Quantifying and mapping digital dermatitis-associated bacteria in lesion and nonlesion body sites and dairy farm environment.

The source of infection of digital dermatitis (DD), an infectious lameness condition, is still uncertain. In this cross-sectional study, we aimed to identify potential reservoirs of DD bacteria in dairy cattle body sites with different stages of the disease and farm environments. We collected skin swabs from 85 dairy cows from 5 herds, 3 with and 2 without DD, from foot, hock, and udder cleft skin (with lesions or not), saliva, urine, and feces. We also obtained environmental samples. Real-time quantitative PCR targeted Treponema phagedenis, Treponema medium, Treponema pedis, Porphyromonas levii, Bacteroides pyogenes, Fusobacterium necrophorum, and Fusobacterium mortiferum. Digital dermatitis-associated Treponema spp. were exclusively detected in DD-affected herds in DD-foot and other skin lesions, healthy skin, saliva, and environmental samples. In contrast, the non-Treponema spp. were found in samples from both DD-negative and affected herds. As expected, DD lesions had higher bacterial loads than healthy skin. Interestingly, similar counts were observed in udder cleft lesions, indicating a potential opportunistic behavior on compromised skin. None of the targeted species were detected in fecal samples, but P. levii, B. pyogenes, and F. necrophorum were detected in urine. All 7 species were detected in saliva, although in low quantities. No associations were observed between the presence of each bacterial species in DD lesions and urine; however, there was an association between the presence of DD-Treponema spp. in lesions and saliva, hock, and udder skin. Feces and urine do not seem to be a DD bacteria primary source, but saliva and other skin lesions may play a role. Longitudinal studies would improve our understanding of DD-associated bacteria's transient or persistent presence in these sites. Investigating the sources of DD-associated bacteria will guide future interventions to minimize bacterial shedding and transmission, ultimately more effectively reducing bacterial load, transmission, and sources of infection in dairy herds.

Identification of bovine-associated coagulase-negative staphylococci by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a direct transfer protocol.

This study evaluated matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) for the identification of bovine-associated coagulase-negative staphylococci (CNS), a heterogeneous group of different species. Additionally, we aimed to expand the MALDI-ToF MS database with new reference spectra as required to fill the gaps within the existing commercial spectral library. A total of 258 isolates of CNS were used in the study, covering 16 different CNS species. The majority of the isolates were previously identified by rpoB gene sequencing (n = 219), and the remainder were identified by sequencing of 16S rRNA, hsp60, or both rpoB and hsp60. The genotypic identification was considered the gold standard identification. All MALDI-ToF MS identifications were carried out using the direct transfer method. In a preliminary evaluation (n = 32 isolates; 2 of each species) with the existing commercial database, MALDI-ToF MS showed a typeability of 81% (26/32) and an accuracy of 96% (25/26). In the main evaluation (n = 226 isolates), MALDI-ToF MS with the existing commercial Biotyper (Bruker Daltonics Inc., Billerica, MA) database achieved a typeability of 92.0% (208/226) and an accuracy of 99.5% (207/208). Based on the assessment of the existing commercial database and prior knowledge of the species, a total of 13 custom reference spectra, covering 8 species, were created and added to the commercial database. Using the custom reference spectra expanded database, isolates were identified by MALDI-ToF MS with 100% typeability and 100% accuracy. Whereas the MALDI-ToF MS manufacturer's cutoff for species-level identification is 2.000, the reduction of the species level cutpoint to ≥1.700 improved the species-level identification rates (from 64 to 92% for the existing commercial database) when classifying CNS isolates. Overall, MALDI-ToF MS using the direct transfer method was shown to be a highly reliable tool for the identification of bovine-associated CNS.

Dairy farms testing positive for Mycobacterium avium ssp. paratuberculosis have poorer hygiene practices and are less cautious when purchasing cattle than test-negative herds.

