Dataset Information

Growth of Staphylococcus epidermidis on the Surface of Teatcups from Milking Parlours.

ABSTRACT: The growth of two Staphylococcus epidermidis isolates (one biofilm-forming and one not) on teatcups for cattle (made of rubber) or sheep (made of silicone) were assessed in nine multiplicates for 24 h post-smearing on the teatcup surface. Staphylococci were smeared on an area of 0.0003142 m2 on the material and their growth and expansion further on were monitored for 24 h. There were no differences in the frequency of recoveries between the two isolates (p > 0.82 for all comparisons). There were more recoveries from sheep teatcups than from cattle teatcups: 1280/1728 (74.1%) versus 942/1728 (54.5%), for both isolates (p < 0.0001). Significance was observed only 6 h to 15 h after smearing (p < 0.0001 for all comparisons). The median speed of linear dissemination of the isolates was 0.00000021 m s-1 on cattle teatcups and 0.00000033 m s-1 on sheep teatcups (p < 0.0001). The increased growth and faster expansion of staphylococci on silicone teatcups raise important points from a clinical viewpoint. The model could be used in the testing of staphylococcal growth in the material of milking parlours in various conditions.

SUBMITTER: Katsarou EI

PROVIDER: S-EPMC8071573 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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Growth of Staphylococcus epidermidis on the Surface of Teatcups from Milking Parlours.

Katsarou Eleni I EI Katsafadou Angeliki I AI Karakasidis Theodoros T Chatzopoulos Dimitris C DC Vasileiou Natalia G C NGC Lianou Daphne T DT Mavrogianni Vasia S VS Petinaki Efthymia E Fthenakis George C GC

Microorganisms 20210415 4

The growth of two Staphylococcus epidermidis isolates (one biofilm-forming and one not) on teatcups for cattle (made of rubber) or sheep (made of silicone) were assessed in nine multiplicates for 24 h post-smearing on the teatcup surface. Staphylococci were smeared on an area of 0.0003142 m2 on the material and their growth and expansion further on were monitored for 24 h. There were no differences in the frequency of recoveries between the two isolates (p > 0.82 for all ...[more]

PMID: 33921135

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Project description:The objectives of the present study were as follows: (a) to describe the prevalence of irregular findings on teatcups in milking parlours with dairy sheep and goats after the post-milking cleaning procedures had been completed, (b) to associate staphylococcal isolation from teatcups with the presence with irregular findings and (c) to identify predictors of the presence of irregular findings on teatcups. The teatcups in the milking parlour of 255 sheep and 66 goat farms were macroscopically evaluated for the presence of irregular findings immediately after the completion of cleaning of the parlour. In total, 1115 and 303 teatcups, respectively, were assessed. A detailed interview with the farmer served to record the characteristics of the milking parlour and obtain information about husbandry and health management variables in the farm. Teatcups with macroscopically evident irregular findings were observed in 150 milking parlours (46.7%). Overall, 593 teatcups (41.8%) were found with macroscopically evident irregular findings. Dirt, milk residues and cracks or tears were recorded in the teatcups of 90.0%, 36.0% and 12.7% of parlours with irregular findings. Staphylococci were more frequently isolated from teatcups with irregular findings than from undamaged ones: from 37.4% (222/593) versus 12.8% (106/825). They were more frequently isolated from teatcups with milk residues (39.4%) and teatcups with dirt (39.0%). Via multivariable analysis, the following three variables emerged as significant predictors of presence of teatcups with irregular findings: the daily number of milking sessions, month into the lactation period at sampling and number of available milking units per animal position. The study provides, for the first time internationally, an appraisal of the frequency of problematic teatcups in the milking parlours of small ruminant farms. The analysis of predictors has provided a focus for specific management aspects, where interventions need to be performed, in order to improve the situation in farms with problems. The consequent increased staphylococcal burden on teatcups with irregular findings points to the increased risk of intramammary infections in such cases.

