Biofilm-producing ability of Staphylococcus aureus isolates from Brazilian dairy farms

This study aimed to investigate the in silico biofilm production ability of Staphylococcus aureus strains isolated from milking parlor environments on dairy farms from São Paulo, Brazil. The Staph. aureus isolates were obtained from 849 samples collected on dairy farms, as follows: milk from individual cows with subclinical mastitis or history of the disease (n = 220); milk from bulk tank (n = 120); surfaces of milking machines and utensils (n = 389); and milk handlers (n = 120). Thirty-one Staph. aureus isolates were obtained and categorized as pulsotypes by pulsed-field gel electrophoresis and submitted to assays for biofilm formation on polystyrene, stainless steel, rubber, and silicone surfaces. Fourteen (45.2%) pulsotypes were considered producers of biofilm on the polystyrene microplate assay, whereas 13 (41.9%) and 12 (38.7%) pulsotypes were biofilm producers on stainless steel and rubber, respectively. None of the pulsotypes evaluated produced biofilms on silicone. Approximately 45% of Staph. aureus pulsotypes isolated from different sources on dairy farms showed the ability to produce biofilms in at least one assay, indicating possible persistence of this pathogen in the milking environment. The potential involvement of Staph. aureus in subclinical mastitis cases and its occurrence in milk for human consumption emphasize the need to improve hygiene practices to prevent biofilm formation on the farms studied.     

Bactericidal and antibiofilm properties of Rumex japonicus Houtt. on multidrug-resistant Staphylococcus aureus isolated from milk

Multidrug-resistant (MDR) Staphylococcus aureus and its biofilm formation have been challenging to control in milk and dairy industries. Biofilms formed by Staph. aureus may result in the failure of antibacterial agents and disinfectants to penetrate the biofilm in an attempt to control contamination. Novel natural antibacterial agents are required to combat MDR bacteria and biofilms. In this study, we evaluated the bactericidal, antibiofilm, and antimotility effects of Rumex japonicus Houtt. (RJH) extract on MDR Staph. aureus isolated from milk. The RJH extract exhibited good antibacterial activity against MDR strains with minimum inhibitory concentrations (MIC) ranging from 0.78 to 6.25 mg/mL and minimum bactericidal concentrations ranging from 3.125 to 12.5 mg/mL. The extract showed strong inhibition of biofilm formation (81.9%) at sub-MIC value and eradication of biofilm at higher concentrations. The motility of Staph. aureus was effectively blocked by the extract. Major compounds emodin, chrysophanol, and physcion were identified in RJH extract using HPLC-linear trap quadrupole (LTQ)/Orbitrap-mass spectrometry. The extract was nontoxic to human epithelial cell lines such as Caco-2 and HT-29 cell lines at concentrations ranging from 0.1 to 0.5 mg/mL, and from 0.1 to 0.75 mg/mL, respectively. These findings suggest that RJH extract could be an alternative to synthetic preservatives in milk and dairy products.     

Attachment and biofilm formation of Pseudomonas fluorescens PSD4 isolated from a dairy processing line

The effects of different nutrient sources and temperatures on attachment and biofilm formation of Pseudomonas fluorescens PSD4, a dairy isolate, were studied. Initial adherence and attachment capabilities among different strains were studied using microtitre plate assays. Biofilm development was observed using confocal microscopy. Strongly adherent cells were seen in protein rich media. Citrate as a carbon source enhanced biofilm formation. Glucose did not favor biofilm development. Psychrotrophic P. fluorescens PSD4 formed strongly adherent biofilms having high metabolic activities at low temperatures. P. fluorescens PSD4 with spoilage potential was capable of forming strong biofilms in dairy processing environments. Biofilm formation was influenced by nutrient availability and growth conditions. These factors should be considered for design of effective anti-biofilm strategies.     

