Invited review: Prevalence, risk factors, treatment, and barriers to best practice adoption for lameness and injuries in dairy cattle-A narrative review.

Lameness and leg injuries are both painful and prevalent across the dairy industry, and are a major welfare concern. There has been a considerable amount of research focused on investigating the risk factors associated with lameness and injuries and how they might be prevented and treated. The objectives of this narrative review were to summarize herd-level prevalence estimates, risk factors, strategies for prevention, control, and treatment of these conditions, and the barriers to best practice adoption for lameness and injuries on dairy farms. There is a relatively high within-herd prevalence of lameness on dairy farms globally, with a recent systematic review estimating the mean prevalence at 22.8%. Similarly, there is a relatively high prevalence of hock injuries, with within-herd estimates ranging from 12% to 81% of cows affected. Knee and neck injuries have been reported to be less common, with 6% to 43% and 1% to 33%, respectively. Numerous risk factors have been associated with the incidence of lameness, notably housing (e.g., access to pasture, bedding depth, bedding type, flooring type, stall design), management (e.g., stall cleanliness, frequency of trimming, holding times, stocking density), and cow-level (e.g., body condition, parity, injured hocks) factors. Risk factors associated with hock injuries can be similarly classified into housing (e.g., bedding type and depth, outdoor access, parlor type, stall design), management (e.g., bedding depth, cleanliness), and cow (e.g., parity, days in milk, lameness) factors. Key preventative approaches for lameness include routine preventative and corrective hoof trimming, improving hoof cushioning and traction through access to pasture or adding rubber flooring, deep-bedded stalls, sand bedding, ensuring appropriate stocking densities, reduced holding times, and the frequent use of routine footbaths. Very little research has been conducted on hock, knee, and neck injury prevention and recovery. Numerous researchers have concluded that both extrinsic (e.g., time, money, space) and intrinsic (e.g., farmer attitude, perception, priorities, and mindset) barriers exist to addressing lameness and injuries on dairy farms. There are many diverse stakeholders in lameness and injury management including the farmer, farm staff, veterinarian, hoof trimmer, nutritionist, and other advisors. Addressing dairy cattle lameness and injuries must, therefore, consider the people involved, as it is these people who are influencing and implementing on-farm decisions related to lameness prevention, treatment, and control.

Investigating Ontario dairy farmers motivations and barriers to the adoption of biosecurity and Johne's control practices.

For the control of Johne's disease (JD), management practices to minimize disease transmission must be implemented and maintained. Once infected, animals will enter a latent phase and will typically only manifest clinical symptoms years later. As young calves are the main susceptible group on farm, the observed effects of management practices geared toward minimizing their exposure to infective material may not be realized until years later. This delayed feedback limits the sustained implementation of JD control practices. Although quantitative research methods have demonstrated changes to management practices as well as their association with changes to JD prevalence, dairy farmers can offer insights into the current challenges relating to JD implementation and control. Thus, this study aims to use qualitative methods and in-depth interviews (n = 20) with Ontario dairy farmers who had previously been engaged in a Johne's control program to explore their motivations and barriers to the implementation of JD control practices and general herd biosecurity. A thematic analysis using inductive coding was completed generated the following 4 overarching themes: (1) the hows and whys of Johne's control, (2) barriers to general herd biosecurity, (3) barriers to Johne's control, and (4) overcoming barriers. Farmers no longer believed JD was an issue on their farm. Johne's was low on their list of concerns due to little public discourse, absence of animals displaying clinical signs, and no financial support for diagnostic testing. Producers who were still actively engaged in JD control cited animal and human health as their primary motivations. Financial support, targeted education, and promoting engagement through discourse may help encourage producers to reconsider their participation in JD control. Government and industry collaboration with producers may help to develop more effective biosecurity and disease control programs.

Management practices associated with Johne's bulk tank milk ELISA positivity.

