Emerging Foodborne Pathogen Research Articles

Genetic diversity and potential transmission of Escherichia albertii in a poultry breeding rural village.

Escherichia albertii is an emerging foodborne pathogen causing diarrhea. Though various animals, especially poultry, serve as reservoirs, the transmission of E. albertii among reservoirs and the risk to humans remain unclear. This study investigated an E. albertii infected infant with poultry exposure and collected samples from contact persons, poultries, and environment to better understand the transmission dynamics of E. albertii. One E. albertii isolate from contact person, seven isolates from poultries, and six isolates from environment were recovered, respectively. Whole genome sequencing analysis showed that eight strains derived from poultry or environment and classified as ST4633, shared great similarity (cgSNP ≤ 20). However, the patient-derived strain ESA311 had a cgSNP difference of 1165 with human strain ESA339, and differed from poultry and environmental strains (cgSNP range 2417 to 14997), suggesting a distant relatedness. Whole genome phylogeny showed several human-derived E. albertii strains were clustered with those from animal origins. Our results suggested that family-breeding poultry constituted a possible reservoir of E. albertii, with the environment acting as a crucial vector for the spread of these bacteria, posing a risk to humans. Further poultry surveillance is needed to elucidate public health risks associated with E. albertii infection.

Conditional expression of flagellar motility, curli fimbriae, and biofilms in Shiga toxin- producing Escherichia albertii.

Escherichia albertii is an emerging foodborne pathogen. We previously reported that some avian Shiga toxin-producing E. albertii strains exhibited higher or comparable cytotoxicity in Vero-d2EGFP cells with several enterohemorrhagic E. coli (EHEC) outbreak strains. To better understand the environmental persistence of this pathogen, comparative genomics and phenotypic assays were applied to assess adhesion capability, motility, and biofilm formation in E. albertii. Among the 108 adherence-related genes, those involved in biogenesis of curli fimbriae, hemorrhagic E. coli pilus, type 1 fimbriae, and Sfm fimbriae were conserved in E. albertii. All 20 E. albertii strains carried a complete set of primary flagellar genes that were organized into four gene clusters, while five strains possessed genes related to the secondary flagella, also known as lateral flagella. Compared to EHEC strain EDL933, the eight chemotaxis genes located within the primary flagellar gene clusters were deleted in E. albertii. Additional deletion of motility genes flhABCD and motBC was identified in several E. albertii strains. Swimming motility was detected in three strains when grown in LB medium, however, when grown in 5% TSB or in the pond water-supplemented with 10% pigeon droppings, an additional four strains became motile. Although all E. albertii strains carried curli genes, curli fimbriae were detected only in four, eight, and nine strains following 24, 48, and 120 h incubation, respectively. Type 1 fimbriae were undetectable in any of the strains grown at 37°C or 28°C. Strong biofilms were detected in strains that produced curli fimbriae and in a chicken isolate that was curli fimbriae negative but carried genes encoding adhesive fimbriae K88, a signature of enterotoxigenic E. coli strains causing neonatal diarrhea in piglets. In all phenotypic traits examined, no correlation was revealed between the strains isolated from different sources, or between the strains with and without Shiga toxin genes. The phenotypic variations could not be explained solely by the genetic diversity or the difference in adherence genes repertoire, implying complex regulation in expression of various adhesins. Strains that exhibited a high level of cytotoxicity and were also proficient in biofilm production, may have potential to emerge into high-risk pathogens.

Comparison of CRISPR typing and conventional molecular methods for distinguishing Laribacter hongkongensis isolates from fish, frogs and humans

High-resolution and efficient typing for Laribacter hongkongensis (L. hongkongensis) is essential for epidemiological investigation of such emerging foodborne pathogens. Clustered regularly interspaced short palindromic repeats (CRISPR) typing is an innovative molecular method that shows great promise for L. hongkongensis typing. Here, we explored the CRISPR typing method by combining CRISPR1 and CRISPR2 loci to characterize a collection of 109 L. hongkongensis isolates from humans and animals and compared it to current molecular methods such as pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The results showed that all three methods have high discriminatory power (diversity index was 0.9902 for PFGE, 0.9663 for CRISPR and 0.9562 for MLST); strong congruence was observed between them (Rand index was 0.969 between CRISPR and PFGE, 0.953 between CRISPR and MLST, 0.958 between PFGE and MLST). CRISPR typing could well distinguish the isolates in the same STs or PFGE profiles, and the genetic information contained by the CRISPR array is useful for deep phylogenetic typing. We demonstrate that rapid CRISPR typing is a practical genetic fingerprinting tool with high resolution, comparable ease of use and lower cost, ability to track the source of various groups of L. hongkongensis strains and indication of genetic characteristics.