Biofilm formation of Staphylococcus aureus dairy isolates representing different genotypes

The objective of this study was to compare the biofilm-forming capabilities of different genotypes of Staphylococcus aureus dairy isolates from Switzerland and northern Italy, including Staph. aureus genotype B (GTB) and methicillin-resistant Staph.aureus (MRSA). We hypothesized that biofilm formation might be more pronounced in the contagious GTB isolates compared with other genotypes affecting individual animals. Twenty-four dairy isolates, including 9 MRSA, were further characterized by genotyping by using ribosomal spacer PCR, spa typing, biofilm formation under static and dynamic conditions, and scanning electron microscopy. The GTB isolates (n = 6) were more able to form biofilms than other genotypes at 37°C and at 20°C after 48 and 72 h of incubation in the static assay using polystyrene microtiter plates. This result was supported by scanning electron micrographs showing a GTB isolate producing strong biofilm with extracellular matrix in contrast to a genotype C isolate. Furthermore, none of the MRSA isolates formed strong biofilms in the static assay. However, some MRSA produced low or moderate amounts of biofilm depending on the applied conditions. Under dynamic conditions, a much more diverse situation was observed. The ability of GTB isolates to be strong biofilm formers was not observed in all cases, emphasizing the importance of growth conditions for the expression of biofilm-related genes. No specific genotype, spa type, or MRSA isolate could be categorized significantly into one level of biofilm formation. Nineteen percent of isolates behaved similarly under static and dynamic conditions. The results of this study expand our knowledge of different dairy-related Staph. aureus subtypes and indicate the benefit of genotyping when biofilms are studied.     

The influence of temperature on the adhesion of mixed cultures of Staphylococcus aureus and Escherichia coli to polypropylene

The adhesion of Staphylococcus aureus and Escherichia coli, in mixed cultures, to polypropylene surfaces was evaluated at 12°C and 30°C. The micro-organisms were isolated from a chicken carcass and cultured in an aqueous extract, prepared from the same carcass, for the production of biofilms on polypropylene coupons. Adhered cells were counted by epifluorescence microscopy with acridine orange staining. Escherichia coli adhered in greater numbers to the coupons than S. aureus at both temperatures. Staphylococcus aureus adhered better at 12°C than at 30°C, while the reverse was true for E. coli. At 30°C, there was no increase in the number of adherent cells ofS. aureus over 8 h, while E. coli increased from a median of 5·0–19·0 per microscope field. At 12°C, the major increase in adherent cell numbers for both species occurred between 2 and 4 h, so that leaving cleaning until 8 h, as is common, would not result in greatly increased biofilms, 2-hourly cleansing is clearly unrealistic. However, total adherent cell numbers were the same at 12° and 30°C between 4 and 6 h incubation. Hence it seems that reduced temperature has little to offer for restricting biofilm formation on polypropylene work surfaces in a well-run food processing plant.     

Coagulase-negative staphylococci species affect biofilm formation of other coagulase-negative and coagulase-positive staphylococci

Coagulase-negative staphylococci (CNS) are considered to be commensal bacteria in humans and animals, but are now also recognized as etiological agents in several infections, including bovine mastitis. Biofilm formation appears to be an important factor in CNS pathogenicity. Furthermore, some researchers have proposed that CNS colonization of the intramammary environment has a protective effect against other pathogens. The mechanisms behind the protective effect of CNS have yet to be characterized. The aim of this study was to evaluate the effect of CNS isolates with a weak-biofilm phenotype on the biofilm formation of other staphylococcal isolates. We selected 10 CNS with a weak-biofilm phenotype and 30 staphylococcal isolates with a strong-biofilm phenotype for this study. We measured biofilm production by individual isolates using a standard polystyrene microtiter plate assay and compared the findings with biofilm produced in mixed cultures. We confirmed the results using confocal microscopy and a microfluidic system with low shear force. Four of the CNS isolates with a weak-biofilm phenotype (Staphylococcus chromogenes C and E and Staphylococcus simulans F and H) significantly reduced biofilm formation in approximately 80% of the staphylococcal species tested, including coagulase-positive Staphylococcus aureus. The 4 Staph. chromogenes and Staph. simulans isolates were also able to disperse pre-established biofilms, but to a lesser extent. We also performed a deferred antagonism assay and recorded the number of colony-forming units in the mixed-biofilm assays on differential or selective agar plates. Overall, CNS with a weak-biofilm phenotype did not inhibit the growth of isolates with a strong-biofilm phenotype. These results suggest that some CNS isolates can negatively affect the ability of other staphylococcal isolates and species to form biofilms via a mechanism that does not involve growth inhibition.     

Formation and dispersal of biofilms in dairy substrates

The formation and dispersion of biofilms in the dairy industry is a problem because it increases cross‐contamination, affecting the shelf life of the products and their safety. The objective of this study was to evaluate the influence of different dairy substrates (cows’ milk and whey protein) on the formation and dispersion of Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus in two biofilm systems (mono‐species and multi‐species) on stainless steel at 25 °C. The dominant behaviour of E. faecalis occurred in most of the tests on mono‐species and multi‐species biofilms. A greater dispersion of biofilm cells was observed in skimmed milk.     

