Genomics-based epidemiology of bovine Mycoplasma bovis strains in Israel.

BACKGROUND: Mycoplasma bovis is an important etiologic agent of bovine mycoplasmosis affecting cattle production and animal welfare. In the past in Israel, M. bovis has been most frequently associated with bovine respiratory disease (BRD) and was rarely isolated from mastitis. This situation changed in 2008 when M. bovis-associated mastitis emerged in Israel. The aim of this study was to utilize whole genome sequencing to evaluate the molecular epidemiology and genomic diversity of M. bovis mastitis-associated strains and their genetic relatedness to M. bovis strains isolated from BRD in local feedlot calves and those imported to Israel from different European countries and Australia. RESULTS: Phylogeny based on total single nucleotide polymorphism (SNP) analysis of 225 M. bovis genomes clearly showed clustering of isolates on the basis of geographical origin: strains isolated from European countries clustered together and separately from Australian and Chinese isolates, while Israeli isolates were found in the both groups. The dominant genotype was identified among local mastitis-associated M. bovis isolates. This genotype showed a close genomic relatedness to M. bovis strains isolated from calves imported to Israel from Australia, to original Australian M. bovis strains, as well as to strains isolated in China. CONCLUSIONS: This study represents the first comprehensive high-resolution genome-based epidemiological analysis of M. bovis in Israel and illustrates the possible dissemination of the pathogen across the globe by cattle trade.

The National Mastitis Council: A Global Organization for Mastitis Control and Milk Quality, 50 Years and Beyond.

The National Mastitis Council was founded in 1961 based on the desire of a forward-thinking group of individuals to bring together "all forces of organized agriculture in the United States to combat, through every practical device, the mastitis threat to the Nation's health and food safety". What started as a small organization focused on mastitis of dairy cattle in the United States has grown into a global organization for mastitis and milk quality. Over the last 50-plus years the concerted efforts of the membership have led to the synthesis and dissemination of a considerable body of knowledge regarding udder health, milk quality, and food safety which has improved dairy cattle health and well-being and farm productivity.

Mycoplasma mastitis in cattle: To cull or not to cull.

Bovine mastitis caused by mycoplasmas, in particular Mycoplasma bovis, is a major problem for milk production and animal welfare in large dairy herds in the USA and a serious, although sporadic, disease in Europe and the Middle East. It causes severe damage to the udder of cattle and is largely untreatable by chemotherapy. Mycoplasma mastitis has a distinct epidemiology and a unique set of risk factors, the most important of which is large herd size. The disease is often self-limiting, disappearing within months of outbreaks, sometimes without deliberate intervention. Improved molecular diagnostic tests are leading to more rapid detection of mycoplasmas. Typing tests, such as multi-locus sequence typing, can help trace the source of outbreaks. An approach to successful control is proposed, which involves regular monitoring and rapid segregation or culling of infected cows. Serious consideration should be given by owners of healthy dairy herds to the purchase of M. bovis-free replacements. Increased cases of disease could occur in Europe and Israel if the trend for larger dairy herds continues.

Pulsed-field gel electrophoresis patterns of Mycoplasma isolates from various body sites in dairy cattle with Mycoplasma mastitis.

OBJECTIVE: To determine whether Mycoplasma strains typically associated with mastitis in dairy cattle can be isolated from body sites other than the mammary gland. DESIGN: Prospective clinical trial. ANIMALS: 7 Holstein cows in various stages of lactation with intramammary Mycoplasma infection. PROCEDURE: Milk samples, antemortem swab specimens from various body sites, and postmortem swab and tissue specimens were submitted for Mycoplasma culture. Pulsed-field gel electrophoresis (PFGE) was performed on chromosomal digests of all Mycoplasma isolates. Isolates with the same number and size of chromosomal digest bands were considered to be of the same type. RESULTS: For each cow, all isolates obtained from milk, mammary gland parenchyma, and supramammary lymph nodes had the same PFGE pattern. All cows had at least 1 isolate from nonmammary system tissues that had the same PFGE pattern as isolates from the mammary system. Overall, 44 of the 70 (63%) Mycoplasma isolates obtained from body sites other than mammary system sites had the same PFGE pattern as did mammary system isolates. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirmed our hypothesis that Mycoplasma strains isolated from the milk of dairy cattle with Mycoplasma mastitis frequently have PFGE patterns identical to those for strains isolated from other body sites, suggesting that there is at least a potential for internal transmission of Mycoplasma organisms.

