Coagulase-positive Staphylococci Research Articles

Zoonotic Staphylococcus spp. among domestic animals in Ukraine: antibiotic resistance and diagnostic approaches

Staphylococcus pseudintermedius and S. aureus are common infectious agents with zoonotic potential, capable of acquiring antibiotic resistance and causing hospital-acquired infections. This study aimed to assess the prevalence of S. pseudintermedius among dogs in Ukraine and explore the possibility of using chromogenic media for accurate identification and differentiation of S. aureus and S. pseudintermedius. The data obtained indicate a higher prevalence of nasal carriage (44.8%) of Staphylococcus spp. in healthy dogs compared to ear carriage (28.5%). Among the isolated strains in nasal swabs, S. pseudintermedius was most often detected (24.1%), S. aureus was detected less often (5.2%). Among the samples taken from sick dogs, 36.5% were found to be Staphylococcus spp., of which 20.5% contained S. pseudintermedius and 4.5% S. aureus. Among the studied cats, 36.8% were found to contain Staphylococcus spp., S. pseudintermedius – 10.5%, and S. aureus – 5.3%. Antibiotic resistance profiles were different: S. pseudintermedius was resistant to the combination of trimethoprim-sulfamethoxazole, as well as to penicillin and erythromycin, while S. aureus was resistant to penicillin. It should be noted that only one coagulase-positive S. aureus isolate obtained from healthy animals was resistant to oxacillin. The use of chromogenic media demonstrated the possibility of successful differentiation of S. aureus and S. pseudintermedius based on differences in culture properties. In addition, various combinations of microbiological tests were evaluated, resulting in the development of two schemes that included specific media and additional reactions to improve identification accuracy. The combination of CHROMagar™ Orientation with the coagulase detection reaction demonstrated higher sensitivity (100%) for the detection of S. pseudintermedius compared to the combination of mannitol salt agar (MSA) with the coagulase detection reaction (75%). In addition, the PCR reaction was valuable for the definitive identification of coagulase-positive staphylococci (CoPS). These comprehensive results shed light on the prevalence, species distribution, antibiotic resistance patterns, and diagnostic approaches for S. aureus in dogs and cats in Ukraine, emphasizing the importance of developing reliable surveillance and control strategies.

Resistance and Biofilm Production Profile of Potential Isolated from Kpètè-Kpètè Used to Produce Traditional Fermented Beer.

This study aimed to characterize the pathogenicity of bacteria isolated from the starter of two traditional beers produced and consumed in Benin. After standard microbial identification, species were identified by specific biochemical tests such as catalase, coagulase, and API 20 E. Antibiotic sensitivity was tested according to the French Society of Microbiology Antibiogram Committee. The crystal violet microplate technique evaluated the biofilm production and conventional PCR was used to identify genes encoding virulence and macrolide resistance. According to our data, the traditional starter known as kpètè-kpètè that is used to produce beer is contaminated by Enterobacteriaceae and staphylococci species. Thus, 28.43% of the isolated bacteria were coagulase-negative staphylococci (CNS), and 10.93% coagulase-positive staphylococci (CPS). Six species such as Klebsiella terrigena (1.38%), Enterobacter aerogens (4.14%), Providencia rettgeri (5.51%), Chryseomonas luteola (6.89%), Serratia rubidae (15.16%), and Enterobacter cloacae (27.56%) were identified among Enterobacteriaceae. Those bacterial strains are multi-resistant to conventional antibiotics. The hight capability of produced biofilms was recorded with Enterobacter aerogens, Klebsiella terrigena (100%), Providencia rettgeri (75%), and Staphylococcus spp (60%). Enterobacter cloacae (4%) and coagulase-negative Staphylococcus (5.55%) harbor the macrolide resistance gene. For other strains, these genes were not detected. Foods contaminated with bacteria resistant to antibiotics and carrying a virulence gene could constitute a potential public health problem. There is a need to increase awareness campaigns on hygiene rules in preparing and selling these traditional beers.

