Correction: Jarzynka, S. et al. Combination of High-Pressure Processing and Freeze-Drying as the Most Effective Techniques in Maintaining Biological Values and Microbiological Safety of Donor Milk. Int. J. Environ. Res. Public Health 2021, 18, 2147.

There was an error in the original publication [...].

Comparison of Phenotype and Genotype Virulence and Antimicrobial Factors of Salmonella Typhimurium Isolated from Human Milk.

Salmonella is a common foodborne infection. Many serovars belonging to Salmonella enterica subsp. enterica are present in the gut of various animal species. They can cause infection in human infants via breast milk or cross-contamination with powdered milk. In the present study, Salmonella BO was isolated from human milk in accordance with ISO 6579-1:2017 standards and sequenced using whole-genome sequencing (WGS), followed by serosequencing and genotyping. The results also allowed its pathogenicity to be predicted. The WGS results were compared with the bacterial phenotype. The isolated strain was found to be Salmonella enterica subsp. enterica serovar Typhimurium 4:i:1,2_69M (S. Typhimurium 69M); it showed a very close similarity to S. enterica subsp. enterica serovar Typhimurium LT2. Bioinformatics sequence analysis detected eleven SPIs (SPI-1, SPI-2, SPI-3, SPI-4, SPI-5, SPI-9, SPI-12, SPI-13, SPI-14, C63PI, CS54_island). Significant changes in gene sequences were noted, causing frameshift mutations in yeiG, rfbP, fumA, yeaL, ybeU (insertion) and lpfD, avrA, ratB, yacH (deletion). The sequences of several proteins were significantly different from those coded in the reference genome; their three-dimensional structure was predicted and compared with reference proteins. Our findings indicate the presence of a number of antimicrobial resistance genes that do not directly imply an antibiotic resistance phenotype.

Surface Hygiene Evaluation Method in Food Trucks as an Important Factor in the Assessment of Microbiological Risks in Mobile Gastronomy.

Street food outlets are characterised by poor microbiological quality of the food and poor hygiene practices that pose a risk to consumer health. The aim of the study was to evaluate the hygiene of surfaces in food trucks (FT) using the reference method together with alternatives such as PetrifilmTM and the bioluminescence method. TVC, S. aureus, Enterobacteriaceae, E. coli, L. monocytogenes, and Salmonella spp. were assessed. The material for the study consisted of swabs and prints taken from five surfaces (refrigeration, knife, cutting board, serving board, and working board) in 20 food trucks in Poland. In 13 food trucks, the visual assessment of hygiene was very good or good, but in 6 FTs, TVC was found to exceed log 3 CFU/100 cm2 on various surfaces. The assessment of surface hygiene using various methods in the food trucks did not demonstrate the substitutability of culture methods. PetrifilmTM tests were shown to be a convenient and reliable tool for the monitoring of mobile catering hygiene. No correlation was found between the subjective visual method and the measurement of adenosine 5-triphosphate. In order to reduce the risk of food infections caused by bacteria in food trucks, it is important to introduce detailed requirements for the hygiene practices used in food trucks, including techniques for monitoring the cleanliness of surfaces coming into contact with food, in particular cutting boards and work surfaces. Efforts should be focused on introducing mandatory, certified training for food truck personnel in the field of microbiological hazards, appropriate methods of hygienisation, and hygiene monitoring.

Factors Influencing Biofilm Formation by Salmonella enterica sv. Typhimurium, E. cloacae, E. hormaechei, Pantoea spp., and Bacillus spp. Isolated from Human Milk Determined by PCA Analysis.

Bacteria enter milk during poor hygiene practices and can form a biofilm on surfaces that come into contact with human milk. The presence of a biofilm increases the risk of infections among newborns as bacteria protected by biofilm are resistant to washing and disinfection processes. The formation of the biofilm depends on the microbial species, environmental conditions, and the specific materials colonized. The aim of this study is to analyze the effects of factors such as temperature, incubation time, and initial cell concentration on biofilm formation by pathogenic bacteria isolated from human milk on model hydrophobic polystyrene surfaces. Model studies confirm that pathogenic bacteria appearing in human milk as a result of cross-contamination tend to form a biofilm. The majority of isolates formed biofilm at both 25 and 37 °C after 12 h at 1 × 103 CFU/mL inoculum count. Multivariate principal component analysis (PCA) showed that at lower temperatures, biofilm formation by bacterial isolates was the main determinant of biofilm formation, other factors were less important; however, at 37 °C, time was a factor in biofilm formation. The model research performed underlines the importance of maintaining the proper hygiene of rooms, surfaces, and devices for expressing, storing, and preparing mothers' milk and powdered infant formula (PIF) in facilities responsible for feeding newborns and premature babies.

