A Survey of Antibiotic Resistance in Micrococcaceae Isolated from Italian Dry Fermented Sausages

The transfer of bacteria that are resistant to antimicrobial agents or resistance genes from animals to humans via the food chain is increasingly a problem. Therefore, it is important to determine the species and the numbers of bacteria involved in this phenomenon. For this purpose, 148 strains of microstaphylococci were isolated from three types of Italian dry fermented sausages. Eight of 148 strains belonged to the genera Kocuria and Micrococcus. The remaining 140 strains belonged to 11 different species of the genus Staphylococcus. The species most frequently isolated was Staphylococcus xylosus, followed by Staphylococcus saprophyticus and Staphylococcus aureus. Antibiotic resistance levels differed among the species and depended on the strain origin. Microstaphylococci were generally susceptible to beta-lactams, but 12 strains were resistant to methicillin, 8 were resistant to oxacillin, and 9 were resistant to penicillin G. No resistance was observed for aminoglicosides and cephalosporines. Many strains were resistant to sulfonamide, colistin suphate, tetracyclin, and bacitracin. Two strains of S. aureus, four strains of S. xylosus, and one strain of Staphylococcus sciuri were able to grow in the presence of 8 microg of vancomycin per g, but all strains were susceptible to teicoplanin. Twenty-two microstaphylococci were resistant to at least five of the tested antibiotics. The multiresistant strain S. aureus 899 was unaffected by eight antibiotics, including vancomycin and methicillin, indicating that a more prudent use of antibiotics in animal husbandry and better hygienic conditions during production should be encouraged because they can play a major role in reducing the incidence of such multiresistant microorganisms and the possible spread of the genetic elements of their resistance

Effects of Starter Cultures and Type of Casings on the Microbial Features and Volatile Profile of Fermented Sausages

In the literature, the effect of the type of casing on fermented sausages is quite unexplored, while several studies are focused on the impact of starter cultures. Therefore, this paper studied the effect of three commercial starter cultures and two casings (natural or collagen) on Italian fermented sausages. Physico-chemical parameters (aw, pH, weight loss), microbiota, aroma profile and sensory analysis were evaluated. Results showed that collagen casings promoted a higher reduction of pH and weight loss. Concerning the microbiota, samples with natural casing had higher counts of lactic acid bacteria, while yeast proliferation was promoted in those with collagen. Regardless of the starters and casings applied, levels of enterococci and Enterobacteriaceae were low (≤2 log CFU/g). The aroma profile was significantly affected by casing: despite the starter applied, the presence of collagen casing favoured acid accumulation (mainly acetate and butanoate) and reduction of ketones. Sensory analysis highlighted significant differences only for odour, colour intensity and sourness. The differences observed suggest that collagen casings may provide a greater availability of oxygen. Overall, casings rather than starter cultures impact the microbial and sensorial features of fermented sausages

Numerical models of mass transfer during ripening and storage of salami

Ripening, in the dry sausages manufacturing process, has an influence over the main physical, chemical and microbiological transformations that take place inside these products and that define the final organoleptic properties of dry sausages. A number of study about the influence of ripening conditions on the main chemical and microbiological characteristics of dry sausages is available today. All these studies indicate that the final quality and safety standards achieved by the sausage manufacturing process can be considered to be strictly dependent from the specific ripening conditions. The water diffusion inside a seasoned sausage is surely an aspect of primary importance with regard to the quality of final product. As a consequence the aim of this research was to develop two parametric numerical models, concerning the moisture diffusion physics, describing salami ripening and storage. Mass transfer equations inside the sausage volume were numerically solved using a finite element technique. A first model describes diffusion phenomena occurring inside the salami and the exchange phenomena involving the surface of the product and the environment. After the ripening, the salami are stored in waterproof packaging, consequently an additional model able to describe also the evaporation and condensation phenomena occurring between the salami surface and the air in the package, was developed. The moisture equilibrium between salami surface and conservation atmosphere is mainly ruled by the temperature changes during storage. Both models allow to analyze the history of the moisture content inside the salami and are parametrised on product size and maturation/storage conditions. The models were experimentally validated, comparing the numerical outputs of the simulations with experimental data, showing a good agreement

Lactobacillus paracasei 4341 as adjunct culture to enhance flavor in short ripened Caciotta-type cheese

Caciotta is the name used to define a type of Italian semi-hard cheese Caciotta-type cheese. Due to the short ripening time, pasteurization is necessary to eliminate the potential pathogenic bacteria, which may be present in raw milk, causing also the reduction of ripened cheese flavor. The purpose of this research was to evaluate the effect of a selected wild Lactobacillus paracasei strain experimentally used as adjunct culture to enhance the flavour formation in a short-ripened caciotta-type cheese. An integrated polyphasic approach was used to compare the experimental and control Caciotta produced in a company located in Emilia Romagna region (Italy). It was demonstrated how the L. paracasei 4341 was able to develop in curd and cheese interacting with the acidifying commercial starter. The main acidifying starter species, were differently affected by the presence of the adjunct culture. Streptococcus thermophilus shown comparable behavior in all cheese-making step of control and experimental Caciotta, while Lactobacillus delbrueckii subsp bulgaricus, growth was slowed down by the presence of the adjunct culture during the whole ripening time. The higher amount of volatile compounds and organic acids due to the adjunct L. paracasei 4341 lead to a clear differentiation of the experimental Caciotta respect to the control, in terms of aromatic profile, color, texture and sensorial perception

Listeria monocytogenes sensitivity to antimicrobial treatments depends on cell origin