Epidemiologic study of intramammary infections with Staphylococcus aureus during a control program in nine commercial dairy herds.

OBJECTIVE: To determine the epidemiologic pattern of intramammary infections (IMIs) with Staphylococcus aureus during implementation of a control program in 9 commercial dairy herds. DESIGN: Cohort study. ANIMALS: 1,651 lactating cows and 53,098 quarter milk samples. PROCEDURES: Nine herds located in different regions of Italy were enrolled. Control of S. aureus infections followed the general principles of contagious mastitis control and was based on precise diagnostic procedures and strict control and segregation of infected cows. All lactating cows in each herd were tested, and those free of S. aureus IMI were enrolled as the cohorts. Further additions to the cohort group were cows and heifers free of S. aureus IMI, as determined from aseptically collected milk samples taken approximately 7 and 14 days after calving. RESULTS: After the ninth month of the program, incidence decreased to < 2 new IMIs/100 cow-months in 7 of the herds. At the end of the study, 8 of 9 herds had an incidence of < or = 1 new IMI/100 cow-months. Heifers were most at risk of developing an IMI. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that control of S. aureus IMIs can be achieved by use of a control program based on segregation and use of hygienic and therapeutic protocols. Analysis of incidence and identification of risk factors in a herd could avoid the possible shortcomings of the program, maximizing the probability of success.

Patterns of mycoplasma shedding in the milk of dairy cows with intramammary mycoplasma infection.

OBJECTIVE: To determine patterns of mycoplasma shedding in the milk of dairy cows with intramammary mycoplasma infection. DESIGN: Prospective clinical trial. ANIMALS: 10 Holstein cows with intramammary mycoplasma infection. PROCEDURE: Milk samples were collected from each cow daily for 28 days and plated on mycoplasma agar to evaluate shedding patterns. To determine whether enrichment improved recovery of organisms, some samples were also inoculated in mycoplasma enrichment medium and incubated for 4 days prior to plating. Somatic cell count (SCC) was determined in samples collected weekly. RESULTS: Mycoplasma organisms were not isolated from 81 of 280 (29%) composite milk samples, but > 10(6) colonies/mL were obtained from 151 (54%). Similarly, mycoplasma organisms were not isolated from 433 of 1,008 (43%) quarter milk samples, but > 10(6) colonies/mL were obtained from 392 (39%). For 71 of 104 (68%) samples, mycoplasma organisms were isolated both following direct plating and following enrichment; for 24 of 104 (23%), mycoplasma organisms were isolated only following enrichment; and for 9 of 104 (9%), mycoplasma organisms were isolated only after direct plating. There was a linear correlation between logarithm of the SCC and logarithm of the number of colony-forming units of mycoplasma per milliliter of milk for composite and quarter milk samples. CONCLUSIONS AND CLINICAL RELEVANCE: Shedding of organisms was inconsistent in dairy cows with intramammary mycoplasma infection, increasing the risk of misdiagnosis if multiple milk samples are not tested.

Characterization of fatty acid modifying enzyme activity in staphylococcal mastitis isolates and other bacteria.

BACKGROUND: Fatty acid modifying enzyme (FAME) has been shown to modify free fatty acids to alleviate their bactericidal effect by esterifying fatty acids to cholesterol or alcohols. Although it has been shown in previous studies that FAME is required for Staphylococcus aureus survival in skin abscesses, FAME is poorly studied compared to other virulence factors. FAME activity had also been detected in coagulase-negative staphylococci (CNS). However, FAME activity was only surveyed after a bacterial culture was grown for 24 h. Therefore if FAME activity was earlier in the growth phase, it would not have been detected by the assay and those strains would have been labeled as FAME negative. RESULTS: Fifty CNS bovine mastitis isolates and several S. aureus, Escherichia coli, and Streptococcus uberis strains were assayed for FAME activity over 24 h. FAME activity was detected in 54% of CNS and 80% S. aureus strains surveyed but none in E. coli or S. uberis. While some CNS strains produced FAME activity comparable to the lab strain of S. aureus, the pattern of FAME activity varied among strains and across species of staphylococci. All CNS that produced FAME activity also exhibited lipase activity. Lipase activity relative to colony forming units of these CNS decreased over the 24 h growth period. No relationship was observed between somatic cell count in the milk and FAME activity in CNS. CONCLUSIONS: Some staphylococcal species surveyed produced FAME activity, but E. coli and S. uberis strains did not. All FAME producing CNS exhibited lipase activity which may indicate that both these enzymes work in concert to alter fatty acids in the bacterial environment.