Johne's disease (JD) control is often based on the culling of positive animals and the adoption of management practices that minimize exposure of young stock to the pathogen (Mycobacterium avium ssp. paratuberculosis). Throughout 2010 to 2013, the province of Ontario, Canada, instituted a voluntary Johne's control program consisting of whole-herd testing and risk assessment. The JD risk assessment evaluated 5 management areas to characterize herd JD risk. Using a modified milk ELISA technique with an optical density cut-off of 0.089, province-wide bulk tank milk (BTM) testing was used to assess the prevalence of JD high-risk herds at the end of the control program and again 4 yr after its completion. Approximately 71% of Ontario bulk tanks were classified as positive in 2017 compared with roughly 46% in 2013. In 2019, the same JD risk assessment used in the original program was readministered on 180 Ontario dairy farms. Using this cross-sectional approach, logistic regression models were built using data from the original program risk assessment and follow-up risk assessment as well as the BTM ELISA results to determine management factors associated with the control of JD. We demonstrated that management of the maternity area is an important factor in the control of Johne's disease. Although it is believed that the highest risk group for JD infection is calves under 6 mo, the cleanliness scores of older heifers and their exposure to mature cow manure was significantly associated with JD control; farms with highly contaminated weaned and bred heifers and those that had exposure to mature cow manure were more likely to be unsuccessful in their JD control efforts. Careful management of young calves appears to be important for JD control, and this management should continue even after calves have left the maternity area.

Descriptive analysis of the changes in Johne's disease management practices on Ontario dairy farms through repeat risk assessment.

Johne's disease (JD) control is often based on the culling of positive animals and the adoption of management practices that minimize exposure of calves to the pathogen Mycobacterium avium ssp. paratuberculosis (MAP). From 2010 to 2013, Ontario, Canada, instituted a voluntary Johne's control program consisting of whole-herd testing and a Risk Assessment and Management Plan (RAMP). The RAMP consisted of 38 questions that evaluated 5 different management areas to characterize herd risk for MAP introduction and within-herd spread. The RAMP produced a numerical score for each area, with higher scores associated with higher risk. The RAMP focused on animal purchases, calving management, calf management, and heifer and cow cleanliness and management. In the summer of 2019, the RAMP was repeated on 180 farms that had participated in the JD program of 2010 to 2013 and had bulk tank milk ELISA results from 2013 and 2017. This cross-sectional study demonstrated that many producers changed management practices over the 4- to 7-year period. Producers changed their cattle buying practices, with a reduction in purchasing from multiple sources and more herds refraining from buying in animals. However, overall scores were higher in 2019 than in 2013. The 2019 RAMP indicated that fewer farms were utilizing individual calving pens in 2019 than in 2013 (13% vs. 26%), yet more farms had policies in place to deal with sick or suspect JD cows entering the maternity area (92% vs. 74%). Management changes occurred over time, some of which represent increased risk (crowded maternity pens) and others decreased risk (closed herd, protocols in place for JD-positive cows) for MAP introduction and transmission. These results highlight the importance of frequent risk assessments and the documentation of changes to management practices on-farm as a means to assess herd disease risk more accurately.

Promoting farm advisor engagement and action toward the improvement of dairy cattle lameness.

Lameness represents a significant management challenge in the dairy industry worldwide. In addition to farmers, many advisors are involved in lameness management, including veterinarians, hoof trimmers, and nutritionists. These advisors support farmers through lameness prevention practices, treatments, and advice, but little is known about how advisors view others' roles and the extent to which they work together. This study used qualitative participatory methods to facilitate the formation of lameness advisory groups among 13 advisors (4 veterinarians, 5 hoof trimmers, and 4 nutritionists), with the aims of promoting advisors' engagement with one another and documenting their perceptions of the benefits and challenges of the research activities through semi-structured interviews and thematic analysis. Participants shared predominantly positive views toward the project and voiced appreciation about the quality of discussions and the opportunity to connect and build relationships with other advisors. Participants reported improved communication with others via sharing reports and farm information, as well as increased confidence in reaching out to other advisors. Reported challenges included time constraints and discomfort in leading the lameness advisory group meetings. Difficulties were also found in bringing issues to farmers' attention when they might not align with the farmers' goals or priorities, as well as participants' questioning other advisors' knowledge and intentions. This study describes a promising avenue of facilitating advisor engagement, although more work is needed to determine whether such engagement translates to a reduction of lameness on farms.

