actA Gene Research Articles

Effects of hydroxyl radical on Listeria monocytogenes in fresh-cut iceberg lettuce (Lactuca sativa var. iceberg): Survival, membrane permeability, microstructure and virulence gene expression

Contamination of fresh-cut vegetables with Listeria monocytogenes is a food safety issue of particular concern. However, the inhibitory effect of hydroxyl radicals on Listeria monocytogenes in fresh-cut iceberg lettuce is not clear. This paper expounded the control effect and mechanism of hydroxyl radicals on Listeria monocytogenes in fresh-cut iceberg lettuce. Results showed that hydroxyl radicals (·OH) treated lettuce for 20 min could effectively inhibit the number of Listeria monocytogenes, and better maintain the sensory quality of fresh-cut lettuce. Hydroxyl radicals (·OH) increased cell membrane permeability and UV-absorbing substances by destroying the cell membrane integrity of Listeria monocytogenes; and attenuated the virulence of Listeria monocytogenes by down-regulating the expression of the virulence-related genes actA and recA; further affecting the survival and proliferation of Listeria monocytogenes in iceberg lettuce. This paper proposed an effective method to inhibit Listeria monocytogenes in fresh-cut lettuce, which will provide technical support and theoretical basis for its application in the fresh-cut lettuce industry.

Multidrug-resistant Escherichia coli isolated from free-range chickens in the Caatinga biome.

Antimicrobial-resistant Escherichia coli is a global health challenge from a One Health perspective. However, data on its emergence in the Caatinga biome are limited. This biome is exclusive to the Brazilian Northeast and offers unique epidemiological conditions that can influence the occurrence of infectious diseases and antimicrobial resistance. In this study, the carriage proportion, antimicrobial susceptibility, and population structure of cephalosporin-resistant E. coli were assessed in 300 cloacal swab samples of free-range chickens from three Brazilian states covered by the Caatinga biome. The results showed that 44 (14.7%) samples were positive for cephalosporin-resistant E. coli, and Paraíba state had the highest frequency of isolates (68.2%). Genes encoding cephotaximase-Munich or ampicillin class C (AmpC) enzymes were identified in 30 (68.2%) and 8 (18.2%) isolates, respectively, comprising 31 E. coli isolates. Overall, molecular typing by genome restriction using XbaI endonuclease followed by pulsed-field gel electrophoresis revealed four clusters from two properties of Paraíba state composed by extended-spectrum β-lactamase-producing and AmpC-producing E. coli carrying blaCTX-M-1-like and blaMIR-1/ACT-1 genes and belonging to different phylogenetic groups. There is a need to control antimicrobial resistance while taking into account the genetic diversity of the strains and their implications for animal and public health, especially in free-range chickens reared in the Brazilian Caatinga biome.

Effects of copper, and aluminium in ionic, and nanoparticulate form on growth rate and gene expression of Setaria italica seedlings

This study aims to determine the effects of copper, copper oxide nanoparticles, aluminium, and aluminium oxide nanoparticles on the growth rate and expression of ACT-1, CDPK, LIP, NFC, P5CR, P5CS, GR, and SiZIP1 genes in five days old seedling of Setaria italica ssp. maxima, cultivated in hydroponic culture. Depending on their concentration (ranging from 0.1 to 1.8 mg L−1), all tested substances had both stimulating and inhibiting effects on the growth rate of the seedlings. Copper and copper oxide-NPs had generally a stimulating effect whereas aluminium and aluminium oxide-NPs at first had a positive effect but in higher concentrations they inhibited the growth. Treating the seedlings with 0.4 mg L−1 of each tested toxicant was mostly stimulating to the expression of the genes and reduced the differences between the transcript levels of the coleoptiles and roots. Increasing concentrations of the tested substances had both stimulating and inhibiting effects on the expression levels of the genes. The highest expression levels were usually noted at concentrations between 0.4 and 1.0 mg/L of each metal and metal nanoparticle, except for SiZIP1, which had the highest transcript amount at 1.6 mg L−1 of Cu2+ and at 0.1–0.8 mg L−1 of CuO-NPs, and LIP and GR from the seedling treated with Al2O3-NPs at concentrations of 0.1 and 1.6 mg L−1, respectively.