Colonization of methicillin-resistant Staphylococcus species in healthy and sick pets: prevalence and risk factors

BackgroundThe characterization of staphylococcal species that colonize pets is important to maintain animal health and to minimize the risk of transmission to owners. Here, the prevalence of Staphylococcus spp. and methicillin resistance was investigated in canine and feline isolates, and risk factors of staphylococcal colonization were determined. Pets were examined and separated into four groups: (1) healthy dogs, (2) healthy cats, and (3) dogs and (4) cats with clinical signs of bacterial infections of skin, mucous membranes, or wounds. Specimens were collected by a veterinary physician from six anatomic sites (external ear canal, conjunctival sacs, nares, mouth, skin [groin], and anus). In total, 274 animals (cats n = 161, dogs n = 113) were enrolled.ResultsStaphylococcus species were highly diverse (23 species; 3 coagulase-positive and 20 coagulase-negative species), with the highest variety in healthy cats (19 species). The most frequent feline isolates were S. felis and S. epidermidis, while S. pseudintermedius was the most prevalent isolate in dogs. Risk factors of staphylococcal colonization included the presence of other animals in the same household, medical treatment within the last year, and a medical profession of at least one owner. Methicillin resistance was higher in coagulase-negative (17.86%) compared to coagulase-positive (1.95%) staphylococci. The highest prevalence of methicillin-resistant CoNS colonization was observed in animals kept in homes as the most common (dogs and cats).ConclusionsThe association of methicillin-resistant CoNS colonization with animals most often chosen as pets, represents a high risk of transmission between them and owners. The importance of nosocomial transmission of CoNS was also confirmed. This information could guide clinical decisions during the treatment of veterinary bacterial infections. In conclusion, the epidemiologic characteristics of CoNS and their pathogenicity in pets and humans require further research.

From Hospital to Community: Exploring Antibiotic Resistance and Genes Associated with Virulence Factor Diversity of Coagulase-Positive Staphylococci.

Coagulase-positive staphylococcus (CoPS), including methicillin-resistant Staphylococcus aureus (MRSA), poses a global threat. The increasing prevalence of MRSA in Saudi Arabia emphasizes the need for effective management. This study explores the prevalence of virulence-associated genes and antibiotic resistance patterns in CoPS. Nasal swabs from 200 individuals were collected, and standard protocols were used for the isolation, identification, and characterization of CoPS and coagulase-negative staphylococci (CoNS). Additionally, antimicrobial susceptibility testing and PCR were conducted. Bacterial growth was observed in 58.5% of participants, with 12% positive for CoPS and 30% positive for CoNS. Hospital personnel carriers showed a significantly higher proportion of CoNS compared with non-hospital personnel carriers. Non-hospital personnel CoPS strains displayed higher sensitivity to oxacillin than hospital personnel strains. Cefoxitin exhibited the highest sensitivity among β-lactam antibiotics. All isolates were sensitive to trimethoprim/sulfamethoxazole, rifampin, and quinupristin. Polymerase chain reaction analysis detected methicillin resistance genes in both non-hospital and hospital personnel MRSA strains. The coa and spa genes were prevalent in MRSA isolates, while the Luk-PV gene was not detected. A high prevalence of CoPS and CoNS was observed in both non-hospital and hospital personnel carriers. Occupational risk factors may contribute to the differences in the strain distribution. Varying antibiotic susceptibility patterns indicate the effectiveness of oxacillin and cefoxitin. Urgent management strategies are needed due to methicillin resistance. Further research is necessary to explore additional virulence-associated genes and develop comprehensive approaches for CoPS infection prevention and treatment in Saudi Arabia.

Efects of Pasteurized Sheep's Milk Use on Production and Ripening of Siirt Herby Cheese

Siirt herby cheese is a varietty of local cheeses, and raw sheep’s and goat’s milk are used in its production. The curds obtained by curdling the milk obtained in the highlands is sold in the city centre during the day or the next day. Siirt herby cheese is produced from the retail curds. Three different cheese samples form retail curd, raw sheep’s and pasteurized sheep’s milks were made. The samples were ripened at 4°C for 120 d. Hygienic quality of raw milk supplied from a farm and raw milk curd bought from local retailers did not have adequate hygiene. Coagulase positive staphylococci were 3.11±0.6 log10 CFU/ml in raw milk, then it increased to 6.6±0.5 log10 CFU/g in the curd before salting. Pasteurization of milk was decreased the total count of the coagulase positive staphylococci below the maximum permissible value (103 CFU/g) in the regulation on Microbiological Criteria of the Turkish Food Codex. These samples were only contained >103 CFU/g coagulase positive staphylococci at the 120th d of ripening. The pasteurization of milk allowed to obtain cheese that meets the desired hygienic criteria. In addition, sufficient acidity and pH development were observed in pasteurized dairy cheeses even though starter culture was not used. It was found that the traditional cheese production using both retail curd and raw milk may not be suitable for producing cheese in accordance with the official standards and regulations. It was concluded that in the case of the production of these cheeses, it is not appropriate to offer them for sale before the completion of the legal ripening period of 120 d. It was deemed necessary to conduct further research in order to draw up a production diagram including the parameters of the Siirt herby cheese production process.