A large panel of chicken cells are invaded in vivo by Salmonella Typhimurium even when depleted of all known invasion factors.

Poultry are the main source of human infection by Salmonella. As infected poultry are asymptomatic, identifying infected poultry farms is difficult, thus controlling animal infections is of primary importance. As cell tropism is known to govern disease, our aim was therefore to identify infected host-cell types in the organs of chicks known to be involved in Salmonella infection and investigate the role of the three known invasion factors in this process (T3SS-1, Rck and PagN). Chicks were inoculated with wild-type or isogenic fluorescent Salmonella Typhimurium mutants via the intracoelomic route. Our results show that liver, spleen, gall bladder and aortic vessels could be foci of infection, and that phagocytic and non-phagocytic cells, including immune, epithelial and endothelial cells, are invaded in vivo in each organ. Moreover, a mutant defective for the T3SS-1, Rck and PagN remained able to colonize organs like the wild-type strain and invaded non-phagocytic cells in each organ studied. As the infection of the gall bladder had not previously been described in chicks, invasion of gall bladder cells was confirmed by immunohistochemistry and infection was shown to last several weeks after inoculation. Altogether, for the first time these findings provide insights into cell tropism of Salmonella in relevant organs involved in Salmonella infection in chicks and also demonstrate that the known invasion factors are not required for entry into these cell types.

Murine AML12 hepatocytes allow Salmonella Typhimurium T3SS1-independent invasion and intracellular fate.

Numerous studies have demonstrated the key role of the Salmonella Pathogenicity Island 1-encoded type III secretion system (T3SS1) apparatus as well as its associated effectors in the invasion and intracellular fate of Salmonella in the host cell. Several T3SS1 effectors work together to control cytoskeleton networks and induce massive membrane ruffles, allowing pathogen internalization. Salmonella resides in a vacuole whose maturation requires that the activity of T3SS1 subverts early stages of cell signaling. Recently, we identified five cell lines in which Salmonella Typhimurium enters without using its three known invasion factors: T3SS1, Rck and PagN. The present study investigated the intracellular fate of Salmonella Typhimurium in one of these models, the murine hepatocyte cell line AML12. We demonstrated that both wild-type Salmonella and T3SS1-invalidated Salmonella followed a common pathway leading to the formation of a Salmonella containing vacuole (SCV) without classical recruitment of Rho-GTPases. Maturation of the SCV continued through an acidified phase that led to Salmonella multiplication as well as the formation of a tubular network resembling Salmonella induced filaments (SIF). The fact that in the murine AML12 hepatocyte, the T3SS1 mutant induced an intracellular fate resembling to the wild-type strain highlights the fact that Salmonella Typhimurium invasion and intracellular survival can be completely independent of T3SS1.

Dairy farmer, hoof trimmer, and veterinarian perceptions of barriers and roles in lameness management.

Lameness is a leading animal welfare concern in the dairy industry. Multiple stakeholders are involved in lameness management on a dairy farm, including farmers, hoof trimmers, and veterinarians. This study sought to explore perceptions of lameness, perceptions of roles in lameness management, and barriers to improved lameness management in these groups. Fourteen homogeneous focus groups were held in Minnesota, Wisconsin, and New York from April 2017 to March 2020; 5 with farmers (n = 31), 4 with hoof trimmers (n = 32), and 5 with veterinarians (n = 25). The 1-h facilitated discussions were audio-recorded, transcribed verbatim, and common themes identified through thematic analysis. Lameness was perceived by participants as a complex health problem and one in which the connections between pathogenesis, facilities, and management were not always well understood or easy to change. The complexity of the problem encompassed the lack of agreement on a definition of lameness, normalization to its signs, and the interconnectedness of lameness with other health and management issues. These issues appeared to contribute to resignation by participants that lameness was inevitable. Despite shared concerns about lameness among these groups, respondents reported a lack of communication, especially between hoof trimmers and veterinarians. Participants also voiced a desire to work together more productively, with hoof trimmers and veterinarians valuing the ability to deliver a consistent message to farmers. These findings suggest a need for increased efforts to facilitate collaboration between farmers, hoof trimmers, and veterinarians to improve lameness management on dairy farms.

