Intramammary infections and teat canal colonization with coagulase-negative staphylococci after postmilking teat disinfection: species-specific responses.

Coagulase-negative staphylococci (CNS) are the most common pathogens associated with intramammary infections (IMI) in dairy cows. We hypothesized that postmilking teat disinfection would reduce microbial colonization of the teat canal and thus reduce the prevalence of IMI caused by certain CNS species. The efficacy of iodine postmilking teat dip was tested against CNS colonization of the teat canal, and incidence of IMI was measured. Using an udder-half model, 43 Holstein cows at the Washington State University Dairy were enrolled in the trial; postmilking teat dip was applied to one udder-half, treatment (TX), and the remaining half was an undipped control (CX). Teat canal swabbing and mammary quarter milk samples were taken in duplicate once a week for 16 wk for microbial culture. Isolates from agar cultures were presumptively identified as CNS and then speciated using PCR-RFLP and agarose gel electrophoresis. Colonization of the teat canal and IMI by CNS were assessed. Thirty CNS IMI were diagnosed and the number of new IMI in CX quarters (21) was significantly greater than that in TX mammary quarters (9). The majority of CNS IMI were caused by Staphylococcus chromogenes (30%) and Staphylococcus xylosus (40%), and the latter were appreciably reduced by teat dip. Except for S. xylosus, an association was observed between teat canal colonization and IMI by all CNS species in this study, in which the majority of IMI were preceded by teat canal colonization. The total number of CNS IMI was greater for CX group cows compared with TX group cows. However, the effect of disinfection on IMI did not appear to be the same for all CNS species.

Salmonella Cerro isolated over the past twenty years from various sources in the US represent a single predominant pulsed-field gel electrophoresis type.

Salmonella Cerro prevalence in US dairy cattle has increased significantly during the past decade. Comparison of 237 Salmonella isolates collected from various human and animal sources between 1986 and 2009 using pulsed-field gel electrophoresis, antimicrobial resistance typing, and spvA screening, showed very limited genetic diversity, indicating clonality of this serotype. Improved subtyping methods are clearly needed to analyze the potential emergence of this serotype. Our results thus emphasize the critical importance of population-based pathogen surveillance for the detection and characterization of potentially emerging pathogens, and caution to critically evaluate the adequacy of diagnostic tests for a given study population and diagnostic application.

Incidence and transmission of Mycoplasma bovis mastitis in Holstein dairy cows in a hospital pen: A case study.

The objective was to determine the incidence and transmission of mycoplasma mastitis in the hospital pen in a dairy herd of 650 lactating cows after a hospital pen was established following an outbreak of this disease. Mycoplasma mastitis status was monitored for 3 months through repeated collection of milk samples from cows with clinical mastitis (CM) and from bulk tank milk. During the outbreak 13 cows were diagnosed with Mycoplasma bovis CM, 1 cow with Mycoplasma sp. mastitis and 8 cows showed signs of arthritis, 3 of which were confirmed as having M. bovis arthritis. M. bovis isolates from cows with CM, arthritis and bulk tank milk had indistinguishable chromosomal digest pattern fingerprints. Incidence rates of M. bovis CM cases in the milking and hospital pens were 0.01 and 1.7 cases per 100 cow-days at risk. Approximately 70% of cows with M. bovis CM became infected within 12 days of entering the hospital pen. Transmission of M. bovis in the hospital pen occurred as 3 episodes. Each episode corresponded to the introduction of a cow with M. bovis CM from a milking pen. Evidence indicates that cows with M. bovis CM from milking pens were the source of transmission of the disease in the hospital pen and thus their presence in the hospital pen appeared to be a risk factor for transmission of M. bovis mastitis in this single case study herd.

The prevalence of multidrug resistance is higher among bovine than human Salmonella enterica serotype Newport, Typhimurium, and 4,5,12:i:- isolates in the United States but differs by serotype and geographic region.