Methicillin Resistance of Food-Borne Biofilm-Forming Staphylococci

The aim of this study was to evaluate the biofilm-forming ability and the resistance of planktonic cells and biofilm to methicillin (MIC and MBC, and log CFU/cm2 reduction, respectively). Thirty-four isolates were used, including coagulase-positive Staphylococcus (CPS) and coagulase-negative Staphylococcus (CNS), recovered from ready-to-eat (RTE) foods. Biofilm-forming ability was based on enumeration of viable cells from biofilms formed on three types of surfaces, namely stainless steel, polystyrene, and glass. Thirteen of the thirty-four isolates (38.2%) were methicillin-resistant (MIC higher than 4 µg/mL). Staphylococcus aureus (CPS) and Staphylococcus saprophyticus (CNS) were the species that showed the highest percentage of resistance (50% and 71.4%, respectively). Staphylococcus warneri was the only species susceptible to methicillin. In 20 of the 34 isolates, the MBC value was twice the MIC value. The CPS (S. aureus) and the CNS (the other species) were not significantly different (p > 0.05) on biofilm-forming ability on the three surfaces used. However, when comparing the averages obtained for the biofilm-forming ability of the isolates, the values obtained (log CFU/cm2) in polystyrene were significantly higher (p < 0.05) than those obtained with the isolates on stainless steel. On the other hand, data from biofilm-forming ability of the isolates on glass and stainless steel are positively and significantly correlated with each other (r = 0.54; p = 0.02). It was not possible to determine the concentration of methicillin that promotes biofilm removal since log reductions were less than 3 log CFU/cm2. The results of this study indicate that foodborne CPS and CNS can form biofilms on different types of material. As these biofilms are resistant to high concentrations of methicillin, their occurrence in food environments and their spread to medical settings can result in staphylococcal food poisoning or, in the worst-case scenario, septicemia, respectively. Good hygiene and good manufacturing practices (GHP and GMP) are therefore mandatory to prevent contamination with Staphylococcus spp.

Physical and microbiological characterisation of artisanal cheese and isolation, identification, and in vitro evaluation of Lactobacillus strains with probiotic potential and antibiotic susceptibility

AbstractThe aim of this study was to isolate lactic acid bacteria (LAB) from artisanal cheeses and evaluate their probiotic potential and antibiotic susceptibility under in vitro conditions. Cheeses obtained at different maturation times were analysed for moisture and lipid contents, as well as for the presence of various microorganisms, including coagulase positive staphylococci, Salmonella spp., Escherichia coli, Listeria monocytogenes, filamentous fungi, yeasts, total mesophilic bacteria, and LAB. After identification, the selected LAB were subjected to human gastrointestinal tract (HGT) conditions to evaluate their survival rates. Of the 18 Lactobacillus strains isolated, 11 survived the HGT test and presented γ-haemolysis. No resistance was observed against antibiotics. Lactobacillus fermentum C1a, C1b, C1c, and C1f, as well as Lactobacillus paracasei C1d, C1e, and C1g, were identified as potential starter cultures for the food industry.

