Serratia Liquefaciens Research Articles

Multi-functional PGPR Serratia liquefaciens confers enhanced resistance to lead stress and bacterial blight in soybean (Glycine max L.)

Lead toxicity and bacterial blight disease caused by Pseudomonas savastanoi pv. glycinea have destructive impacts on soybean growth and productivity. Plant growth-promoting rhizobacteria have been used as an eco-friendly approach for augmenting crop growth and stress resistance. The current study investigated the efficacy of Serratia liquefaciens ZM6 strain in enhancing soybean resistance to lead (Pb) stress and bacterial blight. Two pot experiments were performed. In the first pot experiment, soybean plants were inoculated with S. liquefaciens ZM6 and grown under variable Pb stress levels (0, 200 and 400µM of Pb(NO3)2). In the second experiment, S. liquefaciens-inoculated soybean plants were infected with P. savastanoi pv. glycinea, and disease severity was assessed two weeks post infection. The results revealed that S. liquefaciens strain resisted Pb stress up to 400µM Pb(NO3)2 and exhibited the highest levels of solubilized phosphate, solubilized zinc, siderophore, indole acetic acid, exopolysaccharide, trehalose and antioxidant enzymes at 400µM Pb compared to the other treatments. Moreover, Pb stress (200 and 400µM) significantly decreased the growth, yield, nutrient uptake, gas exchange, and contents of chlorophyll, soluble proteins, sugars, and phenolics of soybean plants. Pb stress also induced the levels of proline, glycine betaine, Pb, oxidative stress markers, antioxidant enzymes, ascorbate, glutathione and expression of stress-responsive genes (CAT, APX, POD, Fe-SOD, CHS7, CHI1A, PAL, IFS2, P5CS and WRKY54) in soybean plants. On the other hand, S. liquefaciens application markedly boosted the growth, yield and levels of nutrients, gas-exchange, chlorophyll, osmolytes, antioxidant enzymes and expression of stress-tolerant genes of Pb-stressed soybean plants. The bacterial inoculation significantly diminished oxidative stress indicators and Pb content in stressed plants. Inoculation of soybean plants with S. liquefaciens also caused significant reductions in blight disease symptoms in P. savastanoi pv. glycinea-infected plants, indicating the efficiency of this strain in controlling harmful blight disease. Overall, this study demonstrated S. liquefaciens ZM6 effectiveness in enhancing soybean resistance to Pb stress and bacterial blight.

Reuterin Enhances the Efficacy of Peracetic Acid Against Multi-species Dairy Biofilm.

Biofilms may contain pathogenic and spoilage bacteria and can become a recurring problem in the dairy sector, with a negative impact on product quality and consumer health. Peracetic acid (PAA) is one of the disinfectants most frequently used to control biofilm formation and persistence. Though effective, it cannot be used at high concentrations due to its corrosive effect on certain materials and because of toxicity concerns. The aim of this study was to test the possibility of PAA remaining bactericidal at lower concentrations by using it in conjunction with reuterin (3-hydroxypropionaldehyde). We evaluated the efficacy of PAA in pure form or as BioDestroy®, a PAA-based commercial disinfectant, on three-species biofilms formed by dairy-derived bacteria, namely Pseudomonas azotoformans PFlA1, Serratia liquefaciens Sl-LJJ01, and Bacillus licheniformis Bl-LJJ01. Minimum inhibitory concentrations of the three agents were determined for each bacterial species and the fractional inhibitory concentrations were then calculated using the checkerboard assay. The minimal biofilm eradication concentration (MBEC) of each antibacterial combination was then calculated against mixed-species biofilm. PAA, BioDestroy®, and reuterin showed antibiofilm activity against all bacteria within the mixed biofilm at respectively 760ppm, 450ppm, and 95.6mM. The MBEC was lowered significantly to 456ppm, 337.5ppm, and 71.7mM, when exposed to reuterin for 16h followed by contact with disinfectant. Combining reuterin with chemical disinfection shows promise in controlling biofilm on food contact surfaces, especially for harsh or extended treatments. Furthermore, systems with reuterin encapsulation and nanotechnologies could be developed for sustainable antimicrobial efficacy without manufacturing disruptions.

Role of the Pseudomonas koreensis BB2.A.1 and Serratia liquefaciens BB2.1.1 Bacterial Strains in Maize Trace Metal Stress Management.

