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1.
Hig. aliment ; 36(294): e1071, jan.-jun. 2022. ilus
Artigo em Português | VETINDEX | ID: biblio-1363221

RESUMO

A pesquisa teve o objetivo de avaliar as condições higiênicas e sanitárias de comercialização do queijo de coalho por vendedores ambulantes na praia de Copacabana, por meio de inspeção visual e análises microbiológicas. Para o estudo foram coletadas quinze amostras de cinco diferentes ambulantes e estas foram encaminhadas para o Centro Estadual de Pesquisa em Qualidade de Alimentos da Empresa de Pesquisa Agropecuária do Estado do Rio de Janeiro (PESAGRO-RIO/CEPQA) onde foram submetidas às análises microbiológicas. O resultado da avaliação visual das condições higiênico-sanitárias demonstrou 100% de não conformidades em relação à adoção das Boas Práticas de Fabricação por parte dos manipuladores. Os resultados das contagens de Estafilococos coagulase positiva e de Coliformes termotolerantes se apresentaram acima do limite preconizado pela legislação vigente em 75% e 6,7% das amostras respectivamente. Em 25% das amostras foi verificada a presença de Salmonella spp. O estudo demonstrou que o queijo de coalho vendido por ambulantes da praia de Copacabana estava impróprio para o consumo podendo representar risco à saúde dos consumidores.(AU)


The research aimed to evaluate the hygienic and sanitary commercialized conditions of coalho cheese by street vendors on Copacabana beach, through visual inspection and microbiological analysis. For conducting the study, fifteen samples were collected from five different street vendors and these were analyzed at the State Center for Research in Food Quality of the Agriculture Research Company of the State of Rio de Janeiro (PESAGRO-RIO/CEPQA). The result of the visual assessment of hygienic-sanitary conditions demonstrated 100% of non-conformities in relation to the adoption of Good Manufacturing Practices by the manipulators. In counts of coagulase positive Staphylococcus and Thermotolerant Coliforms were above the limit recommended by the current legislation in 75% and 6.7% of the samples, respectively. Salmonella spp. was present in 25% of the analyses.The study showed that the coalho cheese sold by street vendors on Copacabana beach was unfit for consumption and could pose a risk to consumer health.(AU)


Assuntos
Salmonella/isolamento & purificação , Queijo/microbiologia , Coagulase/isolamento & purificação , Coliformes/isolamento & purificação , Brasil , Padrão de Identidade e Qualidade para Produtos e Serviços
2.
Food Microbiol ; 105: 104023, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35473976

RESUMO

Canastra Cheese is one of the most commercialized artisanal cheeses in Brazil and intrinsic characteristics of its production, such as the use of raw milk and natural whey starter cultures as well short ripening time on wooden shelves, offer risk of contamination by a plethora of microorganisms. Here, we used 16 S rRNA gene amplicon sequencing approach to characterize the bacterial communities from Canastra cheese processing environments and final products, accessing cheesemaking facilities with distinct profiles of Food Safety Management Systems (FSMS), in order to estimate whether differences in microbial composition and diversity could also be observed between the two sampled groups of facilities. Our results revealed that the diversity of bacterial communities in the processing environments was much higher than that observed for cheeses, with greater discrepancy for facilities with inadequate FSMS. Additionally, in facilities with inadequate FSMS the bacterial communities from environments, especially hand surfaces and ripening wooden shelves, were similar to those during processing and finished cheese. These evidences highlight the importance of implementing and maintaining FSMS in the facilities, in order to assure quality and safety of Canastra cheese, but also the stability and economic viability of the Canastra cheese production chain.


Assuntos
Queijo , Bactérias/genética , Queijo/microbiologia , Laticínios , Análise de Perigos e Pontos Críticos de Controle , Gestão da Segurança
3.
Microb Biotechnol ; 15(5): 1404-1421, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35393728

RESUMO

Ethical, environmental and health concerns around dairy products are driving a fast-growing industry for plant-based dairy alternatives, but undesirable flavours and textures in available products are limiting their uptake into the mainstream. The molecular processes initiated during fermentation by lactic acid bacteria in dairy products is well understood, such as proteolysis of caseins into peptides and amino acids, and the utilisation of carbohydrates to form lactic acid and exopolysaccharides. These processes are fundamental to developing the flavour and texture of fermented dairy products like cheese and yoghurt, yet how these processes work in plant-based alternatives is poorly understood. With this knowledge, bespoke fermentative processes could be engineered for specific food qualities in plant-based foods. This review will provide an overview of recent research that reveals how fermentation occurs in plant-based milk, with a focus on how differences in plant proteins and carbohydrate structure affect how they undergo the fermentation process. The practical aspects of how this knowledge has been used to develop plant-based cheeses and yoghurts is also discussed.