Communication preferences and social media engagement among Canadian dairy producers.

The objective of this study was to determine communication preferences of dairy producers in Canada. A secondary objective was to evaluate social media engagement of dairy producers. A survey was administered to Canadian dairy producers between March and April, 2015 to collect information on current management practices on their farms. A total of 1,373 Canadian dairy producers responded to the survey, representing a response rate of 12%. The survey consisted of 192 questions; however, only questions regarding producer demographics, importance of information sources, and internet and social media use were evaluated in this study. The primary outcome variables of interest included use of the internet to access dairy information, importance of different sources of information about dairy herd health and management, and use of online search engines and social media platforms. For each outcome, logistic regression analyses were used to investigate associations between the outcome and demographic variables. Veterinarians were viewed as a "very important" source of information by the majority of respondents (79%), whereas milk recording and dairy producer organizations were viewed as a "very important" source of information by 36% of respondents. Other producers (46%) and magazines or newspapers (51%) were commonly viewed as an "important" source of information. Online search engines were commonly used by respondents (94%). Social media was viewed as less important, and had mixed levels of use. YouTube (70%), Facebook (63%), and Twitter (18%) were the most commonly used social media platforms. Eighty percent of Twitter users reported using the platform to interact with and obtain or share information about herd health management online, which was the highest reported interactivy regarding herd health among all social media platforms. This exploratory study offers insight into the communication preferences of Canadian dairy producers and can be used to facilitate future communication strategies aimed at engaging rural farming audiences across Canada.

Passive immunity and colostrum management practices on Ontario dairy farms and auction facilities: A cross-sectional study.

Failed transfer of passive immunity (FTPI) in dairy calves has substantial health consequences, but there have been no recently published estimates of the prevalence of FTPI on Ontario dairy farms. The major objectives of this cross-sectional study were to determine the level of FTPI in dairy calves across the province of Ontario and to determine colostrum management practices used. A convenience sample of 109 dairy farms in Ontario were visited from June to August 2019, along with 4 auction facilities from January to February 2019. At the single visit to each dairy farm, an in-person questionnaire was administered, and all calves between 24 h and 9 d of age had a blood sample collected to determine the levels of serum total protein (STP). At the auction facility, a blood sample was collected on all calves that arrived on the day of the visit. The dairy farms were distributed throughout Ontario, and the farms milked on average 151 dairy cows. Of the 444 calves sampled on Ontario dairy farms, 46% were male, 54% were female, and the mean weight and age at sampling were 46.3 kg and 4.4 d, respectively. The mean STP of calves sampled on the dairy farm was 5.69 g/dL (standard deviation = 0.78), and 107 calves (24%) had FTPI. The likelihood of FTPI was not associated with sex (female vs. male, risk ratio = 0.89, 95% confidence interval = 0.64-1.24), age (per day, risk ratio = 1.02, 95% confidence interval = 0.94-1.11), or weight (per kg, risk ratio = 0.98, 95% confidence interval = 0.96-1.01). In contrast, at the auction facilities, 386 male dairy calves were sampled and had a mean STP of 5.79 g/dL (standard deviation = 0.83). Using a cut point of ≥5.2 g/dL, 91 calves (24%) had FTPI. From the in-person questionnaire administered on the dairy farms, it was reported that the median time to first feeding of colostrum was 2.5 h (range = 0-12 h). The farms reported providing a mean of 3.3, 2.9, and 3.4 L of colostrum from 0 to 6 h after birth, 6 to 12 h after birth, and 12 to 24 h after birth, respectively. Ninety-one farms (83%) fed colostrum from the dam of the calf as their predominant source of colostrum. The most common method used to deliver the first feeding of colostrum was a nipple bottle (89 farms, 82%). Twenty-seven farms (25%) reported managing colostrum differently for male calves, which included a different colostrum source, use of poorer quality colostrum, a smaller quantity of colostrum, a longer time from birth to feeding, and generally having a lower focus on colostrum management for male calves. The prevalence of FTPI on Ontario dairy farms appears to have decreased since previous estimates, and substantial room for improvement remains. Although no overall differences were found in FTPI between male and female calves, differential reported colostrum management by sex indicates this may be a risk for male calves on a proportion of Ontario dairy farms.