Salmonella represents an important zoonotic pathogen worldwide, but the transmission dynamics between humans and animals as well as within animal populations are incompletely understood. We characterized Salmonella isolates from cattle and humans in two geographic regions of the United States, the Pacific Northwest and the Northeast, using three common subtyping methods (pulsed-field gel electrophoresis [PFGE], multilocus variable number of tandem repeat analysis [MLVA], and multilocus sequence typing [MLST]). In addition, we analyzed the distribution of antimicrobial resistance among human and cattle Salmonella isolates from the two study areas and characterized Salmonella persistence on individual dairy farms. For both Salmonella enterica subsp. enterica serotypes Newport and Typhimurium, we found multidrug resistance to be significantly associated with bovine origin of isolates, with the odds of multidrug resistance for Newport isolates from cattle approximately 18 times higher than for Newport isolates from humans. Isolates from the Northwest were significantly more likely to be multidrug resistant than those from the Northeast, and susceptible and resistant isolates appeared to represent distinct Salmonella subtypes. We detected evidence for strain diversification during Salmonella persistence on farms, which included changes in antimicrobial resistance as well as genetic changes manifested in PFGE and MLVA pattern shifts. While discriminatory power was serotype dependent, the combination of PFGE data with either MLVA or resistance typing data consistently allowed for improved subtype discrimination. Our results are consistent with the idea that cattle are an important reservoir of multidrug-resistant Salmonella infections in humans. In addition, the study provides evidence for the value of including antimicrobial resistance data in epidemiological investigations and highlights the benefits and potential problems of combining subtyping methods.

Multilocus variable-number tandem-repeat analysis and plasmid profiling to study the occurrence of blaCMY-2 within a pulsed-field gel electrophoresis-defined clade of Salmonella enterica serovar Typhimurium.

Salmonella enterica serovar Typhimurium circulating in food animal populations and carrying resistance to antimicrobial agents represents a human health risk. Recently, a new clade of S. Typhimurium, WA-TYP035/187, was reported in cattle and humans in the Pacific Northwest, United States of America. The objective of this study was to describe a possible mechanism of acquisition of expanded-spectrum cephalosporin resistance in this clade. Ceftazidime resistance increased steadily among WA-TYP035/187 isolates, from 0% (0/2) in 1999 to 77.8% (28/36) in 2006 (chi2 for linear trend, P value of <0.001). Among 112 bovine-source and 18 human-source isolates, 49 (43.8%) and 12 (66.7%) were resistant to ceftazidime, respectively. Multiple-locus variable-number tandem-repeat analysis (MLVA) and plasmid profiling suggested that resistance was acquired by multiple independent genetic events within the WA-TYP035/187 clade. Given the lack of an obvious reservoir in species other than cattle and a parallel rise in ceftiofur resistance in the bovine-specific serovar Salmonella enterica serovar Dublin in the same time frame and region, selection pressure due to the use of the expanded-spectrum cephalosporin drug ceftiofur in cattle is a likely factor driving the increasing cephalosporin resistance of WA-TYP035/187.

Association between an outbreak strain causing mycoplasma bovis mastitis and its asymptomatic carriage in the herd: a case study from Idaho, USA.

The objective of this study was to determine the association between mycoplasma mastitis and colonization of mycoplasma organisms at body sites of asymptomatic carriers. The investigation was done in a dairy herd with a first outbreak of mycoplasma mastitis. Milk and swab solution specimens from accessible mucosal surfaces of body sites from cows and replacements were sampled at quarterly intervals (Herd Samplings 1-4). Samples were cultured and Mycoplasma spp. were isolated, speciated and fingerprinted. During Herd Sampling 1 two cows with mycoplasma bovis mastitis were identified and all swabbing solutions of body site samples from 18 of 84 cows and 36 of 77 replacements were positive to Mycoplasma bovis and fingerprinted as the same strain. A case of clinical M. bovis mastitis developed during Herd Sampling 3. During Herd Samplings 2-4, 4 lactating cows and 12 replacements were positive to M. bovis at various body sites with 4 different strains. Three isolates of Mycoplasma californicum were found from swabbing solutions of three cows during Herd Samplings 3 and 4. Only one strain of M. bovis caused mastitis although four strains were isolated from body sites of animals. Isolation of M. bovis from a body site never preceded mastitis. No lactating cow developed mastitis during Herd Sampling 4 although some animals were colonized with the organism. It appears that during the initial outbreak of M. bovis mastitis colonization of body sites by the outbreak strain may be common. However, the prevalence of colonization subsides and colonization does not appear to precede mastitis.