Project description:There is a paucity of information regarding staphylococcal populations on teatcups of milking parlours in sheep and goat farms. The objectives were to describe the populations of staphylococci on teatcups in milking parlours in sheep or goat farms in two field investigations throughout Greece and to potentially associate the findings with the use of anti-staphylococcal mastitis vaccinations in the farms visited during the two investigations. In a cross-sectional (255 sheep and 66 goat farms across Greece) and a longitudinal (12 sheep farms, four samplings, throughout lactation) study, swab samples were collected from 1418 teatcups (upper and lower part) for staphylococcal recovery, identification and assessment of biofilm-formation. A total of 328 contaminated teatcups (23.1%) were found in 105 sheep (41.2%) and 35 goat (53.0%) farms. Staphylococci were more frequently recovered from the upper than the lower part of teatcups: 269 versus 139 teatcups, respectively. After identification, 253 staphylococcal isolates were found: Staphylococcus aureus, Staphylococcus equorum, Staphylococcus lentus, and Staphylococcus capitis predominated. Of these isolates, 87.4% were biofilm-forming. The proportion of contaminated teatcups was smaller in farms where vaccination against anti-staphylococcal mastitis in general or vaccination specifically against mastitis caused specifically by biofilm-forming staphylococcal strains was applied, 19.7% or 10.9%, respectively, versus 25.5% in farms without vaccination. In the longitudinal study, contaminated teatcups were identified in 28 (58.3%) sampling occasions, with staphylococci being recovered more frequently from their upper part. The same species as in the cross-sectional study predominated. Of these isolates, 61.9% were biofilm-forming. In farms where vaccination against mastitis caused specifically by biofilm-forming staphylococcal strains was applied, the proportion of contaminated teatcups was smaller: 20.4% versus 48.3% in farms without vaccination. There were no differences in proportions of contaminated teatcups between sampling occasions. In conclusion, the great majority of staphylococci recovered from teatcups of milking parlours in sheep and goat farms included biofilm-forming isolates. Reduced staphylococcal isolation was noted in farms where anti-staphylococcal vaccination was performed; this was possibly the effect of reduced excretion of staphylococci in the milk of vaccinated animals.

Project description:The objectives of this work are (a) to describe staphylococci on the teatcups of milking parlours in goat farms and identify predictors for the presence of staphylococcal isolates on the teatcups, (b) to evaluate relationships with total bacterial counts and somatic cell counts in bulk-tank milk, and (c) to establish patterns of susceptibility to antibiotics for the staphylococcal isolates and identify predictors for the recovery of resistant isolates. In a cross-sectional study of 66 goat farms across Greece, swab samples were collected from 303 teatcups (upper and lower part) for staphylococcal recovery, identification, and assessment of biofilm formation. Details regarding health management on the farms (including conditions in the milking parlour) and the socio-demographic characteristics of farmers were collected by means of a structured questionnaire. A total of 87 contaminated teatcups (28.7%) were found on 35 goat farms (53.0%). Staphylococci were more frequently recovered from the upper than the lower part of teatcups: 73 versus 43 teatcups, respectively. After identification, 67 staphylococcal isolates (i.e., excluding similar isolates) were recovered from the teatcups; Staphylococcus aureus, Staphylococcus capitis, and Staphylococcus equorum predominated. Of these isolates, 82.1% were biofilm-forming. In multivariable analysis, the annual incidence of clinical mastitis in the herd emerged as the only significant factor associated with the isolation of staphylococci from the teatcups. Of the 67 isolates, 23 (34.3%) were resistant to at least one antibiotic, and 14 (22.4%) were multi-resistant. Resistance was found most commonly against penicillin and ampicillin (22.4% of isolates), fosfomycin (17.9%), clindamycin (14.9%), erythromycin, and tetracycline (13.4%). In multivariable analysis, the annual incidence of clinical mastitis in the herd and the use of detergent for parlour cleaning emerged as significant factors associated with the isolation of staphylococci resistant to antibiotics.