Plant-growth-promoting rhizobacteria (PGPR), in addition to their well-known direct effects on plant growth and development, have been reported to be effective in plant abiotic (trace metal, drought, etc.) and biotic (phytopathogens, insects, etc.) stress management. PGPRs are involved in shaping the fate of trace metals in the rhizosphere and plants and thus may also reduce trace metal stress in plants. The aims of our study were to isolate and select indigenous trace-metal-resistant PGP strains and investigate their effects on maize germination and early development. The roles of the two selected strains, Pseudomonas koreensis and Serratia liquefaciens isolated from trace-metal-contaminated soil were investigated to mitigate trace metal stress in 21-day-old Zea mays seedlings. In the present study, 13 bacterial strains were isolated and screened for PGP traits under normal and trace metal stress conditions. The effect of two selected strains was further studied on plant experiments. The germination process, plant growth parameters (length, weight, dry matter content), photosynthetic activity, GPOX activity, trace metal accumulation, and translocation in microbes inoculated Cd (0.5 mM), Zn (1 mM), and Cd + Zn (0.1 + 0.5 mM) treated maize plants was studied. Our results revealed that trace metal toxicity, in terms germination and growth parameters and antioxidant enzyme activity, was enhanced upon inoculation with Pseudomonas koreensis BB2.A.1. Chlorophyll content and accumulation studies showed enhanced results following inoculation with Serratia liquefaciens BB2.1.1. Therefore, both bacterial strains possessed beneficial traits that enabled them to reduce metal toxicity in maize.

Effect of Essential Oils and Dried Herbs on the Shelf Life of Fresh Goat Lump Cheese.

In recent years, the use of natural preservatives in food products has gained significant attention due to their potential health benefits and effectiveness. A standardized microbiological analysis was conducted on Slovak farm-produced lump goat cheese samples to determine the antibacterial activity of dry herbs and essential oils added to vacuum-packed goat cheese. We employed five dried herbs and five essential oils derived from the same plants. The microbiological quality of 145 fresh and vacuum-packed goat cheese samples was assessed. The number of coliform bacteria, total viable count, lactic acid bacteria, and microscopic filamentous fungi were examined in raw cheese samples stored for 12 days at 4 °C. All cheese samples were vacuum-packed (control samples were packed without vacuum). This study evaluated the potential benefits of using essential oils and dried herbs from thyme (Thymus serpyllum L.), black pepper (Piper nigrum L.), clove (Eugenia caryophyllus Thunb.), mint (Mentha × piperita L.), and basil (Ocimum basilicum L.) as preservatives. The essential oils were obtained from Hanus Ltd., Nitra, Slovakia, and were applied at a concentration of 2%. The dried herbs were obtained from Popradský čaj (Poprad, Slovakia) and Mäspoma Ltd. (Zvolen, Slovakia). The results showed that all microorganism groups were significantly reduced in cheese samples following the application of essential oils throughout the entire storage period. During the preservation of cheese samples in polyethylene bags used for vacuum packing food, Lactococcus garvieae, L. lactis, Enterobacter cloacae, and Serratia liquefaciens were the most frequently isolated microbiota. Essential oils and dried herbs demonstrated antimicrobial potential during the storage of vacuum-packed goat cheese.

Biological Characterization and Genomic Analysis of Three Novel Serratia- and Enterobacter-Specific Virulent Phages.

Due to the high microbiological contamination of raw food materials and the increase in the incidence of multidrug-resistant bacteria, new methods of ensuring microbiological food safety are being sought. One solution may be to use bacteriophages (so-called phages) as natural bacterial enemies. Therefore, the aim of this study was the biological and genomic characterization of three newly isolated Serratia- and Enterobacter-specific virulent bacteriophages as potential candidates for food biocontrol. Serratia phage KKP_3708 (vB_Sli-IAFB_3708), Serratia phage KKP_3709 (vB_Sma-IAFB_3709), and Enterobacter phage KKP_3711 (vB_Ecl-IAFB_3711) were isolated from municipal sewage against Serratia liquefaciens strain KKP 3654, Serratia marcescens strain KKP 3687, and Enterobacter cloacae strain KKP 3684, respectively. The effect of phage addition at different multiplicity of infection (MOI) rates on the growth kinetics of the bacterial hosts was determined using a Bioscreen C Pro growth analyzer. The phages retained high activity in a wide temperature range (from -20 °C to 60 °C) and active acidity values (pH from 3 to 12). Based on transmission electron microscopy (TEM) imaging and whole-genome sequencing (WGS), the isolated bacteriophages belong to the tailed bacteriophages from the Caudoviricetes class. Genomic analysis revealed that the phages have linear double-stranded DNA of size 40,461 bp (Serratia phage KKP_3708), 67,890 bp (Serratia phage KKP_3709), and 113,711 bp (Enterobacter phage KKP_3711). No virulence, toxins, or antibiotic resistance genes were detected in the phage genomes. The lack of lysogenic markers indicates that all three bacteriophages may be potential candidates for food biocontrol.

