Identification of canine osteoarthritis using an owner-reported questionnaire and treatment monitoring using functional mobility tests.

OBJECTIVES: To investigate the diagnostic value of an owner-completed canine osteoarthritis screening checklist to help identify previously undiagnosed osteoarthritis cases, and assess their response to carprofen treatment by monitoring pain and functional mobility. MATERIALS AND METHODS: Dogs (n=500) whose owners reported ≥1 positive response to the osteoarthritis checklist were examined to identify dogs with previously undiagnosed osteoarthritis. Eligible dogs (n=133) were evaluated for pain and video mobility analysis by Helsinki Chronic Pain Index and visual analogue scale scores, respectively, following carprofen treatment, administered for 30 days (n=95) or up to 120 days (n=38). Dogs were filmed at clinics performing activities (walking, jogging, sitting/lying, walking up and down stairs), and scored at days 0, 30 and 120 using visual analogue scale by an independent blinded expert. RESULTS: A diagnosis of osteoarthritis was confirmed by a veterinarian in 38% (188 of 500) of dogs. Balance of sensitivity and specificity across the original group of nine screening questions was optimised to approximately 88 and 71%, respectively, after elimination of three questions. Pain measured by Helsinki Chronic Pain Index and functional mobility improved over time in response to treatment with carprofen. Mean ability scores for activities significantly improved between days 0 and 30 for walking, jogging, sitting/lying and walking down stairs, and days 0 and 120 for sitting/lying and walking up stairs. CLINICAL SIGNIFICANCE: More osteoarthritis cases were identified in study dogs than previous prevalence estimates, indicating the screening checklist's potential to help identify for further evaluation cases that could otherwise remain undiagnosed. Improvements in function were demonstrated after carprofen treatment.

Factors associated with septic arthritis of the distal interphalangeal joint in beef cattle: A case-control study.

Lameness in cattle is a welfare concern and is associated with important economic losses in beef cattle operations. Infection of the distal interphalangeal joint (DIJ) results in severe lameness and if left untreated carries a poor prognosis. Diagnosis of DIJ infection is challenging in the field. The aim of this study was to determine factors associated with septic arthritis of the DIJ in beef cattle. Thirty-nine beef cattle with a complaint of single-foot lameness were used in this study. History and lameness examination data were recorded and analyzed. Radiographic changes and cytology of synovial fluid of the affected DIJ were used to define the presence (cases) or absence (controls) of DIJ infection. Asymmetric swelling at the coronary band of the affected foot and a lameness score of ≥4/5 significantly increased the odds (odds ratio [OR]=63.2 and OR=120, respectively) of diagnosis of septic arthritis of the DIJ in beef cattle with a single-foot lameness compared to cattle with no asymmetry of the coronary band or lameness scores <3. Routine lameness examination findings could be used in the field to rapidly recognize infection of the DIJ in lame beef cattle.

Meta-analysis of the effects of laidlomycin propionate, fed alone or in combination with chlortetracycline, compared with monensin sodium, fed alone or in combination with tylosin, on growth performance, health, and carcass outcomes in finishing steers in North America.

The objective of this research was to use data from multiple studies to comprehensively quantify the effects of feeding 1) laidlomycin propionate (LP), alone and/or in combination with chlortetracycline, compared with 2) monensin sodium (MS), alone and/or in combination with tylosin, at commercially approved dosages, on ADG, DMI, feed efficiency (FE), mortality, and carcass characteristics (HCW and liver abscesses). A secondary objective was to explore potential sources of heterogeneity among the comparative effectiveness studies. A systematic review of peer-reviewed literature and industry reports was used to identify studies that included direct comparisons of these treatments in finishing steers in North America. Random-effects meta-analysis models of performance, carcass, and health-related outcomes were fitted with extracted data, consisting of a total of 17 data sets comprising a total of 135 pens and 13,603 steers. Results showed that pens of steers fed LP had increased ADG (live and carcass adjusted), DMI, and HCW compared with those fed monensin ( < 0.05). However, liver abscesses were more common in LP-fed cattle than in MS-fed cattle ( < 0.05), presumably because of differences in the concurrently fed antimicrobials. No significant effects ( > 0.05) were identified for FE or for health-related outcomes (overall and cause-specific mortality). There was a substantial amount of heterogeneity in outcomes among studies, and when pen size and type of production setting were included in mixed-effects meta-regression models, they accounted for only a small proportion of the between-study heterogeneity found in the meta-analysis models. Therefore, caution should be exercised when interpreting summary estimates in the presence of substantial heterogeneity. However, these results provide comprehensive information on the comparative effects of different ionophores across multiple studies and multiple years, states, and production settings. These unique results can enable quantitative and informed decisions by potential end users of these feed additives that are widely used in the U.S. beef industry for reducing the costs of beef production through enhanced cattle performance.