Assuntos
Queijo , Produtos Fermentados do Leite , Lactobacillales , Queijo/microbiologia , Laticínios , Fermentação , Aromatizantes/metabolismo , Lactobacillales/genética , Lactobacillales/metabolismo , Iogurte/microbiologia
4.
Food Microbiol ; 104: 103979, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35287808

RESUMO

The behaviour of Listeria monocytogenes was investigated in soft pasteurized milk cheese elaborated with different salt concentrations (1.17 and 0.30% w/w) and in cured raw sheep milk cheese over storage up to 189 days at different isothermal conditions. Commercial 25-g cheese samples were inoculated with a 4-strain cocktail of L. monocytogenes (serovars 4b, 1/2a, 1/2b and 1/2c) at approximately 104 CFU/g. The inoculated samples were stored at 4 and 22 °C and withdrawn at proper intervals for L. monocytogenes enumeration. The prevalence of the different serovar strains of L. monocytogenes was characterized on soft cheese samples over storage at 4 °C using multiplex PCR. Salt reduction did not affect the survival of L. monocytogenes in soft cheeses and a maximum of 1-log reduction was observed in both regular and low-salt cheeses after 189 days of storage at 4 °C. The pathogen showed greater survival capacity in both soft and cured cheeses during storage at 4 °C compared to the storage at 22 °C, where more than 2.5 log reductions were computed. The fate of L. monocytogenes was described through a Weibull model fitted to survival data. The time required for a first tenfold reduction of the L. monocytogenes population (δ) at 4 °C is around 150 days in soft and 72 days in cured cheeses. At 22 °C, the estimated δ values are at least 60% lower in both cheese types. Among the four L. monocytogenes serovars present in the inoculated cocktail, the serovar 4b strain was the most sensitive to refrigerated storage, while the prevalence of serovar 1/2c strain increased over time in soft cheeses. Overall, the data obtained in this study help to deepen knowledge into factors affecting L. monocytogenes behaviour on cheeses and evidenced the variability between serovars in terms of survival capacity, which may be considered when performing microbial risk assessments.


Assuntos
Queijo , Armazenamento de Alimentos , Listeria monocytogenes , Animais , Queijo/análise , Queijo/microbiologia , Microbiologia de Alimentos , Listeria monocytogenes/classificação , Listeria monocytogenes/genética , Listeria monocytogenes/fisiologia , Ovinos , Temperatura , Fatores de Tempo
5.
Food Microbiol ; 104: 104006, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35287824

RESUMO

Pink discoloration defect can cause economic losses for cheese producers due to the impossibility to sell the defected cheese, but few knowledge is currently available on the causes of this defect. To gain more insight on the causes that lead to the formation of pink discoloration in Pecorino Toscano cheese with the Protected Designation of Origin (PDO) status, the bacterial community in defected and not defected cheese was characterized by high-throughput sequencing of bacterial 16S rRNA gene. The bacterial community in the defected cheese significantly differed compared to the control. The relative abundance of the genera Acidipropionibacterium, Enterococcus, Escherichia/Shigella, Lactobacillus, Lentilactobacillus and Propionibacterium was higher in the cheese with pink discoloration defect. The concentration of short chain fatty acids and of lactic acid in cheese was measured and a shift towards the production of propionate in the cheese with pink discoloration defect was observed. Furthermore, the possible involvement of microbially produced vitamin B12 in the formation of pink discoloration was not supported by the data, since a tendency to a lower concentration of vitamin B12 was measured in the defected cheese compared to the control.