Short communication: Biofilm production characterization of mecA and mecC methicillin-resistant Staphylococcus aureus isolated from bovine milk in Great Britain

Staphylococcus aureus is an important cause of contagious intramammary infection in dairy cattle, and the ability to produce biofilm is considered to be an important virulence property in the pathogenesis of mastitis. The aim of this study was to characterize the biofilm formation capacity of methicillin-resistant Staph. aureus (MRSA), encoding mecA or mecC, isolated from bulk tank milk in Great Britain. For this purpose, 20 MRSA isolates were grown on microtiter plates to determine the biofilm production. Moreover, the spa-typing and the presence of the intercellular adhesion genes icaA and icaD were analyzed by PCR. All MRSA isolates tested belonged to 9 spa-types and were PCR-positive for the ica genes; 10 of them (50%) produced biofilm in the microtiter plate assay. This is also the first demonstration of biofilm production by mecC MRSA.     

Removal of meat biofilms from surfaces by ultrasounds combined with enzymes and/or a chelating agent

A curved ultrasonic transducer was devised to standardise biofilm removal for hygiene testing in internal or curved food contact surfaces. Meat biofilms made with Escherichia coli and Staphylococcus aureus on stainless steel sheets were studied. Ultrasounds (10 s at 40 kHz) alone failed to completely remove biofilms: 49 ± 5% and 39 ± 5% recovery rates were obtained for E. coli and S. aureus biofilms, respectively. A combined treatment, which involved the application of ultrasounds to EDTA and/or in enzymes solutions, allowed to remove up to 75 ± 4% and 100 ± 15% of E. coli and S. aureus biofilms, respectively. This application was in agreement with an industrial control i.e. a combined treatment: ultrasound generation in enzymes preparation restricted to an active chamber area with a fast and good reproducible recovery.Industrial relevanceThe biofilm phenomenon has been under intensive research for several years in food industry. A curved ultrasonic transducer was devised to standardise biofilm removal for hygiene testing in internal or curved food contact surfaces. This apparatus uses the mechanical effects of ultrasonic cavitation produced at 40 KHz (10 s) for the non-destructive detection of biofilms in food processing equipment. We report the utilisation of a combined treatment, which involved the application of ultrasounds to EDTA and/or in enzymes solutions on meat biofilms made with E. coli and S. aureus on stainless steel sheets. This application was in agreement with an industrial control i.e. a combined treatment: ultrasound generation in EDTA and/or enzymes preparation restricted to an active chamber area with a fast and good reproducible recovery.     

Detecting Staphylococcus aureus in milk from dairy cows using sniffer dogs

Fast and accurate identification of disease-causing pathogens is essential for specific antimicrobial therapy in human and veterinary medicine. In these experiments, dogs were trained to identify Staphylococcus aureus and differentiate it from other common mastitis-causing pathogens by smell. Headspaces from agar plates, inoculated raw milk samples, or field samples collected from cows with Staphylococcus aureus and other mastitis-causing pathogens were used for training and testing. The ability to learn the specific odor of Staphylococcus aureus in milk depended on the concentration of the pathogens in the training samples. Sensitivity and specificity for identifying Staphylococcus aureus were 91.3 and 97.9%, respectively, for pathogens grown on agar plates; 83.8 and 98.0% for pathogens inoculated in raw milk; and 59.0 and 93.2% for milk samples from mastitic cows. The results of these experiments underline the potential of odor detection as a diagnostic tool for pathogen diagnosis.     

Biofilm Formation of Staphylococcus aureus on Various Surfaces and Their Resistance to Chlorine Sanitizer

This study investigated the effect of material types (polystyrene, polypropylene, glass, and stainless steel) and glucose addition on Staphylococcus aureus biofilm formation, and the relationship between biofilm formation measured by crystal violet (CV) staining and the number of biofilm cells determined by cell counts was studied. We also evaluated the efficacy of chlorine sanitizer on inhibiting various different types of S. aureus biofilms on the surface of stainless steel. Levels of biofilm formation of S. aureus were higher on hydrophilic surfaces (glass and stainless steel) than on hydrophobic surfaces (polypropylene and polystyrene). With the exception of biofilm formed on glass, the addition of glucose in broth significantly increased the biofilm formation of S. aureus on all surfaces and for all tested strains (P ≤ 0.05). The number of biofilm cells was not correlated with the biomass of the biofilms determined using the CV staining method. The efficacy of chlorine sanitizer against biofilm of S. aureus was not significantly different depending on types of biofilm (P > 0.05). Therefore, further studies are needed in order to determine an accurate method quantifying levels of bacterial biofilm and to evaluate the resistance of bacterial biofilm on the material surface.     