Multiplex Quantitative PCR Assays for the Detection and Quantification of the Six Major Non-O157 Escherichia coli Serogroups in Cattle Feces.

Shiga toxin-producing Escherichia coli (STEC) serogroups O26, O45, O103, O111, O121, and O145, called non-O157 STEC, are important foodborne pathogens. Cattle, a major reservoir, harbor the organisms in the hindgut and shed them in the feces. Although limited data exist on fecal shedding, concentrations of non-O157 STEC in feces have not been reported. The objectives of our study were (i) to develop and validate two multiplex quantitative PCR (mqPCR) assays, targeting O-antigen genes of O26, O103, and O111 (mqPCR-1) and O45, O121, and O145 (mqPCR-2); (ii) to utilize the two assays, together with a previously developed four-plex qPCR assay (mqPCR-3) targeting the O157 antigen and three virulence genes (stx1, stx2, and eae), to quantify seven serogroups and three virulence genes in cattle feces; and (iii) to compare the three mqPCR assays to a 10-plex conventional PCR (cPCR) targeting seven serogroups and three virulence genes and culture methods to detect seven E. coli serogroups in cattle feces. The two mqPCR assays (1 and 2) were shown to be specific to the target genes, and the detection limits were 4 and 2 log CFU/g of pure culture-spiked fecal samples, before and after enrichment, respectively. A total of 576 fecal samples collected from a feedlot were enriched in E. coli broth and were subjected to quantification (before enrichment) and detection (after enrichment). Of the 576 fecal samples subjected, before enrichment, to three mqPCR assays for quantification, 175 (30.4%) were quantifiable (≥4 log CFU/g) for at least one of the seven serogroups, with O157 being the most common serogroup. The three mqPCR assays detected higher proportions of postenriched fecal samples (P > 0.01) as positive for one or more serogroups compared with cPCR and culture methods. This is the first study to assess the applicability of qPCR assays to detect and quantify six non-O157 serogroups in cattle feces and to generate data on fecal concentration of the six serogroups.

Insecticidal sugar baits for adult biting midges.

The mixing of an insecticide with sugar solution creates an oral toxin or insecticidal sugar bait (ISB) useful for reducing adult insect populations. The ability of ISBs to kill the biting midge Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae), a vector of bluetongue virus, epizootic hemorrhagic disease and vesicular stomatitis viruses, was tested. The commercial insecticide formulations (percentage active ingredient) tested included bifenthrin, cyfluthrin, deltamethrin, permethrin, dinotefuran, imidacloprid, thiamethoxam and spinosad. Mortality rates were determined for various concentrations of commercial formulations (0.01, 0.05, 0.1, 1, 2 and 3%) and observed at 1, 4, 10 and 24 h post-exposure to the ISB. In the first set of assays, laboratory-reared midges were fed sugar ad libitum and then exposed to insecticide-treated sugar solutions to measure mortality. The second assay assessed competitive feeding: midges were provided with a control sugar solution (10% sucrose) in one vial, and a sugar and insecticide solution in another. Pyrethroid treatments resulted in the greatest mortality in the first hour at the lowest concentrations and spinosad consumption resulted in the least mortality. Biting midges were not deterred from feeding on the 1% ISB solutions despite the presence of an insecticide-free alternative source of sugar.

Sugar-feeding status alters biting midge photoattraction.

The biting midge Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae) transmits pathogens to both livestock and wildlife. Biting midge surveillance relies heavily on light traps for collection; however, little is known about the light spectra preferences of C. sonorensis midges. A light assay arena was constructed and light-emitting diodes (LEDs) of various light spectra were used as light sources to evaluate midge photoattraction. A comparison of responses to light spectra indicated the highest proportions of C. sonorensis were attracted to ultraviolet (UV) light and that midges differentiated 10-nm differences in wavelength. Stronger intensities of UV light resulted in greater attraction. Midges exhibited both sugar-seeking and escape behaviours under different conditions of sugar supplementation before and during the experiment. These behaviours occurred with lights of 355 nm and 365 nm in wavelength. Based on the results of this study, the attraction of C. sonorensis to light traps can be improved through the use of bright LEDs at 355 nm or 365 nm.