Introduction of new multidrug-resistant Salmonella enterica strains into commercial dairy herds.

A longitudinal observational study of 59 dairy herds was conducted in Washington State to estimate the rate of introduction of new multidrug-resistant (MDR) Salmonella enterica strains onto commercial dairy herds. Samples were collected on these herds over 7 visits separated by intervals of 2 to 4 mo over a period of 15 to 21 mo. Samples were cultured for Salmonella spp. and serogroup, serovar, and antimicrobial susceptibility patterns were identified for MDR Salmonella isolates. Fingerprinting generated by pulsed-field gel electrophoresis (PFGE) using XbaI restriction enzyme digestion generated genotyping profiles for all MDR isolates identified in the study. The rate of new MDR Salmonella strain introduction was 0.9 per herd-year (95% confidence interval: 0.6-1.4). The rates for the most commonly introduced MDR Salmonella serovars were 0.4/herd-year for Typhimurium, 1.2/herd-year for Newport, and 0.1/herd-year for Dublin. Thirty-three of 59 herds (56%) had at least one new MDR Salmonella introduction during the study period. The number of new MDR Salmonella strains acquired by dairy herds ranged from zero to 8. Thirteen of the 59 herds had a history of clinical salmonellosis. Among these 13 herds, 6 herds acquired new MDR Salmonella strains, although these strains were different than historical clinical strains. These data indicate that acquisition of new MDR Salmonella strains by dairy herds was a common event in participating herds, although the number of strains introduced varied greatly among herds.

The role of animal movement, including off-farm rearing of heifers, in the interherd transmission of multidrug-resistant Salmonella.

Fifty-nine commercial dairy farms were sampled 7 times over 15 to 21 mo to determine the role of animal movement, including off-farm rearing of heifers, in the interherd transmission of multidrug-resistant (MDR) Salmonella spp. Farm management data were collected by on-site inspections and questionnaires on herd management practices before and after the study. Forty-four percent (26/59) of herds did not acquire any new MDR Salmonella strains. The number of newly introduced MDR Salmonella strains acquired by the remaining 56% (33/59) of herds ranged from 1 to 8. Logistic regression models indicated that off-farm heifer raising, including contract heifer raising where heifers commingle with cattle from other farms [commingled heifers, odds ratio (OR) = 8.9, 95% confidence interval (CI): 2.4, 32.80], and herd size per 100-animal increment (herd size, OR = 1.04, 95% CI, 1.01, 1.05) were significantly associated with the introduction of new MDR Salmonella strains. The negative binomial regression similarly revealed that commingled heifers [relative risk (RR) = 2.3, 95% CI: 1.1, 4.7], herd size per 100 animals (RR = 1.02, 95% CI, 1.01, 1.03), and a history of clinical salmonellosis diagnosed before the study (RR = 2.5, 95% CI, 1.3, 5.0) were significantly associated with the number of new MDR Salmonella strains that were introduced. Factors not associated with the introduction of new MDR Salmonella strains were housing of heifers and cows in the same close-up pen, a common hospital-maternity pen, and the number of purchased cattle. This study highlights the role of animal movement in the interherd transmission of MDR Salmonella spp.

Short communication: The effect of centrifugation and resuspension on the recovery of Mycoplasma species from milk.

