Molecular epidemiology and transmission dynamics of S.uberis bovine clinical mastitis

Mastitis remains one of the most common, costly and intractable diseases affecting the dairy cattle industry worldwide. In spite of concerted efforts meaningful progress in reducing the incidence of mastitis has been limited over the past thirty years by our partial understanding of the epidemiology of key pathogens, such as S.uberis. The aim of this thesis was to investigate the epidemiology of S.uberis mastitis by analysis of the population structure and transmission dynamics of clinical mastitis patterns within and between commercial dairy herds in England and Wales. In Chapter 3 Multi Locus Sequence Typing (MLST) was used to describe the sub-species (sequence type) genetic heterogeneity of S.uberis isolates collected from a longitudinal study of clinical mastitis on 52 farms. The clinical cases were classified according the pattern of occurrence of the sequence types in the herds. The findings suggested that a small subset of MLST sequence types were disproportionally important in the epidemiology of S.uberis mastitis, with cow-to-cow transmission of S.uberis, potentially occurring in the majority of herds in the United Kingdom; this may be the most important route of S.uberis transmission in approximately one third of herds. In Chapter 4 cow and herd level variables, including monthly recorded milk constituents, yield and parity were evaluated against the clinical case classifications defined in Chapter 3. The temporal relationships between clinical cases and classifications were also evaluated as potential predictors of transmission dynamics within a herd. The findings indicated that the time interval between clinical cases classified by genotype as potentially contagious transmission was significantly shorter than that between successive mastitis clinical cases attributed to environmental transmission. The distribution of clinical cases throughout lactation also indicated a higher proportion of potentially contagious isolates were cultured from clinical cases originating from lactation rather than the dry period compared to those attributed to environmental transmission. In Chapter 5 Matrix Assisted Laser Desorption (MALDI) mass spectroscopy was used to generate spectral profiles of S.uberis isolates cellular composition. Spectral profiles were used successfully as an alternative method of discriminating between clinical mastitis isolates associated with contagious transmission from those associated with and environmental origin of infection defined in Chapter 3. The findings of this chapter demonstrated marked variation between herds in the spectral profiles of isolates from the same clinical case classification. In Chapter 6 selected isolates of S.uberis associated with contagious transmission and persistent intramammary infections were sequenced using next generation technology and compared by core genome multi locus sequence typing (cgMLST) to evaluate the discriminatory capacity of the MLST and MALDI. The results from this chapter confirmed the importance of the herd unit in the genotypic population structure of S.uberis suggested by the results of Chapter 5 and also support the results of Chapter3 which suggest ‘low grade’ contagious transmission of S.uberis is superimposed on a ubiquitous, environmental S.uberis mastitis pattern in many herds This thesis demonstrated that the S.uberis population is complex with variation at the bacterial, cow and herd level suggesting different patterns of disease and bacterial evolution occurring in different circumstances. Categorisation of sub-species of S.uberis within herds as being transmitted via contagious or environmental routes appears to be eminently achievable using modern, high throughput technologies; this could lead to a step change in mastitis control