An Evaluation of the Hygiene Practices of Polish Street Food Vendors in Selected Food Trucks and Stands.

Today, street food vending is becoming a dynamically developing food industry in Europe, including Poland. Lifestyle changes and socioeconomic factors, especially among young consumers, make it a convenient food alternative, even in countries without this tradition. The aim of the study was to evaluate hygiene conditions and practices in selected street food facilities in Poland. The study was carried out in accordance with an observation checklist developed on the basis of the hygienic requirements of the European Union. The study covered 550 randomly selected street food vendors in Poland in 10 cities. The hygiene of street food outlets was assessed in three aspects: ensuring proper production conditions, hygiene of production and distribution, and hygiene of personnel. The evaluation of street food outlets showed that the level of hygiene was not fully acceptable. A small percentage of the examined objects satisfactorily fulfilled the requirements of the production conditions, production and distribution hygiene, and staff hygiene. The proposed hygienic checklist for quick daily monitoring of street food outlets can be used to perform internal and external inspections. It seems that specific codes of conduct for European street foods facilities are necessary.

Effect of Herbal Addition on the Microbiological, Oxidative Stability and Sensory Quality of Minced Poultry Meat.

The study aimed to assess the effect of herbal additions with antioxidant properties (pepper, thyme and oregano) on the microbiological and oxidative stability as well as the sensory quality of minced poultry meat. Meatballs treatments without additives and treatments with the addition of three types of spices in two forms-dry spices and industrial extracts were examined. Popular seasoning additives of oregano (Origanum vulgare), thyme (Thymus vulgaris) and black pepper (Piper nigrum) at 0.3% of dry herbal or 0.003% as industrial extract were added to minced meat. The microbiological, chemical and sensory tests were performed at specified times and storage temperature. Based on the studied criteria, products maintained constant and adequate quality by up to 10 days while stored in 4 °C. In the case of all tested samples, the overall sensory quality began to deteriorate after 10 days of storage. The preservative role of herbs and extracts in meat products during processing and storage was observed. Oregano and black pepper in both forms maintained good microbial quality and showed their inhibitory effects on the growth of psychrotrophic bacteria. It was observed that dried herbs revealed a stronger antioxidant effect than additives in the form of extracts. The studied dried herbs played an antioxidant, antimicrobial and preservative role in meat products during processing and storage.

Cluster Analysis Classification of Honey from Two Different Climatic Zones Based on Selected Physicochemical and of Microbiological Parameters.

The geographical origin of honey affects its composition, which is of key importance for the health-promoting properties and safety of the product. European regulations clearly define the physicochemical requirements for honey that determine the microbiological quality. On the other hand, legislation abolishes microbiological criteria. In the study 40 honey samples originating from two different climatic zones were analyzed. The water content, pH, water activity analysis and the microbiological quality of honey samples have been tested using the reference plate method (total viable count, yeast and molds, lactic acid bacteria, Bacillus spp.). The cluster classification showed that total viable count of bacteria could be used as a measure alternative to the count of Bacillus spp. and 70% of honeys from the tropical climate zone had different microbiological quality than honeys from the temperate climate zone but still under the level 3.0 log cfu/g. The study has revealed that geographical origin of honey may significantly affect the quality and safety of honey. It was considered that water content can be the most informative and handy marker of the microbiological quality of honeys. Analysis of lactic acid bacteria showed temperate climate zone honeys as a source of beneficial bacteria in the diet.

Combination of High-Pressure Processing and Freeze-Drying as the Most Effective Techniques in Maintaining Biological Values and Microbiological Safety of Donor Milk.

