Molecular fingerprinting confirms extensive cow-to-cow intra-herd transmission of a single Mycobacterium bovis strain

In this study we have characterized M. bovis isolates from a herd of cattle in Uvalde, Texas in which 52 of the 193 animals selected at random in 1994 from a herd of 331 were caudal fold skin-test positive. Thirty-two of 52 skin-test positive cattle had gross lesions at slaughter, and isolations of M. bovis were made from 29 animals. The herd was comprised of Red Devon cattle purchased between 1978 and 1980 (n = 26) and breeding bulls (n = 3) introduced at later times, and all were tuberculosis test negative at the time of purchase. Other animals were natural additions (offspring) of these cattle. One additional animal, a Holstein present on the ranch at the time of purchase in 1976, was retained to nurse orphaned and weak calves. Using several molecular fingerprinting techniques we have verified a clonal relationship among the M. bovis isolates consistent with infection originating with a single strain. The molecular fingerprint patterns demonstrate the stability of the profiles despite persistence and spread of the organism within the herd for two decades and confirms their use in epidemiological tracing.     

Molecular epidemiology of bovine tuberculosis in wild animals in Spain: a first approach to risk factor analysis

In human tuberculosis (Mycobacterium tuberculosis), molecular epidemiology has accurately indicated the risk factors involved in active transmission of the disease, by comparing individuals whose isolates belong to a cluster with patients whose strains are considered unique. Nevertheless, this application has not been used in bovine tuberculosis (Mycobacterium bovis). Our study describes the integration of epidemiological data into molecular classification data on M. bovis isolates. These were isolated from wild ungulates in Extremadura (western Spain) with the objective of detecting the risk factors linked to the association of strains in clades, which are indicators of the active spread of the disease. The molecular markers used were spoligotyping + VNTR typing (loci: VNTR 2165, VNTR 2461, VNTR 0577, VNTR 0580, VNTR 3192 VNTR 2163a and VNTR 2163b) on a population of 59 M. bovis strains isolated from deer (Cervus elaphus), 112 from wild boar (Sus scrofa), six from bovines, 28 from pigs and 2 from goats (n = 207). Epidemiological variables included the animal species from which the strain was isolated, pathological condition of the host (incipient lesion, early and late generalisation), date of sampling (during or after the reproductive period) and hunting season. Bivariant analysis was used to establish the risk factors connected to the association of strains and later, the variables were evaluated by means of logistic regression. Molecular typing grouped a total of 131 strains (64.21%) in 28 clusters and 76 isolates shows unique profiles. The association of strains was connected to the appearance of macroscopic lesions during the reproductive period (O.R. 4.80; 95% CI 1.09–22.99, P < 0.005), showing a possible higher transmission during the courting period. This happened mainly during the last hunting season analysed (2002–2003, O.R. 3.69; 95% CI 1.27–11.9, P < 0.05), clashing with the time of higher prevalence of the disease in wild ungulates. Active spread was not connected to any species in particular, or to any concrete pathological condition.     

Survival of Mycobacterium avium subsp. paratuberculosis in bovine monocyte-derived macrophages is not affected by host infection status but depends on the infecting bacterial genotype

In this study we investigated the ability of different Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) strains to survive in bovine monocyte-derived macrophages (MDMs) of cows naturally infected with M. paratuberculosis and control cows. We tested the hypotheses that infection status of cows affects macrophage killing ability and that survival of M. paratuberculosis in macrophages is dependent on the strain. Peripheral blood mononuclear cells (PBMC) were obtained from Johne's disease-positive (n = 3) and age and stage of lactation matched Johne's disease-negative (n = 3) multiparious cows. Following differentiation, MDMs were challenged in vitro with four M. paratuberculosis strains of different host specificity (cattle and sheep). Two hours and 2, 4, and 7 days after infection, ingestion, and intracellular survival of M. paratuberculosis strains were determined by fluorescence microscopy. There was no effect of the origin of MDMs (Johne's disease-positive or control animals) on phagocytosis, survival of bacteria, or macrophage survival. In contrast, important strain differences were observed. These findings suggest that some M. paratuberculosis strains interfere more successfully than others with the ability of macrophages to kill intracellular pathogens which may make it important to include strain typing when designing control programs.     