Hepatitis E Virus RNA Detection in Liver and Muscle Tissues Sampled from Home Slaughtered Domestic Pigs in Central Italy.

Hepatitis E virus is a worldwide emerging foodborne pathogen; raw or undercooked meats and liver pork products can cause infection through the orofecal route. In Central-Southern Italy, small traditional farming method, associated with the possibility of environmental sharing with wild species, can facilitate HEV diffusion and persistence. The aim of this study was to determine HEV genotype and subtype in Marche region from home slaughtered domestic pigs involved in small and traditional food chains. A total of 236 liver and muscle tissues and 6 pooled salami samples were screened. Laboratory workflow started with homogenization, followed by RNA extraction. Nested reverse transcription PCR and qRT-PCR were used to amplify specific parts of overlapping open reading frames belonging to the HEV genome. A total of 42/236 (17.79%) liver and 8/236 (3.39%) diaphragm specimens were positive; none of the pooled salami specimens showed positive HEV signal. The discovered HEV3c presented high nucleotide similarities with ones amplified from wild boar populations hunted in the same province. Extensive farming methods and environmental sharing with wild animal species support cross-infection infections, as observed in the present study. Although salami resulted negative for HEV RNA detection, the effects of food technologies on viral loads remain unclear. Therefore, further scientific investigations coupled with efficacious standardized laboratory procedures will be the next challenge.

A metagenomics-based workflow for the detection and genomic characterization of GBS in raw freshwater fish.

The need for a rapid and accurate food microbiological testing method is apparent for a timely and effective foodborne outbreak response. This is particularly relevant for emerging foodborne pathogens such as Group B Streptococcus (GBS) whose associated food safety risk might be undercharacterized. By using GBS in raw freshwater fish as a case example, this study describes the development of a metagenomics-based workflow for rapid food microbiological safety testing and surveillance. This study can inform as a working model for various foodborne pathogens in other complex food matrices, paving the way for future methodological development of metagenomics for food microbiological safety testing.

Occurrence of Helicobacter pullorum in Retail Chicken Meat: A One-Health Approach to Consumer Health Protection.

Helicobacter pullorum is an emerging foodborne pathogen that commonly colonizes the gastrointestinal tract of poultry, causing gastroenteritis. It has been related to several clinically important infections, including colitis and hepatitis, inflammatory bowel disease, recurrent diarrhea, and bacteremia in the human population. The bacterium may be transmitted to humans through undercooked poultry meat. In order to investigate the occurrence of H. pullorum in raw retail chicken meat (thighs and breasts), we analyzed 240 samples: 120 chicken thigh and 120 chicken breast samples. The samples were analyzed by means of an isolation protocol using Steele and McDermott's modified filtration technique on Brucella agar supplemented with 5% of defibrinated sheep's blood. The presumptive colonies were biochemically identified and analyzed using a previously described conventional PCR test based on the 16S rRNA gene. In total, 35% of analyzed samples were positive using the microbiological protocol and 45% were positive by PCR. These results suggest that H. pullorum can be transmitted to humans through the handling and consumption of raw poultry meat, representing a risk for food business operators and consumers. Efforts to control H. pullorum in broiler meat should prioritize the implementation of stringent hygienic practices across all stages of the food chain, from the farm to the consumer.

Gas-Selective Catalytic Regulation by a Newly Identified Globin-Coupled Sensor Phosphodiesterase Containing an HD-GYP Domain from the Human Pathogen Vibrio fluvialis.