BACKGROUND: Human milk banks have a pivotal role in provide optimal food for those infants who are not fully breastfeed, by allowing human milk from donors to be collected, processed and appropriately distributed. Donor human milk (DHM) is usually preserved by Holder pasteurization, considered to be the gold standard to ensure the microbiology safety and nutritional value of milk. However, as stated by the European Milk Banking Association (EMBA) there is a need to implement the improvement of the operating procedure of human milk banks including preserving and storing techniques. AIM: The purpose of this study was to assess the effectiveness and safety of the selected new combination of methods for preserving donor human milk in comparison with thermal treatment (Holder pasteurization). METHODS: We assessed (1) the concentration of bioactive components (insulin, adiponectin, leptin, activity of pancreatic lipase, and hepatocyte growth factor) and (2) microbiological safety in raw and pasteurized, high-pressure processed and lyophilization human breast milk. RESULTS: The combination of two techniques, high-pressure processing and freeze-drying, showed the best potential for preserving the nutritional value of human milk and were evaluated for microbiological safety. Microbiological safety assessment excluded the possibility of using freeze-drying alone for human milk sample preservation. However, it can be used as a method for long-term storage of milk samples, which have previously been preserved via other processes. CONCLUSION: The results show that high-pressure treatment is the best method for preservation that ensures microbiological safety and biological activity but subsequent freeze-drying allowed long-term storage without loss of properties.

Effect of Nonthermal Processing on Human Milk Bactericidal Activity Against Escherichia coli.

Nonthermal methods are more efficient at preserving various biological properties of human milk, as compared with holder pasteurization (HoP), which is the most common preservation method. This study was performed to assess the effects of nonthermal processing on bactericidal activity against Escherichia coli in human milk. Milk samples obtained from the Regional Human Milk Bank in Warsaw at Holy Family Hospital were processed by HoP, irradiated with ultraviolet-C (UV-C) for 5, 10, and 15 minutes (6720 J/L each minute), subjected to 2 variations of high-pressure processing (HPP): 450 MPa for 15 minutes and 200 MPa for 10 minutes + 400 MPa for 10 min, with a 10-minutes break. The samples were then evaluated by a bactericidal assay (raw untreated human milk was used as a control). The bactericidal capacity after HoP was preserved in 12.1% of samples, showing a significant reduction in bactericidal properties compared with in raw milk (P < 0.05). The differences between samples preserved by nonthermal methods and raw milk were not significant (P > 0.05). Nonthermal methods of human milk treatment better preserve the bactericidal capacity compared with holder pasteurisation. Those alternative technologies to HoP can be proposed after further investigation for milk processing for Human Milk Banks facilities.

225Ac- and 213Bi-Substance P Analogues for Glioma Therapy.

Within the last decades, there has been no major improvement in treatment of patients with glioma, especially with glioblastoma multiforme (GBM) which is related to specific features of this tumor type, such as heterogeneity at the macroscopic, microscopic and genetic level, the infiltrative nature of tumors and the obstacle of the brain-blood barrier which limits the accessability of most drugs. The current standard of care is surgical resection, followed by radio- and chemotherapy. After first-line treatment of the primary lesion, tumor recurrence is diagnosed in virtually all GBM patients. Treatment of tumor recurrence represents a challenging clinical task. Surgical resection to relief symptoms of mass effect and/or salvage chemotherapy are often considered as last therapeutic option. A new treatment option is urgently needed. Targeted alpha therapy with an intratumoral injection of 213Bi-DOTA-Substance P (SP) or 225Ac-DOTAGA-Substance P has been introduced into the therapeutic armamentarium of recurrent GBM. There are many advantages of using SP such as very high prevalence of increased NK-1 expression in GBM cells, regardless of the degree of malignancy, and expression of the NK-1 receptor system not only on the membrane of cancer cells but also strong expression of NK1 receptors within the tumor neovasculature suggesting concomitant targeting of vascular and neoplastic structures. Radioisotopes with different physical properties, mainly beta-emitting metallic radionuclides, were implemented for brain tumor treatment. Based on their radiophysical properties, however, alpha emitters exhibit more promising properties. In investigator-initiated phase I and II studies, targeted alpha therapy using Bi-213/Ac-225 radiolabeled Substance P for malignant gliomas compare favorably with standard therapy, with the limitation that no large controlled series have so far been generated. Further development should focus on the improvement of the biological and chemical properties of the compound and the application by dedicated catheter systems to improve the intratumoral distribution of the radiopharmaceutical within growth and infiltrative zone of these glial neoplasms.