Gene Expression and Phenotypic Assessment of Egg Quality across Developmental Stages of Atlantic Cod throughout the Spawning Season.

Egg quality in fishes is commonly determined by fertilisation success and cleavage patterns as a phenotypic outcome of underlying regulatory mechanisms. Although these phenotypic estimators of egg quality are useful in farming conditions, these "good quality" egg batches do not always translate to good larval growth and survival. The identification of genes involved in embryonic development may help find links between genetic factors of maternal origin and egg quality. Herein, the relative expression of seven stage-specific developmental genes of Atlantic cod was analysed using quantitative PCR to understand the function during embryogenesis and its relationship with egg quality. Genes ccnb2 and pvalb1 showed significant differential expression between developmental stages and significant upregulation from blastula and somite stages, respectively. The comparison of spawning batches showed that the relative gene expression of genes ccnb2, acta, tnnt3 and pvalb1 was significantly higher from the middle of the spawning season where phenotypic quality estimators establish the best egg quality. Moreover, a positive significant correlation was observed between quality estimators based on egg morphology and the genetic expression of genes acta and acta1 during somitogenesis. This study suggests that the combination of quality estimators, genetics and batch timing could help optimise reproductive protocols for commercial stocks of Atlantic cod.

High Prevalence of Virulence-Associated Genes and Length Polymorphism in actA and inlB Genes Identified in Listeria monocytogenes Isolates from Meat Products and Meat-Processing Environments in Poland.

Listeria monocytogenes is a human pathogen that has the ability to cause listeriosis, a disease with possible fatal outcomes. The typical route of infection is ingestion of the bacteria with contaminated food. In this study, 13 virulence-associated genes were examined with PCR in the genomes of 153 L. monocytogenes isolates collected from meat products and processing environments in Poland. All isolates possessed genes from LIPI-1-hly, actA, plcA, plcB and mpl-as well as four internalins: inlA, inlB, inlC, inlJ. Invasion-associated protein iap, as well as genes prfA and sigB, encoding regulatory proteins, were also detected in all isolates. Gene flaA, encoding flagellin, was detected in 113 (74%) isolates. This was the only gene that was not detected in all isolates, as its presence is serotype-dependent. Gene actA showed polymorphism with longer and shorter variants in PCR amplicons. Two isolates were characterized by truncated inlB genes, lacking 141 bp in their sequence, which was confirmed by gene sequencing. All isolates were positive in hemolysis assays, proving the synthesis of functional PrfA and Hly proteins. Four genotypes of L. monocytogenes based on actA polymorphism and two genotypes based on inlB polymorphism were distinguished within the isolates' collection.

Molecular detection of Ovine listeric abortion in Nineveh governorate, Iraq

Listeriosis is a important abortive diseases in sheep , and is considered one of the most risky bacterial zoonotic disease worldwide.The study was carried out in 50 sheep flocks placed in the Nineveh governorate, Iraq during November and December 2022, a total of 300 specimens of blood , abomasum content and brain ( 100 each)were collected from local breed ovine aborted fetues(in the last stage of gestation) were obtained to be tested for molecular detection of Listeria monocytogenes . L monocytogenes DNA was detected in a total of 61(20.3%) specimens , distributed as: 35(57.4%) ,15(24.6% and 11(18.0%) strains from fetal brain, abomasum and blood specimens respectively using direct genus-specific conventional polymerase chain reaction (prfA gene) C- PCR. Two L. monocytogenes strains ( HMB1 listeriolysin, HMB 2 listeriolysin ) deposited in GenBank under accession numbers LC769365.1,and LC769366.1. All L. monocytogenes strains were positive for three genes (InlJ, InlA, and hlyA) except actA gene was detected in 46 (75.4%) strains. In conclusion , L. monocytogenes is one of the important causal agents of abortion in sheep flocks in Nineveh province , Iraq, and most of the brain specimens were positive for listeric infection compared to the other specimens.