Globin-coupled sensors constitute an important family of heme-based gas sensors, an emerging class of heme proteins. In this study, we have identified and characterized a globin-coupled sensor phosphodiesterase containing an HD-GYP domain (GCS-HD-GYP) from the human pathogen Vibrio fluvialis, which is an emerging foodborne pathogen of increasing public health concern. The amino acid sequence encoded by the AL536_01530 gene from V. fluvialis indicated the presence of an N-terminal globin domain and a C-terminal HD-GYP domain, with HD-GYP domains shown previously to display phosphodiesterase activity toward bis(3',5')-cyclic dimeric guanosine monophosphate (c-di-GMP), a bacterial second messenger that regulates numerous important physiological functions in bacteria, including in bacterial pathogens. Optical absorption spectral properties of GCS-HD-GYP were found to be similar to those of myoglobin and hemoglobin and of other bacterial globin-coupled sensors. The binding of O2 to the Fe(II) heme iron complex of GCS-HD-GYP promoted the catalysis of the hydrolysis of c-di-GMP to its linearized product, 5'-phosphoguanylyl-(3',5')-guanosine (pGpG), whereas CO and NO binding did not enhance the catalysis, indicating a strict discrimination of these gaseous ligands. These results shed new light on the molecular mechanism of gas-selective catalytic regulation by globin-coupled sensors, with these advances apt to lead to a better understanding of the family of globin-coupled sensors, a still growing family of heme-based gas sensors. In addition, given the importance of c-di-GMP in infection and virulence, our results suggested that GCS-HD-GYP could play an important role in the ability of V. fluvialis to sense O2 and NO in the context of host-pathogen interactions.

Antimicrobial Blue Light (aBL) as a potential tool to reduce bacterial spoilage in the fishery chain

Along the fishery chain, a high amount of fish is lost for the activity of spoilage microorganisms originating from the environment, human handling and fish themselves. Different techniques are conventionally used to reduce the growth of bacteria: from cold temperature and icing to high concentration of salts, from drying to natural antimicrobial compounds. In this study, the antimicrobial Blue Light (aBL) was considered as an innovative tool. In particular, the irradiation with light at 410 nm inhibited the growth of most bacteria isolated from skin samples of anchovies and sardines chosen for their worldwide commercial importance. Bacterial strains showed a different sensitivity to light treatment: the ones isolated from anchovy were more sensitive than those from sardine. Investigations were performed on Aeromonas bestiarum, an emerging foodborne pathogen. Upon irradiation with light at 410 nm (200 J/cm2), a statistically significant decrease of 3 log units was observed. The same fluence rate successfully inhibited the biofilm formation of A. bestiarum, and disrupted 50 % of the adherent biomass of a 24-h old biofilm. The irradiation of Staphyococcus vitulinus compromised its viability and the associated proteolytic activity known to contribute to meat spoilage. In vivo experiments showed that aBL caused a remarkable decrease (at least 50 %) of viable counts of bacteria from anchovy and sardine skin samples conserved at 4 °C for one day. In conclusion, these results support the potential use of blue light in reducing the growth of skin microorganisms potentially responsible for loss of food safety, quality and decrease of storage life.

Antibiotic-Resistant Arcobacter spp. in commercial and smallholder farm animals in Asante Akim North Municipality, Ghana and Korogwe Town Council, Tanzania: a cross-sectional study