Genotyping of Mycobacterium bovis by geographic location within Mexico

The spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 Mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of Mexico. Our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). Results from the genetic analysis indicate that M. bovis isolates cannot be grouped by geographic location due to a wide range of genetic types involved in dairy cattle infections. Isolates even from the same herd showed different spoligotypes but some isolates from different region had similar genetic patterns. Genetic typing without epidemiologic information does not seem to be a plausible method to trace back animals to source of origin to detect and eliminate sources of infection.     

Serological reactivity to Mycobacterium bovis protein antigens in cattle.

The serological response to 12 purified Mycobacterium bovis antigens were examined in an ELISA assay. These antigens included the majority of M. bovis protein antigens described to date and in most cases they were very similar to the M. tuberculosis antigens of the same molecular mass.

The purified antigens were tested against sera from M. bovis infected cattle, M. bovis culture-negative cattle from infected herds and animals infected with related microorganisms, mainly other mycobacterial species. All the antigens gave strong reactions with at least some sera from the M. bovis infected group and showed cross-reactivity with some of the sera from the other two groups. The antigen with the highest specificity reacted strongly with only 60% of the M. bovis infected sera. Antigens that reacted with most or all of the M. bovis infected sera also gave the highest cross-reactivity with sera from the other two groups. These results indicate that a serological test based on any one or a combination of these antigens, without removal of the cross-reacting epitopes, would be unsatisfactory.     

Management factors associated with Babesia bovis seroprevalence in cattle from eastern Yucatán, Mexico.

The effect of management on the seroprevalence of Babesia bovis was studied in 399 Bos indicus cattle 1–2 years old from 92 farms in the eastern Yucatán, México. The management factors studied were: farm-type, production system, herd size, farm size, stocking density, vector control, dipping interval, type of dipping, type of acaricide and cattle introduction to the farm. A cross-sectional study was carried out (2-stage cluster sampling). The number of serum samples was proportionally distributed according to the number of farms in the nine locations of eastern Yucatán, México (399 animals from 92 farms). Antibody activity to B. bovis was tested using an indirect ELISA. The farms with a seroprevalence ≤75% were considered as cases and those with seroprevalence >75% were considered as controls. The variables with p ≤ 0.20 were included in fixed effects logistic regression. The seroprevalence of the zone was 73.8% (66.3–81.3%). The following risk factors were found: Stocking density (<1 head/ha, OR = 4.04, CI (OR) = 1.20–13.62) and dipping interval (>60 days, OR = 5.07 CI (OR) = 1.26–20.48).     

新疆石河子地区牛支原体的分离鉴定及药物敏感性分析

【目的】确定引起新疆石河子地区集约化牛场常发性肺炎的主要病原同时进行病原的体外药物敏感性分析。【方法】采集有典型咳嗽、流涕症状的牛鼻拭子10份和病死牛肺脏组织1份,用牛支原体特异性引物进行PCR检测,将检测为阳性的样本进行病原培养纯化,对纯化后的分离株菌落进行形态学观察、Dienes染色、生化试验及16S rRNA测序和进化分析,通过测定颜色变化单位(CCU)测定分离株生长曲线,并对分离株进行药物敏感性试验。【结果】PCR结果显示,10份鼻拭子中检测出7份牛支原体阳性样本,1份病死牛肺脏组织也检测为阳性;在涂有肺脏组织研磨液培养液的PPLO固体培养基上长出针尖状的菌落,纯化后分离株菌落形态为典型的煎蛋状;Dienes染色可见明显的深蓝色中心脐;生化试验结果显示,分离株不水解明胶、精氨酸、七叶苷,不发酵乳糖、葡萄糖和甘露醇,不分解尿素,可还原氯化三苯基四氮唑;16S rRNA测序结果显示,分离株与牛支原体国际标准株PG45相似性为99.7%,与国内牛支原体地方流行株XBY01、Ningxia-1、NM2012、Tibet-10的相似性最高,均为99.9%;生长曲线测定结果显示,分离株在培...     