Enhancement of Biogas (Methane) Production from Cow Dung Using a Microbial Electrochemical Cell and Molecular Characterization of Isolated Methanogenic Bacteria

Biogas has long been used as a household cooking fuel in many tropical counties, and it has the potential to be a significant energy source beyond household cooking fuel. In this study, we describe the use of low electrical energy input in an anaerobic digestion process using a microbial electrochemical cell (MEC) to promote methane content in biogas at 18, 28, and 37 °C. Although the maximum amount of biogas production was at 37 °C (25 cm3), biogas could be effectively produced at lower temperatures, i.e., 18 (13 cm3) and 28 °C (19 cm3), with an external 2 V power input. The biogas production of 13 cm3 obtained at 18 °C was ~65-fold higher than the biogas produced without an external power supply (0.2 cm3). This was further enhanced by 23% using carbon-nanotubes-treated (CNT) graphite electrodes. This suggests that the MEC can be operated at as low as 18 °C and still produce significant amounts of biogas. The share of CH4 in biogas produced in the controls was 30%, whereas the biogas produced in an MEC had 80% CH4. The MEC effectively reduced COD to 42%, whereas it consumed 98% of reducing sugars. Accordingly, it is a suitable method for waste/manure treatment. Molecular characterization using 16s rRNA sequencing confirmed the presence of methanogenic bacteria, viz., Serratia liquefaciens and Zoballella taiwanensis, in the inoculum used for the fermentation. Consistent with recent studies, we believe that electromethanogenesis will play a significant role in the production of value-added products and improve the management of waste by converting it to energy.

Vairimorpha (Nosema) ceranae can promote Serratia development in honeybee gut: an underrated threat for bees?

The genus Serratia harbors opportunistic pathogenic species, among which Serratia marcescens is pathogenic for honeybees although little studied. Recently, virulent strains of S. marcescens colonizing the Varroa destructor mite's mouth were found vectored into the honeybee body, leading to septicemia and death. Serratia also occurs as an opportunistic pathogen in the honeybee's gut with a low absolute abundance. The Serratia population seems controlled by the host immune system, but its presence may represent a hidden threat, ready to arise when honeybees are weakened by biotic and abiotic stressors. To shed light on the Serratia pathogen, this research aims at studying Serratia's development dynamics in the honeybee body and its interactions with the co-occurring fungal pathogen Vairimorpha ceranae. Firstly, the degree of pathogenicity and the ability to permeate the gut epithelial barrier of three Serratia strains, isolated from honeybees and belonging to different species (S. marcescens, Serratia liquefaciens, and Serratia nematodiphila), were assessed by artificial inoculation of newborn honeybees with different Serratia doses (104, 106, and 108 cells/mL). The absolute abundance of Serratia in the gut and in the hemocoel was assessed in qPCR with primers targeting the luxS gene. Moreover, the absolute abundance of Serratia was assessed in the gut of honeybees infected with V. ceranae at different development stages and supplied with beneficial microorganisms and fumagillin. Our results showed that all tested Serratia strains could pass through the gut epithelial barrier and proliferate in the hemocoel, with S. marcescens being the most pathogenic. Moreover, under cage conditions, Serratia better proliferates when a V. ceranae infection is co-occurring, with a positive and significant correlation. Finally, fumagillin and some of the tested beneficial microorganisms could control both Serratia and Vairimorpha development. Our findings suggest a correlation between the two pathogens under laboratory conditions, a co-occurring infection that should be taken into consideration by researches when testing antimicrobial compounds active against V. ceranae, and the related honeybees survival rate. Moreover, our findings suggest a positive control of Serratia by the environmental microorganism Apilactobacillus kunkeei in a in vivo model, confirming the potential of this specie as beneficial bacteria for honeybees.