Phenotypic and Genotypic Virulence Potentials of Listeria monocytogenes Isolated from Different Food Samples in Yola

Aim: the aim of this work is to determine the Phenotypic and Genotypic Virulence Potentials of Listeria monocytogenes Isolated from Different Food Samples in Yola. Study Design: the study was a randomized study; the samples (sources of organisms) were collected at random from different market and shops within the study area. Place and Duration of Study: the study was carried out in Yola, the capital of Adamawa State, north eastern Nigeria. Methodology: phenotypic virulence potentials of the isolates was determined by haemolytic activity on 5% sheep blood agar and biofilm formation ability, using Christensen’s tube method and microtiter plate method for the qualitative and quantitative biofilm formation respectively. Genotypic virulence determination was done by polymerase chain reaction amplification of plcB, hlyA and actA genes. Results: All the isolates (100%) were beta haemolytic on 5% sheep blood agar and all the isolates were biofilm formers but with different degree of biofilm formation abilities. Genotypically all the isolates (100%) indicated the presence of all the three virulence genes amplified. Conclusion: food samples isolates analyzed are potentially virulence pathogenic of L. monocytogenes that can pose food borne listeriosis to high risk individuals in the study area.

Virulence properties of Listeria monocytogenes isolated from meat and meat contact surfaces in a slaughterhouse.

Listeria monocytogenes is a ubiquitous microorganism that is isolated from a variety of sources such as soil, water, decaying vegetation, sewage, animal feeds, silage, farm environments and food-processing environments. This study aimed to determine the prevalence, serogroups, biofilm formation, virulence factor genes, and genetic relationships of L. monocytogenes strains isolated from beef meat and meat contact surfaces obtained from a slaughterhouse in Burdur, Turkey. In this study, a total of 179 beef meat and meat contact surface samples were analyzed for the presence of L. monocytogenes by polymerase chain reaction (PCR). Out of a total of 179 beef meat and meat contact surface samples, 83 (46.37%) were found to be contaminated with L. monocytogenes, with the highest incidence (53.01%) occurring in beef meat. In the present study, most of the isolated strains belonged to serogroups IIB and IVB (lineage I). The L. monocytogenes strain also contained monoA-B, prfA, plcA, plcB, mpl, hlyA, actA, gtcA, dltA, Fri, flaA, InlA, InlC, InlJ, and iap genes. Biofilm formation was not determined in the tested samples at pH 5.5 and different temperatures (4°C, 10°C, 25°C, and 37°C). However, strong biofilm formation was observed in 6.45% (2/31) of the strains at pH 7.0 after 48 h incubation at 37°C, and in 3.22% (1/31) of the strains at pH 7.0 after 48 h incubation at 4°C and 10°C. Pulsed-field gel electrophoresis (PFGE) results showed that L. monocytogenes isolates were clonally related, and cross-contamination was present. In addition, PFGE results also revealed that AscI had more distinguishing power than the ApaI restriction enzyme. These results indicate that L. monocytogenes detected from meat and meat contact surfaces in the slaughterhouse pose a potential risk to public health.

Apple crown and collar canker and necrosis caused by Cytospora balanejica sp. nov. in Iran

Apple is the most important fruit tree in West Azarbaijan province of Iran. In a survey of apple orchards, a disease with crown and collar canker and necrosis symptoms was observed in three young apple orchards in Urmia, affecting 15% and 1% of ‘Red Delicious’ and ‘Golden Delicious’ cultivars, respectively. A fungus with typical characteristics of the asexual morph of Cytospora was regularly isolated from the diseased tissues. Morphological characteristics and phylogenetic analyses inferred from the combined dataset of the ITS-rDNA, parts of LSU, tef1-α, rpb2, and act1 genes revealed that the isolates represent a new species of Cytospora, described herein as Cytospora balanejica sp. nov.. The pathogenicity of all isolates was confirmed on apple cv. ‘Red Delicious’ based on Koch’s postulates. Also, the reaction of 12 other apple cultivars was assessed against five selected isolates with the highest virulence. The results showed that except for cv. ‘Braeburn’, which did not produce any symptoms of the disease, the other 11 cultivars showed characteristic disease symptoms including sunken and discolored bark and wood. The mean length of the discolored area was different among the 11 so-called susceptible cultivars, hence cvs. ‘M4’ and ‘Golden Delicious’ showed the highest and the lowest lesion length, respectively. Moreover, the aggressiveness of the five tested isolates was different, and the isolates BA 2-4 and BA 3-1 had the highest and lowest aggressiveness, respectively. Based on our observations on the potential ability of the fungus to cause disease on young and actively growing apple trees, it will be a serious threat to apple cultivation and industry.