Features of snail expertise

Snails are a delicacy that people have been eating for centuries. Products made from this meat contain few calories per 100 grams and fill the human body with useful elements. They can be prepared by cooking, namely escargot with various fillings (Burgundy snails, Catalan snails, Asian snails, snails with porcini mushrooms), and also, now a new type of product containing m gastropod mollusks – puffs with snails. If determining the quality and nutritional value of meat, it is necessary to determine indicators, if there is doubt about its suitability, which characterize the chemical composition, freshness, as well as the determination of microbiological contamination. The purpose of this work was to determine the features and practical aspects of examination of gastropod molluscs – snails. The studies highlighted in the scientific publications of Ukrainian scientists are significant, but they relate to the determination of the degree of freshness of snail meat by various methods and the organization of snail farming in various ways. Snails are a delicacy, dietary product of high commercial value and belong to the few types of food products. They are subject to mandatory veterinary and sanitary examination. A special feature of examination of snails is microbiological and bacteriological types of control to establish their general safety. In the course of research, first of all, the organoleptic indicators of molluscs, which reflect their freshness, are determined. Next, they conduct research on the presence of pathogenic microflora, determine the presence of radionuclides, conduct bacteriological research. In addition to the mollusk itself, the water with which they are irrigated, as well as the soil – the permanent habitat of snails – are also examined for the presence of helminth eggs. It was established that it is necessary to carry out laboratory control of each batch of snail meat for the content of microorganisms, such as MAFAnM, BGKP (coliforms), coagulase-positive staphylococcus (S. aureus), bacteria of the genus Proteus, pathogenic microorganisms, including salmonella, Listeria monocytogenes, radionuclides – Cs-137 and Sr-90 and helminths (nematodes).

Identification of the skin microbiota of healthy dogs and those with pyoderma

Among the diseases of dogs, pyoderma is a frequent reason for referral to veterinary clinics. To effectively treat pyoderma and develop new strategies for preventing this disease, it is necessary to study the causes of formation and the conditions for the manifestation of pathogenic properties in causative agents. The purpose of the study will be to identify the microbiota isolated from the healthy skin of dogs and those with pyoderma. Washes were collected from the skin of various areas in clinically healthy dogs and dogs with pyoderma. The material was sown on selective media for the isolation of specific genera of microorganisms; after cultivation, pure cultures were identified according to the methods generally accepted in microbiological practice (cultural, tinctorial, morphological, and biochemical tests), according to Bergi's determinant. In addition, the following test systems were used: “STAPHY-test 16”, “En-coccus-test 36”, and “NEFERM test-24”. It was established that coagulase-negative and coagulase-positive species of staphylococci belong to the representatives of the autochthonous microbiota of the skin of clinically healthy dogs, which are isolated from different areas in 100 and 60–68.5 % of cases, respectively—the coagulase-positive species S. pseudintermedius and the coagulase-variable species S. schleiferi subsp. Coagulants occupy up to 65 % of the central niche, and СNS accounts for up to 30 % of all identified staphylococci. The causative agent of pyoderma in monoculture was S. pseudintermedius in 85.8 % of cases. In 57.1 % of cases, association with pyoderma was represented by СPS and Escherichia coli, and microbial association with СPS and Pseudomonas aeruginosa was detected 4.0 times less often – 14.3 % of cases. In 28.6 % of cases, microbial associations included СPS, Proteus in combination with intestinal or Pseudomonas aeruginosa. On the share of coagulase-positive staphylococci S. pseudintermedius, S. aureus, and coagulase-variable subspecies S. schleiferi subsp. Coagulants account for the central part of the staphylococcal microbiota (93.1 %), identified from inflammatory skin foci in pyoderma. At the same time, the species S. pseudintermedius significantly prevailed over other species; in particular, its share accounted for 78.4 % of the staphylococci isolated, which is 16.0 times more than S. aureus and 8.0 times more than S schleiferi subsp. coagulants So, in the etiology and pathogenesis of pyoderma in dogs, a significant role is played by СPS, in particular, the species S. pseudintermedius.

Evaluation of vacuum packaging for extending the shelf life of Sardinian fermented sausage