Assuntos
Queijo , Microbiota , Queijo/microbiologia , Lactobacillaceae/genética , Lactobacillus/genética , RNA Ribossômico 16S/genética
6.
Comp Immunol Microbiol Infect Dis ; 84: 101785, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35276464

RESUMO

Sanitary-hygienic failures in cheese making can pose health risks to consumers. This study aimed to identify multiresistant pathogens in different production stages of artisanal goat coalho cheese in Brazil and characterize their phenotypic and genotypic resistance. Eleven properties in the state of Pernambuco, Brazil, participated in the study. Samples were obtained from different stages of production and the humans involved. The samples obtained were submitted to microbiological culture, then all the isolated microorganisms were submitted to the Matrix Associated Laser Desorption-Ionization - Time of Flight technique for the microbiological identification of the species. Subsequently, Staphylococcus spp., Enterococcus spp. and Macrococcus caseolyticus were subjected to polymerase chain reaction to search for resistance genes and disc diffusion technique to evaluate the resistance profile. A total of 111 isolates were obtained and 31 species were identified, with the frequency of Staphylococcus spp. (62.20%; 69/111), Enterococcus spp. (11.60%; 13/111), Macrococcus caseolyticus (10%; 11/111), Bacillus spp. (3.60%; 4/111), Enterobacter spp. (3.60%; 4/111), Aureobasidium pullulans (1.80%; 2/111), Corynebacterium camporealensis (1.80%; 2/111), Issatchenkia occidentalis (1.80%; 2/111), Kocuria kristinae (1.80%; 2/111), Aerococcus viridans (0.90%; 1/111) and Filifactor villosus (0.90%; 1/111). Phenotypic and genotypic resistance was also detected with the occurrence of 15.90% (7/44) of the mecA gene, 4% (1/25) vanA, and 4% (1/25) vanB in Staphylococcus spp. and 20% (2/10) vanB in and Enterococcus spp. Emerging multiresistant pathogens are present in the production chain of artisanal goat cheese and humans, who exert an important role in disseminating these bacteria with imminent risks to human health.


Assuntos
Queijo , Animais , Brasil/epidemiologia , Queijo/análise , Queijo/microbiologia , Enterococcus/genética , Cabras , Staphylococcaceae , Staphylococcus
7.
Foodborne Pathog Dis ; 19(5): 316-323, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35263183

RESUMO

Bacteria develop resistance to antibiotics naturally, but the inappropriate and widespread use of antibiotics in humans and animals has made antimicrobial resistance one of the biggest threats to modern medicine. Raw milk cheese can represent an important source of antimicrobial resistance. Thus, the objective of this study was to evaluate the prevalence and sensitivity of Escherichia coli isolated from artisanal cheese made from raw milk produced in Minas Gerais, Brazil. E. coli counts were determined using the most probable number method. An antibiogram was performed using the disk diffusion method, following the protocol described by the Brazilian Committee on Antimicrobial Susceptibility Testing (BrCAST) for 14 antibiotics of nine classes. E. coli was detected in 35 (71.4%) of the samples, with populations between 0.56 to 4.87 log (NMP/g) of cheese. The presence of E. coli resistant to multiple antimicrobials was more frequent in cheeses, with an E. coli population below the levels established by regulatory limits. Only four samples (11.4%) had all E. coli isolates susceptible to the 14 antimicrobials evaluated. The results showed the heterogeneity of antimicrobial resistance in E. coli between the producing regions of Minas artisanal cheese. Multidrug resistance was detected in 29% of the E. coli isolates and in almost 40% (38.8%) of the cheese samples. The frequency of multidrug-resistant (MDR) isolates was different between the production regions (p < 0.05). The presence of MDR E. coli in cheese from region D was 14, 4, and 20 times more likely than in cheese from regions A, B, and C, respectively. A multiple antibiotic resistance index of 0.200 predicted the presence of MDR E. coli in raw milk artisanal cheese with 99% probability. In conclusion, artisanal cheese can act as sources of MDR E. coli to colonize the human gastrointestinal tract.


Assuntos
Queijo , Animais , Antibacterianos/farmacologia , Queijo/microbiologia , Escherichia coli , Testes de Sensibilidade Microbiana , Leite/microbiologia
8.
Int J Food Microbiol ; 368: 109618, 2022 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-35279452