The occurrence of Staphylococcus aureus on a farm with small-scale production of raw milk cheese

In recent years, the small-scale production of raw milk products has increased in Norway, and there is some concern that such foods may pose a risk of staphylococcal food poisoning to consumers. The aim of the study was to evaluate potential sources of contamination of raw milk cheese with Staphylococcus aureus on a bovine dairy farm with small-scale production. Samples for bacteriological analyses (n = 144) were collected from the animals, the environment, processing equipments, from humans, and from cheeses at various stages of production. Staphylococcus aureus was isolated from 10 of 11 cows, the farmer, equipment, the environment, and the cheese. Seventy-five Staph. aureus isolates were genotyped by pulsed-field gel electrophoresis, tested for enterotoxin (SE) production by reversed passive latex agglutination, for SE genes by multiplex polymerase chain reaction, and for penicillin resistance by the cloverleaf method. Five different pulsotypes were identified and SE gene fragments were identified in 11 isolates, but no isolates produced SE or were penicillin resistant. Staphylococcus aureus was found throughout the farm, and appeared to be spread with the milk to the environment, equipment, and to products. One pulsotype dominated and was identified from most sample sites on the farm. Raw milk products are vulnerable to contamination with Staph. aureus. Strategies to reduce the occurrence of Staph. aureus in bulk milk are of particular importance on farms where milk is used for raw milk products.     

An improved method to culture Staphylococcus aureus from bovine milk

Staphylococcus aureus is an important udder pathogen often associated with subclinical mastitis in dairy cows. Identification of Staph. aureus-positive udder quarters and cows is an important part of control programs to reduce spread of Staph. aureus within and between dairy herds. Therefore, accurate and easy-to-perform culturing methods of Staph. aureus in milk are needed. In the present study, 8 methods for isolation of Staph. aureus in bovine milk samples were investigated. The methods involved different culturing volumes, enrichment, incubation, and freezing processes as well as sedimentation and use of the Mastistrip cassette (SVA, Uppsala, Sweden). Three different sets of milk samples were collected, and 6, 5, and 4 methods were used in each subset of samples. Our results indicate an increased probability of detecting Staph. aureus in milk samples when a simple incubation step (37°C for 18 h) without additives was included before culturing. Using this incubation method, the number of Staph. aureus-positive udder quarters and cows increased by 50 and 29%, respectively, compared with using the standard method of direct culturing of 10 μL of milk. The improved method may be especially useful for detection of low concentrations of Staph. aureus in milk; for example, when screening herds for Staph. aureus.     

Resistance of pathogenic bacteria on the surface of stainless steel depending on attachment form and efficacy of chemical sanitizers

Various bacteria including food spoilage bacteria and pathogens can form biofilms on different food processing surfaces, leading to potential food contamination or spoilage. Therefore, the survival of foodborne pathogens (Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, Staphylococcus aureus, Cronobacter sakazakii) in different forms (adhered cells, biofilm producing in TSB, biofilm producing at RH 100%) on the surface of stainless steel and stored at various relative humidities (RH 23%, 43%, 68%, 85%, and 100%) at room temperature for 5 days was investigated in this study. Additionally, the efficacy of chemical sanitizers (chlorine-based and alcohol-based commercial sanitizers) on inhibiting various types of biofilms of E. coli O157:H7 and S. aureus on the surface of stainless steel was investigated. The number of pathogens on the surface of stainless steel in TSB stored at 25 °C for 7 days or RH 100% at 25 °C for 7 days was significantly increased and resulted in the increase of 3 log₁₀ CFU/coupon after 1 day, and these levels were maintained for 7 days. When stainless steel coupons were stored at 25 °C for 5 days, the number of pathogens on the surface of stainless steel was significantly reduced after storage at RH 23%, 43%, 68%, and 85%, but not at 100%. When the bacteria formed biofilms on the surface of stainless steel in TSB after 6 days, the results were similar to those of the attached form. However, levels of S. aureus and C. sakazakii biofilms were more slowly reduced after storage at RH 23%, 43%, 68%, and 85% for 5 days than were those of the other pathogens. Formation of biofilms stored at RH 100% for 5 days displayed the highest levels of resistance to inactivation. Treatment with the alcohol sanitizer was very effective at inactivating attached pathogens or biofilms on the surface of stainless steel. Reduction levels of alcohol sanitizer treatment ranged from 1.91 to 4.77 log and from 4.35 to 5.35 log CFU/coupon in E. coli O157:H7 and S. aureus, respectively. From these results, the survival of pathogens contaminating the surfaces of food processing substrates such as stainless steel varied depending on RH and attachment form. Also, alcohol-based sanitizers can be used as a potential method to remove microbial contamination on the surfaces of utensils, cooking equipment, and other related substrates regardless of the microbial attached form.     