Potential public health hazards related to consumption of poultry contaminated with antibiotic resistant Listeria monocytogenes in Egypt

Listeria monocytogenes is an important foodborne pathogen that incorporated into many serious infections in human especially immunocompromised individuals, pregnant women, the elderly, and newborns. The consumption of food contaminated with such bacteria is considered a source of potential risk for consumers. Therefore, a total of 250 poultry purchased in highly popular poultry stores besides 50 swabs from workers hands in the same stores, in Mansoura City had been tested for the L. monocytogenes prevalence, virulence genes, and antibiotic resistance profile illustrating the health hazards from such poultry. The L. monocytogenes were recovered from 9.6% of poultry samples while not detected from workers hand swabs. The antimicrobial susceptibility of 24 L. monocytogenes strains against 24 antibiotics of seven different classes revealed high susceptibility rates to erythromycin (79.17%), streptomycin (66.67%), gentamycin (66.67%), vancomycin (58.33%), chloramphenicol (58.33%) and cefotaxime (41.67%). The majority (79.2%) of L. monocytogenes were classified as multidrug resistant strains with high resistance to tetracyclines and β-lactams antibiotics while 16.7% of the strains were categorized as extensively resistant ones. The iap virulence-specific determination gene had been detected in all recovered L. monocytogenes isolates while 83.33 and 70.83% of the isolates harbored hylA and actA genes. In addition, the study confirmed the capability of most L. monocytogenes isolates for biofilm formation by moderate to strong production and the quantitative risk assessment illustrated the risk of developing listeriosis as the risk value exceeded 100. The current results illustrate that poultry meat can be a source of pathogenic antibiotic resistant strains that may cause infection with limited or no treatment in immunosuppressed consumers via the food chain.

Detection of Some Virulence Factors of Listeria Species Isolated from Chicken and their Products

The current study was conducted to determine the occurrence of Listeria species in live chicken and some chicken products (fillets, burger and luncheon). In addition, determine the antibiogram profile of Listeria strains and detection of some virulence genes of Listeria monocytogenes which recovered from the examined samples. The study included 100 samples including cloacal swabs from broiler chicken (n =25) from 5 different broiler chicken farms beside as well as chicken products (25 samples each of chicken fillets, burger and luncheon) that were randomly purchased from supermarkets. Isolation and identification of Listeria was performed according to the standard protocol provided by ISO (2017) using Listeria selective agar (Oxford formulation) with Listeria selective supplement then the suspected colonies were identified using a panel of biochemical tests. It was recorded that the recovery rate of Listeria species from the examined samples was 24, 20, 8 and 16%, in cloacal swabs, chicken fillets, burger and luncheon, respectively with an overall incidence of 17%. In addition, biochemical identification of the recovered Listeria isolates revealed the presence of L. monocytogenes, L. innocua, L. ivanovii, L. seeligeri and L. grayi with various rates where L. monocytogenes scored the highest rate 29.4% (5 isolates). Antimicrobial susceptibility of Listeria strains (n=17) clarified that the highest antibiotic resistance was recorded when using streptomycin (82.4%) followed by Neomycin (76.5%) then Doxycycline and Cefotaxime (52.9%) while the lowest resistance was noticed when using Amikacin (11.8%) then Ciprofloxacin (35.3%). Finally, molecular detection of virulence genes in the identified L. monocytogenes strains (n= 5) clarified the detection of iap and actA genes in the five isolates while hylA gene was detected in three isolates only. At the end, results obtained by this study emphasize a significant occurrence of resistant Listeria in chicken and chicken products including processed animal-originated foods commonly purchased by the public in including; chicken fillets, burger and luncheon. Seriously, the presence of such resistant bacteria endangers human health by making routine antibiotic courses useless in the treatment of such infections. Finally, data proved that retail foods of animal origin pose a risk to human health.

Thyridiumlauri sp. nov. (Thyridiaceae, Thyridiales): a new pathogenic fungal species of bay laurel from Italy.