BackgroundArcobacter species are considered emerging foodborne pathogens that can potentially cause serious infections in animals and humans. This cross-sectional study determined the frequency of potentially pathogenic Arcobacter spp. in both commercial and smallholder farm animals in Ghana and Tanzania. A total of 1585 and 1047 (poultry and livestock) samples were collected in Ghana and Tanzania, respectively. Selective enrichment media, along with oxidase and Gram testing, were employed for isolation of suspected Arcobacter spp. and confirmation was done using MALDI-TOF MS. Antibiotic susceptibility was assessed through disk diffusion method and ECOFFs were generated, for interpretation, based on resulting inhibition zone diameters.ResultsThe overall Arcobacter frequency was higher in Ghana (7.0%, n = 111) than in Tanzania (2.0%, n = 21). The frequency of Arcobacter in commercial farms in Ghana was 10.3% (n/N = 83/805), while in Tanzania, it was 2.8% (n/N = 12/430). Arcobacter was detected in only 3.6% (n/N = 28/780) of the samples from smallholder farms in Ghana and 1.5% (n/N = 9/617) of the samples from Tanzania. For commercial farms, in Ghana, the presence of Arcobacter was more abundant in pigs (45.1%, n/N = 37/82), followed by ducks (38.5%, n/N = 10/26) and quails (35.7%, n/N = 10/28). According to MALDI-TOF-based species identification, Arcobacter butzleri (91.6%, n/N = 121/132), Arcobacter lanthieri (6.1%, n/N = 8/132), and Arcobacter cryaerophilus (2.3%, n/N = 3/132) were the only three Arcobacter species detected at both study sites. Almost all of the Arcobacter from Ghana (98.2%, n/N = 109/111) were isolated during the rainy season. The inhibition zone diameters recorded for penicillin, ampicillin, and chloramphenicol allowed no determination of an epidemiological cut-off value. However, the results indicated a general resistance to these three antimicrobials. Multidrug resistance was noted in 57.1% (n/N = 12/21) of the Arcobacter isolates from Tanzania and 45.0% (n/N = 50/111) of those from Ghana. The type of farm (commercial or smallholder) and source of the sample (poultry or livestock) were found to be associated with multi-drug resistance.ConclusionsThe high levels of MDR Arcobacter detected from farms in both countries call for urgent attention and comprehensive strategies to mitigate the spread of antimicrobial resistance in these pathogens.

Genomic and Phenotypic Characterization of Shiga Toxin-Producing Escherichia albertii Strains Isolated from Wild Birds in a Major Agricultural Region in California.

Escherichia albertii is an emerging foodborne pathogen. To better understand the pathogenesis and health risk of this pathogen, comparative genomics and phenotypic characterization were applied to assess the pathogenicity potential of E. albertii strains isolated from wild birds in a major agricultural region in California. Shiga toxin genes stx2f were present in all avian strains. Pangenome analyses of 20 complete genomes revealed a total of 11,249 genes, of which nearly 80% were accessory genes. Both core gene-based phylogenetic and accessory gene-based relatedness analyses consistently grouped the three stx2f-positive clinical strains with the five avian strains carrying ST7971. Among the three Stx2f-converting prophage integration sites identified, ssrA was the most common one. Besides the locus of enterocyte effacement and type three secretion system, the high pathogenicity island, OI-122, and type six secretion systems were identified. Substantial strain variation in virulence gene repertoire, Shiga toxin production, and cytotoxicity were revealed. Six avian strains exhibited significantly higher cytotoxicity than that of stx2f-positive E. coli, and three of them exhibited a comparable level of cytotoxicity with that of enterohemorrhagic E. coli outbreak strains, suggesting that wild birds could serve as a reservoir of E. albertii strains with great potential to cause severe diseases in humans.

Cronobacter spp. Isolated from Quick-Frozen Foods in China: Incidence, Genetic Characteristics, and Antibiotic Resistance.

Cronobacter spp. are emerging foodborne pathogens that cause severe diseases. However, information on Cronobacter contamination in quick-frozen foods in China is limited. Therefore, we studied the prevalence, molecular characterization, and antimicrobial susceptibility of Cronobacter in 576 quick-frozen food samples collected from 39 cities in China. Cronobacter spp. were found in 18.75% (108/576) of the samples, and the contamination degree of the total positive samples was 5.82 MPN/g. The contamination level of frozen flour product samples was high (44.34%). Among 154 isolates, 109 were C. sakazakii, and the main serotype was C. sakazakii O1 (44/154). Additionally, 11 serotypes existed among four species. Eighty-five sequence types (STs), including 22 novel ones, were assigned, indicating a relatively high genetic diversity of the Cronobacter in this food type. Pathogenic ST148, ST7, and ST1 were the main STs in this study. ST4, epidemiologically related to neonatal meningitis, was also identified. All strains were sensitive to cefepime, tobramycin, ciprofloxacin, and imipenem, in which the resistance to cephalothin was the highest (64.94%).Two isolates exhibited multidrug resistance to five and seven antimicrobial agents, respectively. In conclusion, these findings suggest that the comparatively high contamination level of Cronobacter spp. in quick-frozen foods is a potential risk warranting public attention.