Fecal shedding of non-O157 serogroups of Shiga toxin-producing Escherichia coli in feedlot cattle vaccinated with an Escherichia coli O157:H7 SRP vaccine or fed a Lactobacillus-based direct-fed microbial.

The objectives of this study were to determine whether fecal shedding of non-O157 Shiga toxin-producing Escherichia coli (STEC) in feedlot cattle was affected by the use of an E. coli O157:H7 vaccine or a direct-fed microbial (DFM) and whether the shedding of a particular non-O157 STEC serogroup within feces was associated with shedding of O157 or other non-O157 STEC serogroups. A total of 17,148 cattle in 40 pens were randomized to receive one, both, or neither (control) of the two interventions: a vaccine based on the siderophore receptor and porin proteins (E. coli SRP vaccine, two doses) and a DFM product (low-dose Bovamine). Fresh fecal samples (30 samples per pen) were collected weekly from pen floors for four consecutive weeks beginning approximately 56 days after study allocation. DNA extracted from enriched samples was tested for STEC O157 and non-O157 serogroups O26, O45, O103, O111, O121, and O145 and for four major virulence genes (stx1, stx2, eae, and ehxA) using an 11-gene multiplex PCR assay. Generalized linear mixed models were used to analyze the effects of treatments and make within-sample comparisons of the presence of O-serogroup-specific genes. Results of cumulative prevalence measures indicated that O157 (14.6%), O26 (10.5%), and O103 (10.3%) were the most prevalent STEC O serogroups. However, the vaccine, DFM, or both had no significant effect (P > 0.05) on fecal prevalence of the six non-O157 STEC serogroups in feedlot cattle. Within-sample comparisons of the presence of STEC serogroup-specific genes indicated that fecal shedding of E. coli O157 in cattle was associated with an increased probability (P < 0.05) of fecal shedding of STEC O26, O45, O103, and O121. Our study revealed that neither the E. coli O157:H7 vaccine, which reduced STEC O157 fecal shedding, nor the DFM significantly affected fecal shedding of non-O157 STEC serogroups, despite the fact that the most prevalent non-O157 STEC serogroups tended to occur concurrently with O157 STEC strains within fecal samples.

Spatial epidemiology of Escherichia coli O157:H7 in dairy cattle in relation to night roosts Of Sturnus vulgaris (European Starling) in Ohio, USA (2007-2009).

The goal of our study was to use spatial scan statics to determine whether the night roosts of European starlings (Sturnus vulgaris) act as point sources for the dissemination of Escherichia coli O157:H7 among dairy farms. From 2007 to 2009, we collected bovine faecal samples (n = 9000) and starling gastrointestinal contents (n = 430) from 150 dairy farms in northeastern Ohio, USA. Isolates of E. coli O157:H7 recovered from these samples were subtyped using multilocus variable-number tandem repeat analysis (MLVA). Generated MLVA types were used to construct a dendrogram based on a categorical multistate coefficient and unweighted pair-group method with arithmetic mean (UPGMA). Using a focused spatial scan statistic, we identified statistically significant spatial clusters among dairy farms surrounding starling night roosts, with an increased prevalence of E. coli O157:H7-positive bovine faecal pats, increased diversity of distinguishable MLVA types and a greater number of isolates with MLVA types from bovine-starling clades versus bovine-only clades. Thus, our findings are compatible with the hypothesis that starlings have a role in the dissemination of E. coli O157:H7 among dairy farms, and further research into starling management is warranted.

Hierarchical Bayesian modeling of heterogeneous variances in average daily weight gain of commercial feedlot cattle.