Laurusnobilis is an important Mediterranean tree and shrub native to Italy that is also commercially grown as spice and ornamental plant. Field surveys conducted since 2021 in Sicily (Italy) revealed that bay laurel plants in urban and private gardens and nurseries were severely affected by symptoms of stem blight and internal necrosis, which were associated with ambrosia beetle entry holes in the bark and internal wood galleries. The occurring ambrosia beetle was identified as Xylosandruscompactus, an invasive wood-boring pest previously reported from Sicily. Investigation of fungi from symptomatic tissues primarily resulted in the isolation of Thyridium-like colonies. The main symbiont of X.compactus, Ambrosiellaxylebori, was also isolated from infested plants. Phylogenetic analyses of a combined matrix of ITS, LSU, act1, rpb2, tef1, and tub2 gene regions revealed that the isolated Thyridium-like colonies represent a new fungal species within the genus Thyridium. Based on both phylogeny and morphology, the new isolated fungus is described as Thyridiumlauri sp. nov. Moreover, two recently described species, Phialemoniopsishipposidericola and Phialemoniopsisxishuangbannaensis, are transferred to the genus Thyridium due to the confirmed synonymy of both genera, as supported by molecular phylogenies. Pathogenicity test conducted on potted plants demonstrated that T.lauri is pathogenic to bay laurel, causing internal necrosis and stem blight. The new species was consistently re-isolated from the symptomatic tissue beyond the inoculation point, thereby fulfilling Koch's postulates. This study represents the first report of a new pathogenic fungus, T.lauri, causing stem blight and internal necrosis of bay laurel plants and associated with infestation of the invasive ambrosia beetle X.compactus.

Efficacy of disinfectant and bacteriophage mixture against planktonic and biofilm state of Listeria monocytogenes to control in the food industry

Fresh produce and animal-based products contaminated with Listeria monocytogenes have been the main cause of listeriosis outbreaks for many years. The present investigation explored the potential of combination treatment of disinfectants with a bacteriophage cocktail to control L. monocytogenes contamination in the food industry. A mixture of 1 minimal inhibitory concentration (MIC) of disinfectants (sodium hypochlorite [NaOCl], hydrogen peroxide [H2O2], and lactic acid [LA]) and multiplicity of infection (MOI) 100 of phage cocktail was applied to both planktonic cells in vitro and already-formed biofilm cells on food contact materials (FCMs; polyethylene, polypropylene, and stainless steel) and foods (celery and chicken meat). All the combinations significantly lowered the population, biofilm-forming ability, and the expression of flaA, motB, hlyA, prfA, actA, and sigB genes of L. monocytogenes. Additionally, in the antibiofilm test, approximately 4 log CFU/cm2 was eradicated by 6 h treatment on FCMs, and 3 log CFU/g was eradicated within 3 days on celery. However, <2 log CFU/g was eradicated in chicken meat, and regrowth of L. monocytogenes was observed on foods after 5 days. The biofilm eradication efficacy of the combination treatment was proven through visualization using scanning electron microscopy (SEM) and confocal microscopy. In the SEM images, the unusual behavior of L. monocytogenes invading from the surface to the inside was observed after treating celery with NaOCl+P or H2O2 + P. These results suggested that combination of disinfectants (NaOCl, H2O2, and LA) with Listeria-specific phage cocktail can be employed in the food industry as a novel antimicrobial and antibiofilm approach, and further research of L. monocytogenes behavior after disinfection is needed.

Listeria monocytogenes, a silent foodborne pathogen in Ecuador.

Listeria monocytogenes is a foodborne pathogen that can produce serious, even fatal, infections. Among other foods, it can be found in unpasteurized dairy and ready-to-eat products. Surveillance of L. monocytogenes is of great interest since sources of infection are difficult to determine due to the long incubation period, and because the symptoms of listeriosis are similar to other diseases. We performed a genomic study of L. monocytogenes isolated from fresh cheeses and clinical samples from Ecuador. Sixty-five isolates were evaluated and sequenced, 14 isolates from cheese samples and 20 from clinical listeriosis cases from the National Institute of National Institute of Public Health Research, and 31 isolates from artisanal cheese samples from 8 provinces. All isolates exhibited heterogeneous patterns of the presence of pathogenicity islands. All isolates exhibited at least 4 genes from LIPI-1, but all references (26 L. monocytogenes closed genomes available in the NCBI database) showed the complete island, which encompasses 5 genes but is present in only two Ecuadorian isolates. Most isolates lacked gene actA. Genes from LIPI-2 were absent in all isolates. LIPI-3 and LIPI-4 were present in only a few references and isolates. With respect to the stress survival islets, our samples either presented SSI-1 or SSI-F2365, except for one isolate that presented SSI-F2365 and also one gene from SSI-1. None of the samples presented SSI-2. The predominant ST (sequence type) was ST2 (84.62% 55/65), and the only ST found in food (93.33% 42/45) and clinical samples (65% 13/20). Isolates were not grouped according to their sampling origin, date, or place in a phylogenetic tree obtained from the core alignment. The presence of ST2 in food and clinical samples, with high genomic similarity, suggests a foodborne infection risk linked to the consumption of fresh cheeses in Ecuador.