Distinct behavior of bovine-associated staphylococci species in their ability to resist phagocytosis and trigger respiratory burst activity by blood and milk polymorphonuclear leukocytes in dairy cows

Mastitis affects a high proportion of dairy cows and is still one of the greatest challenges faced by the dairy industry. Staphylococcal bacteria remain the most important cause of mastitis worldwide. We investigated how distinct staphylococcal species evade some critical host defense mechanisms, which may dictate the establishment, severity, and persistence of infection and the outcome of possible therapeutic and prevention interventions. Thus, the present study investigated variations among distinct bovine-associated staphylococci in their capability to resist phagocytosis and to trigger respiratory burst activity of blood and milk polymorphonuclear neutrophil leukocytes (PMNL) in dairy cows. To do so, PMNL of 6 primiparous and 6 multiparous dairy cows were used. A collection of 38 non-aureus staphylococci (NAS) and 12 Staphylococcus aureus were included. The phagocytosis and intracellular reactive oxygen species (ROS) production by blood and milk PMNL were analyzed by flow cytometry. Phagocytosis, by both blood and milk PMNL, did not differ between S. aureus and NAS as a group, although within-NAS species differences were observed. Staphylococcus chromogenes (a so-called milk-adapted NAS species) better resisted phagocytosis by blood PMNL than the so-called environmental (i.e., Staphylococcus fleurettii) and opportunistic (i.e., Staphylococcus haemolyticus) NAS species. Otherwise, S. haemolyticus was better phagocytosed by blood PMNL than S. aureus, S. fleurettii, and S. chromogenes. No influence of the origin of the isolates within the staphylococci species in the resistance to phagocytosis by blood and milk PMNL was found. Overall, both S. aureus and NAS did not inhibit intracellular ROS production in blood and milk PMNL. Non-aureus staphylococci induced fewer ROS by milk PMNL than S. aureus, which was not true for blood PMNL, although species-specific differences in the intensity of ROS production were observed. Staphylococcus chromogenes induced more blood PMNL ROS than S. fleurettii and S. haemolyticus, and as much as S. aureus. Conversely, S. chromogenes induced fewer milk PMNL ROS than S. aureus. The origin of the isolates within the staphylococci species did not affect the ROS production by blood and milk PMNL. In conclusion, our study showed differences in staphylococci species in evading phagocytosis and triggering ROS production, which may explain the ability of some staphylococci species (i.e., S. aureus and S. chromogenes) to cause persistent infection and induce inflammation.     

Molecular characterisation and biofilm production in Staphylococcusaureus isolates from the dairy production chain in Northern Italy

Staphylococcus aureus is able to produce enterotoxins causing staphylococcal food poisoning, and is frequently harboured by dairy products. Also, S. aureus is able to form biofilm in the production environment, enhancing the risk of food contamination. The ability of 49 S. aureus isolates from the dairy production chain to form biofilm aim was tested, and their genetic diversity in terms of population structure and presence of genes involved in biofilm formation or enterotoxins production was explored. The majority of the genotypes found were generally bovine associated; however, some have been also reported frequently in human clinical cases. Two isolates were methicillin-resistant. In total, 38.7% of the isolates were biofilm producers, and among them 47.3%, 42.1% and 10.5% exhibited weak, moderate, or strong biofilm-forming ability, respectively. In total 68% of the biofilm producing isolates were also positive for enterotoxins genes, raising concerns for consumer safety.     