Salsiccia sarda or Sardinian fermented sausage is a traditional dry-fermented sausage included in the list of traditional food products of Sardinia (Italy). At the request of some producing plants, the possibility of extending the shelf life of the vacuum-packed product up to 120 days was evaluated. Manufacturing of 90 samples, representing 3 different batches of Sardinian fermented sausage was carried out in two producing plants (A and B). In the packaged product and subsequently every 30 days for four months (T0, T30, T60, T120), the following analyses were conducted on all samples: physicochemical characteristics, total aerobic mesophilic count, Enterobacteriaceae count, detection of Listeria monocytogenes, Salmonella spp., mesophilic lactic acid bacteria, and coagulase-positive Staphylococci. Moreover, surfaces in contact and surfaces not in contact with food were sampled in both producing plants. Sensory profile analysis was also performed for every analysis time. At the end of the extended shelf life, pH values were equal to 5.90±0.11 (producing plant A) and 5.61±0.29 (producing plant B). Water activity mean values at T120 were 0.894±0.02 (producing plant A) and 0.875±0.01 (producing plant B). L. monocytogenes was detected in 73.3% (33/45) of the samples from producing plant A, with mean levels of 1.12±0.76 log10 CFU/g. In producing plant B, L. monocytogenes was never detected. Enterobacteriaceae were detected in 91.1% (41/45) of samples in producing plant A with mean values of 3.15±1.21 log10 CFU/g, and in 35.5% (16/45) samples in producing plant B samples with mean values of 0.72±0.86 log10 CFU/g. Salmonella and Staphylococcus aureus were never detected. Regarding environmental samples, the sites that were most contaminated by L. monocytogenes were the bagging table (contact surface) and processing room floor drains (non-contact surface) with a prevalence of 50% each (8/16 positive samples for both sampling sites). Sensory analysis results showed that at T30 the overall sensory quality was at its highest; moreover, the visual-tactile aspect, the olfactory characteristics, the gustatory aspects, and the texture showed significant differences in samples throughout the shelf life, with a decreased intensity at 120 days of storage. Overall, the quality and sensory acceptance of the vacuum-packed Sardinian fermented sausage were not affected until 120 days of shelf-life. However, the possible contamination by L. monocytogenes calls attention to the hygienic management of the entire technological process. The environmental sampling was confirmed as a useful verification tool during control.

Prevalence Study of Biofilm Marker ica D Gene among Coagulase Positive Staphylococcus of Mastitic Origin

Bovine Mastitis is an inflammation of the mammary gland, usually due to microbial infection, which causes economic loss to dairy producer every year. Staphylococcus is one of the most common pathogens responsible for contagious mastitis in bovines. The aim of this study was to identify coagulase positive Staphylococci isolated from bovine subclinical mastitis. In vitro biofilm forming ability of the isolates, an important virulence factor of mastitogen and prevalence of biofilm marker icaD gene among the isolates. In the present study, 70 isolates were biochemically identified as S. aureus. Biochemically characterized strains of S. aureus were screened for biofilm formation. Phenotypic detection of biofilm formation of S.aureus were done by two methods, namely, Congo Red Agar (CRA) method and Tube Method (TM). The frequency of biofilm formation in S. aureus strains was 28.57 per cent on CRA plates while 71.42 per cent were biofilm non-producers. The frequency of biofilm formation in S. aureus strains was 38.57 per cent in total as shown by tube method while 61.42 per cent were biofilm non-producers. Employing PCR, molecular detection technique of icaD gene was carried out. In this study, 72.85 per cent S. aureus strains were found to be positive for icaD gene which produced an amplicon of 381bp.

Cheilitis in an atopic dermatitis patient associated with co-infection of Staphylococcus pseudintermedius and Staphylococcus aureus

BackgroundAtopic dermatitis (AD) is an inflammatory skin condition distinguished by an activated Th2 immune response. The local skin microbial dysbiosis is a contributing factor to the development of AD. The pathogenic coagulase-positive Staphylococcus aureus is the primary species responsible for the progression of AD. Even though Staphylococcus pseudintermedius is an animal-origin pathogen, it is increasingly becoming a source of concern in human diseases. As another coagulase-positive Staphylococci, it is crucial to pay more attention to S. pseudintermedius isolated from the lesion site.ResultsIn our investigation, we presented a case of cheilitis in a patient with atopic dermatitis (AD). We utilized culture and next-generation genomic sequencing (NGS) to identify the bacteria present on the skin swabs taken from the lip sites both prior to and following treatment. Our findings indicated that the predominant bacteria colonizing the lesion site of AD were S. pseudintermedius and S. aureus, both of which were eradicated after treatment. The Multi-locus sequence typing (MLST) of S. pseudintermedius and S. aureus demonstrated coordinated antibiotic susceptibility, with ST2384 and ST22 being the respective types. Although the skin abscess area resulting from S. pseudintermedius infection was significantly smaller than that caused by S. aureus in mice, the expression of cytokines interleukin-4 (IL-4) and interleukin-5 (IL-5) were significantly higher in the S. pseudintermedius-infected mice.ConclusionsThe S. pseudintermedius strain isolated from the lesion site of the AD patient exhibited a higher expression of IL-4 and IL-5 when colonized on mouse skin, as compared to S. aureus. This observation confirms that S. pseudintermedius can effectively induce the Th2 response in vivo. Our findings suggest that animal-origin S. pseudintermedius may play a role in the development of AD when colonized on the skin, emphasizing the importance of taking preventive measures when in contact with animals.