Prevalence of antimicrobial resistance among Salmonella on midwest and northeast USA dairy farms

The objective of this study was to determine the prevalence of antimicrobial resistance among Salmonella isolated from dairy herds in New York, Minnesota, Michigan, and Wisconsin, USA. Serogroup and antimicrobial susceptibility characteristics were determined for Salmonella from cattle and environmental samples collected during August 2000–October 2001 as part of a longitudinal study where 129 herds were visited at 2-month intervals. Salmonella isolates were tested (using a broth microdilution method) for susceptibility to amoxicillin/clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim/sulfamethoxazole. Of the 1506 isolates tested for minimum inhibitory concentrations to these 14 antimicrobial agents, 81.2% were pan-susceptible and for most herds (81.6%) the predominant antimicrobial resistance pattern was pan-susceptible. At least 1 Salmonella isolate resistant to 5 or more antimicrobial agents was found on 23.6% of herds. This resistance phenotype was most common among serogroups B and E1 and among samples from calves and farmer-designated sick cows. Resistant samples most frequently exhibited resistance to tetracycline, streptomycin, and/or ampicillin. No samples were resistant to ceftriaxone (though 13 were in the intermediate range), and very few samples were resistant to ciprofloxacin (n = 1), nalidixic acid (n = 5), or trimethoprim/sulfamethoxazole (n = 7).     

Evaluation of MPB70, bovine PPD and lipoarabinomannan as antigens in ELISA for the serodiagnosis of bovine tuberculosis

The performance of the secretory protein MPB70 of Mycobacterium bovis, bovine PPD, and lipoarabinomannan (LAM) were evaluated as antigens in ELISA for detection of tuberculosis (TB) infected cattle. Sera were from 120 M. bovis infected cattle and 223 cattle from a TB free herd. ELISA results were analyzed using receiver operating characteristic (ROC) curves in relation to culture results. The areas under the three ROC curves were 71 ± 49% SE (MPB70), 71 ± 27% SE (bovine PPD), and 56 ± 4% SE (LAM).     

Bacterial flora and risk of infection of the ovine teat duct and mammary gland throughout lactation

We collected samples of teat duct material and mammary secretion from ewes in three farms (flock A, polyparous n = 7; flock B, polyparous n = 6, primiparous n = 4; flock C, polyparous n = 4): 14 samples immediately after lambing (before sucking of lambs), 244 samples during the suckling period and 156 samples during the milking period. Conventional bacteriological techniques were used. The results were modeled using survival analysis, initially by the Kaplan–Meier method and then by the Cox Proportional Hazards method. Then, we calculated the minimum true risk of an “at-risk” teat or mammary gland being infected and analyzed these data with STATA using the GLLAMM program for Generalised Linear Latent and Mixed Models. During the suckling period, bacteria were isolated from 52 (21%) duct material and 19 (8%) secretion samples; respective results for the milking period were 20 (13%) and 9 (6%). There was an increased risk of duct rather than secretion samples being infected (P < 0.001). There was a significant difference among flocks in isolating bacteria from duct (P < 0.01) or secretion (P < 0.001) samples during suckling period, but not during hand-milking period (P > 0.4 and 0.1, respectively). There were no differences between isolation of bacteria from duct (P > 0.5) or secretion (P > 0.7) samples among primiparous and polyparous animals. Most bacterial isolates were staphylococci. Persistent isolation of the same bacterial species from duct material samples obtained from a particular ewe was recorded with five Staphylococcus spp. and two Mannheimia haemolytica isolates. The results indicate that infections of the teat duct can take place easily; however, not all infections result to infection of the mammary gland. The results support experimental evidence that defence mechanisms of the healthy teat are able to limit the infection. Maintenance of healthy teats contributes to effective defence mechanisms, and coupled with minimal infections of the teat duct, would contribute to the prevention of mastitis in ewes.     

Analyses of lipopolysaccharides, outer membrane proteins and DNA fingerprints reveal intraspecies diversity in Moraxella bovis isolated in Argentina

Intra-specific diversity within Moraxella bovis was investigated analysing DNA fingerprints, outer membrane proteins (OMP) and lipopolysaccharides (LPS) profiles. Three collection strains and 57 isolates of M. bovis, collected during 3 years from cattle with infectious bovine keratoconjunctivitis (IBK) symptoms, from diverse geographical locations of Argentina, were examined. The LPS and OMP profiles were studied through SDS–PAGE analysis and genotype was determined by PCR-DNA fingerprinting. Genotyping identified five DNA types while analysis of LPS and OMP profiles identified three rough LPS types and three OMP types among the 60 isolates of M. bovis including the three collection strains. None of the three methods employed to assess diversity was discriminating when used alone because the degree of heterogeneity in each group of surface structures was limited, but when data of each typing method were combined, 15 distinct subgroups were determined. This subgrouping was clearly able to differentiate isolates of the same genotype. These typing methods appear to be useful to assess different aspects of the disease such as the diversity within a population of M. bovis associated to epidemic conditions, track the causal agent in an outbreak of the disease, monitoring vaccination programs and studies on virulence.     