New Achievements in High-Pressure Processing to Preserve Human Milk Bioactivity.

High-pressure processing (HPP) is a non-thermal technology that is being increasingly applied in food industries worldwide. It was proposed that this method could be used as an alternative to holder pasteurization (HoP; 62.5°C, 30 min) in milk banks but its impact on the immunologic, enzymatic and hormonal components of human milk has not yet been evaluated in detail. The aim of our study was to compare the effects of HPP in variants: (1) 600 MPa, 10 min (2) 100 MPa, 10 min, interval 10 min, 600 MPa, 10 min (3) 200 MPa, 10 min, interval 10 min, 400 MPa, 10 min (4) 200 MPa, 10 min, interval 10 min, 600 MPa, 10 min in temperature range 19-21°C and HoP on the leptin, adiponectin, insulin, hepatocyte growth factor (HGF), lactoferrin and IgG contents in human milk. HoP was done at the Regional Human Milk Bank in Warsaw at the Holy Family Hospital on S90 Eco pasteurizer (Sterifeed, Medicare Colgate Ltd). Apparatus U4000/65 (Unipress Equipment, Poland) was used for pascalization. Milk samples were obtained from women during 2-6 weeks of lactation. Post-treatment culture showed no endogenous bacterial contamination in any tested option. Concentrations of selected components were determined using ELISA tests. The level of all analyzed components were significantly decreased by HoP: leptin 77.86%, adiponectin 32.79%, insulin 32.40%, HGF 88.72%, lactoferrin 60.31@.%, IgG 49.04%. All HPP variants caused an increase in leptin concentration, respectively (1) 81.79% (2) 90.01% (3) 86.12% (4) 47.96%. Retention of insulin after HPP was (1) 88.20% (2) 81.98% (3) 94.76% (4) 90.31% HGF (1) 36.15% (2) 38.81% 97.15% (3) 97.15% (4) 43.02%, lactoferrin (1) 55.78% (2) 57.63% (3) 78.77% (4) 64.75%. Moreover, HPP variant as 200 + 400 MPa preserved IgG (82.24%) better than HoP and resulted not statistically significant change of adiponectin level (38.55%) compare to raw milk. Our results showed that HPP leads to preservation of adipokines, growth factor, and lactoferrin, IgG much better or comparable with HoP.

NK cell count and glucotransporter 4 (GLUT4) expression in subjects with type 2 diabetes and colon cancer.

BACKGROUND: Type 2 diabetes (T2D) and colon cancer (CC) are numbered among the most common diseases in the world. The decreased activity of natural killer (NK) cells previously revealed in both mentioned pathological states may be correlated with impaired expression of GLUT4 as the major insulin-dependent glucose transporter in these cells. METHODS: The aim of this study was to evaluate GLUT4 expression and NK cells number in subjects with T2D and/or CC in comparison with control group. We evaluated 78 individuals divided into four groups: (1) patients with CC and T2DM, (2) patients with CC, (3) patients with T2DM (4) healthy control. GLUT4 expression on the surface of NK cells was measured using flow cytometry and phenotyping of NK cell was performed by immunofluorescent method. RESULTS: Subjects with diabetes had the highest GLUT4 expression (21.35 ± 7.2 %) in comparison with other groups (P < 0.01). The mean values of GLUT4 expression in group with CC and in patients with both T2D and CC were similar (1.4 ± 0.4 % vs 1.5 ± 1.0 %; respectively). These values were significantly lower than in control group (12.6 ± 2.9 %; P < 0.01). In patients with T2D and CC the number of NK cells (20.15 ± 6.6 %) was significantly higher than in other groups, i.e. in group with T2D (14.08 ± 5.7 %), in group with CC (9.21 ± 3.6 %) and in control group (9.48 ± 4.7 %), respectively (P < 0.01). CONCLUSIONS: It seems that there is a need to pay more attention to the high incidence of colon cancer among patients with type 2 diabetes. Decreased GLUT4 expression observed on NK cells in patients with colon cancer may be responsible for dysfunction of these cells and the higher carcinogenic risk in type 2 diabetic subjects.