Biofilm formation by Campylobacter jejuni in controlled mixed-microbial populations

This study was to screen the ability of biofilm formation by Campylobacter jejuni strains found in New Zealand, and investigate the biofilm growth of C. jejuni in a controlled mixed-microbial population that includes five different bacteria. The ability of C. jejuni to form a biofilm in monoculture and mixed-microbial populations was measured in a laboratory assay using a microtiter plate screening assay. The optical density of the biofilm and cell growth from mixed-microbial populations was converted to a Biofilm Formation Index (BFI). This index was used to standardize the biofilm formation in the mixed-microbial populations. High BFI was observed for Enterococcus faecalis (2.30) and Staphylococcus simulans (3.75) when they were grown with C. jejuni multilocus sequence type ST-474: a dominant poultry and human-associated type in New Zealand. C. jejuni cells were recovered from most of the biofilms containing E. faecalis and/or S. simulans. These results suggest that E. faecalis and S. simulans may play a role in biofilm formation in the poultry environment as both of these microorganisms are found in poultry processing environments and were able to form a biofilm in association with C. jejuni under microaerobic conditions. Understanding the relationships among C. jejuni, E. faecalis and S. simulans in poultry processing plants and farms may help in the design of strategies to reduce the reservoir of contamination of these bacteria and reduce the incidence of campylobacteriosis.     

Bacterial biofilm in silver-impregnated contact lens cases

PurposeThis study investigated the efficacy of pre-conditioning lens cases on bacterial biofilm formation and removal.MethodsSilver impregnated (MicroBlock / ProGuard™ & i-Clean) and control storage cases were pre-conditioned for 24 h with their respective multipurpose solutions (MPDSs). Cases were then inoculated with 2 ml of 10⁶ CFU/mL of ocular isolates of either P. aeruginosa or S. aureus and incubated for 48 h. Cases were subsequently disinfected (4−6 hours) as per the manufacturer’s recommended disinfecting time (MRDT) followed by the recommended case hygiene procedures - recapping wet (MicroBlock / ProGuard™ cases only) or rinse and air-dry or rinse, tissue-wipe and air dry (mechanical disruption). Surviving bacteria were enumerated using standard techniques.ResultsPre-conditioning the MicroBlock / ProGuard™ cases with MPDS significantly reduced biofilm formation (-1.1 log₁₀ CFU, p < 0.01 for P. aeruginosa & -1.3 log₁₀, p < 0.001, CFU for S. aureus) compared to the i-Clean lens cases. Maintaining the MicroBlock / ProGuard™ lens cases wet after the MRDT resulted in partial removal of bacterial biofilms (-2.9 log₁₀ CFU, p < 0.001 for P. aeruginosa and -2.6 log₁₀ CFU, p < 0.001 for S. aureus). Air-drying of all three types of lens storage cases after MRDT significantly reduced the bacterial biofilm (-5.4 log₁₀ CFU, p < 0.001 for P. aeruginosa and -3.5 log₁₀ CFU, p < 0.001 for S. aureus). Mechanical disruption produced the greatest reduction in the levels of bacterial biofilm in all 3 types of lens cases tested (-6.8 log₁₀ CFU, p < 0.001 for P. aeruginosa and -4.5 log₁₀ CFU, p < 0.001 for S. aureus). Synergi MPDS was significantly better than AQuify MPDS in removing bacterial biofilm from all 3 lens case types for case hygiene treatments with an air-drying step.ConclusionPre-conditioning of silver-impregnated ProGuard™ lens cases inhibited initial bacterial biofilm formation. Synergi MPDS was more effective than AQuify MPDS in removing bacterial biofilm in silver impregnated cases and tissue-wiping significantly improved biofilm removal.     

Short communication: Lack of intramammary niche recolonization during a sanitation program for the contagious mastitis pathogen Staphylococcus aureus genotype B

In Switzerland, sanitation programs of dairy herds infected with the contagious mastitis pathogen Staphylococcus aureus genotype B (GTB) have been established for several years. In recent years, Streptococcus uberis and non-aureus staphylococci have emerged as the bacteria most frequently isolated from bovine milk samples. The latter cause subclinical mastitis, and some species are more persistent or pathogenic than others. The present study aimed to investigate the developments in the intramammary colonization spectrum of 5 dairy herds undergoing a sanitation program for Staph. aureus GTB. We collected single-quarter milk samples aseptically from all lactating cows at 3-mo intervals during the sanitation period; after classical bacteriological analysis, MALDI-TOF mass spectrometry was used to identify the isolates to the species level. Non-aureus staphylococci were found to be the bacterial group most frequently occurring on the selected farms, with Staphylococcus chromogenes and Staphylococcus xylosus being predominant. The present study demonstrated that GTB-infected cows treated with antibiotics lacked systematic recolonization with other bacteria during herd sanitation for the contagious Staph. aureus GTB.