Isolation of emerging human pathogens and foodborne pathogens in clinical cases of infections from dogs and cats admitted to a veterinary clinic in northern Portugal

Nowadays, dogs and cats are considered family members, translating into increased proximity to humans. Studies have shown that 40-60% of owners have very close interactions e.g., sharing the same dish, face lickings, or sleeping on the same bed. This promotes the exchange of microorganisms between species. Therefore, an One Health approach should be applied to encompass both human and animal health problems. In this study, 37 cases of companion animals (54.1% dogs and 45.9% cats) in a veterinary clinic in northern Portugal were analysed, and urinary and eye infections, respiratory conditions, and dermatological problems were confirmed. According to sample type, 43.2% were from urine, 13.5% from ear exudate, 8.1% from skin exudate, 8.1% from trachealexudate, 2.7% from vaginal exudate, 2.7% from conjunctiva exudate, 2.7% from purulent exudate, 2.7% from bronchoalveolar exudate, 2.7% from ocular exudate, and 2.7% from blood. In all, 21 pathogen species were isolated, of which Pantoea agglomerans, Cronobacter sakazakii, Leclercia adecarboxylata, Sphingomonas paucimobilis are currently considered emerging agents of human infection. The occurrence of these species was 2.4% for Leclercia adecarboxylata (one young male cat), 2.4% for Pantoea agglomerans (one adult male dog), 2.4% for Cronobacter sakazakii (one adult male dog), and 2.4% for Sphingomonas paucimobilis (one adult male dog). Leclercia adecarboxylata is a gram-negative bacillus of the Enterobacteriaceae family. It is a ‘novel’ rare human pathogen, mostly affecting immunocompromised individuals or causing polymicrobial infections in immunocompetent patients. Cronobacter sakazakii is an emerging foodborne pathogen that causes necrotizing enterocolitis and bacteremia in humans, Pantoea agglomerans is associated with a hospital-acquired infection, mostly in immunocompromised individuals with a fatal outcome. Sphingomonas paucimobilis is an emerging opportunistic bacterium with a particular tropism toward bones and soft tissues. The intimate relationship between humans and companion animals presents a potential risk for the transmission of zoonotic pathogens. Therefore, more research based on an One Health approach should be performed to more accurately determine the occurrence and incidence of emerging agents between species.

Characterization and application of a lytic jumbo phage ZPAH34 against multidrug-resistant Aeromonas hydrophila.

Aeromonas hydrophila is an emerging foodborne pathogen causing human gastroenteritis. Aeromonas species isolated from food such as seafood presented multidrug-resistance (MDR), raising serious concerns regarding food safety and public health. The use of phages to infect bacteria is a defense against drug-resistant pathogens. In this study, phage ZPAH34 isolated from the lake sample exerted lytic activity against MDR A. hydrophila strain ZYAH75 and inhibited the biofilm on different food-contacting surfaces. ZPAH34 has a large dsDNA genome of 234 kb which belongs to a novel jumbo phage. However, its particle size is the smallest of known jumbo phages so far. Based on phylogenetic analysis, ZPAH34 was used to establish a new genus Chaoshanvirus. Biological characterization revealed that ZPAH34 exhibited wide environmental tolerance, and a high rapid adsorb and reproductive capacity. Food biocontrol experiments demonstrated that ZPAH34 reduces the viable count of A. hydrophila on fish fillets (2.31 log) and lettuce (3.28 log) with potential bactericidal effects. This study isolated and characterized jumbo phage ZPAH34 not only enriched the understanding of phage biological entity diversity and evolution because of its minimal virion size with large genome but also was the first usage of jumbo phage in food safety to eliminate A. hydrophila.

Antibiotic Resistance in Selected Emerging Bacterial Foodborne Pathogens-An Issue of Concern?