Mycobacterium avium ssp. paratuberculosis (MAP), the causative agent of Johne's disease, is present on most dairy farms in Alberta, causing economic losses and presenting a potential public health concern. The objective of this cross-sectional study was to identify risk factors for Alberta dairy herds being MAP-positive based on environmental samples (ES). Risk assessments were conducted and ES were collected on 354 Alberta dairy farms (62% of eligible producers) voluntarily participating in the Alberta Johne's Disease Initiative. In univariate logistic regression, risk factors addressing animal and pen hygiene, as well as the use of feeding equipment to remove manure and manure application on pastures, were all associated with the number of positive ES. Furthermore, based on factor analysis, risk factors were clustered and could be summarized as 4 independent factors: (1) animal, pen, and feeder contamination; (2) shared equipment and pasture contamination; (3) calf diet; and (4) cattle purchase. Using these factor scores as independent variables in multivariate logistic regression models, a 1-unit increase in animal, pen, and feeder contamination resulted in 1.31 times higher odds of having at least 1 positive ES. Furthermore, a 1-unit increase in cattle purchase also resulted in 1.31 times the odds of having at least 1 positive ES. Finally, a 100-cow increase in herd size resulted in an odds ratio of 2.1 for having at least 1 positive ES. In conclusion, cleanliness of animals, pens, and feeders, as well as cattle purchase practices, affected risk of herd infection with MAP. Therefore, improvements in those management practices should be the focus of effective tools to control MAP on dairy farms.

Short communication: Evaluation of sampling socks for detection of Mycobacterium avium ssp. paratuberculosis on dairy farms.

Mycobacterium avium ssp. paratuberculosis (MAP) causes Johne's disease, a production-limiting disease in cattle. Detection of infected herds is often done using environmental samples (ES) of manure, which are collected in cattle pens and manure storage areas. Disadvantages of the method are that sample accuracy is affected by cattle housing and type of manure storage area. Furthermore, some sampling locations (e.g., manure lagoons) are frequently not readily accessible. However, sampling socks (SO), as used for Salmonella spp. testing in chicken flocks, might be an easy to use and accurate alternative to ES. The objective of the study was to assess accuracy of SO for detection of MAP in dairy herds. At each of 102 participating herds, 6 ES and 2 SO were collected. In total, 45 herds had only negative samples in both methods and 29 herds had ≥1 positive ES and ≥1 positive SO. Furthermore, 27 herds with ≥1 positive ES had no positive SO, and 1 herd with no positive ES had 1 positive SO. Bayesian simulation with informative priors on sensitivity of ES and MAP herd prevalence provided a posterior sensitivity for SO of 43.5% (95% probability interval=33-58), and 78.5% (95% probability interval=62-93) for ES. Although SO were easy to use, accuracy was lower than for ES. Therefore, with improvements in the sampling protocol (e.g., more SO per farm and more frequent herd visits), as well as improvements in the laboratory protocol, perhaps SO would be a useful alternative for ES.

Short communication: Herd-level prevalence of Mycobacterium avium ssp. paratuberculosis is not associated with participation in a voluntary Alberta Johne's disease control program.

Johne's disease (JD) control programs for dairy farms have the general objective of reducing both cow- and herd-level prevalence of Mycobacterium avium ssp. paratuberculosis (MAP). An important aspect of many programs is herd testing for MAP to determine the infection status of participating farms. However, it is uncertain whether MAP herd-level prevalence on farms voluntarily participating in a JD control program is different from that on nonparticipating farms. Therefore, the aim was to compare MAP infection status of participants and nonparticipants in the Alberta Johne's Disease Initiative (AJDI), a voluntary JD control program initiated in 2010 in Alberta, Canada. Between September 2012 and August 2013, environmental fecal samples were collected from 93 randomly selected farms not enrolled in the AJDI. Additionally, 81 farms that initially enrolled in the AJDI during the same time interval were also sampled. Samples were collected from 6 defined locations on each farm and cultured for MAP. Results were confirmed using conventional IS900 PCR and F 0285 quantitative PCR. Overall, 51% of participating and 51% of nonparticipating farms were identified as being MAP-infected. Furthermore, based on multivariable logistic regression, the number of MAP-positive samples was not associated with AJDI participation (taking herd size into account as a potentially modifying or confounding variable). In conclusion, there was no indication that voluntary participation in the AJDI was associated with herd-level MAP prevalence.

Sampling location, herd size, and season influence Mycobacterium avium ssp. paratuberculosis environmental culture results.