RESUMO

Interactions among microorganisms deeply affect the dynamics of cheese microbial communities and, as a consequence, multiple aspects of cheese quality, from the production of metabolites affecting the taste, aroma and flavour, to body, texture and colour. Understanding and exploiting interactions among beneficial or detrimental microorganisms is therefore key to managing cheese quality. This is true for the simplest systems (fresh cheeses produced from pasteurized milk using defined starters) and the more so for complex, dynamic systems, like surface ripened cheese produced from raw milk, in which a dynamic succession of diverse microorganisms is essential for obtained the desired combination of sensory properties while guaranteeing safety. Positive (commensalism, protocooperation) and negative (competition, amensalism, predation and parasitism) interactions among members of the cheese biota have been reviewed multiple times. However, even if the complex, multidimensional datasets generated by multi-omic approaches to cheese microbiology and biochemistry are ideally suited for the representation of biotic and metabolic interactions as networks, network science concepts and approaches are rarely applied to cheese microbiology. In this review we illustrate concepts relevant to the description of microbial interactions using a network science framework. Then, we briefly review methods used for the inference and analysis of microbial association networks (MAN) and their potential use in the interpretation of the cheese interactome. Finally, since these methods can only be used for mining microbial associations, we review the experimental methods used to confirm the nature of microbial interactions among cheese microbes.


Assuntos
Queijo , Microbiota , Animais , Queijo/microbiologia , Microbiologia de Alimentos , Humanos , Leite/microbiologia , Paladar
9.
Toxins (Basel) ; 14(2)2022 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-35202161

RESUMO

Cheese represents a dairy product extremely inclined to fungal growth and mycotoxin production. The growth of fungi belonging to Aspergillus, Penicillium, Fusarium, Claviceps, Alternaria, and Trichoderma genera in or on cheese leads to undesirable changes able to affect the quality of the final products. In the present investigation, a total of 68 types of commercial and traditional Slovak cheeses were analyzed to investigate the occurrence of fungal metabolites. Altogether, 13 fungal metabolites were identified and quantified. Aflatoxin M1, the only mycotoxin regulated in milk and dairy products, was not detected in any case. However, the presence of metabolites that have never been reported in cheeses, such as tryptophol at a maximum concentration level from 13.4 to 7930 µg/kg (average: 490 µg/kg), was recorded. Out of all detected metabolites, enniatin B represents the most frequently detected mycotoxin (0.06-0.71 µg/kg) in the analyzed samples. Attention is drawn to the lack of data on mycotoxins' origin from Slovak cheeses; in fact, this is the first reported investigation. Our results indicate the presence of fungal mycotoxin contamination for which maximum permissible levels are not established, highlighting the importance of monitoring the source and producers of contamination in order to protect consumers' health.


Assuntos
Queijo/análise , Queijo/microbiologia , Depsipeptídeos/análise , Contaminação de Alimentos/análise , Indóis/análise , Micotoxinas/análise , Metabolismo Secundário , Eslováquia
10.
J Dairy Sci ; 105(4): 2931-2947, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35123784

RESUMO

The yeasts involved in the ripening process of artisanal soft raw ewe milk Protected Designation of Origin (PDO) Torta del Casar and Queso de la Serena cheeses produced in Extremadura, Spain, were isolated throughout their ripening process, strain typed, and characterized for some important technological properties. A total of 508 yeast isolates were obtained and identified by inter-single sequence repeat anchored PCR amplification analysis and subsequent sequencing of the internal transcribed spacer ITS1/ITS2 5.8S rRNA. A total of 19 yeast species representing 8 genera were identified. Debaryomyces hansenii, Pichia kudriavzevii, Kluyveromyces lactis, and Yarrowia lipolytica were the predominant species. We selected 157 isolates, by genotyping and origin, for technological characterization. The evaluation of yeast isolates' growth under stress conditions of cheese ripening showed that 87 presented better performance. Among them, 71 isolates were not able to catabolize tyrosine to produce a brown pigment. Principal component analysis of the biochemical features of these isolates showed that 9 strains stood out, 3 K. lactis strains (2287, 2725, and 1507), 2 Pichia jadinii (1731 and 433), 2 Yarrowia alimentaria (1204 and 2150), Y. lipolytica 2495 and P. kudriavzevii 373. These strains displayed strong extracellular proteolytic activity on skim milk agar as well as an adequate enzymatic profile (strong aminopeptidase and weak protease activity), suggesting their great potential for cheese proteolysis. Extracellular lipolytic activity was mainly restricted to Yarrowia spp. isolates and weakly present in P. kudriavzevii 373 and K. lactis 2725, although enzymatic characterization by API-ZYM (bioMérieux SA) evidenced that all may contribute, at least in part, to the lipolysis process. Moreover, these strains were able to assimilate lactose, galactose, and glucose at NaCl concentrations higher than that usually found in cheese. However, lactate and citrate assimilation were limited to Y. lipolytica 2495, P. kudriavzevii 373, and P. jadinii 433, and may contribute to the alkalinizing process relevant to biochemical processes that take place in the last stages of ripening. By contrast, K. lactis strains showed acidifying capacity and ß-galactosidase activity and may take part in the initial stages of ripening, together with lactic acid bacteria. Thus, considering the technological characteristics studied, the 9 selected strains presented biochemical features well suited to their potential use as adjunct cultures, alone or in combination with autochthonous starter bacteria in the cheesemaking process, to overcome the heterogeneity of these PDO cheeses, preserving their unique sensory characteristics.