Biodegradation of polyurethanes by Staphylococcus warneri and by microbial co-culture

With the increasing production and use of polyurethanes (PUs), it is necessary to develop sustainable techniques for the remediation of plastic pollution. The use of microorganisms capable of biodegrading PUs may be an environmentally desirable solution for controlling these plastic contaminants. To contribute to the discovery of alternatives for the mitigation of plastics in the environment, this study aimed to explore the potential of StaphylococcuswarneriUFV_01.21, isolated from the gut of Galleria mellonellalarvae, for biodegradation of PU in pure culture and microbial co-culture with Serratia liquefaciensL135. S. warneri grew using Impranil® PU as the sole carbon source in pure culture and co-culture. With six days of incubation, the biodegradation of Impranil® in Luria Bertani broth was 96, 88 and 76%, while in minimal medium, it was 58, 54 and 42% for S. warneri, S. liquefaciens, and co-culture, respectively. In addition, S. warneri in pure culture or co-culture was able to biodegrade, adhere and form biofilms on the surfaces of Impranil® disks and poly[4,4′-methylenebis (phenyl isocyanate)-alt-1,4-butanediol/di(propylene glycol)/polycaprolactone] (PCLMDI) films. Scanning electron microscopy also revealed biodegradation by detecting the formation of cracks, furrows, pores, and roughness on the surfaces of inoculated PU, both with pure culture and microbial co-culture. This study is the first to demonstrate the potential of S. warneriin PU biodegradation.

Insight into the spoilage heterogeneity of meat-borne bacteria isolates with high-producing collagenase

Chilled chicken is inevitably contaminated by microorganisms during slaughtering and processing, resulting in spoilage. Cutting parts of chilled chicken, especially wings, feet, and other skin-on products, are abundant in collagen, which may be the primary target for degradation by spoilage microorganisms. In this work, a total of 17 isolates of spoilage bacteria that could secrete both collagenase and lipase were determined by raw-chicken juice agar (RJA) method, and the results showed that 7 strains of Serratia, Aeromonas, and Pseudomonas could significantly decompose the collagen ingredients. The gelatin zymography showed that Serratia liquefaciens (F5) and Pseudomonas saponiphila (G7) had apparent degradation bands around 50 kDa, and Aeromonas veronii (G8) and Aeromonas salmonicida (H8) had a band around 65 and 95 kDa, respectively. The lipase and collagenase activities were detected isolate-by-isolate, with F5 showing the highest collagenase activity. For spoilage ability on meat in situ, F5 performed strongest in spoilage ability, indicated by the total viable counts, total volatile basic nitrogen content, sensory scores, lipase, and collagenase activity. This study provides a theoretical basis for spoilage heterogeneity of strains with high-producing collagenase in meat.

Biological characterization and complete genome analysis of the newly isolated Serratia liquefaciens phage vB_SlqS_ZDD2.

A novel lytic phage named vB_SlqS_ZDD2 was isolated from hospital sewage using the double-layer agar method with Serratia liquefaciens ATCC 27592 as the host. BLASTn analysis showed that the genome sequence of phage vB_SlqS_ZDD2 did not resemble any other phages in the NCBI database. Phenotype and phylogeny analysis indicated that this phage might be a new member of the class Caudoviricetes. Phage vB_SlqS_ZDD2 has a dsDNA genome of 49,178 bp with 55% GC content and has 73 open reading frames. This phage exhibited strong lytic activity and a wide range of pH (3-12) and temperature tolerance (below 70℃).

Proliferation of psychrotrophic bacteria in cold-stored platelet concentrates.