Mycobacterium avium subspecies paratuberculosis (MAP), the etiologic agent of Johne's disease, a chronic progressive enteritis, is a common pathogen on dairy farms. Environmental sampling is frequently used to detect MAP-infected herds, because it does not require sample collection from individual animals. The objectives were to determine (1) location-specific odds of MAP-positive environmental sampling results and whether season or herd size affect results; (2) whether season and herd size affect the odds of collection of samples from certain locations; and (3) whether sample-set composition affects the odds of a positive set. Herd veterinarians, producer organization staff, and University of Calgary staff collected 5,588 samples on dairy farms in Alberta and Saskatchewan. Samples from sick-cow and calving pens and samples from dry-cow housing had lower odds of testing MAP-positive than lactating cow-pen samples (odds ratio=0.3 and 0.4, respectively). Samples collected from bedding packs and manure piles were less frequently MAP-positive than those collected from alleyways and lagoons, whereas samples collected in spring and summer more often tested MAP-positive than those collected in winter. Sample sets collected in summer more often included samples from all locations than samples collected in winter; therefore, we inferred that collectors had difficulties accessing certain areas in winter. Substitution of sample locations with others had minor effect on the sensitivity of sample sets containing 6 samples. However, set composition had an effect on the sensitivity of sample sets containing only 2 samples. In that regard, whereas sets with 2 manure-storage-area samples detected 81% of farms with at least one positive environmental sample, sets with only dry, sick, or calving-pen samples detected only 59%. Environmental samples should be collected from areas where manure from numerous cows accumulates and can be well mixed (e.g., alleyways and manure lagoons). Collection of samples should be performed in spring or summer when locations are easier to access and samples have higher odds for testing MAP-positive.

Factors affecting management changes on farms participating in a Johne's disease control program.

Modern Johne's disease programs aim to control Mycobacterium avium ssp. paratuberculosis (MAP) infection through implementation of management practices that reduce the probability of MAP introduction and within-herd transmission on dairy farms. Success of these programs depends on whether weaknesses in management are corrected through implementation of management improvements. The objectives of this study were, therefore, to (1) assess whether scores in risk-assessment (RA) questions predicted suggestions for management changes for the upcoming year; and (2) determine factors as assessed in an RA that motivated producers to make management improvements and assess whether management improvements were influenced by previously received test results. The RA determining on-farm management related to MAP introduction and transmission were conducted annually by herd veterinarians on 370 dairy farms participating in the Alberta Johne's Disease Initiative. A maximum of 3 management changes that the farmer and the veterinarian agreed upon were recorded in a management plan. The MAP infection status of the herds was assessed through culture of 6 environmental samples. Whereas a management change was proposed for only 4% of questions with scores 1 or 2 (low risk), a change was proposed for 19% of questions with scores >2 [high risk; odds ratio (OR)=11.4]. Improvement in RA question scores was more likely between the first and second annual RA than between the second and third RA (OR=1.6). Farms with >3 culture-positive environmental samples collected in the previous year were more likely to improve their management than environmental sample culture-negative farms (OR=1.3). In conclusion, proposed management changes were oriented toward previously identified weaknesses in management practices, suggesting that the RA was properly used to design targeted management suggestions. Furthermore, improvements in management were not randomly distributed among farms participating in the control program. Instead, knowledge of MAP infection status of a herd, suggestions for management improvements, and duration of participation all influenced implementation of management improvements.

Factors associated with participation of Alberta dairy farmers in a voluntary, management-based Johne's disease control program.

The Alberta Johne's Disease Initiative (AJDI) is a voluntary, management-based prevention and control program for Johne's disease (JD), a wasting disease in ruminants that causes substantial economic losses to the cattle industry. Despite extensive communication about the program's benefits and low cost to participating producers, approximately 35% of Alberta dairy farmers have not enrolled in the AJDI. Therefore, the objective was to identify differences between AJDI nonparticipants and participants that may influence enrollment. Standardized questionnaires were conducted in person on 163 farms not participating and 61 farms participating in the AJDI. Data collected included demographic characteristics, internal factors (e.g., attitudes and beliefs of the farmer toward JD and the AJDI), external factors (e.g., farmers' JD knowledge and on-farm goals and constraints), as well as farmers' use and influence of various information sources. Nonparticipants and participants differed in at least some aspects of all studied categories. Based on logistic regression, participating farms had larger herds, higher self-assessed knowledge of JD, better understanding of AJDI details before participation, and used their veterinarian more often to get information about new management practices and technologies when compared with nonparticipants. In contrast, nonparticipants indicated that time was a major on-farm constraint and that participation in the AJDI would take too much time. They also indicated that they preferred to wait and see how the program worked on other farms before they participated.