Antibiotic resistance (AR) and multidrug resistance (MDR) have been confirmed for all major foodborne pathogens: Campylobacter spp., Salmonella spp., Escherichia coli and Listeria monocytogenes. Of great concern to scientists and physicians are also reports of antibiotic-resistant emerging food pathogens-microorganisms that have not previously been linked to food contamination or were considered epidemiologically insignificant. Since the properties of foodborne pathogens are not always sufficiently recognized, the consequences of the infections are often not easily predictable, and the control of their activity is difficult. The bacteria most commonly identified as emerging foodborne pathogens include Aliarcobacter spp., Aeromonas spp., Cronobacter spp., Vibrio spp., Clostridioides difficile, Escherichia coli, Mycobacterium paratuberculosis, Salmonella enterica, Streptocccus suis, Campylobacter jejuni, Helicobacter pylori, Listeria monocytogenes and Yersinia enterocolitica. The results of our analysis confirm antibiotic resistance and multidrug resistance among the mentioned species. Among the antibiotics whose effectiveness is steadily declining due to expanding resistance among bacteria isolated from food are β-lactams, sulfonamides, tetracyclines and fluoroquinolones. Continuous and thorough monitoring of strains isolated from food is necessary to characterize the existing mechanisms of resistance. In our opinion, this review shows the scale of the problem of microbes related to health, which should not be underestimated.

First report of enterotoxigenic Staphylococcus argenteus as a foodborne pathogen

Staphylococcal enterotoxins preformed in food are the causative agents of staphylococcal food poisoning outbreaks (SFPO). In this study we characterised in depth two coagulase-positive non-pigmented staphylococci involved in two independent outbreaks that occurred in France. While indistinguishable from Staphylococcus aureus using PCR methods and growth phenotype comparisons, both isolates were identified as Staphylococcus argenteus by whole genome sequencing. The genomes were analysed for the presence of enterotoxin genes, whose expression was determined in laboratory medium and, for the first time, in artificially-contaminated milk samples by using liquid chromatography-mass spectrometry and ELISA methods. The concentration measured for the SEB toxin in milk (0.67 ng/ml) was comparable to concentrations reported for other types of enterotoxins behind SFPO. From a collection of publicly available genomes, we performed an unprecedented systematic investigation of the enterotoxin gene set of S. argenteus, including variants and pseudogenes. The most prevalent genes were sex, followed by sel26, sel27 and sey. The egc cluster was less frequent and most of the time carried a dysfunctional seg gene. Our results shed light on the enterotoxigenic properties of S. argenteus, and emphasize the importance in monitoring of S. argenteus as an emerging foodborne pathogen.

Thermal inactivation kinetics of uropathogenic Escherichia coli in sous-vide processed chicken breast

Chicken is a suspected reservoir of uropathogenic Escherichia coli (UPEC), resulting in foodborne urinary tract infections (UTIs). Sous-vide ready-to-eat (RTE) food products may be associated with microbial hazards due to the low-temperature long-time (LTLT) process. However, little is known regarding the survival of UPEC during sous-vide cooking. The aim of this study was to evaluate the heat resistance of UPEC in chicken breast during sous-vide processing and establish predictive inactivation models. Chicken breast samples were inoculated with a four-strain cocktail of UPEC, including reference strains from UTI patients and chicken isolates. The inoculated samples, with or without 3% NaCl solution for marination, were vacuum sealed in bags, immersed in a temperature-controlled water bath, and cooked at 50 °C, 55 °C, 60 °C, and 63 °C. The change in survival of populations of UPEC was fitted with the linear and Weibull inactivation models to obtain the survival curves at different temperatures; the D- and z-values were also calculated. The goodness-of-fit was evaluated using the root mean square error (RMSE), sum of squared errors (SSE), adjusted R2, and Akaike information criterion (AIC). The results showed that the linear model with tail was better than the Weibull model in terms of fitting performance. With the addition of salt marinade, D-values at 50 °C, 55 °C, 60 °C, and 63 °C determined by the linear model with tail decreased from 299.78 to 166.93 min, 16,60 to 13.87 min, 4.06 to 3.05 min, and 1.05 to 0.87 min, respectively, compared with the controls. The z-values of control and salt-marinated samples were 6.14 °C and 5.89 °C, respectively. The model developed for predicting UPEC survival under sous-vide cooking was validated using an additional survival curve at 58 °C. The validation results showed that the RMSE was 0.122 and 0.133 log CFU/g, and the proportion of relative error was 0.875 and 0.750 in the acceptable prediction zones for the control and salt-marinated samples, respectively. In conclusion, the heat resistance of an emerging foodborne pathogen, UPEC, in sous-vide processed chicken breast was revealed for the first time. Our results showed that salt marinade (3% NaCl) increases the heat sensitivity of UPEC during the sous-vide processing. The developed survival functions based on the linear model with tail can be applied to control the thermal lethality of UPEC.