Project description:Using live microbes as therapeutic candidates is a strategy that has gained traction across multiple therapeutic areas. In the skin, commensal microorganisms play a crucial role in maintaining skin barrier function, homeostasis, and cutaneous immunity. Alterations of the homeostatic skin microbiome are associated with a number of skin diseases. Here, we present the design of an engineered commensal organism, Staphylococcus epidermidis, for use as a live biotherapeutic product (LBP) candidate for skin diseases. The development of novel bacterial strains whose growth can be controlled without the use of antibiotics or genetic elements conferring antibiotic resistance enables modulation of therapeutic exposure and improves safety. We therefore constructed an auxotrophic strain of S. epidermidis that requires exogenously supplied d-alanine. The S. epidermidis NRRL B-4268 Δalr1 Δalr2 Δdat strain (SEΔΔΔ) contains deletions of three biosynthetic genes: two alanine racemase genes, alr1 and alr2 (SE1674 and SE1079), and the d-alanine aminotransferase gene, dat (SE1423). These three deletions restricted growth in d-alanine-deficient medium, pooled human blood, and skin. In the presence of d-alanine, SEΔΔΔ colonized and increased expression of human β-defensin 2 in cultured human skin models in vitro. SEΔΔΔ showed a low propensity to revert to d-alanine prototrophy and did not form biofilms on plastic in vitro. These studies support the potential safety and utility of SEΔΔΔ as a live biotherapeutic strain whose growth can be controlled by d-alanine.IMPORTANCE The skin microbiome is rich in opportunities for novel therapeutics for skin diseases, and synthetic biology offers the advantage of providing novel functionality or therapeutic benefit to live biotherapeutic products. The development of novel bacterial strains whose growth can be controlled without the use of antibiotics or genetic elements conferring antibiotic resistance enables modulation of therapeutic exposure and improves safety. This study presents the design and in vitro evidence of a skin commensal whose growth can be controlled through d-alanine. The basis of this strain will support future clinical studies of this strain in humans.

Project description:Nasopharyngeal carriage of staphylococci spreads potentially pathogenic strains into (peri)oral regions and increases the chance of cross-infections. Some laboratory strains can also move rapidly on hydrated agar surfaces, but the biological relevance of these observations is not clear. Using soft-agar [0.3% (wt/vol)] plate assays, we demonstrate the rapid surface dispersal of (peri)oral isolates of Staphylococcus aureus and Staphylococcus epidermidis and closely related laboratory strains in the presence of mucin glycoproteins. Mucin-induced dispersal was a stepwise process initiated by the passive spreading of the growing colonies followed by their rapid branching (dendrites) from the colony edge. Although most spreading strains used mucin as a growth substrate, dispersal was primarily dependent on the lubricating and hydrating properties of the mucins. Using S. aureus JE2 as a genetically tractable representative, we demonstrate that mucin-induced dendritic dispersal, but not colony spreading, is facilitated by the secretion of surfactant-active phenol-soluble modulins (PSMs) in a process regulated by the agr quorum-sensing system. Furthermore, the dendritic dispersal of S. aureus JE2 colonies was further stimulated in the presence of surfactant-active supernatants recovered from the most robust (peri)oral spreaders of S. aureus and S. epidermidis. These findings suggest complementary roles for lubricating mucins and staphylococcal PSMs in the active dispersal of potentially pathogenic strains from perioral to respiratory mucosae, where gel-forming, hydrating mucins abound. They also highlight the impact that interspecies interactions have on the co-dispersal of S. aureus with other perioral bacteria, heightening the risk of polymicrobial infections and the severity of the clinical outcomes.ImportanceDespite lacking classical motility machinery, nasopharyngeal staphylococci spread rapidly in (peri)oral and respiratory mucosa and cause cross-infections. We describe laboratory conditions for the reproducible study of staphylococcal dispersal on mucosa-like surfaces and the identification of two dispersal stages (colony spreading and dendritic expansion) stimulated by mucin glycoproteins. The mucin type mattered as dispersal required the surfactant activity and hydration provided by some mucin glycoproteins. While colony spreading was a passive mode of dispersal lubricated by the mucins, the more rapid and invasive form of dendritic expansion of Staphylococcus aureus and Staphylococcus epidermidis required additional lubrication by surfactant-active peptides (phenol-soluble modulins) secreted at high cell densities through quorum sensing. These results highlight a hitherto unknown role for gel-forming mucins in the dispersal of staphylococcal strains associated with cross-infections and point at perioral regions as overlooked sources of carriage and infection by staphylococci.

Dataset Information

Growth of Staphylococcus epidermidis on the Surface of Teatcups from Milking Parlours.

Publications

Growth of <i>Staphylococcus epidermidis</i> on the Surface of Teatcups from Milking Parlours.

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