Metagenomic, microbiological, chemical and sensory profiling of Caciocavallo Podolico Lucano cheese

In this study, Caciocavallo Podolico Lucano (CPL) cheese was deeply characterized for its bacterial community, chemical composition and sensory aspects. The entire cheese making process (from milk collection to ripened cheese) was performed by strictly applying the traditional protocol for CPL production in four dairy factories (A–D) representative of the production area. The vat made of wood represents the main transformation tool for CPL cheese production and the biofilms hosted onto the internal surfaces of all vats analyzed in this study were dominated by lactic acid bacteria. Total mesophilic microorganisms present in bulk milk (4.7–5.0 log CFU/ml) increased consistently after contact with the wooden vat surfaces (5.4–6.4 log CFU/ml). The application of Illumina sequencing technology identified barely 18 taxonomic groups among processed samples; streptococci and lactobacilli constituted the major groups of the wooden vat biofilms [94.74–99.70 % of relative abundance (RA)], while lactobacilli dominated almost entirely (94.19–100 % of total RA) the bacterial community of ripened cheeses. Except coagulase positive staphylococci, undesirable bacteria were undetectable. Among chemical parameters, significant variations were registered for unsaturated, monounsaturated, polyunsaturated fatty acids and antioxidant properties (significantly lower for CPL cheeses produced in factory B). The cheeses from factories A, C and D were characterized by a higher lactic acid and persistence smell attributes than factory B. This work indicated that the strict application of CPL cheese making protocol harmonized the main microbiological, physicochemical and sensory parameters of the final cheeses produced in the four factories investigated.

Bacterial colonisation and the effect of a cleaning regime on iPad patient side electronic devices used in a veterinary healthcare setting.

This study aimed to determine the prevalence of clinically relevant bacteria on the surface of hospital-issued iPads and to assess the effectiveness and residual effect of a new cleaning regime using 70% alcohol and 2% chlorhexidine wipes. Hospital-issued iPads were swabbed to determine the presence of clinically relevant organisms. The iPads were wiped using 70% alcohol and 2% chlorhexidine. Further samples were taken 5 mins, 6 h and 12 h after implementation of the cleaning regime. Cultured bacteria were tested for antimicrobial resistance. A total of 25 hospital-issued iPads were analysed. Seventeen iPads (68%) sampled in this study were contaminated. Bacillus species (21%) were the most predominant, followed by Pasteurella species (14%), Acinetobacter species (11%), Eikenella species (11%), beta-haemolytic streptococci (11%), coagulase-positive staphylococci (7%), Escherichia coli (7%), coagulase-negative staphylococci (7%), alpha-haemolytic streptococci (3%), Enterococcus species (4%) and Pseudomonas species (4%). Of the isolated bacteria, 89% were resistant to at least one of the tested antibiotics. Of our isolates, 24 (75%) were resistant to clindamycin. After the cleaning regime, there was no bacterial growth from any of the devices at 5 mins, 6 h and 12 h despite repetitive use within the hospital. A variety of nosocomial pathogens, including antibiotic resistant pathogens, were isolated from the iPads. Cleaning with 70% alcohol and 2% chlorhexidine wipes is recommended every 12 h during use, between patient contacts and after witnessed contamination. A variety of nosocomial pathogens, including antibiotic-resistant pathogens with potential devastating effects on both human and animal health, were isolated from the iPads. Infection prevention strategies related to the devices should be employed in a hospital setting.

Caracterização microbiológica de duas espécies de peixes neotropicais oriundas de área quilombola maranhense, Brasil