Clarithromycin-, and metronidazole-resistant Helicobacter pylori isolated from raw and ready-to-eat meat in Mansoura, Egypt

Helicobacter pylori (H. pylori), classified as a class-I carcinogen, is one of the leading medical pathogens of global concern associated mainly with the development of gastric adenocarcinomas and gastric mucosa-associated lymphoid tissue lymphomas; nevertheless, its prevalence in food especially meat and meat products is not fully covered. Additionally, the resistance of H. pylori towards clarithromycin is increasing worldwide and consider the leading cause of H. pylori treatment failure. Thus, the present study was designed to determine the prevalence, molecular characterization, and antimicrobial resistance profiles of clarithromycin-, and metronidazole-resistant H. pylori isolated from raw and ready-to-eat meat samples retailed in Mansoura city, Egypt. Among the 250 samples tested, H. pylori were molecularly confirmed in 40.8 % (49/120) of raw meat products and in 29.2 % (38/130) of ready-to-eat meat products. Precisely, 53.3 % (32/60), 56.7 % (17/30), 40 % (8/20), 55 % (11/20), 60 % (12/20), 13.3 % (4/30), and 15 % (3/20) of raw ground beef, beef burger, beef burger sandwiches, beef shawarma sandwiches, beef kofta sandwiches, beef luncheon, and beef sausage sandwiches, respectively were positive for H. pylori. Of the 204 biochemically-identified H. pylori isolates, 53.9 % (110/204) were molecularly confirmed by PCR through the detection of glmM, cagA, or vacA genes, which were detected at an incidence of 95.5 % (105/110), 77.3 % (85/110), and 20.9 % (23/110) among the isolates, respectively. The antimicrobial sensitivity testing revealed that all of the 110 (100 %) molecularly-confirmed H. pylori isolates were multidrug-resistant (MDR; resistant to four or more antibiotics). Interestingly, 100 % and 61.8 % of H. pylori isolated from raw and ready-to-eat meat were resistant to metronidazole and clarithromycin, respectively which consider alarming results as metronidazole and clarithromycin are the mainstay antibiotics in the treatment of H. pylori infections. Additionally, 94.5 %, 94.5 %, 24.5 %, 23.6 %, and 13.6 % of isolates were resistant to vancomycin, sulphamethoxazole-trimethoprim, imipenem, levofloxacin, and nitrofurantoin, respectively. The widespread contamination of examined raw and ready-to-eat meat product samples with MDR H. pylori isolates could constitute a tremendous public health hazard. Further studies concerning the prevalence and possible methods of elimination of H. pylori in different food categories distributed in the various provinces in Egypt as well as in other countries is required for a better understanding the H. pylori as an emerging foodborne pathogen.

Profile of Aliarcobacter spp. from edible giblets: Genetic diversity, antibiotic resistance, biofilm formation