Variability in ADG of feedlot cattle can affect profits, thus making overall returns more unstable. Hence, knowledge of the factors that contribute to heterogeneity of variances in animal performance can help feedlot managers evaluate risks and minimize profit volatility when making managerial and economic decisions in commercial feedlots. The objectives of the present study were to evaluate heteroskedasticity, defined as heterogeneity of variances, in ADG of cohorts of commercial feedlot cattle, and to identify cattle demographic factors at feedlot arrival as potential sources of variance heterogeneity, accounting for cohort- and feedlot-level information in the data structure. An operational dataset compiled from 24,050 cohorts from 25 U. S. commercial feedlots in 2005 and 2006 was used for this study. Inference was based on a hierarchical Bayesian model implemented with Markov chain Monte Carlo, whereby cohorts were modeled at the residual level and feedlot-year clusters were modeled as random effects. Forward model selection based on deviance information criteria was used to screen potentially important explanatory variables for heteroskedasticity at cohort- and feedlot-year levels. The Bayesian modeling framework was preferred as it naturally accommodates the inherently hierarchical structure of feedlot data whereby cohorts are nested within feedlot-year clusters. Evidence for heterogeneity of variance components of ADG was substantial and primarily concentrated at the cohort level. Feedlot-year specific effects were, by far, the greatest contributors to ADG heteroskedasticity among cohorts, with an estimated ∼12-fold change in dispersion between most and least extreme feedlot-year clusters. In addition, identifiable demographic factors associated with greater heterogeneity of cohort-level variance included smaller cohort sizes, fewer days on feed, and greater arrival BW, as well as feedlot arrival during summer months. These results support that heterogeneity of variances in ADG is prevalent in feedlot performance and indicate potential sources of heteroskedasticity. Further investigation of factors associated with heteroskedasticity in feedlot performance is warranted to increase consistency and uniformity in commercial beef cattle production and subsequent profitability.

A multivariable assessment quantifying effects of cohort-level factors associated with combined mortality and culling risk in cohorts of U.S. commercial feedlot cattle.

Economic losses due to cattle mortality and culling have a substantial impact on the feedlot industry. Since criteria for culling may vary and may affect measures of cumulative mortality within cattle cohorts, it is important to assess both mortality and culling when evaluating cattle losses over time and among feedlots. To date, there are no published multivariable assessments of factors associated with combined mortality and culling risk. Our objective was to evaluate combined mortality and culling losses in feedlot cattle cohorts and quantify effects of commonly measured cohort-level risk factors (weight at feedlot arrival, gender, and month of feedlot arrival) using data routinely collected by commercial feedlots. We used retrospective data representing 8,904,965 animals in 54,416 cohorts from 16 U.S. feedlots from 2000 to 2007. The sum of mortality and culling counts for each cohort (given the number of cattle at risk) was used to generate the outcome of interest, the cumulative incidence of combined mortality and culling. Associations between this outcome variable and cohort-level risk factors were evaluated using a mixed effects multivariable negative binomial regression model with random effects for feedlot, year, month and week of arrival. Mean arrival weight of the cohort, gender, and arrival month and a three-way interaction (and corresponding two-way interactions) among arrival weight, gender and month were significantly (P<0.05) associated with the outcome. Results showed that as the mean arrival weight of the cohort increased, mortality and culling risk decreased, but effects of arrival weight were modified both by the gender of the cohort and the month of feedlot arrival. There was a seasonal pattern in combined mortality and culling risk for light and middle-weight male and female cohorts, with a significantly (P<0.05) higher risk for cattle arriving at the feedlot in spring and summer (March-September) than in cattle arriving during fall, and winter months (November-February). Our results quantified effects of covariate patterns that have been heretofore difficult to fully evaluate in smaller scale studies; in addition, they illustrated the importance of utilizing multivariable approaches when quantifying risk factors in heterogeneous feedlot populations. Estimated effects from our model could be useful for managing financial risks associated with adverse health events based on data that are routinely available.

Association of wild bird density and farm management factors with the prevalence of E. coli O157 in dairy herds in Ohio (2007-2009).

Our objective was to determine the role that European starlings (Sturnus vulgaris) play in the epidemiology of Escherichia coli O157:H7 in dairy cattle. We visited 150 dairy farms in Ohio twice during summer and fall months from 2007 to 2009. Fresh faecal pats from 30 lactating cows were collected during each visit. Information on farm management and environmental variables was gathered through a questionnaire administered to the farm owner. The number of starlings observed on the farm was also recorded. Approximately 1% of dairy cattle and 24% of farms were positive for E. coli O157. Risk factors associated with the presence of E. coli O157 in faecal pats included contact between adult cattle and calves, types or number of ventilation and manure management systems and number of birds per milking cow.

Associations between the distance traveled from sale barns to commercial feedlots in the United States and overall performance, risk of respiratory disease, and cumulative mortality in feeder cattle during 1997 to 2009.