Assuntos
Queijo , Animais , Candida , Queijo/microbiologia , Microbiologia de Alimentos , Leite/microbiologia , Ovinos , Leveduras
11.
Molecules ; 27(3)2022 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-35164362

RESUMO

The aim of this study was to use local LAB cultures for the production of organic acid-rennet cheeses from unpasteurized cow's milk. Under industrial conditions, three types of cheese were produced, i.e., traditionally with acid whey (AW), with starter culture L. brevis B1, or with starter culture L. plantarum Os2. Strains were previously isolated from traditional Polish cheeses. Chemical composition, physico-chemical, microbiological, and sensory studies during 2 months of storage were carried out. As a result of this research, it was found that the basic composition was typical for semi-hard, partially skimmed cheeses. Mainly saturated fatty acids were detected. The cheeses were rich in omega-3, -6, and -9 fatty acids and conjugated linoleic acid (CLA), and were characterized by good lipid quality indices (LQI). All of the cheeses were characterized by a high number of lactic acid bacteria, with Enterobacteriaceae, yeast, molds, and staphylococci contaminants, which is typical microbiota for unpasteurized milk products. Water activity, pH, and total acidity were typical. A lower oxidation-reduction potential (ORP) of cheeses with the addition of strains and stability of the products during storage were observed. The B1 and Os2 cheeses were lighter, less yellow, had a more intense milk and creamy aroma, were softer, moister, and more elastic than AW cheese. The research results indicate the possibility of using environmental LAB strains in the production of high-quality acid-rennet cheeses, but special attention should be paid to the production process due to the microbiological quality of the cheeses.


Assuntos
Queijo/análise , Queijo/microbiologia , Microbiologia de Alimentos/métodos , Lactobacillales/fisiologia , Proteínas do Leite/metabolismo , Leite/química , Animais , Bovinos , Feminino
12.
BMC Microbiol ; 22(1): 48, 2022 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-35130830

RESUMO

BACKGROUND: Next-generation sequencing (NGS) methods and especially 16S rRNA gene amplicon sequencing have become indispensable tools in microbial ecology. While they have opened up new possibilities for studying microbial communities, they also have one drawback, namely providing only relative abundances and thus compositional data. Quantitative PCR (qPCR) has been used for years for the quantification of bacteria. However, this method requires the development of specific primers and has a low throughput. The constraint of low throughput has recently been overcome by the development of high-throughput qPCR (HT-qPCR), which allows for the simultaneous detection of the most prevalent bacteria in moderately complex systems, such as cheese and other fermented dairy foods. In the present study, the performance of the two approaches, NGS and HT-qPCR, was compared by analyzing the same DNA samples from 21 Raclette du Valais protected designation of origin (PDO) cheeses. Based on the results obtained, the differences, accuracy, and usefulness of the two approaches were studied in detail. RESULTS: The results obtained using NGS (non-targeted) and HT-qPCR (targeted) show considerable agreement in determining the microbial composition of the cheese DNA samples studied, albeit the fundamentally different nature of these two approaches. A few inconsistencies in species detection were observed, particularly for less abundant ones. The detailed comparison of the results for 15 bacterial species/groups measured by both methods revealed a considerable bias for certain bacterial species in the measurements of the amplicon sequencing approach. We identified as probable origin to this PCR bias due to primer mismatches, variations in the number of copies for the 16S rRNA gene, and bias introduced in the bioinformatics analysis. CONCLUSION: As the normalized microbial composition results of NGS and HT-qPCR agreed for most of the 21 cheese samples analyzed, both methods can be considered as complementary and reliable for studying the microbial composition of cheese. Their combined application proved to be very helpful in identifying potential biases and overcoming methodological limitations in the quantitative analysis of the cheese microbiota.