In this study we investigated how cell origin could affect the efficacy of an antimicrobial treatment (mild heating combined with terpenoids) in Listeria monocytogenes Scott A, considering cells from: 1. single colony, 2. glycerol stock, 3. cold adapted culture, and 4. fresh culture in stationary phase. After treatment, culturability on BHI medium and viability assessed by flow cytometry were evaluated. Our results showed that the cell origin significantly impacted viability and culturability of L. monocytogenes towards antimicrobial treatment. The mild heat treatment combined or not with terpenoids mainly affected culturability rather than viability, although the culturability of cells from single colony was less impacted. Therefore, to mimic the worst scenario, these latter were selected to contaminate Gorgonzola rind and roast beef slices and we evaluated the ability of L. monocytogenes cells to recover their culturability (on ALOA agar medium) and to growth on the food matrix stored at 4 °C for 7 days. Our results suggest that only Gorgonzola rind allowed a partial recovery of the culturability of cells previously heated in presence or not of terpens. In conclusion, we found a connection between the cell history and sensitivity toward an antimicrobial treatment, underlying the importance to standardize the experimental procedures (starting from the cells to be used in the assay) in the assessment of cell sensitivity to a specific treatment. Finally, our study clearly indicated that VBNC cells can resuscitate under favorable conditions on a food matrix, becoming a threat for consumer’s health

Microbial Spoilage of Traditional Goose Sausages Produced in a Northern Region of Italy

Recently, during the ripening of goose sausage, a defect consisting of ammonia and vinegar smell was noticed. The producer of the craft facility, located in Lombardia, a Northern region of Italy, asked us to identify the cause of that defect. Therefore, this study aimed to identify the potential responsible agents for the spoilage of this lot of goose sausages. Spoilage was first detected by sensory analysis using the "needle probing" technique; however, the spoiled sausages were not marketable due to the high ammonia and vinegar smell. The added starter culture did not limit or inhibit the spoilage microorganisms, which were represented by Levilactobacillus brevis, the predominant species, and by Enterococcus faecalis and E. faecium. These microorganisms grew during ripening and produced a large amount of biogenic amines, which could represent a risk for consumers. Furthermore, Lev. brevis, being a heterofermentative lactic acid bacteria (LAB), also produced ethanol, acetic acid, and a variation in the sausage colour. The production of biogenic amines was confirmed in vitro. Furthermore, as observed in a previous study, the second cause of spoilage can be attributed to moulds which grew during ripening; both the isolated strains, Penicillium nalgiovense, added as a starter culture, and P. lanosocoeruleum, present as an environmental contaminant, grew between the meat and casing, producing a large amount of total volatile nitrogen, responsible for the ammonia smell perceived in the ripening area and in the sausages. This is the first description of Levilactobacillus brevis predominance in spoiled goose sausage

Insights into the Metabolomic Diversity of Latilactobacillus sakei

: Latilactobacillus sakei (L. sakei), widely used as a starter culture in fermented sausages, is a species adapted to meat environments. Its ability to survive for a long time in such products is due to the exploitation of different metabolic pathways to gain energy (hexose and pentose sugar fermentation, amino acids catabolism, etc.). Since L. sakei demonstrates high phenotypic and metabolic strain biodiversity, in this work, a metabolomic approach was used to compare five strains of different origins. They were cultivated in a defined medium with glucose or ribose at two concentrations, and analyzed through nuclear magnetic resonance (1H-NMR) spectroscopy to monitor amino acid consumptions and accumulation of organic acids and aroma compounds. The results showed that all the strains were able to use arginine, especially when cultivated with ribose, while serine was consumed mainly in the presence of glucose. Aroma compounds (i.e., diacetyl and acetoin) were mainly accumulated in samples with ribose. These aspects are relevant for starter cultures selection, to confer specific features to fermented sausages, and to optimize the fermentations. Moreover, the use of 1H-NMR allowed the fast identification of different classes of compounds (without derivatization or extraction procedures), providing a powerful tool to increase the knowledge of the metabolic diversity of L. sakei

Pedunculated Angiomyofibroblastoma of the Vulva: Case Report and Review of the Literature

Angiomyofibroblastoma (AMFB) is a rare benign mesenchymal tumour that occurs almost exclusively in the vulvovaginal region of women but can also occur occasionally in the inguinoscrotal region of men. It is a well-circumscribed lesion that clinically is often thought to represent a Bartholin's gland cyst and usually does not form a pedunculated mass. To our knowledge, only five cases of vulvar AMFB with pedunculated mass have been reported in the English literature and all cases involving the labia majora and middle-aged women. We report the first case of pedunculated AMFB of the vulva occurring in a young woman of 21 years old and involving the left labia minora. After excluding the most common diseases, pedunculated AMFB should be part of differential diagnosis in the workup of any pedunculated vulvar mass even in young women with a lesion involving the labia minora. We reviewed the literature and summarized all reported cases

Survival, growth, and biogenic amine production of Enterococcus faecium FC12 in response to extracts and essential oils of Rubus fruticosus and Juniperus oxycedrus

Enterococci are lactic acid bacteria (LAB) usually found as food contaminants in fermented products such as cheeses and fermented sausages. Due to their antibiotic resistance, the presence of virulence factors, and the ability to produce biogenic amines (BAs), the determination of these bacteria is crucial to assure food quality and safety. BAs production and consequent accumulation in foods can cause toxicological eects on human health. Plant phenolic compounds are promising alternatives to chemical preservatives and reflect consumers’ demand for “green” solutions. In this study, the antimicrobial eect of blackberry (Rubus fruticosus) leaves and prickly juniper (Juniperus oxycedrus) needles, both as phenolic extracts (PE) and essential oils (EO), were evaluated against Enterococcus faecium FC12, a known tyramine-producing strai

A survey of yeasts in traditional sausages of southern Italy

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