Platelet concentrates (PC) are stored at 20-24°C to maintain platelet functionality, which may promote growth of contaminant bacteria. Alternatively, cold storage of PC limits bacterial growth; however, data related to proliferation of psychotrophic species in cold-stored PC (CSP) are scarce, which is addressed in this study. Eight laboratories participated in this study with a pool/split approach. Two split PC units were spiked with ~25 colony forming units (CFU)/PC of Staphylococcus aureus, Klebsiella pneumoniae, Serratia liquefaciens, Pseudomonas fluorescens and Listeria monocytogenes. One unit was stored under agitation at 20-24°C/7 days while the second was stored at 1-6°C/no agitation for 21 days. PC were sampled periodically to determine bacterial loads. Five laboratories repeated the study with PC inoculated with lyophilized inocula (~30 CFU/mL) of S. aureus and K. pneumoniae. All species proliferated in PC stored at 20-24°C, reaching concentrations of ≤109 CFU/mL by day 7. Psychrotrophic P. fluorescens and S. liquefaciens proliferated in CSP to ~106 CFU/mL and ~105 CFU/mL on days 10 and 17 of storage, respectively, followed by L. monocytogenes, which reached ~102 CFU/mL on day 21. S. aureus and K. pneumoniae did not grow in CSP. Psychrotrophic bacteria, which are relatively rare contaminants in PC, proliferated in CSP, with P. fluorescens reaching clinically significant levels (≥105 CFU/mL) before day 14 of storage. Cold storage reduces bacterial risk of PC to levels comparable with RBC units. Safety of CSP could be further improved by implementing bacterial detection systems or pathogen reduction technologies if storage is beyond 10 days.

Impact of Irrigation Water on the Bacteriological Quality of Lettuce (Lactuca sativa) in the Market Garden Zone of Niamey, Identification of Enterobacteria

The urban community of Niamey is characterized by a prodigious development of urban and peri-urban agriculture. Lettuce (Lactuca sativa) is one of the main crops grown in these urban and peri-urban areas. The aim of this study is to assess the impact of irrigation water on the bacteriological quality of lettuce produced in the market gardening zone of the Niamey city. Three (3) market garden sites were studied (Harobanda (site 1), Gamkallé (site 2) and Gounti yéna (site 3). Eighteen (18) lettuce samples and twenty-nine (29) irrigation water samples were taken from the market garden sites and subjected to microbiological analysis using conventional methods appropriate to each indicator. Thus, five (5) indicators of contamiantion were searched, Total Mesophilic Aerobic Flora (TMAF), total coliforms (TC), faecal coliforms (FC), enterobacteria (Ent), Faecal streptococci (FS), Escherichia coli (E. coli) and Clostridium perfringens (CP). The results showed that irrigation water samples from site 1 were highly contaminated with Total Coliforms (TC) and Faecal Coliforms (FC) (3.65±7.88.106 and 1.41±3.04. 106 CFU/100ml respectively), those from site 2 by Faecal Streptococci (FS) (1.19±2.03.105 CFU/100ml) and from site 3 by Enterobacteria (Ent) (1.61±2.37.108 CFU/100ml) and Escherichia coli (8.22±11.86.105 CFU/100ml). Thus, lettuce samples from site 1 are more contaminated with TC (5.32±8.32.106 CFU/g) and Ent (3.10±0.95.107 CFU/g). Faecal coliforms (1.35±1.13.106 CFU/g) and faecal streptococci (5.30±8.21.105 CFU/g) predominate on the site2 and finally site3 is more contaminated with E. coli (2.43±0.38.105 CFU/g). However, enterobacteria species such as Citrobacter freundii, Enterobacter cloacae, Enterobacter sakazakii, Escherichia coli1, Klebsiella pneumoniae ssp ozaenae, Pantoea spp 1, Proteus mirabilis, Raoultella terrigena, Salmonella enterica arizonae, Salmonella spp, Serratia liquefaciens, Serratia marcescens and Serratia odorifera1 were identified in irrigation water samples and lettuce at these sites. In fact, the quality of the irrigation water is not suitable for irrigation, and strongly influences the contamination of the lettuce produced by the bacteria. Consumption of this lettuce without the minimum of precautions could prove dangerous to consumers.

Antivirulence effects of cell-free culture supernatant of endophytic bacteria against grapevine crown gall agent, Agrobacterium tumefaciens, and induction of defense responses in plantlets via intact bacterial cells