Assuntos
Bactérias/genética , Queijo/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Microbiota/genética , RNA Ribossômico 16S/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Bactérias/classificação , Bactérias/isolamento & purificação , Biologia Computacional , DNA Bacteriano/genética , Ensaios de Triagem em Larga Escala/métodos , Análise de Sequência de DNA
13.
J Dairy Sci ; 105(3): 2069-2081, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35033338

RESUMO

Traditionally, starter cultures for Cheddar cheese are combinations of Lactococcus lactis and Lactococcus cremoris. Our goal was to compare growth and survival of individual strains during cheesemaking, and after salting and pressing. Cultures used were 2 strains of L. lactis (SSM 7605, SSM 7436) and 2 strains of L. cremoris (SSM 7136, SSM 7661). A standardized Cheddar cheese make procedure was used that included a 38°C cook temperature and salting levels of 2.0, 2.4, 2.8, 3.2, and 3.6% from which were selected cheeses with salt-in-moisture levels of 3.5, 4.5, and 5.5%. Vats of cheese were made using each strain on its own as biological duplicates on different days. Starter culture numbers were enumerated by plate counting during cheesemaking and after 6 d storage at 6°C. Flow cytometry with fluorescent staining by SYBR Green and propidium iodide was used to determine the number of live and dead cells in cheese at the different salt levels. Differences in cheese make times were strain dependent rather than species dependent. Even with correction for average culture chain length, cheeses made using L. lactis strains contained ∼4 times (∼0.6 log) more bacterial cells than those made using L. cremoris strains. Growth of the strains used in this study was not influenced by the amount of salt added to the curd. The higher pH of cheeses with higher salting levels was attributed to those cheeses having a lower moisture content. Based on flow cytometry, ∼5% of the total starter culture cells in the cheese were dead after 6 d of storage. Another 3 to 19% of the cells were designated as being live, but semipermeable, with L. cremoris strains having the higher number of semipermeable cells.


Assuntos
Queijo , Lactococcus lactis , Animais , Queijo/microbiologia , Lactococcus , Cloreto de Sódio , Cloreto de Sódio na Dieta
14.
Braz J Microbiol ; 53(1): 303-316, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34661886

RESUMO

The biodiversity and succession of lactic acid bacteria (LAB) involved in the production and storage of Brazilian buffalo mozzarella cheese were evaluated. The isolates were characterized by Gram staining and catalase test, by the ability to grow at different conditions: temperatures, pH, concentrations of NaCl, and production of CO2 from glucose. The biodiversity and succession of 152 LAB isolated during cheese production were evaluated by 16S rRNA gene sequencing, Random Amplified Polymorphic DNA (RAPD-PCR), and Restriction Fragment Length Polymorphism (RFLP-PCR) techniques. Most of the strains grow well at 30 °C and are tolerant to 6.5% of NaCl, and in general, the best pH for growing was 9.6. Leuconostoc mesenteroides, Lacticaseibacillus casei, Limosilactobacillus fermentum, and Enterococcus sp. were prevalent and present in almost all steps of production. The LAB strains are typically found in the traditional Italian cheese, except the Leuconostoc citreum species. Sixty clusters were obtained by RAPD-PCR with 85% of similarity (114 isolates) while most of the LAB was clustered with 100% of similarity by the RFLP-PCR technique. The applied techniques enabled a valuable elucidation of the LAB biodiversity and succession, contributing to a better understanding of the specific microbial cultures with a technological aptitude of this cheese.


Assuntos
Queijo , Microbiota , Animais , Biodiversidade , Búfalos , Queijo/microbiologia , Microbiologia de Alimentos , Leite/microbiologia , RNA Ribossômico 16S/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico
15.
J Food Prot ; 85(2): 278-286, 2022 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-34669925