Herd-level risk factors for Salmonella enterica subsp. enterica in U.S. market pigs

Midwest U.S. herds (n = 63) were studied to identify risk factors for harboring Salmonella enterica among slaughter-weight pigs. Samples collected on farms (feces) and at slaughter (distal colonic content, cecal content and ileocolic lymph nodes) were cultured using conventional means. Approximately 15 pigs were studied per herd, for a total of 3754 samples. The proportion of pigs positive in one or more samples was calculated for each herd. Herd characteristics were described by a combination of interview and written survey. Logistic regression was used to detect relationships between the detection of Salmonella and potential herd-level risk factors. The mean individual pig prevalence was 5% for feces, 4% for distal colonic content, 15% for ileocolic lymph nodes, and 17% for cecal contents. One or more Salmonella isolates were detected in at least one sample type in every herd. The five most common serovars were S. Agona, S. Derby, S. Schwarzengrund, S. Typhimurium and S. Senftenberg, with 25 additional serovars detected. Salmonella prevalence estimates were positively correlated among all samples except distal colonic content and ileocolic lymph nodes. Pigs with culture positive fecal samples were at increased odds of being detected positive for each of the slaughter-collected samples examined, namely distal colonic content (OR = 30.5), ileocolic lymph nodes (OR = 12.9) and cecal content (OR = 23.2). Herds with positive fecal sample(s) had increased odds of having positive cecal content (OR > 1.5), distal colonic content (OR = 15.3) and ileocolic lymph nodes (OR = 12.7). Pigs from herds with at least some bowl drinkers had eight-fold higher odds of testing Salmonella positive than did pigs from herds with only nipple drinkers. Pigs from herds with only dry feeders had five-fold higher odds of testing Salmonella positive when compared with pigs from herds with combinations of wet/dry style feeders. Interventions at these two points should be considered when designing growing pig facilities to reduce Salmonella shedding.     

Studies on vertical transmission of Trichinella spp. in experimentally infected ferrets (Mustela putorius furo), foxes (Vulpes vulpes), pigs, guinea pigs and mice

Vertical transmission of Trichinella spiralis was evaluated in ferrets (n = 21), foxes (n = 11), pigs (n = 12), guinea pigs (n = 16), and mice (n = 41). The placental barrier to be crossed by migratory Trichinella larvae varies structurally in different animal species. Ferrets and foxes have an endotheliochorial placenta structure, guinea pigs and mice a haemochorial, and pigs an epitheliochorial placenta. The non-encapsulating Trichinella pseudospiralis larvae have an extended muscle migration prior to entering a muscle cell. To evaluate if T. pseudospiralis was more likely to be transmitted to offspring, an additional group of foxes (n = 11) infected with T. pseudospiralis was included. Two different dose levels were used for ferrets, pigs, guinea pigs, and mice. In pigs and guinea pigs, infection was given at different times of the gestation period. Vertical transmission, measured as recovery of muscle larvae in the offspring, was demonstrated in both ferrets groups, in all four guinea pig groups, and in the high dose mouse group, but not in any fox or pig groups.     

Risk factors for herd-level bovine-tuberculosis seropositivity in transhumant cattle in Uganda

We investigated the prevalence and risk factors to positive herd-level tuberculin reactivity between October 2003 to May 2004 to bovine tuberculosis (BTB) in the four transhumant districts of Uganda: three districts (Karamoja region) of nomadic transhumance cattle rearing (30 superherds and 1522 cattle), and one district (Nakasongola) of fixed-transhumance (7 herds and 342 cattle). We used the comparative intradermal skin-test, sampled 50 animals per superherd/herd, and considered herd positive if there was at least one reactor. Of the 30 superherds under nomadic transhumance, 60% (95% CI 41.4, 79) were tuberculin-test positive; of the 7 fixed herds, 14.3% (95% CI −20.7, 49.2) were tuberculin test positive. The true herd prevalence was estimated at 46.6%. Many risk factors were collinear. The final multivariable logistic-regression model included: recent introductions from market (OR = 3.4; 95% CI 1.1, 10.3), drinking water form mud holes during dry season (OR = 49; 95% CI 9.1, 262), and the presence of monkeys (OR = 0.08; 95% CI 0.0, 0.6) or warthogs (OR = 0.1; 95% CI 0.0, 0.3). No association was found between herd size or number of herd contacts with reactors; it was probably masked by the effect of high between-herd interactions. Provision of water from mud holes in dry river beds and introductions of new animals are risk factors that might be targeted to control BTB in transhumance areas.     