BackgroundCrown gall disease caused by Agrobacterium tumefaciens is a very destructive affliction that affects grapevines. Endophytic bacteria have been discovered to control plant diseases via the use of several mechanisms. This research examined the potential for controlling crown gall by three endophytic bacteria that were previously isolated from healthy cultivated and wild grapevines including Pseudomonas kilonensis Ba35, Pseudomonas chlororaphis Ba47, and Serratia liquefaciens Ou55.ResultAt various degrees, three endophytic bacteria suppressed the populations of A. tumefaciens Gh1 and greatly decreased the symptoms of crown gall. Furthermore, biofilm production and motility behaviors of A. tumefaciens Gh1were greatly inhibited by the Cell-free Culture Supernatant (CFCS) of endophytic bacteria. According to our findings, CFCS may reduce the adhesion of A. tumefaciens Gh1 cells to grapevine cv. Rashe root tissues as well as their chemotaxis motility toward the extract of the roots. When compared to the untreated control, statistical analysis showed that CFCS significantly reduced the swimming, twitching, and swarming motility of A. tumefaciens Gh1. The findings demonstrated that the endophytic bacteria effectively stimulated the production of plant defensive enzymes including superoxide dismutase (SOD), polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia lyase (PAL), and total soluble phenols at different time intervals in grapevine inoculated with A. tumefaciens Gh1. The Ba47 strain markedly increased the expression levels of defense genes associated with plant resistance. The up-regulation of PR1, PR2, VvACO1, and GAD1 genes in grapevine leaves indicates the activation of SA and JA pathways, which play a role in enhancing resistance to pathogen invasion. The results showed that treating grapevine with Ba47 increased antioxidant defense activities and defense-related gene expression, which reduced oxidative damage caused by A. tumefaciens and decreased the incidence of crown gall disease.ConclusionThis is the first study on how A. tumefaciens, the grapevine crown gall agent, is affected by CFCS generated by endophytic bacteria in terms of growth and virulence features. To create safer plant disease management techniques, knowledge of the biocontrol processes mediated by CFCS during microbial interactions is crucial.

Ecological features of trace elements tolerant microbes isolated from sewage sludge of urban wastewater treatment plant

Worldwide wastewater treatment plants generate enormous amounts of sewage sludge, and their further disposal depends on the treatment technologies applied and spontaneously occurring microbiological processes. From different ages urban sewage sludge, 12 strains of bacteria with simultaneous tolerance to two or more trace elements: Co, Ni, Cu, Zn, Cd and Pb at concentration of 3-5 mmol were isolated and identified by PCR of target genes and Sanger sequencing methods. The isloated metal(loids) tolerant strains belong to the species, i.e., Serratia fonticola, Rhodococcus qingshengii, Pseudomonas fragi, Pseudomonas extremaustralis, Pseudomonas cedrina, Stenotrophomonas maltophilia, Serratia liquefaciens and Citrobacter freundii. The ecological features of the isolated strains were studied. The optimal growth temperatures for most strains was 15–30°C at pH range of 5–9, although some strains grew at 7°C (Pseudomonas fragi SS0-4, Serratia fonticola SS0-9 and Serratia fonticola SS12-11). Satisfactory growth of two strains (Serratia fonticola SS0-1and Citrobacter freundii SS60-12) was noted in an acidic medium at pH 4. Most of the strains grew in the NaCl concentration range of 1–5%. The isolated bacteria resistant to high concentrations of trace elements can be used for the effective mineralization of sewage sludge and for the decontamination of wastewater.

Deciphering the Mechanism by Which Carbon Dioxide Extends the Shelf Life of Raw Milk: A Microbiomics- and Metabolomics-Based Approach.

Microbial community succession in raw milk determines its quality and storage period. In this study, carbon dioxide (CO2) at 2000 ppm was used to treat raw milk to investigate the mechanism of extending the shelf life of raw milk by CO2 treatment from the viewpoint of microbial colonies and metabolites. The results showed that the shelf life of CO2-treated raw milk was extended to 16 days at 4 °C, while that of the control raw milk was only 6 days. Microbiomics analysis identified 221 amplicon sequence variants (ASVs) in raw milk, and the alpha diversity of microbial communities increased (p < 0.05) with the extension of storage time. Among them, Pseudomonas, Actinobacteria and Serratia were the major microbial genera responsible for the deterioration of raw milk, with a percentage of 85.7%. A combined metagenomics and metabolomics analysis revealed that microorganisms altered the levels of metabolites, such as pyruvic acid, glutamic acid, 5'-cmp, arginine, 2-propenoic acid and phenylalanine, in the raw milk through metabolic activities, such as ABC transporters, pyrimidine metabolism, arginine and proline metabolism and phenylalanine metabolism, and reduced the shelf life of raw milk. CO2 treatment prolonged the shelf life of raw milk by inhibiting the growth of Gram-negative aerobic bacteria, such as Acinetobacter guillouiae, Pseudomonas fluorescens, Serratia liquefaciens and Pseudomonas simiae.