High herd-level prevalence of Mycobacterium avium subspecies paratuberculosis in Western Canadian dairy farms, based on environmental sampling.

Mycobacterium avium subspecies paratuberculosis (MAP) causes chronic progressive enteritis in ruminants. The pathogen is present in most countries with modern dairy production, causing substantial economic losses for the industry. The objectives of this study were to estimate dairy herd prevalence of MAP in the Western Canadian provinces of Alberta and Saskatchewan, and to determine whether herd size and housing system (tie-stall versus freestall or loose housing) affected the risk of a herd testing positive for MAP. Six environmental samples were collected on 360 Alberta farms (60% of registered producers) and on 166 Saskatchewan dairy farms (99%). In total, 47% of the sampled farms in Alberta and 53% of the sampled farms in Saskatchewan had at least one environmental sample that was MAP culture positive and were, therefore, defined as infected. Sensitivity of environmental sampling was estimated using 3 subsequent annual tests performed on 82 farms. Because laboratory protocols were continuously improved throughout the project, the sensitivity increased over time. Therefore, a mean of the sensitivity estimates weighted on sampling year was constructed; this resulted in sensitivities of 68 and 69% for Alberta and Saskatchewan, respectively. Implementing those estimates in an approximate Bayesian computation model resulted in a true herd prevalence of 68% (95% probability interval: 60-80%) for Alberta and 76% (95% probability interval: 70-85%) for Saskatchewan. Herds with >200 cows had 3.54 times higher odds of being environmental sample positive and had more positive samples than herds with <50 cows (neither province nor housing system affected those results). In conclusion, the majority of Alberta and Saskatchewan dairy farms were infected with MAP and larger herds were more often MAP positive than smaller herds.

Predicting sensitivity of repeated environmental sampling for Mycobacterium avium subsp. paratuberculosis in dairy herds using a Bayesian latent class model.

Between-herd transmission of Mycobacterium avium subsp. paratuberculosis (MAP) by subclinically infected cattle is an important risk which can hamper effective control of paratuberculosis. Knowledge of herd status would substantially reduce this risk; MAP positive farms can be detected with environmental sampling. The objective of this study was to compare cumulative sensitivities of annual environmental sampling with two or four samples per sampling event without knowledge of true herd status and to calculate the number of sampling events to achieve a cumulative sensitivity of at least 0.9. Data from three repeated sampling events in two study populations, one with 55 herds (two samples/event) and another with 30 herds (four samples/event) including test results, herd and sample characteristics and prior prevalence estimates, were derived from the Alberta Johne's Disease Initiative (Alberta, Canada). A recursive Bayesian latent class model was used to predict the cumulative sensitivity of repeated environmental sampling events. A sampling scheme with four samples per sampling event had a higher cumulative sensitivity than an alternative scheme with two samples. To achieve a cumulative sensitivity of at least 0.9 with 95% probability, eight sampling events with two environmental samples per set, or four sampling events with four samples per set were required. Further model assessment demonstrated that these results can only be generalized to cattle populations with a similar within-herd prevalence to those studied here (approximately 0.08). Nonetheless, these results could help predict herd-level prevalence in cattle populations after environmental testing and provide information regarding the uncertainty behind status estimates for herds repeatedly tested using environmental samples.

Determining the IgG concentrations in bovine colostrum and calf sera with a novel enzymatic assay.