RESUMO

ABSTRACT: Cheese made with unpasteurized milk has been associated with outbreaks of illness. However, there are limited data on the prevalence of Shiga toxin-producing Escherichia coli (STEC) in these products and a lack of clarity over the significance of E. coli as a general indicator of hygiene in raw milk cheeses. The aim of this study was to provide further data to address both of these issues, as well as assessing the overall microbiological quality of raw milk cheeses available to consumers in England. A total of 629 samples of cheese were collected from retailers, catering premises, and manufacturers throughout England. The majority (80%) were made using cow's milk, with 14% made from sheep's milk and 5% from goat's milk. Samples were from 18 different countries of origin, with the majority originating from either the United Kingdom (40%) or France (35%). When interpreted against European Union microbiological criteria and United Kingdom guidance, 82% were considered to be of satisfactory microbiological quality, 5% were borderline, and 12% were unsatisfactory. Four samples (0.6%) were potentially injurious to health due to the isolation of STEC from one, >104 CFU/g of coagulase-positive staphylococci in two, and >100 CFU/g of Listeria monocytogenes in the fourth sample. Indicator E. coli and Listeria species were detected more frequently in soft compared with hard cheese. Higher levels of indicator E. coli were significantly associated with a greater likelihood of detecting Shiga toxin genes (stx1 and/or stx2).


Assuntos
Queijo , Escherichia coli Shiga Toxigênica , Animais , Bovinos , Queijo/microbiologia , Qualidade de Produtos para o Consumidor , Feminino , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Leite/microbiologia , Ovinos
16.
Appl Environ Microbiol ; 88(2): e0193921, 2022 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-34757819

RESUMO

The aim of this study was to investigate the temporal stability of microbial contamination during cheddar cheese production by examining patterns of nonstarter bacteria in 60-day aged cheddar collected from the start and end of 30 consecutive production days. Further, we explored the source of these temporal microbial variations by comparing microbial communities in the aged cheese to those on food contact surfaces from a piece of cheesemaking equipment previously identified as a major source of nonstarter bacteria in the same processing environment. 16S rRNA metabarcoding and culture-based sequencing methods identified two Streptococcus sequence variants significantly associated with the end of the production day in both the aged cheese and the cheese processing environment. Closer inspection of these sequence variants in the aged cheese over the 40-day sampling period revealed sinusoidal-like fluctuations in their relative ratios, which appeared to coincide with the Lactococcus starter rotation schedule. These results demonstrate that the microbial composition of finished cheese can vary according to the timing of processing within a production day. Further, our results demonstrate that time-of-day microbial differences in cheese can result from bacterial growth on food contact surfaces and that the composition of these microbial differences is subject to change day-to-day and may be linked to routine changes in the Lactococcus starter culture. IMPORTANCE Long production schedules used in modern cheese manufacturing can create circumstances that support the growth of microorganisms in the cheese processing environment. This work demonstrates that this growth can lead to significant changes in the microbial quality of aged cheese produced later in the production day. Further, we demonstrate that the dominant bacteria associated with these microbial changes throughout production are subject to change between days and might be influenced by specific cheese manufacturing practices. These findings improve understanding of microbial contamination patterns in modern food manufacturing facilities, thereby improving our ability to develop strategies to minimize quality losses due to microbial spoilage.


Assuntos
Queijo , Microbiota , Bactérias/genética , Queijo/microbiologia , Lactococcus , RNA Ribossômico 16S/genética
17.
J Food Prot ; 85(3): 484-493, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-34855936

RESUMO

ABSTRACT: The consumption of cheese in the People's Republic of China is increasing rapidly. Little is known about the microbiota, the presence of antibiotic-resistant bacteria, or the distribution of antibiotic resistance genes (ARGs) in commercially produced cheeses sold in China. This information is important for evaluating quality and safety. This study was conducted using 16S rRNA gene sequencing to assess the metagenomics of 15 types of cheese. Fourteen bacterial genera were detected, and Lactococcus, Lactobacillus, and Streptococcus were dominant based on number of sequence reads. Multidrug-resistant lactic acid bacteria (i.e., resistant to two or more types of antibiotic) were isolated from most of the types of cheese. Of these isolates, 100 and 91.7% were resistant to streptomycin and sulfamethoxazole, respectively, and genes involved in acquired resistance to streptomycin (strB) and sulfonamides (sul2) were detected with high frequency. To analyze the distribution of ARGs in the cheeses overall, 309 ARGs from eight categories and nine transposase genes were profiled. A total of 169 ARGs were detected in the 15 cheeses; their occurrence and abundance varied significantly between cheeses. Our study revealed diverse bacteria and ARGs in cheeses sold in China. The risks associated with multidrug resistance among dominant lactic acid bacteria are of great concern.