Aliarcobacter spp. are recognized as emerging foodborne pathogens and consumption of foods contaminated with them can be a hazard to human and animal health. This study was conducted to investigate the prevalence of Aliarcobacter spp. in edible internal organs of different animal species from retail markets and giblet sellers. Additionally, this study was focused on the antimicrobial resistance, virulence profiles, biofilm-forming capabilities, and phylogenetic relationships of obtained isolates. A total of 270 samples were analyzed from which, 28 (10.4 %) were isolated as Aliarcobacter spp. by conventional methods. Within the 28 Aliarcobacter spp. isolates, 17 (60.7 %) were identified as A. butzleri, 10 (35.7 %) were A. cryaerophilus and one (3.5 %) was A. skirrowii by PCR method. The disc diffusion method showed that the highest resistance rate of Aliarcobacter spp. was seen against oxacillin (78.5 %), and 20 (71.4 %) out of the 28 isolates exhibited multidrug resistance (MDR). Out of the 28 isolates, mviN, pldA, tlyA, and hecB virulence genes were detected in 85.7 %, 46.4 %, 46.4 %, and 3.5 %, respectively, but irgA, Cj1349, ciaB, cadF, and hecA genes were not detected. According to the microplate test, 27 (96.4 %) isolates had weak biofilm ability while one A. cryaerophilus isolate (3.6 %) exhibited strong biofilm formation. ERIC-PCR band patterns suggested that isolated Aliarcobacter spp. from giblets, have different contamination sources. The presence of pathogenic and multidrug-resistant Aliarcobacter spp. in food poses a potential risk to public health and control measures throughout the food chain are necessary to prevent the spread of these strains.

A Serbian endemic plant „Nepeta rtanjensis“ Diklić et Milojević – chemical composition and biological activity

Nepeta rtanjensis Diklić et Milojević as an endemic and critically endangered plant in Serbia is the object of great scientific interest. N. rtanjensis is a rare plant of a limited distribution its application in traditional medicine is practically unknown. However, bioactive potential of 4aα,7α,7aβ-nepetalactone, the main compound from volatile fraction, as well as rosmarinic and 3-O-caffeoylquinic acid from non-volatile fraction is great. Scientists found that this plant possesses good antimicrobial activities against Aspergillus sp., Bacillus cereus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Listeria monocytogenes, Micrococcus flavus, Penicillium sp., Pseudomonas aeruginosa, Salmonella sp. and Staphylococcus aureus. N. rtanjensis can be used preserving foods and beverages as well as for prevention and treatment of diseases caused by common and emerging food borne pathogens. Furthermore, antioxidant, anticancer (cervix, breast and colon carcinoma, lung adenocarcinoma and myelogenous leukemia cells) and immunomodulating activity are also noted. However, toxicity in zebrafish model was not observed, so N. rtanjensis can be classified as safe for human health. Furthermore, phytotoxic potential against Ambrosia artemisiifolia as well as activity against plant pathogens (Alternaria sp., Cladosporium cladosporoides, Bipolaris spicifera and Trichoderma viridae) are also noted. There is potential fordeveloping novel biopesticide formulations.

Occurrence of Listeria spp. in Soft Cheese and Ice Cream: Effect of Probiotic Bifidobacterium spp. on Survival of Listeria monocytogenes in Soft Cheese.

Listeria monocytogenes is one of the most important emerging foodborne pathogens. The objectives of this work were to investigate the incidence of Listeria spp. and L. monocytogenes in soft cheese and ice cream in Assiut city, Egypt, and to examine the effect of some probiotic Bifidobacterium spp. (Bifidobacterium breve, Bifidobacterium animalis, or a mixture of the two) on the viability of L. monocytogenes in soft cheese. The existence of Listeria spp. and L. monocytogenes was examined in 30 samples of soft cheese and 30 samples of ice cream. Bacteriological analyses and molecular identification (using 16S rRNA gene and hlyA gene for Listeria spp. and L. monocytogenes, respectively) were performed on those samples. Additionally, Bifidobacterium spp. were incorporated in the making of soft cheese to study their inhibitory impacts on L. monocytogenes. Out of 60 samples of soft cheese and ice cream, 25 samples showed Listeria spp., while L. monocytogenes was found in only 2 soft cheese samples. Approximately 37% of soft cheese samples (11 out of 30) had Listeria spp. with about 18.0% (2 out of 11) exhibiting L. monocytogenes. In ice cream samples, Listeria spp. was presented by 47% (14 out of 30), while L. monocytogenes was not exhibited. Moreover, the addition of B. animalis to soft cheese in a concentration of 5% or combined with B. breve with a concentration of 2.5% for each resulted in decreasing L. monocytogenes efficiently during the ripening of soft cheese for 28 d. Listeria spp. is widely found in milk products. Probiotic bacteria, such as Bifidobacterium spp., can be utilized as a natural antimicrobial to preserve food and dairy products.