BACKGROUND: Immune protection in newborn calves relies on a combination of the timing, volume and quality of colostrum consumed by the calf after birth. Poor quality colostrum with inadequate immunoglobulin concentration contributes to failed transfer of passive immunity in calves, leading to higher calf morbidity and mortality. Therefore, estimating colostrum quality and ensuring the transfer of passive immunity on farm is of critical importance. Currently, there are no on-farm tools that directly measure immunoglobulin content in colostrum or serum. The aim of this study was to apply a novel molecular assay, split trehalase immunoglobulin G assay (STIGA), to directly estimate immunoglobulin content in dairy and beef colostrum and calf sera, and to examine its potential to be developed as on-farm test. The STIGA is based on a split version of trehalase TreA, an enzyme that converts trehalose into glucose, enabling the use of a common glucometer for signal detection. In a first study, 60 dairy and 64 beef colostrum and 83 dairy and 84 beef calf sera samples were tested with STIGA, and the resulting glucose production was measured and compared with radial immunodiffusion, the standard method for measuring immunoglobulin concentrations. RESULTS: Pearson correlation coefficients between the methods were determined and the sensitivity, specificity, and accuracy of the test were calculated for different colostrum quality and failed transfer of passive immunity cut-off points. The correlations of the STIGA measured by colorimetric enzymatic reaction compared to radial immunodiffusion for dairy and beef colostrum were 0.72 and 0.73, respectively, whereas the correlations for dairy and beef sera were 0.9 and 0.85, respectively. Next, STIGA was tested in a blinded study with fresh colostrum and serum samples where the correlation coefficient was 0.93 and 0.94, respectively. Furthermore, the performance of STIGA followed by glucometer readings resulted in correlations with radial immunodiffusion of 0.7 and 0.85 for dairy and beef colostrum and 0.94 and 0.83 for dairy and beef calf serum. CONCLUSIONS: A split TreA assay was validated for measurement of the immunoglobulin content of colostrum and calf sera using both a lab-based format and in a more user-friendly format compatible with on-farm testing.

Missing pieces of the puzzle to effectively control digital dermatitis.

Since the first report of bovine digital dermatitis (DD) in 1974, there is a large body of the literature published; however, effective prevention and control of the disease remain elusive. Although many aspects of the pathogenesis of DD have been investigated, even some of the most basic questions such as the aetiology of this disease remain under debate. Treponema spp. have been strongly associated with DD lesions and occur in abundance in advanced lesions; however, efforts to induce disease with pure cultures of these organisms have been largely underwhelming and inconsistent. Furthermore, although the disease has been presented for several decades, there is limited scientific evidence regarding effective treatment of DD. Apparent discrepancies between effectiveness in vitro and in vivo have challenged the scientific community to identify new potential treatment options. With no treatment resulting in a 100% cure rate, the current expectation is manageable control, but prospects for the eradication of the disease are unlikely using current approaches. In order to develop more effective approaches to control DD on-farm, there is a critical need for a deeper understanding regarding the causation, ecology, transmission and treatment of this disease. In this article, we attempt to provide insights into specific research needs related to DD in order to assist the industry, researchers, pharmaceutical companies and research sponsors with decision-making and identified research gaps.

Knowledge gaps that hamper prevention and control of Mycobacterium avium subspecies paratuberculosis infection.

In the last decades, many regional and country-wide control programmes for Johne's disease (JD) were developed due to associated economic losses, or because of a possible association with Crohn's disease. These control programmes were often not successful, partly because management protocols were not followed, including the introduction of infected replacement cattle, because tests to identify infected animals were unreliable, and uptake by farmers was not high enough because of a perceived low return on investment. In the absence of a cure or effective commercial vaccines, control of JD is currently primarily based on herd management strategies to avoid infection of cattle and restrict within-farm and farm-to-farm transmission. Although JD control programmes have been implemented in most developed countries, lessons learned from JD prevention and control programmes are underreported. Also, JD control programmes are typically evaluated in a limited number of herds and the duration of the study is less than 5 year, making it difficult to adequately assess the efficacy of control programmes. In this manuscript, we identify the most important gaps in knowledge hampering JD prevention and control programmes, including vaccination and diagnostics. Secondly, we discuss directions that research should take to address those knowledge gaps.

Invasion of Salmonella enteritidis in avian intestinal epithelial cells in vitro is influenced by short-chain fatty acids.

Fermentation reactions in the caeca of chickens, the predominant place for Salmonella colonization, result in high concentrations of short-chain fatty acids (SCFA). Thus Salmonella bacteria are in close contact with SCFA during their life cycle. A study was carried out to analyse the effects of SCFA on invasion of Salmonella enteritidis in an avian intestinal epithelial cell line. Preincubation of S. enteritidis for 4 h in growth media supplemented with various concentrations of propionate or butyrate resulted in decreased invasion compared to bacteria, preincubated in nonsupplemented media, and to bacteria, preincubated in media supplemented with formate or acetate. Incubation of the S. enteritidis bacteria in media supplemented with mixtures of SCFA mimicking the in vivo caecal concentrations resulted in increased invasion compared with butyrate-exposed bacteria, but equal invasion compared with nonexposed bacteria. Increasing the butyrate concentration in these mixtures did not modify invasion compared with the original mixtures.

