar fermentation tests, API 50 CHL tests, and API ID 32 C tests suggest the presence of four different LAB phenotypes and five different yeast phenotypes. 16S rRNA and 26S rRNA sequencing identified the LAB as Lacticaseibacillus paracasei and the yeast isolates as Saccharomyces cerevisiae, respectively. Multilocus sequence typing (MLST) of the L. paracasei isolates indicated that they had identical genotypes at the MLST loci, to L. paracasei subsp. paracasei IBB 3423 or L. paracasei subsp. paracasei F19. This study provides new insights into the microbial composition of the traditional sourdough Parāroa Rēwena starter culture.

Pathogens isolated from bovine clinical mastitis and their antimicrobial resistance

This study aimed to isolate aerobic and microaerophilic bacteria from mastitis milk samples, as well as to determine their antibiotic resistance. A total of 196 bovine mastitis milk samples were tested by standard bacteriological methods and with API identification test kits. Antimicrobial susceptibility testing was performed by the Kirby-Bauer disk diffusion method. The results revealed that the predominant isolate was S. aureus, with an isolation rate of 28%, followed by Streptococcus spp. (27%) and E. coli (19%). Isolation rates for Corynebacterium spp., Mycoplasma spp., and Pseudomonas aeruginosa were 11%, 6%, and 4%, respectively. Compared to the bacteria mentioned above, lower percentages were observed for Trueperella pyogenes (2%), Pasteurella multocida (2%), and Klebsiella pneumoniae (1%). A broad evaluation of antimicrobial resistance showed that the pathogens were resistant to tetracycline (68.63%), oxytetracycline (41.57%), ampicillin (39.08%), ceftiofur (38.1%), cephalexin (32.26%), penicillin (31.25%), amoxicillin/clavulanic acid (24.53%), enrofloxacin (24.44%), gentamycin (23.68%), and trimethoprim/sulfamethoxazole (22.09%). This study demonstrated that the sources of bacteria isolated from mastitis bovine milk samples were both contagious and environmental. More importantly, the present results demonstrate a critically high antimicrobial resistance in dairy cattle. For instance, E. coli isolates showed a crucial resistance to commonly used and recommended antimicrobials, including ceftiofur (100%), cephalexin (83.33%), and tetracycline (94.44%). The results of this study may provide valuable information about clinical aspects of bovine mastitis infections and current antimicrobial resistance levels in dairy cattle.

Assessment of microbiological and chemical quality of springwater in Riverston of Knuckles mountain range in Sri Lanka

Safe and readily available water is important for public health, whether it is used for drinking, domestic use, food production or recreational purposes. The main objectives of this study were to assess the microbiological and chemical quality of spring water in Riverston, situated in the Knuckles Mountain range, Sri Lanka and to identify bacteria isolated from spring water. Water samples were collected from ten springs from different locations. Microbiological and chemical analysis were carried out according to standard protocols. Isolated bacteria were identified using biochemical tests and API identification system. According to the results, Total coliforms (TC) (ranged from 0-27 per 100 ml) and Fecal coliforms (FC) (ranged from 38-326 per 100 ml) bacteria were detected in all water samples tested, and the detected numbers exceeded permitted levels for drinking water. There are four TC species, <em>viz; Escherichia vulneris, Serratia marcescens</em>, <em>Serratia liquefaciens </em>and <em>Proteus mirabilis </em>and one FC species, <em>viz; Escherichia coli </em>(dominant species), were identified during the study. All chemical parameters tested were within the permitted levels. This study reveals that spring water in Riverston, Knuckles Mountain Range is not a safe drinking water source. Hence, it is important to take necessary precautions, especially as spring waters from these areas are consumed by many and is the main source water for the bottling industry in Sri Lanka.