Ocorrência de enterobactérias e taxa de resistência a antimicrobianos em passeriformes do gênero Sporophila provenientes do tráfico de animais silvestres

Abstract We investigate the occurrence of enterobacteria and antimicrobial resistance in passerines of the genus Sporophila seized from illegal trafficking. A total of 35 birds, apparently healthy and from illegal breeding, were sent to the Laboratory of Ornithological Studies at the State University of Ceará (Fortaleza, Ceará, Brazil) by environmental agencies. Cloacal swabs were collected from each bird and subjected to traditional microbiological processing, using standard cultivation and bacterialidentification techniques. We conducted antimicrobial susceptibility testing using the disc-diffusion method. Twenty-three birds (65.7%) exhibited enterobacteria, with Escherichia coli (28.6%) and Serratia liquefaciens (25.7%) being the most common. Antimicrobial resistance in total isolates was observed most frequently in relation to ciprofloxacin (28.1%), followed by tetracycline (25.0%) and enrofloxacin (18.8%). The rate of resistance to Serratia liquefaciens was the highest (i.e., 66.6% of isolates). The birds’ cloacal samples revealed several members of the Enterobacterales order, with E. coli and S. liquefaciens being the most prevalent. We also observed antimicrobial resistance to several drugs, as well as cases of multi-resistance. Antimicrobial resistance is clearly present in wild birds resulting from illegal keeping. These findings highlight the importance of considering antimicrobial-resistant bacteria in release programs to prevent these microorganisms from dispersing into the environment.

Identification of bacteria species among Enterobacteriaceae found in confectionery cakes

The goal of this research was to identify which Enterobacteriaceae species are present in cakes from confectionery establishments and whether these are species that can be considered pathogenic for humans and what their harmful impact on human health could be. The sampling of confectionery cakes was carried out in the area of the City of Zagreb, and the samples were analysed for the presence of Enterobacteriaceae in accordance with the HRN ISO 21528-2:2017 standard. In cases where the samples contained the number of Enterobacteriaceae greater than 102 CFU/g, the MALDI-TOF method was used to identify individual species of enterobacteria. The results of the analyses determined the presence of 10 Enterobacteriaceae species: Enterobacter kobei, Enterobacter cloacae, Pantoea agglomerans, Serratia liquefaciens, Enterobacter asburiae, Klebsiella oxytoca, Buttiauxella gaviniae, Buttiauxella warmboldiae, Raoultella and Cedecea neteri. All species of Enterobacteriaceae determined in this research, according to the literature, were previously isolated from food or water, and all species except Buttiauxella warmboldiae were isolated from humans and the environment. However, for none of the species of Enterobacteriaceae determined in this research a literature reference to support food as a vehicle in case of human illness was found.

Occurrence of Carbapenem-Resistant Enterobacteriaceae (Cre) Abhorring Bla Imp, Bla Kpc And Bla Oxa-48 Genes Recovered From Ready -To- Dispose Poultry Wastes In Osogbo, Osun State

The uncontrolled use of antimicrobials in animal husbandry such as poultry for prophylaxis reasons is to increase output for monetary gains which have led to several acquired community infections with high mortality and morbidity rates. This study is however aimed at investigating the occurrence, prevalence, and molecular detection of carbapenemase genes of carbapenem-resistant Enterobacteriaceae (CRE) bacteria isolated from poultry liters in Osogbo, Osun State Nigeria. Twenty ready-to-pack poultry waste samples were plated for the isolation of Enterobacteriaceae on MacConkey and Brilliance Escherichia coli agar. Disc diffusion method was employed for screening carbapenem resistant isolates while resistant genes were detected using carbapenem resistant gene primers. Forty-six Enterobacteriaeceae were isolated namely: Kluyvera ascorbate (28.3%), Klebsiella oxytoca (10.9%), Citrobacter koseri (8.7%), Klebsiella aereogenes, Raoultell (Klebsiella) planticola, Serratia nematodipila (6.5%), Trabulsiella laguamensis, Salmonella spp. (diarizonae), Proteus vulgaris, Citrobacter farmeri (4.3%), of which Edwardsiella tarda, Providencia rustigianii, Citrobacter freundii, Enterobacter cloacae, Cedecea lapagei, Serratia liquefaciens and Dickeyachry santhemi was (2.2%) respectively. 52.2% of the isolates were carbapenem resistant while 92% of the CRE had bla IMP genes. Only one CRE (Trabulsiella guamensis) was positive to bla KPC gene while four 16.6% of the isolates namely: Citrobacter koseri, Raoultell (Klebsiella) planticola, Serratia nematodipila, Citrobacter freundiu and Citrobacter koseri were positive to blaOXA-48 gene. Therefore, this study infers the high prevalence of CRE isolates in poultry litters as a public health concern for the indiscrimate use of carbapenem.&#x0D; Stamford Journal of Microbiology, 2023. Vol. 13, Issue 1, p. 15-20