Assuntos
Queijo , Animais , Bactérias , Queijo/microbiologia , China , Resistência Microbiana a Medicamentos , Humanos , Leite/microbiologia , RNA Ribossômico 16S/genética
18.
Rev Argent Microbiol ; 54(1): 53-56, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-33906777

RESUMO

Listeria monocytogenes is one of the most important bacteria associated with foodborne diseases, soft cheese being an important L. monocytogenes vehicle. In Ecuador, soft cheese is consumed in 84.3% of urban households. We determined the prevalence of L. monocytogenes and serogroups in 260 fresh artisanal soft cheese samples collected in 18 of 24 Ecuadorian provinces. Listeria spp. detection was carried out by culture-dependent and independent methods; 14.23% of samples were positive for L. monocytogenes. Serogroup IVb was found in 83.78% of the food isolates. Serogroups IIb and IIa were present in 8.11% of our isolates. To our knowledge, this is the first report of L. monocytogenes serogroups associated with food in Ecuador; we also found serogroup similarities between cheese isolates and clinical isolates.


Assuntos
Queijo , Listeria monocytogenes , Queijo/microbiologia , Equador/epidemiologia , Microbiologia de Alimentos , Genótipo , Listeria monocytogenes/genética
19.
J Food Prot ; 85(1): 112-121, 2022 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-34324685

RESUMO

ABSTRACT: The objectives of this investigation were (i) to isolate bacteria from various foods (dairy products, fruits, and vegetables) and evaluate their probiotic potential and (ii) to select, identify, and characterize the bacterial strain(s) with the highest probiotic potential. From 14 food samples, 117 bacterial strains were isolated; however, only 42 (T1 to T42) had the correct characteristics (gram positive, coccoid, and bacilliform) and were catalase and oxidase negative to be considered presumptive lactic acid bacteria (LAB). The antagonistic activity of the 42 strains was evaluated against Escherichia coli (O157:H7E09), Listeria monocytogenes (ATCC 19115), Staphylococcus aureus (ATCC 25923), and Salmonella enterica serotype Typhimurium (ATCC 14028). The nine strains with the highest antagonistic activity were recovered from the following foods: pulque (T1), sprouted beans (T26), Ranchero cheese (T30, T31, T32, T33, T35, and T36), and Tenate cheese (T40). The inhibition zones on culture and sensitivity plates were 17.0 ± 1.2 to 19.3 ± 2.8 mm in diameter. Based on the antagonistic activity against pathogenic bacteria and resistance to low pH and bile salts, strain T40 had the highest probiotic potential. A 16S rRNA technique was used to identify strain T40 as Lacticaseibacillus paracasei (renamed from Lactobacillus paracasei in April 2020). This strain had no resistance to ampicillin, gentamicin, erythromycin, and tetracycline. The antagonistic activity was evaluated in situ (fresh cheese) against pathogenic bacteria, supporting the probiotic potential of L. paracasei. Isolates of this LAB recovered from Tenate cheese had characteristics of a probiotic microorganism with high potential for use in food technology.


Assuntos
Queijo , Lactobacillus paracasei , Listeria monocytogenes , Probióticos , Queijo/microbiologia , RNA Ribossômico 16S
20.
Curr Opin Biotechnol ; 73: 164-170, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34474311

RESUMO

A detailed understanding of the microbiome of cheese and dairy products is key to the optimization of flavour, appearance, overall quality and safety. Microorganisms (including bacteria, yeasts, moulds and viruses, especially bacteriophages) from the environment can enter the dairy supply chain at multiple stages with several implications. The ability to track these microorganisms and to understand their function and interaction can be greatly enhanced by the use of high-throughput sequencing. Depending on the specific production technology, dairy products can harbor several strains and antibiotic-resistance genes that can potentially interact with the gut microbiome, once the product is ingested. Milk-associated or cheese-associated microbial communities with their interaction, function and diversity are a key factor for the dairy industry. Multi-omics approaches have been seldom utilized in literature and they need to be further considered. Studying the role, origin, diversity and function of the microbial species involved in the complex system of dairy production can help improve processes in several fields of application. Integrating an extensive sampling procedure with an extensive culture based methodology is necessary. To this end, local producers, and in general stakeholders, should be guided to discover and maintain their microbial diversity. A better management of microbial resources through precision fermentation processes will in turn reduce overall food losses and increase the possibility to use the microbiome in order to increase the local producers' income.


Assuntos
Queijo , Microbiota , Animais , Queijo/microbiologia , Fermentação , Microbiologia de Alimentos , Leite/microbiologia
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