Tubular glands of the isthmus are the predominant colonization site of Salmonella enteritidis in the upper oviduct of laying hens.

Salmonella enterica serovar Enteritidis is the serovar most frequently isolated from chicken eggs. Colonization of the upper oviduct of hens is believed to play an important role in egg contamination. The interaction of S. enteritidis with gland epithelial cells of the isthmus and the magnum was, therefore, studied in vitro and in vivo. In the first experiment, S. enteritidis bacteria were added to confluent monolayers of primary cultures of chicken tubular epithelial cells of the isthmus (ICTEC) or magnum (MCTEC). Intracellular bacteria in ICTEC and MCTEC were confirmed by a gentamicin protection assay. Internalization in the glandular cells was corroborated by confocal scanning microscopy. Although S. enteritidis was able to invade and proliferate intracellularly during 24 h in the cell culture of both segments, this was significantly more so in the ICTEC. In a second experiment, an in vivo loop model was developed for investigation of the invasiveness of S. enteritidis in the oviduct of laying hens. Loops in the isthmus and magnum of laying hens were made under anaesthesia. S. enteritidis was inoculated into each loop. After 1 h, tubular gland cells were isolated from the infected tissue under gentamicin. S. enteritidis invasiveness was measured as the ratio of isolated bacteria per isolated tubular gland cell. This ratio was higher (P < 0.01) in the isthmus (1.3 x 10(-3)) than in the magnum (5.3 x 10(-5)). In a third experiment, laying hens were intravenously infected with 5 x 10(7) cfu S. enteritidis bacteria. The number of intracellular bacteria was counted in the tubular gland cells of the isthmus and magnum after isolation under gentamicin. The ratio of isolated bacteria per isolated tubular gland cell was again significantly higher in the isthmus as compared with in the magnum. In all 3 assays, the tubular gland cells of the isthmus were more heavily invaded than those of the magnum.

Microencapsulated short-chain fatty acids in feed modify colonization and invasion early after infection with Salmonella enteritidis in young chickens.

Short-chain fatty acids (SCFA) are widely used as feed additives in poultry for the control of pathogenic bacteria, such as Salmonella enteritidis. Recently, a new range of products was developed in which SCFA are encapsulated in mineral carriers, resulting in a slow release during the transport of these carriers through the intestinal tract. To test the efficacy of this type of products against early colonization after Salmonella infection in poultry, a challenge experiment with S. enteritidis was performed. Five groups of 20 chickens were given feed with no supplement or feed supplemented with acetic acid (0.24%), formic acid (0.22%), or propionic acid (0.27%) as film-coated microbeads or butyric acid (0.15%) as spray-cooled microcapsules. The 5 groups were challenged with 5 x 10(3) cfu S. enteritidis at d 5 and 6 posthatch, and samples of ceca, liver, and spleen were taken at d 8 and analyzed for the number of colony-forming units of Salmonella per gram of tissue. Feed supplementation with acetic acid, and to a lesser extent formic acid, resulted in an increase of colonization of ceca and internal organs. Birds receiving propionic acid-coated microbeads as feed supplement were colonized with Salmonella to the same extent as controls. Butyric acid-impregnated microbeads in the feed, however, resulted in a significant decrease of colonization by S. enteritidis in the ceca but not in liver and spleen.

The impact of prebiotics and salmonellosis on apparent nutrient digestibility and Salmonella typhimurium var. Copenhagen excretion in adult pigeons (Columba livia domestica).

The effects of lactose or fructo-oligosaccharide (FOS) supplementation on the excretion of salmonellae, apparent digestibilities and excreta consistency were studied. Thirty-two male pigeons (Columba livia domestica) were randomly divided into 4 equal groups: 3 of 4 groups were orally infected with 10(9) Salmonella Typhimurium var. Copenhagen, after being offered a drinking water supplement of 2% FOS, 2% lactose, or no supplement, respectively, for 2 wk. Pigeons in the fourth group were not challenged with S. Typhimurium and remained unsupplemented. Initially, FOS increased water intake, resulting in more watery excreta. After infection, supplementation showed no major effects on S. Typhimurium excretion, nitrogen retention, or apparent nutrient digestibilities, although lactose--and to a lesser extent FOS--improved apparent fiber digestibility during recovery from the S. Typhimurium infection. The excreta consistency of all pigeons returned to normal when recovering from the Salmonella infection. In this trial, neither FOS nor lactose was successful in tempering the negative aspects of Salmonella infection in pigeons. Nevertheless, it should be stated that future investigations should clarify the importance of duration and level of prebiotic supplementation and infection level.