Cultivable Gut Microbiota in Synanthropic Bats: Shifts of Its Composition and Diversity Associated with Hibernation

The role of bats in the global microbial ecology no doubt is significant due to their unique immune responses, ability to fly, and long lifespan, all contributing to pathogen spread. Some of these animals hibernate during winter, which results in the altering of their physiology. However, gut microbiota shifts during hibernation is little studied. In this research, we studied cultivable gut microbiota composition and diversity of Nyctalus noctula before, during, and after hibernation in a bat rehabilitation center. Gut microorganisms were isolated on a broad spectrum of culture media, counted, and identified with mass spectrometry. Linear modeling was used to investigate associations between microorganism abundance and N. noctula physiological status, and alpha- and beta-diversity indexes were used to explore diversity changes. As a result, most notable changes were observed in Serratia liquefaciens, Hafnia alvei, Staphylococcus sciuri, and Staphylococcus xylosus, which were significantly more highly abundant in hibernating bats, while Citrobacter freundii, Klebsiella oxytoca, Providencia rettgeri, Citrobacter braakii, and Pedicoccus pentosaceus were more abundant in active bats before hibernation. The alpha-diversity was the lowest in hibernating bats, while the beta-diversity differed significantly among all studied periods. Overall, this study shows that hibernation contributes to changes in bat cultivable gut microbiota composition and diversity.

Antibacterial activity of biosynthesized zinc oxide nanoparticles using Kombucha extract

Antibacterial resistance is a growing global vital medical problem when the innovation and development of new antibiotics are dwindling. Different nanomaterials were synthesized and developed as safe and effective alternative antimicrobial agents. The current study highlights the effect of the antibacterial activity of newly biosynthesized zinc oxide nanoparticles (ZnO NPs) obtained from Kombucha extract. Production of ZnO NPs was optimized and the synthesized nanoparticles were characterized using UV–visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Zeta potential and transmission electron microscope (TEM) analysis. The optimum conditions for ZnO NPs production were at temperature 30 °C, pH 9 and 25 mM of Zn(NO3)2.6H2O. The ZnO NPs biosynthesis was detected in the extract within 24 h. at 35 °C in dark conditions. The XRD pattern displayed 100, 101, 110, and 103 crystal planes. FTIR spectrum showed bands of 1° and 2° amines at 2922.6 cm−1 and a stretching vibration band of vinyl at 1657.5 cm−1. In addition, the results confirmed the positive charge (19 ± 3 mV) and crystalline nature of spherical-shaped ZnO NPs with an average size of 23 ± 1.5 nm. The antibacterial activity of ZnO NPs was tested against different American-type culture collection strains. ZnO NPs exhibited minimum inhibition concentration (MIC) values of 25 µg/ml against Escherichia coli ATCC25922, 30 µg/ml against Staphylococcus aureus ATCC25923 and Pseudomonas aeruginosa ATCC27853, 35 µg/ml against Serratia liquefaciens OQ071699.1 and S. saprophyticus OQ071703.1 and 40 µg/ml against Lysinibacillus fusiformis OQ071701.1 and Klebsiella pneumoniae ATCC33495. Ultrastructure TEM study of the treated bacteria by MIC of ZnO NPs confirmed their highly toxic action on the bacterial cell wall, resulting in bacterial cell membrane rupture. Treated bacteria appeared as extensively damaged cells with the formation of vacuoles and lipids. The provided approach of ZnO NPs in combination with Kombucha SCOBY has a promising future in overcoming bacterial resistance problems in place of using antibiotics.