Low sensitivity of a single bulk tank milk culture is a major limitation for detection of mycoplasma organisms. We hypothesized that sedimentation of Mycoplasma spp. in a milk sample by centrifugation followed by resuspension in a small volume of fluid before agar plating would increase the ability to detect Mycoplasma spp. compared with direct conventional culture. The experiment was conducted to determine recovery of Mycoplasma spp. from milk as affected by 1) treatment (centrifugation vs. conventional method); 2) 2 species (Mycoplasma bovis and Mycoplasma californicum and 4 strains for each species); and 3) 4 different concentrations of Mycoplasma spp. (1,000, 100, 10, and 1 cfu/mL). A 5-mL portion of mycoplasma suspension from each strain was inoculated into 45 mL of fresh bulk tank milk to achieve concentrations of 1,000, 100, 10, and 1 cfu/mL. Treatment samples were vigorously mixed and centrifuged at 5,000 x g for 30 min. Control samples were vigorously mixed. All samples were plated on modified Hayflick agar. Plates were incubated at 37 degrees C and 5% CO(2) for 5 d. Mean (+/-SE) log(10) mycoplasma counts (cfu/mL) in the treatment groups (1.91 +/- 0.15) were higher than those in the control groups (1.70 +/- 0.16). Recovery of at least 1 mycoplasma colony on agar culture was 100% in both treatment and control groups at high, medium, and low concentrations. At the lowest concentration, recovery of at least 1 mycoplasma colony on agar culture in treatment and control groups was 75% (n = 12/16) and 18.75% (n = 3/16), respectively. Centrifugation of milk followed by suspension in a smaller volume of saline before conventional culture increased the ability to detect mycoplasma microorganisms in the milk sample compared with controls. Recovery by centrifugation appeared best at the lowest concentration where detection of a positive sample was 4 times more likely than when conventional methods were used.

The effect of a shortened dry period on intramammary infections during the subsequent lactation.

Several recent studies have investigated the effect of shortened dry periods on milk production in the subsequent lactation. What is lacking from these studies is an understanding of the effect that a shortened dry period has on udder health. Four herds, 156 cows, were studied to determine if a shortened dry period (30 d) had a negative effect on mammary gland health during the subsequent lactation as opposed to cows assigned to a long, 45 or 60 d, dry period. Cows in 2 herds were assigned to either 30- or 60-d dry periods (group I), whereas cows in the other 2 herds were assigned to either 30- or 45-d dry periods (group II). Intramammary instillation of commercial preparations of cephapirin benzathine, 300 mg (dry cow formulation), was given to cows assigned a 45- or 60-d dry period length protocol, and 200 mg (lactating cow formulation) was administered to cows assigned a 30-d dry period. Differences in response variables to dry period length were compared within group. Cure rates for 60- vs. 30-d dry period cows were 72% (28/39) vs. 81% (30/37) and 74% (25/34) and 73% (27/37) for 45- vs. 30-d dry periods. Differences were not statistically significant for either comparison group. The majority of intramammary infections were caused by the minor pathogens, coagulase-negative staphylococci (n = 102) or Corynebacterium bovis (n = 11). Only 11 cows had intramammary infections by major pathogens. The herd average percentage of new intramammary infections ranged from 6 to 9% and did not differ among herds between treatment groups. Linear somatic cell counts were not significantly affected by dry period length during the first 6 to 7 mo of the subsequent lactation. Milk production did differ between groups. Mature equivalent milk production was greater in group I cows given a 60-d dry period (11,942 +/- 2,059 kg) as opposed to those given a 30-d dry period (10,749 +/- 2,321 kg). Cows given a 45-d dry period did not produce more milk than cows with a 30-d dry period in group II. Although shortening the dry period to 30 d did not have untoward effects on mammary gland health as measured by intramammary infections or milk somatic cell counts, production may be adversely affected when dry periods are shortened to 30 d.