Short communication: Describing mortality and euthanasia practices on Canadian dairy farms.

The objective of this study was to describe mortality rates and euthanasia practices used for cows, heifer calves, and male dairy calves on Canadian dairy farms. An internet survey was administered to Canadian dairy producers between March and April 2015 to collect information on current management practices. Approximately 81% (867/1,076) and 63% (673/1,065) of respondents reported that at least one animal died unassisted and at least one animal was euthanized on the farm in the preceding 12-mo period, respectively. Overall, mean mortality was 8% for cows, and 6 and 2% for preweaning and weaned heifers, respectively. On average, 48, 76, and 89% of all reported mortality events in cows, weaned heifers, and preweaning heifers were recorded as unassisted deaths. Cows that died without assistance were necropsied more often than preweaning heifers dying without assistance; these cows were also necropsied more than cows that were euthanized. Conversely, preweaning heifers that were euthanized were necropsied more frequently than those that died without assistance. Choosing not to perform necropsies on animals that die of unknown causes could represent a missed opportunity to identify cause of death and inform changes to prevent future deaths. The use of a firearm was the most common method for euthanizing cows and heifers (54%) and male dairy calves (51%). Approximately 7% of respondents reported using blunt force to euthanize animals. The use of this method was much higher for euthanasia of male dairy calves (34%) than for heifer calves (7%). Québec respondents euthanized their animals using blunt force more often than farmers from other regions. Further, 53% of respondents using blunt force indicated it was their primary method of calf euthanasia; these results are extremely concerning. Approximately 31% of respondents who reported that they do not use blunt force to euthanize heifers and cows did report using blunt force to euthanize male dairy calves. These results can be used to inform veterinarian-client communication, broader extension tools and programs, and industry policies to improve dairy cattle health and welfare, a goal that is becoming of increasing importance from a consumer assurance perspective.

Canadian National Dairy Study: Describing Canadian dairy producer practices and perceptions surrounding cull cow management.

Farmer decisions surrounding culling have an important effect on the health and welfare of cull cows. The objectives of this study were to describe the self-reported shipment behaviors of Canadian dairy producers and understand farmer perspectives on the factors that were most influential in their decision to cull a cow. A nationwide survey was administered between March and April 2015 that included 192 questions covering producer background information, farm characteristics, biosecurity practices, disease prevalence, calf health, cow welfare, lameness, milking hygiene, reproduction, and Internet and social media use. The survey yielded a 12% response rate; a total of 1,076 respondents (78% of all survey respondents completed the culling section of the survey) were included in this study for analysis. Approximately 80, 51, and 38% of respondents reported shipping at least 1 cow to auction, direct to slaughter, and to another dairy farm in the past 12 mo, respectively. Ability of the cow to remain standing (93% of respondents) and drug withdrawal times (92% of respondents) were identified as the most important factors for consideration when culling cows. The time between culling decision and when the cow was actually transported was longer for lame cows than sick cows; almost 70% of respondents reported that cows culled for illness were typically shipped within 1 wk of culling decision, whereas only 51% of respondents indicated the same was true for lame cows. Last, Canadian dairy producers generally exhibited strong confidence that their culled cows would arrive at slaughter in the same condition as they left, but felt very unsure about knowing the location of their final destination. These results highlight several gaps between producer perceptions and the true situation, and can be used to develop tailored programs and inform policy and regulatory decisions aimed at improving cull cow decisions and cow welfare.