Intermittent long-term shedding and induction of carrier birds after infection of chickens early posthatch with a low or high dose of Salmonella enteritidis.

Poultry are very likely to become infected with Salmonella in the early posthatch period, due to environmental contamination. The purpose of this study was to evaluate the effect of infection dose on the risk of persistent infection in laying hens. In this study, young layer chicks were orally infected with a low (10(2) at 1 d posthatch) or a high dose (10(9) cfu at 1 wk posthatch) of Salmonella Enteritidis. The pattern and duration of fecal shedding was studied for 18 wk. All chickens shed Salmonella early after infection and shed Salmonella intermittently during the whole study period. There were more positive birds in the high-dose group than in the low-dose group in the first weeks following Salmonella exposure. From 10 wk postinfection onwards, however, birds that were orally infected with the low dose of Salmonella Enteritidis shed more Salmonella than the birds that received the high dose. At 18 wk of age, there was no difference in cecal colonization between the treatment groups. It can be concluded that infection of newly hatched chicks with a low dose of Salmonella Enteritidis can lead to persistent infection until onset of lay, hereby excreting Salmonella bacteria intermittently.

Occurrence of Mycobacterium avium subspecies paratuberculosis and Neospora caninum in Alberta cow-calf operations.

Mycobacterium avium subsp. paratuberculosis (MAP) and Neospora caninum (NC) are two pathogens causing important production limiting diseases in the cattle industry. Significant impacts of MAP and NC have been reported on dairy cattle herds, but little is known about the importance, risk factors and transmission patterns in western Canadian cow-calf herds. In this cross-sectional study, the prevalence of MAP and NC infection in southwest Alberta cow-calf herds was estimated, risk factors for NC were identified, and the reproductive impacts of the two pathogens were assessed. Blood and fecal samples were collected from 840 cows on 28 cow-calf operations. Individual cow and herd management information was collected by self-administered questionnaires and one-on-one interviews. Bayesian estimates of the true prevalence of MAP and NC were computed, and bivariable and multivariable statistical analysis were done to assess the association between the NC serological status and ranch management risk factors, and the clinical effects of the two pathogens. Bayesian estimates of true prevalence indicated that 20% (95% probability interval: 8-38%) of herds had at least one MAP-positive cow, with a within-herd prevalence in positive herds of 22% (8-45%). From the Bayesian posterior distributions of NC prevalence, the median herd-level prevalence was 66% (33-95%) with 10% (4-21%) cow-level prevalence in positive herds. Multivariable analysis indicated that introducing purchased animals in the herd might increase the risk of NC. The negative association of NC with proper carcass disposal and presence of horses on ranch (possibly in relation to herd monitoring and guarding activities), may suggest the importance of wild carnivores in the dynamics of this pathogen in the study area. We also observed an association between MAP and NC serological status and the number of abortions. Additional studies should be done to further examine specific risk factors for MAP and NC, assess the consequences on the reproductive performances in cow-calf herds, and evaluate the overall impact of these pathogens on cow-calf operations.

PCR amplification and high-resolution melting curve analysis as a rapid diagnostic method for genotyping members of the Mycobacterium avium­intracellulare complex.

Some of the members of the Mycobacterium avium­intracellulare (MAI) complex are recognized as human pathogens in both immunocompromised and immunocompetent patients. The current molecular methods that are available for genotyping the MAI complex members can be both expensive and technically demanding. In this report, we describe for the first time the application of a real-time PCR and high-resolution melt approach to differentiate between the complex members by targeting a member of the Pro- Pro-Glu gene family, MACPPE24. To this end, reference strains of the M. avium subspecies and Mycobacterium intracellulare were used to optimize the technique. Then, this real-time PCR­high-resolution melt approach was used to distinguish ten M. avium ssp. hominissuis field isolates from the M. intracellulare reference strain.