Serological survey for antibodies to Leptospira in dogs and raccoons in Washington State.

A high number of reported canine leptospirosis cases occurred in Washington State from 2004 to 2006. This prompted a serosurvey of healthy dogs from around the state to determine the distribution of exposure risk and to provide insight into serovar epidemiology in the region. In addition, a convenience sample of sera from injured raccoons was also tested, and clinical serological data from the Washington Animal Disease Diagnostic Laboratory were examined. The proportion of dogs with an antibody titre (>or=1:100) to any serovar was 27/158 (17.1%, 95% CI 11.6-23.9), and that proportion among raccoons was 22/115 (19.1%, 95% CI 12.4-27.5) suggesting that the potential for exposure in Washington state is not uncommon. The most frequently detected serovars in healthy dogs were Autumnalis, Icterohemorrhagiae and Canicola, in clinical canine samples Autumnalis, Bratislava and Pomona were more frequent and in sick or injured raccoons Autumnalis, and Pomona were most frequently detected. Clinical canine serology demonstrated a late summer-fall seasonality that was consistent with other reports. An outbreak of canine leptospirosis occurred during 2004-2006 and was located primarily in western Washington counties, as were three reported human cases in 2005. Canine leptospirosis surveillance is an important tool for detecting human risk of exposure and may provide insights into which serovars are currently of clinical importance.

Associations between bovine, human, and raw milk, and beef isolates of non-O157 Shiga toxigenic Escherichia coli within a restricted geographic area of the United States.

A survey for Shiga toxigenic Escherichia coli in raw milk and beef was conducted within a defined geographic region of the United States. Prevalence rates based on detection of Shiga toxin gene (stx) were 36% for retail beef, 23% for beef carcasses, and 21% for raw milk samples, which were significantly higher than were Shiga toxigenic E. coli isolation rates of 7.5, 5.8, and 3.2%, respectively. Seasonal prevalence differences were significant for stx positivity among ground beef and milk samples. Distribution of stx subtypes among isolates varied according to sample type, with stx1 predominating in milk, stx2 on carcasses, and the combination of both stx1 and stx2 in beef. Ancillary virulence markers eae and ehx were evident in 23 and 15% of isolates, respectively. Pulsed-field gel electrophoresis demonstrated associations between food isolates and sympatric bovine fecal, and human clinical isolates. These data demonstrate that non-O157 Shiga toxigenic E. coli is present in the food chain in the Pacific Northwest, and its risk to health warrants critical assessment.

Effects of prepartum intramammary antibiotic therapy on udder health, milk production, and reproductive performance in dairy heifers.

Preparturient heifers (n = 561) from 9 herds in 6 US states and 1 Canadian province were enrolled in a study to test the hypothesis that prepartum intramammary therapy would cure existing intramammary infections (IMI) and lead to increased milk production, reduced linear somatic cell count (LSCC), and improved reproductive performance. Mammary secretions were collected 10 to 21 d before expected calving from each quarter. Heifers were then assigned by identification number to receive intramammary therapy consisting of infusion of one tube per mammary quarter of a lactating cow commercial antibiotic preparation containing cephapirin or to a nontreated control group. Overall, 34.1% of mammary quarters were infected with a mastitis pathogen before parturition and 63.4% of heifers had at least one mammary quarter infected. The coagulase-negative staphylococci (CNS) caused the majority (74.8%) of prepartum IMI. Coagulase-positive staphylococci, environmental streptococci, and coliforms accounted for 24.5% of prepartum infections. Treatment had a significant effect on the cure rate of infected mammary quarters. Mammary quarters that were infected prepartum and treated with antibiotics had a 59.5% efficacy of cure rate and the percentage reduction in heifers with IMI was 51.9. Control quarters had a spontaneous cure rate of 31.7%. Treatment did not significantly affect milk production or LSCC in the first 200 d of lactation; however, there was a significant treatment by herd interaction for milk production. Quarters cured of either CNS or major pathogens had a lower LSCC in the first 200 d of lactation. No significant effect on services per conception or days open between treatment and control groups was observed. This trial demonstrated that prepartum intramammary antibiotic therapy did reduce the number of heifer IMI postpartum. Milk production, LSCC, and reproductive performance during the first 200 d of the first lactation were not significantly affected by treatment. Given these results, use of prepartum intramammary antibiotic therapy in heifers as a universal strategy to increase milk production in first-lactation dairy cows may not be warranted.