Most beef cattle are transported at least once during their lives, and this potentially stressful practice may affect subsequent health and performance. Limited research is available quantifying the effects of transport on feedlot performance and health, and particularly the risk of bovine respiratory disease complex (BRD), which is the most common disease of weaned calves after arrival to the feedlot. The objective of this retrospective study was to determine potential associations between distance traveled (DTV) during transportation with health (cumulative BRD morbidity and mortality of all causes) and performance (ADG and HCW) parameters in cattle cohorts (n = 14,601) that arrived to 21 U.S. commercial feedlots from 1997 to 2009. Multivariable mixed-effects negative binomial and linear regression models were employed to determine associations between health and performance outcomes with DTV and other cohort-level demographic variables. Cattle were transported a median of 552 km from origin to feedlot with a mean (± SEM) of 698 ± 4.4 km. The mean (±SEM) cumulative BRD morbidity was 4.9% ± 0.01% (median = 1.1%; range: 0 to 100%) whereas the mean (±SEM) cumulative mortality due to all causes was 1.3% ± 0.01% (median = 0.8%; range: 0 to 28.7%). Distance traveled was significantly associated (P < 0.05) with BRD morbidity, overall mortality, HCW and ADG, and its effects were modified by demographic characteristics (i.e., cohort region of origin, mean arrival BW, gender, and the season of the year) of the cohort. Knowledge of the distance traveled during transportation could allow a more precise prediction of cattle feedlot health and performance.

Effects of body weight loss during transit from sale barns to commercial feedlots on health and performance in feeder cattle cohorts arriving to feedlots from 2000 to 2008.

Body weight loss during transport or shrink (SHK) is a common occurrence in feeder cattle that results from a physiological, complex process. Previous studies have assessed the effects of environmental and dietary stressors on transport-associated BW loss; however, data on associations between shrink and subsequent health and performance parameters in feeder cattle are limited. Operational data from 13 U.S. commercial feedlots (n = 16,590 cattle cohorts) were used to quantify how SHK was associated with bovine respiratory disease (BRD) morbidity and overall mortality risks, HCW and ADG in feeder cattle cohorts arriving to feedlots during 2000 to 2008. Multivariable mixed-effects negative binomial and linear regression models were employed to determine these associations while accounting for other cohort-level demographic variables. The median SHK among the study cohorts was 3.0% with a mean (± SEM) of 2.4 ± 0.02%. The mean (± SEM) cumulative BRD morbidity was 10.0% ± 0.09% (median = 5.8%; range 0 to 100%) and the mean (± SEM) overall cumulative mortality was 1.3% ± 0.01% (median = 0.9%; range: 0 to 25.6%). The mean and median number of days on feed of cohorts experiencing initial BRD cases was 143 and 150 d (range = 23 to 288 d). The effects of SHK were significantly (P < 0.05) associated with BRD morbidity, overall mortality, HCW and ADG, and these effects were significantly (P < 0.05) modified by gender, season and mean arrival BW of the cohort. Combining data on BW loss during transport with cohort demographics could allow a more precise prediction of health and performance of feedlot cattle.

Zoonotic bacteria, antimicrobial use and antimicrobial resistance in ornamental fish: a systematic review of the existing research and survey of aquaculture-allied professionals.

Using systematic review methodology, global research reporting the frequency of zoonotic bacterial pathogens, antimicrobial use (AMU) and antimicrobial resistance (AMR) in ornamental fish, and human illness due to exposure to ornamental fish, was examined. A survey was performed to elicit opinions of aquaculture-allied personnel on the frequency of AMU and AMR in ornamental fish. The most commonly reported sporadic human infections were associated with Mycobacterium marinum, while Salmonella Paratyphi B var. Java was implicated in all reported outbreaks. Aeromonas spp. were most frequently investigated (n=10 studies) in 25 studies surveying ornamental fish from various sources. High levels of resistance were reported to amoxicillin, penicillin, tetracycline and oxytetracycline, which was also in agreement with the survey respondents' views. Studies on AMU were not found in our review. Survey respondents reported frequent use of quinolones, followed by tetracyclines, nitrofurans, and aminoglycosides. Recommendations for future surveillance and public education efforts are presented.

Associations between weather conditions during the first 45 days after feedlot arrival and daily respiratory disease risks in autumn-placed feeder cattle in the United States.