Cost-benefit of implementing a participatory extension model for improving on-farm adoption of Johne's disease control recommendations.

The objective of this study was to perform a cost-benefit analysis (CBA) of a participatory extension model, called Ontario Focus Farms (FF), which was designed to facilitate the adoption of on-farm management practices to control Johne's disease (JD) on Ontario (ON) dairy farms. Partial budget models were developed to estimate the annual herd cost of JD on an average 78-cow Ontario dairy herd and the annual herd cost of neonatal calf diarrhea (NCD). With these estimates, a CBA was developed to assess the simulated net benefits of implementing various on-farm management scenarios (i.e., implementing 1, 2, or 3 of the following: calf feeding, maternity pen management, maternity area structure changes), where the benefits represent a reduction in the annual cost of JD and NCD. These models informed the final CBA assessing the net benefits of FF implementation over a 10-yr period. All monetary values are reported in Canadian dollars (Can$; where 1 Can$ = 0.823 US$ at the time of the study). The annual herd cost of JD was estimated to be $3,242 ($41.56/cow), and that of NCD was estimated to be $1,390 ($36/heifer calf). When farms were expected to have both JD and NCD, all scenarios, when implemented over a 10-yr period, yielded positive net benefits ranging from $439 to $2,543 per farm when changes to maternity area structure were combined with calf feeding changes. These effects were sensitive to changes in level of disease (JD and NCD) on the farm, and the costs and effects of making changes. The NPV of making any on-farm change when JD was not present on the farm was negative. Overall, FF implementation yielded positive net benefits of $426,351 or $749,808, depending on whether a veterinarian or non-veterinarian served as the facilitator. The NPV was most sensitive to changes in burden of disease, the cost of implementing changes, and the proportion of FF participants that had JD and NCD on the farm. Benefits of FF implementation are also likely to accrue to veterinarians, as a result of professional facilitator training, and the Ontario dairy industry, as a by-product of improved milk quality and safety; therefore, the true net benefits of FF implementation are likely underestimated. Overall, the FF process should be considered an economically viable program and worthy of investment as part of a JD control strategy, as it demonstrates potential to yield positive net benefits for the Ontario dairy industry.

Exploring dairy producer and veterinarian perceptions of barriers and motivators to adopting on-farm management practices for Johne's disease control in Ontario, Canada.

Motivating uptake of management change recommendations requires knowledge of the barriers and motivators influencing producer behavior. This study investigated dairy producers' and veterinarians' perceptions of the barriers and motivators influencing the adoption of Johne's disease (JD) control recommendations in Ontario, Canada. Eight focus groups, 6 with dairy producers and 2 with veterinarians, were conducted and thematically analyzed. Both producer and veterinarian groups identified physical resources (i.e., time, money, infrastructure) and producer mindset (i.e., perceived priority of JD, perceived practicality of JD control recommendations) as key barriers to adoption. Producers tended to prioritize JD control on their farm based on their lived experiences with JD and their view of the public's concern about JD. Many agreed that JD recommendations should focus on biosecurity more holistically and emphasize the broader health benefits of limiting calf exposure to many fecal-orally transmitted diseases. Producers also highlighted that some recommendations for on-farm change (i.e., keeping a closed herd, buying from low-risk herds) were unrealistic or too difficult to perform and often disrupted their habits or routine. In contrast, veterinarians suggested that most recommendations were practical and are routinely recommended. Participants suggested both extrinsic (i.e., incentives, premiums, penalties and regulations, and extension and communication) and intrinsic (i.e., pride and responsibility) methods for motivating producers. This study highlights the importance of producer mindset in on-farm change and offers insights into the attitudes and perceived barriers influencing on-farm change.

Evaluating the effect of Focus Farms on Ontario dairy producers' knowledge, attitudes, and behavior toward control of Johne's disease.