Comparison of a waterless hand-hygiene preparation and soap-and-water hand washing to reduce coliforms on hands in animal exhibit settings.

Outbreaks of enteric disease associated with exposure to live animals on exhibit have occurred with increasing frequency in recent years. Possibly the most important pathogen causing such outbreaks is Escherichia coli O157:H7, because of the serious illness it can cause. Hand hygiene is consistently protective against disease among persons exposed to animals implicated in these outbreaks. Livestock barns have limited hand-washing facilities, therefore a waterless hand-sanitizing gel would be a potentially preventive measure readily available to visitors and animal exhibitors. This study compared the reduction of bacterial counts on hands of animal exhibitors when soap and water was used or when an ethanol-based hand gel was used after animal handling. Participants were youth and adults involved with showing livestock. The sanitation methods were similar in reducing the total bacteria and coliform counts on the hands of the participants (Wilcoxon rank sum test P values 0.12 and 0.69 respectively).

Effects of storage time and thawing methods on the recovery of Mycoplasma species in milk samples from cows with intramammary infections.

This study was executed to determine the effects of storage and thawing on the viability of Mycoplasma spp. in milk from cows with intramammary infections. The trial was designed using a control sample and seven handling regimens subjected to two methods of thawing. There was a significant treatment effect on the recovery of colony-forming units in milk samples when comparing the control sample with handling regimens 1 through 7. There was a continuous decline in log (10) mean number of cfu/mL recovered. Mean concentrations were 6.29, 4.64, 3.69, 3.01, 1.86, 4.41, 4.13, and 3.18 for control and handling regimens 1 to 7, respectively. To determine the best thawing method, handling regimen 1 through 7 samples were thawed using two methods. On average, more mycoplasma were recovered from milk samples thawed at ambient temperature (4.04 cfu/mL) than milk samples thawed in a 37 degrees C water bath (3.76 cfu/mL). A final comparison was made between individual treatments. With the exception of the handling regimen 5 to 6 pair-wise comparison, all pair-wise comparisons between handling regimens were significantly different. The results of this study indicate that storage and thawing of milk samples is harmful to mycoplasma organisms. Fresh samples should be used to improve detection of Mycoplasma spp. from milk of infected cattle. If frozen samples are used, then length of storage time should be minimized, and thawing milk at ambient temperature will improve recovery of mycoplasma as opposed to thawing in a 37 degrees C water bath.

Longitudinal study of fecal shedding of Escherichia coli O157:H7 in feedlot cattle: predominance and persistence of specific clonal types despite massive cattle population turnover.

Identification of the sources and methods of transmission of Escherichia coli O157:H7 in feedlot cattle may facilitate the development of on-farm control measures for this important food-borne pathogen. The prevalence of E. coli O157:H7 in fecal samples of commercial feedlot cattle in 20 feedlot pens between April and September 2000 was determined throughout the finishing feeding period prior to slaughter. Using immunomagnetic separation, E. coli O157:H7 was isolated from 636 of 4,790 (13%) fecal samples in this study, with highest prevalence earliest in the feeding period. No differences were observed in the fecal or water trough sediment prevalence values of E. coli O157:H7 in 10 pens supplied with chlorinated drinking water supplies compared with nonchlorinated water pens. Pulsed-field gel electrophoresis of XbaI-digested bacterial DNA of the 230 isolates obtained from eight of the pens revealed 56 unique restriction endonuclease digestion patterns (REDPs), although nearly 60% of the isolates belonged to a group of four closely related genetic subtypes that were present in each of the pens and throughout the sampling period. The other REDPs were typically transiently detected, often in single pens and on single sample dates, and in many cases were also closely related to the four predominant REDPs. The persistence and predominance of a few REDPs observed over the entire feeding period on this livestock operation highlight the importance of the farm environment, and not necessarily the incoming cattle, as a potential source or reservoir of E. coli O157:H7 on farms.