First report of Alternaria alternata associated with spot blotch of barley (Hordeum vulgare) in Iraq.

A field survey was conducted during the barley growing season 2019 in the Karbala Province of Iraq. Barley plants with severe leaf spot symptoms were collected from several barley fields. The associated fungus was isolated and purified from those symptomatic leaves. The fungus colonies were aeriform mycelia in grey to black color. Conidia observed that they were light to dark brown and formed different-length chains. The causal agent was confirmed as Altenaria alternata via the PCR amplification and sequencing of the rDNA-ITS region, actA, and tef1 genes. Pathogenicity test showed that A. alternata isolated was pathogenic by producing light to dark brown spots on barley foliage. This is the first record of this pathogen causing Alternaria spot blotch on barley crops in Iraq. Keywords: Alternaria alternata; leaf spot; Hordeum vulgare; Pathogenicity assessment; Phylogeny analysis.

Danielozyma pruni sp. nov., an asexual yeast species isolated from insect frass.

Two strains (NYNU 218101 and NYNU 218104) of an asexual yeast species were isolated from insect frass collected in insect tunnels of red leaf plum trees in the Henan Province, central China. Molecular phylogenetic analysis of the D1/D2 domain of the large subunit rRNA gene and the internal transcribed spacer (ITS) region showed that these two strains belonged to the genus Danielozyma, with Danielozyma litseae as the closest known species. They differed from the type strain of D. litseae by 0.6 % substitutions (three substitutions and one gap) in the D1/D2 domain and by 5.1 % substitutions (19 substitutions and six gaps) in the ITS region, respectively. When compared with the partial ACT1, TEF1 and RPB1 gene sequences, they differed by 3 % (26 substitutions), 2.7 % (25 substitutions) and 9 %(54 substitutions) from D. litseae NRRL YB-3246T in these regions. Physiologically, they also differed from its closest known species D. litseae based on the ability to assimilate inulin and galactitol, as well as to grow in 0.1 % cycloheximide and its inability to ferment maltose and raffinose. In order to classify the two new isolates based on morphological and molecular evidence, we proposed the description of a novel species Danielozyma pruni sp. nov. with strain JCM 35735T as holotype (Mycobank MB 849101).

Investigation on the inactivation effect and mechanism of Listeria monocytogenes in fresh-cut cucumber during storage by ultrasound combined with sodium hypochlorite

Fresh agricultural products are frequently contaminated with Listeria monocytogenes (L. monocytogenes), which threatens consumer health. The mechanism of the inhibitory effect of ultrasound and sodium hypochlorite (US-NaClO) on L. monocytogenes on fresh-cut cucumber remains poorly understood. Therefore, the bactericidal ability and mechanism of US-NaClO treatment on L. monocytogenes were studied on fresh-cut cucumber during storage using various approaches such as determination of intracellular material leakage, scanning electron microscopy, flow cytometry, and expression analysis of virulence genes. The results showed that the number of L. monocytogenes on fresh-cut cucumber was significantly reduced after ultrasound treatment for 5 min in combined with 75 ppm sodium hypochlorite treatment(P < 0.05). The US–NaClO treatment affected cell morphology, impaired cell membrane integrity, increased cell membrane permeability, and reduced the concentration of K+, inorganic phosphate, ATP, proteins, and DNA in bacterial cells, leading to the inactivation of microorganisms. In addition, the US–NaClO treatment downregulated expression of the virulence genes actA, hly, inlA, mpl, pclA, and plcB, thus decreasing the pathogenicity of bacteria. It can avoid contamination by pathogenic bacteria during the production of fresh-cut cucumber, while providing safety assurance for production.