The aim of study is to feature the microbiological quality of two neotropical fish species from the quilombola area of Maranhão State, Brazil. In order to do so, 21 samples of Hoplerythrinus unitaeniatus and 21 samples of Cichlasoma bimaculatum were captured in flooded environment. Collected fish were euthanized in laboratory environment; muscle fragments were removed for microbiological analyses focused on enumerating molds and yeasts, viable strict and facultative mesophilic microorganisms and coagulase-positive staphylococci; on counting total and thermotolerant coliforms; and on investigating Escherichia coli and Salmonella sp. Microbiological results were compared to the Brazilian legislation, which establishes the list of microbiological standards for food products. Among the assessed fish, 9.52% were classified as non-acceptable for human consumption, based on the Salmonella parameter. Enumerated coagulase-positive staphylococci ranged from < 10 to 3.9 x 104 CFU/g; 9.52% of assessed fish were classified as having intermediate standard for human consumption, whereas 4.76% were classified as non-acceptable for such a purpose. E. coli counting ranged from 3.6 to > 1,100MPN/g; 4.76% of assessed fish were classified as having intermediate standard for human consumption, whereas 4.76% were classified as non-acceptable for such a purpose. Total and thermotolerant coliforms’ counting and the enumeration of viable strict and facultative aerobic microorganisms, as well as of molds and yeasts, have evidenced high microbial population rates; this finding suggests poor hygienic conditions at capture site, contaminated raw material and risk of incidence of enteropathogens. This finding has evidenced imbalance in the investigated environment, as well as compromised aquatic biodiversity.

Prevalence and Molecular Characterization of Methicillin-Resistant Staphylococci (MRS) and Mammaliicocci (MRM) in Dromedary Camels from Algeria: First Detection of SCCmec-mecC Hybrid in Methicillin-Resistant Mammaliicoccus lentus.

Dromedary camels are an important source of food and income in many countries. However, it has been largely overlooked that they can also transmit antibiotic-resistant bacteria. The aim of this study was to identify the Staphylococcaceae bacteria composition of the nasal flora in dromedary camels and evaluate the presence of methicillin-resistant Mammaliicoccus (MRM) and methicillin-resistant Staphylococcus (MRS) in dromedary camels in Algeria. Nasal swabs were collected from 46 camels from seven farms located in two different regions of Algeria (M'sila and Ouargla). We used non-selective media to determine the nasal flora, and antibiotic-supplemented media to isolate MRS and MRM. The staphylococcal isolates were identified using an Autoflex Biotyper Mass Spectrometer (MALDI-TOF MS). The mecA and mecC genes were detected by PCR. Methicillin-resistant strains were further analysed by long-read whole genome sequencing (WGS). Thirteen known Staphylococcus and Mammaliicoccus species were identified in the nasal flora, of which half (49.2%) were coagulase-positive staphylococci. The results showed that four out of seven farms were positive for MRS and/or MRM, with a total of 16 isolates from 13 dromedary camels. The predominant species were M. lentus, S. epidermidis, and S. aureus. Three methicillin-resistant S. aureus (MRSA) were found to be ST6 and spa type t304. Among methicillin-resistant S. epidermidis (MRSE), ST61 was the predominant ST identified. Phylogenetic analysis showed clonal relatedness among M. lentus strains, while S. epidermidis strains were not closely related. Resistance genes were detected, including mecA, mecC, ermB, tet(K), and blaZ. An SCCmec type VIII element was found in a methicillin-resistant S. hominis (MRSH) belonging to the ST1 strain. An SCCmec-mecC hybrid element was detected in M. lentus, similar to what was previously detected in M. sciuri. This study highlights that dromedary camels may be a reservoir for MRS and MRM, and that they contain a specific set of SCCmec elements. This emphasizes the need for further research in this ecological niche from a One Health perspective.

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.

The Safety of Ethnic Foods in Campania Region: A Preliminary Study

Ethnic food is produced by an ethnic group-using their familiarity with local ingredients of plants and/or animal origin-and it is consumed in a country other than the country of origin. In Italy, the ethnic food market has expanded over the last three decades. The aim of this study was to evaluate the correct labeling, the microbiological communities and the allergens present in 50 ethnic foods. The visual inspection of labels and microbiological and allergen analyses have been carried out for evaluating their food safety. The visual inspection of labels revealed the absence of labeling in Italian and/or a failure to specify the place of origin. Microbiological analyses showed the absence of pathogens (i.e., Salmonella spp., Listeria monocytogenes and E. coli 0157:H7) in all matrices, but the presence of process hygiene indicator bacteria (total bacterial count, Coagulase-positive Staphylococci, Bacillus cereus, coliforms, yeasts and molds) was found in 37 samples. With regard to allergens, 12 samples were non-compliant for the presence of at least one allergen, while only two products were of species different from those declared on the label. This research highlights the need to increase the control of ethnic foods and also to improve the labeling system by standardizing international regulations.