Removal of hair surrounding the teat and associated bacterial counts on teat skin surface, in milk, and intramammary infections.

The effectiveness of monthly removal of hair surrounding teats on the reduction of teat skin surface bacteria, and the incidence of intramammary infection (IMI), was studied for 10 months in a dairy farm. A split udder design was used where hair was removed on one side, left or right, with the other side serving as a control. Controls and treatment sides were randomly applied in a systematic fashion to 218 cows. Standard milking time pre- and post-milking hygiene practices were applied to all udders during the trial. Collection of teat skin swab solutions preceded aseptic collection of milk samples, performed at monthly intervals, immediately prior to milking. Teat skin bacterial counts did not differ between control and treated teats. Incidences of IMI were similar for treatment when compared with control mammary quarters, as measured by total or by pathogen type. In a second study, the effect of hair removal on the bacterial content of milk was determined using 40 cows. Treatments and allocations were as described. Udder half milk, milk from both mammary quarters of each udder half, was combined and diverted into separate buckets. Buckets were thoroughly cleaned and sanitized between milkings. A portion of bucket milk was collected 24 h after removal of udder hair. The total milk bacterial counts, and counts of psychrotrophs and thermoduric organisms were not reduced by udder hair removal. Results do not suggest that removal of udder hair leads to an improvement in milk quality as determined by milk bacterial content in the herd studied.

Correlation between geographic distance and genetic similarity in an international collection of bovine faecal Escherichia coli O157:H7 isolates.

Evidence from epidemiological and molecular studies of bovine Escherichia coli O157:H7 suggests that strains are frequently transmitted across wide geographic distances. To test this hypothesis, we compared the geographic and genetic distance of a set of international bovine Escherichia coli O157:H7 isolates using the Mantel correlation. For a measure of genetic relatedness, pulsed-field gel electrophoresis of six different restriction enzyme digests was used to generate an average Dice similarity coefficient for each isolate pair. Geographic distance was calculated using latitude and longitude data for isolate source locations. The Mantel correlation between genetic similarity and the logarithm of geographic distance in kilometers was -0.21 (P<0.001). The low magnitude of the Mantel correlation indicates that transmission over long distances is common. The occurrence of isolates from different continents on the same cluster of the dendrogram also supports the idea that Escherichia coli O157:H7 strains can be transferred with considerable frequency over global distances.

Bulk tank milk analysis: factors associated with appearance of Mycoplasma sp. in milk.

Factors associated with the presence of Mycoplasma sp. in bulk tank milk samples were evaluated from 664 herds during 2.25 years. Milk quality components were not strongly related to the presence of Mycoplasma sp. in bulk tank milk. The presence of other contagious mastitis pathogens, Staphylococcus aureus and Streptococcus agalactiae, was also not related to the presence of mycoplasma, suggesting that the aetiology and transmission of mycoplasma mastitis were different from transmission of other contagious mastitis pathogens. The occurrence of the first isolation of mycoplasma from a bulk tank was not correlated to season of the year. Mycoplasma in bulk tank milk samples were more likely to be found in herds shipping more milk, an indirect measure of herd size. This suggests that larger herds are more likely to have mycoplasma mastitis. However, the first appearance of mycoplasma mastitis in a bulk tank sample was followed by a sample without this pathogen in more than 60% of herds. Mycoplasma sp. was not detected in any herd a year after first isolation. These findings suggest that this pathogen could be controlled and eliminated from herds.