High-Pressure Processing-Impacts on the Virulence and Antibiotic Resistance of Listeria monocytogenes Isolated from Food and Food Processing Environments.

High-pressure processing (HPP) is one of the non-thermal methods of food preservation considered to be safe but may cause an increase/decrease in virulence potential and antibiotic resistance. The aim of the present study was to evaluate the survival of L. monocytogenes isolates after high-pressure processing (200 and 400 MPa for 5 min) and to determine changes in phenotypic and genotypic antibiotic resistance and virulence after this treatment. The 400 MPa treatment was shown to be effective in reducing pathogens to safe levels; however, the potential for cell recovery during storage was observed. In addition, studies on changes in virulence indicated possibilities related to a decrease in actA gene expression, overexpression of the hly and osfX gene, and an increase in biofilm-forming ability. The studies on changes in antibiotic resistance of isolates showed that all isolates showing initial susceptibility to lincomycin, fosfomycin, trimethoprim/sulfamethoxazole, and tetracycline became resistant to these antibiotics, which was associated with an increase in the values of minimum inhibitory concentrations. An increase in the expression of antibiotic resistance genes (mainly tetA_1, tetA_3, tetC) was also observed (mainly after the application of 200 MPa pressure), which was isolate dependent. However, it is noteworthy that the induced changes were permanent, i.e., they persisted even after the restoration of optimal environmental conditions. The results presented in our work indicate that the stress occurring during HPP can affect both phenotypic and genotypic changes in the virulence and antibiotic resistance potential of pathogens isolated from food and food processing environments. The potential associated with cell recovery and persistence of changes may influence the spread of virulent isolates of pathogens with increased antibiotic resistance in the food and food processing environment.

Characterization and Antimicrobial Susceptibility Patterns of Listeria monocytogenes from Raw Cow Milk in the Southern Part of Ethiopia

Food safety remains the main health concern in the developing countries. Thus, the major purpose of the present study was to characterize and determine antibiotic susceptibility patterns of Listeria monocytogenes from raw milk samples collected from southern Ethiopia. Two hundred and forty raw cow milk samples were collected from dairy farms and smallholder dairy producers using a simple random sampling technique and analyzed by cultural and multiplex PCR methods. The antimicrobial susceptibility profile of L. monocytogenes was evaluated using the standard disk diffusion method. Over 28% of the samples were found positive for Listeria spp., of which 17 (7.08%) isolates were identified as L. monocytogenes after morphological and biochemical confirmation. The prevalence of L. monocytogenes was 6.02% in Hawassa city, 5.56% in Dale district, and 9.41% in Arsi Negele district. L. monocytogenes was higher in the wet season (9.32%) than in the dry season (4.92%). The gene for Listeria specific 16S rRNA was detected in all the 17 examined isolates, while hlyA and iapA were only found in 11 of them. Furthermore, no isolate was identified to have the prfA, actA, or plcA genes. Antimicrobial resistance profiling revealed that all the L. monocytogenes isolates were resistant to nalidixic acid (100%), followed by erythromycin (88.24%). However, all the L. monocytogenes isolates were sensitive to vancomycin, gentamicin, and sulfamethoxazole. Raw cow milk is a potential source of L. monocytogenes and it poses a threat to human and animal health. Therefore, it is crucial that dairy producers and vendors of raw milk in the study areas should take considerable precautions to prevent Listeria species from contaminating raw fresh milk.

Molecular genetic studies of Listeria monocytogenes isolates from different animal species in Ukraine

The PCR-profiles of 12 archival cultures of listeria isolated from different species of animals and stored in the collection of microorganisms of the NSC IEСVM were determined. It was established that 10 isolates have genes prs, inlB, inlA, inlC, inlJ, actA, plcB, hly, iap, which characterizes them as highly virulent. The L. monocytogenes 61052 isolate did not contain the inlA gene, and the actA and hly genes were not detected in L. monocytogenes Varja. Using PCR, it was also established that 11 studied cultures belong to serotypes 4b, 4d, 4e, as well as one isolate to 1/2a. The obtained data can be used to improve the diagnosis of animal listeriosis in Ukraine due to the creation of highly specific antigens