Data on associations between weather conditions and bovine respiratory disease (BRD) morbidity in autumn-placed feedlot cattle are sparse. The goal of our study was to quantify how different weather variables during corresponding lag periods (considering up to 7 d before the day of disease measure) were associated with daily BRD incidence during the first 45 d of the feeding period based on a post hoc analysis of existing feedlot operational data. Our study population included 1,904 cohorts of feeder cattle (representing 288,388 total cattle) that arrived to 9 US commercial feedlots during September to November in 2005 to 2007. There were 24,947 total cases of initial respiratory disease (animals diagnosed by the feedlots with BRD and subsequently treated with an antimicrobial). The mean number of BRD cases during the study period (the first 45 d after arrival) was 0.3 cases per day per cohort (range = 0 to 53.0), and cumulative BRD incidence risks ranged from 0 to 36% within cattle cohorts. Data were analyzed with a multivariable mixed-effects binomial regression model. Results indicate that several weather factors (maximum wind speed, mean wind chill temperature, and temperature change in different lag periods) were significantly (P < 0.05) associated with increased daily BRD incidence, but their effects depended on several cattle demographic factors (month of arrival, BRD risk code, BW class, and cohort size). In addition, month and year of arrival, sex of the cohort, days on feed, mean BW of the cohort at entry, predicted BRD risk designation of the cohort (high or low risk), cohort size, and the interaction between BRD risk code and arrival year were significantly (P < 0.05) associated with daily BRD incidence. Our results demonstrate that weather conditions are significantly associated with BRD risk in populations of feedlot cattle. Defining these conditions for specific cattle populations may enable cattle health managers to predict and potentially manage these effects more effectively; further, estimates of effects may contribute to the development of quantitative predictive models for this important disease syndrome.

Risk factors associated with Escherichia coli O157:H7 in Ontario beef cow-calf operations.

The aim of this study was to identify farm management factors associated with the prevalence of Escherichia coli O157:H7 among cattle in Ontario beef cow-calf operations. A total of 119 cow-calf operations with more than 50 cows in southern Ontario were visited between June and December 2002. From each farm, 65 fresh fecal samples were collected and cultured for E. coli O157:H7. Colonies of E. coli O157:H7 were isolated using immunomagnetic separation and standard microbiological techniques. Final confirmation of suspected colonies was based on identifying E. coli O157:H7-specific genes by PCR and serotyping of representative isolates. A questionnaire was administered to collect information on farm size, cattle demographics, farm management practices, the presence of other livestock and wildlife, and other aspects of the farm environment. Associations between the prevalence of E. coli O157:H7 in cattle feces and management factors were determined using a multivariable logistic regression model that included random effects for farm and county. The presence of pigs on farm, use of corn silage supplementation in winter, number of times cattle were taken to a show in the previous 12 months and the percentage of cows on farm were significant risk factors for the presence of E. coli O157:H7 in fecal pat samples, after controlling for region and the age group of the sampled animals. These findings highlight the potential roles of biosecurity and avoiding mixed animal agriculture in controlling the prevalence of E. coli O157:H7 in beef cow-calf operations.

Influence of type of culture medium on characterization of Mycobacterium avium subsp. paratuberculosis subtypes.

We evaluated the effects of culture and/or enrichment methods on the selection of Mycobacterium avium subsp. paratuberculosis subtypes. M. avium subsp. paratuberculosis isolates from bovine fecal samples processed using a centrifugation protocol were investigated in both liquid (MGIT ParaTb tubes) and solid (Herrold's egg yolk medium) culture media. For this evaluation, M. avium subsp. paratuberculosis subtyping was based on the sequence variation in two of the most discriminatory short sequence repeat loci, i.e., mononucleotide G and trinucleotide GGT, in isolates from liquid and solid cultures. This study identified the existence of one major predominating fingerprint (>13G-5GGT) in bovine fecal samples, regardless of the type of medium used for isolation. Matched-pair analysis of subtypes showed that 69% of samples presented unique subtypes in the two culture media used, while 31% shared the same G-GGT allele. Furthermore, the liquid culture method appeared to select for a more genotypically diverse subtype population than the solid culture method. The variety of subtypes observed between liquid and solid cultures obtained from the same fecal samples suggests that the culture method could provide a "microbiological" bias and lead to a discrepancy in the growth of M. avium subsp. paratuberculosis strains. In conclusion, this study identified that these two types of culture media determined differential growth of M. avium subsp. paratuberculosis strains and that this should be considered in evaluating detection capabilities of diagnostic tests or interpreting data from molecular epidemiological studies performed using conventional solid fecal culture or automated liquid medium culture methods.