Use of PCR-RFLP to identify Leishmania species in naturally-infected dogs

Tissue imprints on Giemsa stained slides from dogs were used to investigate the presence of Leishmania amastigotes by either optical microscopy (OM) or Polymerase chain reaction (PCR) detection of DNA. Samples from skin, spleen, lymph node, liver and bone marrow from a Leishmaniasis endemic area dogs where Leishmania (Leishmania) chagasi and Leishmania (Viannia) braziliensis are sympatric were studied. Dogs were initially diagnosed by Indirect Immunofluorescence (IIF), as which 39 were IIF positive (≥1:40) and 16 negative. The IIF positive dogs were clinically grouped as symptomatic (n = 15), oligosymptomatic (n = 12) and asymptomatic (n = 12). Although PCR positivity was higher in symptomatic dogs, specially their skin samples, there was no significant difference among clinical groups or organs examined. Ten (62.5%) out of 16 IIF and OM negative animals were positive for PCR in at least one organ. Forty-eight positive PCR amplicons were further submitted to RFLP for Leishmania identification. All dogs were infected with L. (L.) chagasi except one, infected with L. (V.) braziliensis. PCR was more efficient than IIF and OM to diagnose canine visceral Leishmaniasis (CVL), regardless of the organ examined and the clinical form present. The use of PCR together with serology helps determining the extension of sub clinical infection in CVL endemic areas and provides a better estimate of the number of dogs to be targeted for control measures. In conclusion, our data reinforce the need for a specific diagnosis of canine infection in areas where diverse Leishmania species are sympatric and demonstrate that PCR–RFLP can be used to identify Leishmania species in dog tissue imprint stained slides.     

Protein variability among Mycoplasma hyopneumoniae isolates

Sodium-dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to study the protein variability of Mycoplasma hyopneumoniae isolates. Fifty-six M. hyopneumoniae isolates from 6 different countries and 37 different herds were used. From eight herds, more than one isolate was available. All SDS-PAGE patterns of isolates originating from different herds were clearly divergent. Intra-species protein variability was quantified using the reference strain J and seven field strains all obtained from different herds and classified according to virulence. Between the field strains, a variability of 25% was found, while the culture-adapted strain J was clearly divergent and showed 30% variability with the field strains. No clustering according to virulence was obtained, but a protein band of about 181 kDa was present in the two highly virulent isolates whereas this protein band was absent in the moderately and low virulent isolates. Protein patterns of isolates derived from different animals from the same herd, were identical or differed in only a few protein bands. This study clearly indicates that, in agreement with previous studies on genomic diversity of M. hyopneumoniae isolates, proteomic variability within the species is high. Our study did not find clear evidence that more than one M. hyopneumoniae isolate circulates within a herd at a specific time point. The minor differences found between M. hyopneumoniae isolates from the same herd might reflect the organism's ability to alter its proteomic expression profile under field conditions.     

The use of purine derivatives/creatinine ratio in spot urine samples as an index of microbial protein supply in Yerli Kara crossbred cattle