This study evaluated a participatory-based, experiential learning program, Ontario Focus Farms (FF), which aimed to change dairy producer behavior to control Johne's disease (JD) in Ontario, Canada. The goals were to (1) assess the effect of FF on participating dairy producers' knowledge, attitudes, and behavior with regard to JD control; (2) compare changes in these factors among FF participants to changes among a group of nonparticipating dairy producers; and (3) describe the characteristics of producers who made at least one on-farm management change. Pre- and post-FF intervention questionnaires collected data on respondents' knowledge, attitudes, behavior, herd production, and demographic information; before and after JD-risk assessments were used to assess respondents' on-farm risk of JD transmission. Overall, 176 dairy producers participated in the FF process; 39.8% (70/176) of FF and 14.6% (52/357) of control participants responded to both the pre- and postintervention questionnaires. Upon comparison, FF respondents were more likely to be younger, have larger herds, and have higher management scores. The proportion of FF participants who reported making at least one on-farm change (81%) was significantly higher than that of control respondents (38%). Overall, FF respondents significantly changed their risk score in 4 out of 5 risk areas and had an average reduction of 13 points in their overall risk score between before and after risk assessments. Control respondents' risk assessment scores did not significantly change during the study period. In a JD knowledge assessment, FF and control respondents exhibited a moderate knowledge score before the intervention period, with median scores of 75.9% (22/29) in each group. The FF respondents significantly increased their score at the postintervention assessment, with a median of 82.8% (24/29); control-respondent scores did not significantly change. Both FF and control respondents held strong positive attitudes toward JD control and felt a moderate amount of social pressure from veterinarians and industry organizations to make on-farm changes. However, they questioned their ability to effectively control JD on the farm. Last, participating in FF, having a moderate herd management score, having a positive perception about the practicality of on-farm recommendations, and having a singular learning preference were associated with increased odds of making an on-farm change. Overall, the FF process appears to be effective at influencing producer behavior toward implementing on-farm management practices for JD control. Future JD control programs should consider implementing peer-learning extension processes, such as FF, in combination with other extension approaches, to influence producer behavior.

Multiplicity of Salmonella entry mechanisms, a new paradigm for Salmonella pathogenesis.

The Salmonella enterica species includes about 2600 diverse serotypes, most of which cause a wide range of food- and water-borne diseases ranging from self-limiting gastroenteritis to typhoid fever in both humans and animals. Moreover, some serotypes are restricted to a few animal species, whereas other serotypes are able to infect plants as well as cold- and warm-blooded animals. An essential feature of the pathogenicity of Salmonella is its capacity to cross a number of barriers requiring invasion of a large variety of phagocytic and nonphagocytic cells. The aim of this review is to describe the different entry pathways used by Salmonella serotypes to enter different nonphagocytic cell types. Until recently, it was accepted that Salmonella invasion of eukaryotic cells required only the type III secretion system (T3SS) encoded by the Salmonella pathogenicity island-1. However, recent evidence shows that Salmonella can cause infection in a T3SS-1-independent manner. Currently, two outer membrane proteins Rck and PagN have been clearly identified as Salmonella invasins. As Rck mediates a Zipper-like entry mechanism, Salmonella is therefore the first bacterium shown to be able to induce both Zipper and Trigger mechanisms to invade host cells. In addition to these known entry pathways, recent data have shown that unknown entry routes could be used according to the serotype, the host and the cell type considered, inducing either Zipper-like or Trigger-like entry processes. The new paradigm presented here should change our classic view of Salmonella pathogenicity. It could also modify our understanding of the mechanisms leading to the different Salmonella-induced diseases and to Salmonella-host specificity.

Drinking water consumption patterns in Canadian communities (2001-2007).