Factors affecting the microbiological quality and contamination of farm bulk milk by Staphylococcus aureus in dairy farms in Asella, Ethiopia

BackgroundThe determination of the microbiological quality and safety of raw milk and the associated influencing factors at the farm level is very critical given that the quality or safety of subsequent products that are further produced depends on this. Therefore, this study aimed to determine the microbiological quality and safety of bulk milk and identify associated risk factors, and assess the presence/absence of S. aureus in bulk milk with potential contaminating sources in dairy farms in Asella, Ethiopia.ResultsThe geometric means of bacterial counts in farm bulk milk were 5.25 log cfu/ml, 3.1 log cfu/ml and 2.97 log cfu/ml for total bacterial count (TBC), coliform count (CC) and coagulase-positive staphylococci count (CPS), respectively. Of the 50 dairy farms, 66, 88, and 32% had TBC, CC and CPS counts, respectively, that exceeded the standard international limits for raw cow’s milk intended for direct human consumption. TBC tended to increase as CC increased in bulk milk (r = 0.5). In the final regression model, increased TBC, CC and the contamination of farm bulk milk by S. aureus were significantly associated with dirty barns, dirty cows and soiled udder and teats. TBC was higher during the rainy season than during the dry season. The reported practice of washing teats with warm water significantly decreased CC and CPS. The occurrence of S. aureus was significantly (p < 0.05) higher in bulk farm milk (42%) than in pooled udder milk (37.3%), teat swabs (22.5%), milkers’ hand swabs (18%), bulking bucket swabs (16.7%), milking container swabs (14%), and water for cleaning of udder and milkers’ hands (10%). The questionnaire survey result showed widespred raw milk consumption habits, low level of training and poor hygienic milking practices.ConclusionsThis study revealed low-quality bulk farm milk with high bacterial counts and a high occurrence of S. aureus. This indicates the potential food safety risks due to consumption of raw milk or its products. This study suggests awareness creation to dairy farmers and the public on hygienic milk production and heat treatment of milk before consumption.

Clonal relatedness of coagulase-positive staphylococci among healthy dogs and dog-owners in Spain. Detection of multidrug-resistant-MSSA-CC398 and novel linezolid-resistant-MRSA-CC5.

Nasal carriage of coagulase-positive staphylococci (CoPS) in healthy dogs could indicate increased risks of colonization for in-contact people or vice versa. This study determined the nasal carriage rate of CoPS among healthy dogs and in-contact people, their genotypic characteristics and phylogenetic relatedness. Nasal samples were collected from 27 households (34 dogs and 41 humans) in Spain. Staphylococci were identified by MALDI-TOF-MS, their antimicrobial resistance (AMR) genes and spa-types were tested by PCR/sequencing. The relatedness of CoPS from the same households was assessed by core genome single nucleotide polymorphisms (SNPs) analyses. Staphylococcus aureus carriage was found in 34.1% of humans (including one methicillin-resistant S. aureus MRSA-CC5-t2220-SCCmec type-IV2B) and 5.9% of dogs; Staphylococcus pseudintermedius in 2.4% of humans and 32.4% of dogs; while Staphylococcus coagulans was only detected in dogs (5.4%). Remarkably, one human co-carried S. aureus/S. pseudintermedius, while a dog co-carried the three CoPS species. Household density was significantly associated with S. pseudintermedius carriage in households with > than 1 dog and >than 1 human (OR = 18.10, 95% CI: 1.24-260.93, p = 0.034). Closely related (<15 SNPs) S. aureus or S. pseudintermedius were found in humans or dogs in three households. About 56.3% S. aureus carriers (dog or human) harboured diverse within-host spa-types or AMR genotypes. Ten clonal complexes (CCs) were detected among the S. aureus, of which methicillin-susceptible S. aureus-CC398-IEC-type C (t1451 and t571) was the most frequent, but exclusive to humans. S. aureus and S. pseudintermedius isolates harboured resistance genes or mutations associated to 9 classes of antimicrobials including linezolid (G2261A & T1584A point mutations in 23S rDNA). The S. coagulans isolates were susceptible to all antimicrobials. Most of the S. pseudintermedius carried lukS/F-I, siet, and sient genes, and all S. aureus were negative for lukS/F-PV, tst-1, eta and etb genes. Clonally related human-to-human MSSA and dog-to-human MSSP were found. The detection of the MSSA-CC398 clade highlights the need for its continuous surveillance from One Health perspective.