Three experiments were conducted to evaluate the use of purine derivatives (PD)/creatinine ratio in spot urine samples as an index of microbial protein supply in Yerli Kara crossbred cattle (YK-C). In Experiment I, response of daily PD excretion to feed intake in YK-C at state farm was measured. In Experiment II, spot urine sampling techniques was applied at state farm and four YK-C bulls were used. In Experiment III, spot urine sampling technique was applied at small-holder farms. There were significant correlations (R² = 0.99) between PD excretion (mmol/day) and digestible organic matter intake (DOMI) (kg/day) in Experiment I, Y = 12.5 (± 0.5) + 19.7 (± 3.5)X (R² = 0.99, n = 16). The equation obtained from Experiment I could be expressed as: Y = − 2.3 (± 0.3) + 0.953 (± 0.06)X, (R² = 0.99, n = 49) where Y is PD excretion (mmol/day) and X is the PDC index. The PDC index was calculated as the molar concentration ratio of PD to creatinine times the metabolic body weight (kg). The corresponding microbial-N values to PDC index of groups I, II and III in developed banding system are 15–25 g/day. Experimentally estimated DOMI was 2.21 ± 0.15 kg/day. Estimated DOMI of groups I, II, and III were 2.8 ± 0.6, 2.6 ± 0.7 and 2.7 ± 0.7 kg/day, respectively. In conclusion, the PDC index in spot urine samples could be used under similar farm condition as an indicator of microbial protein supply in YK-C cattle. Estimated DOMI from PDC index in spot urine samples under defined field conditions may help the development of feeding strategies for YK-C cattle held by small holders.     

Risk factors for bovine herpesvirus-1 seropositivity

This paper reports the investigation of risk factors for bovine herpesvirus-1-seropositivity, based on a cluster-sample survey of the Belgian cattle population. This serosurvey was carried out in 1998 in 309 randomly selected unvaccinated herds of all types (dairy, mixed and beef) were all bovids (N = 11,284) were sampled.

Older and male cattle had higher seroprevalence. Origin (homebred or purchased) and herd size interacted; for smaller herds (≤50 cattle on the premises), purchase status and larger herd size were risk factors, whereas these effects were not observed for larger herds.     

Diagnosis of amitraz resistance in Boophilus microplus in New Caledonia with the modified Larval Packet Test

The tick Boophilus microplus represents a serious pathological constraint to livestock production in New Caledonia. Cattle ticks are controlled by chemical application of two acaricides that are currently used in New Caledonia; deltamethrin is used at 46% of the cattle production facilities and amitraz at the remaining 54% premises where resistance to deltamethrin has been identified. In 2003, a modified Larval Packet Test (LPT) was used to conduct a survey for amitraz resistance. Ticks were collected from 29 farms, including farms using deltamethrin (n = 8) or amitraz (n = 21). Of eighteen different tick populations, sixteen populations were defined susceptible to amitraz and two populations were considered amitraz-resistant. This is the first report of populations of B. microplus being resistant to amitraz, using the modified LPT in New Caledonia. A thorough survey of tick susceptibility to amitraz in cattle farms of the country should be conducted to assess the presence of amitraz-resistant populations. The emergence of amitraz resistance so soon after its introduction has some important implications for the strategy and organisation of tick control in New Caledonia, and this paper discusses some of the urgent actions that should be undertaken.     

Efficacy of monensin sodium for the reduction of fecal shedding of Mycobacterium avium subsp. paratuberculosis in infected dairy cattle

Reducing the quantity of Mycobacterium avium subsp. paratuberculosis (MAP) being shed by cows with Johne's disease should decrease the risk of spread of this disease to young stock. Previous work has suggested that monensin sodium decreases the pathologic lesions associated with Johne's disease, but the impact on shedding of viable MAP remains unknown. After serologic screening of 32 dairy herds in southwestern Ontario, 228 cows from 13 of these herds were enrolled into a randomized clinical trial. Fecal culture and PCR were used to identify 114 cows as potential fecal shedders, while another 114 cows were enrolled as ELISA negative, herd and parity matched controls. All cows were randomized to receive either a monensin controlled release capsule (CRC) or a placebo capsule. Serial fecal and blood samples were collected for fecal culture and serum ELISA testing over a 98-day period. On day 98 of the study, treatments were switched for all cows continuing in the trial. These remaining cows were followed for another 98 days with a similar sampling protocol. Mixed effect models were used to measure the impact of treatment on the number of colony forming units identified on fecal cultures over time. During the first 98 days of the study, cows treated with a monensin CRC were found to shed 3.4 cfu per tube less than placebo treated cows (P = 0.05). The serum ELISA S/P ratio was reduced by 1.39 units in cows given monensin (P = 0.06). However, treatment with monensin did not reduce the odds of testing positive on serology. Only the cows shedding MAP on day 0 were found to have a reduced odds of testing positive on fecal culture when treated with monensin (OR = 0.27; P = 0.03). Monensin sodium administered to infected animals at 335 mg/day marginally reduced fecal shedding of MAP in mature dairy cattle, but the biological significance of this reduction is unknown.