A pooled analysis of seven cross-sectional studies from Newfoundland and Labrador, Waterloo and Hamilton Regions, Ontario and Vancouver, East Kootenay and Northern Interior Regions, British Columbia (2001 to 2007) was performed to investigate the drinking water consumption patterns of Canadians and to identify factors associated with the volume of tap water consumed. The mean volume of tap water consumed was 1.2 L/day, with a large range (0.03 to 9.0 L/day). In-home water treatment and interactions between age and gender and age and bottled water use were significantly associated with the volume of tap water consumed in multivariable analyses. Approximately 25% (2,221/8,916) of participants were classified as bottled water users, meaning that 75% or more of their total daily drinking water intake was bottled. Approximately 48.6% (4,307/8,799) of participants used an in-home treatment method to treat their tap water for drinking purposes. This study provides a broader geographic perspective and more current estimates of Canadian water consumption patterns than previous studies. The identified factors associated with daily water consumption could be beneficial for risk assessors to identify individuals who may be at greater risk of waterborne illness.

Characterization of isolates of Listeria monocytogenes from sludge using pulsed-field gel electrophoresis and virulence assays.

AIMS: To study the diversity and virulence of Listeria monocytogenes isolated from sludge. METHODS AND RESULTS: A total of 60 isolates of L. monocytogenes from sludge were characterized by serotyping, PFGE typing and using in vitro and in vivo virulence assays. The PFGE patterns were compared with those of food and human isolates to determine whether specific group clones are associated with environmental samples. The 60 isolates gave 44 different combined ApaI/AscI PFGE patterns. The PFGE patterns of most isolates were similar or very similar to those of epidemic isolates. The majority (93%) of isolates were found to be virulent by plaque-forming assay and by mouse virulence assay. CONCLUSIONS: Our findings suggest that L. monocytogenes strains found in non-sanitized sludge are virulent and represent a potential health hazard. Although no case of listeriosis related to sludge spread onto agricultural land has been reported, particular attention to this pathogen is needed. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study dealing with the characterization of L. monocytogenes isolates from non-sanitized sludge samples by molecular typing methods and in vitro and in vivo virulence assays. Our findings provide relevant information for evaluating the health risks associated with spreading sludge onto agricultural land.

A Listeria monocytogenes strain is still virulent despite nonfunctional major virulence genes.

The low-virulence Listeria monocytogenes strains have been previously assigned to 4 phenotypic groups. This study aimed to characterize the A23 strain, which exhibits a pulsed-field gel electrophoresis profile specific to low-virulence strains. This strain has the same causal mutations as the group III strains and a supplementary mutation in the mpl gene, leading to the absence of internalin A expression and the presence of inactive internalin B, phosphatidyl-inositol phospholipase C, and phosphatidylcholine phospholipase C. Despite these mutations in major virulence genes, the A23 strain formed plaques in cell monolayers and contaminated 100% of inoculated mice, suggesting that it evolved from group III strains by acquiring new virulence genes.

Prevalence of low-virulence Listeria monocytogenes strains from different foods and environments.

Various studies have demonstrated variations in the levels of virulence of different L. monocytogenes strains. In our laboratory, a plaque-forming assay followed by subcutaneous footpad inoculation of mice enabled us to estimate the prevalence of the low-virulence strains. This value fell from 16.3% to 1.7% with bacteria collected before 1994 and after 1997 respectively. This could be related to the modification in 1997 of the reference method EN ISO 11 290-1 of Listeria detection which recommended the use of polymyxin-acriflavine-LiCl-ceftazidime-aesculin-mannitol (PALCAM) medium. The aim of this study was to determine whether the percentage of low-virulence strains detected has changed due to the modification of the detection method recommending the use of the ALOA medium. After analyzing 380 L. monocytogenes strains, no increase in the percentage of low-virulence strains could be detected. The prevalence reached only 2.6% (ten of the 380 strains tested). The low virulence of L. monocytogenes strains was not related to rare serotypes and was also observed in serotypes usually involved in human disease. Low-virulence strains were found in dairy, meat, ready-to-eat products and also in the environment, highlighting the absence of one specific source. These results are discussed in terms of detection methods and the definition of low virulence.