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Pasteurella multocida biofilm formation, and the interrelationship of P. multocida with Histophilus somni in a polymicrobial biofilm during bovine respiratory diseasePetruzzi, Briana Lynn 08 February 2018 (has links)
Pasteurella multocida is an important multi-host animal and zoonotic pathogen that is capable of causing respiratory and multi-systemic diseases, bacteremia, and infections resulting from bite wounds. The glycosaminoglycan capsule (CPS) of P. multocida is an essential virulence factor, protecting the bacterium from host defenses. However, chronic infections such as bovine respiratory disease (BRD) and avian cholera may be associated with biofilm formation.
Biofilm formation was inversely related to capsule production (determined by uronic acid and N-acetylglucosamine assays), and was confirmed with capsule-deficient mutants of mucoid strains. Capsule-deficient mutants formed biofilms with a larger biomass that was much thicker and smoother than encapsulated strains.
Gas chromatography-mass spectrometry, nuclear magnetic resonance, and enzymatic digestion demonstrated that the matrix material of the biofilm was composed predominately of a glycogen exopolysaccharide (EPS). Therefore, CPS may interfere with biofilm formation by blocking adherence to a surface or by preventing the EPS matrix to encase large numbers of bacterial cells.
Chemical mutagenesis was performed on P. multocida strain P1059, resulting in isolation of an acapsular mutant designated as P1059-R8. A uridyltransferase encoded by gene P1059_01979 was mutated in such a way that a polar amino acid was changed to a non-polar amino acid near the active site. The protein product of P1059_01979 is important for the biosynthesis of the CPS subunit N-acetylglucosamine. CPS quantification revealed that the subunit glucuronic acid was produced in equal concentrations to the parent, but the CPS subunit N-acetylglucosamine was not detected in the chemical mutant. Biofilm formation in the chemical mutant was significantly higher than in WT P1059 and the capsule-deficient mutant. We hypothesize that P1059_01979 is essential for CPS production in P. multocida serogroup A.
Histophilus somni and Pasteurella multocida cause bovine respiratory disease (BRD) and systemic infections in cattle. Following respiratory infection of calves with H. somni, P. multocida is also often isolated from the lower respiratory tract. Because H. somni normally forms a biofilm during BRD, we suspected that P. multocida may co-exist with H. somni in a polymicrobial biofilm. Interactions between the two species in the biofilm were characterized and quantified by fluorescence in situ hybridization (FISH), and the biofilm matrix of each species examined by fluorescently-tagged lectins (FTL), confocal scanning laser microscopy of in vitro biofilms and bovine pulmonary tissue following dual H. somni and P. multocida infection. FISH and FTL were used to show that P. multocida and H. somni were evenly distributed in the in vitro biofilm, and both species contributed to the polymicrobial biofilm matrix. COMSTAT z-stack image analysis revealed that the average biomass and biofilm thickness of the individual and polymicrobial biofilms were greatest when both species were present. Encapsulated P. multocida isolates not capable of forming a biofilm still formed a polymicrobial biofilm with H. somni, but only the EPS of H. somni could be detected by FTL staining of bovine tissues from which both species were isolated. Bacteria within a biofilm are more quiescent than during planktonic growth and induce less of an inflammatory response, indicating encapsulated P. multocida may take advantage of the H. somni biofilm to persist in the host during less severe, but more chronic, BRD. These results may have important implications for the management of BRD.
Acute avian cholera is associated with encapsulated P. multocida, while chronic and asymptomatic cases of avian cholera are associated with acapsular P. multocida isolates. We hypothesize that biofilm formation is present and an important factor for chronic and asymptomatic avian cholera. Experimental infections of chickens with biofilm deficient P. multocida strain WT X73, proficient biofilm forming P. multocida strain X73ΔhyaD, and proficient biofilm forming clinical isolates 775 and 756 showed that virulence inversely correlated with biofilm formation. Histopathological analysis showed that biofilm forming isolates induced little inflammation in the lungs, heart, and liver, while biofilm deficient isolates induced greater inflammation. Biofilm material was located in pulmonary tissues of chickens diagnosed with chronic avian cholera using FTL staining.. Quantitative real-time PCR for expression of cytokine genes in the spleens of infected chickens indicated that P. multocida induced Th1 and Th17 immune responses during acute and chronic avian cholera. Chickens that succumbed to acute avian cholera after experimental challenge with WT X73 had high levels of INF-ƴ, IL-1β, IL-6, IL-12A, IL-22, IL-17A, and IL-17RA expression in the spleen compared to all other experimental groups. Antibody titers were low, indicating that antibodies may be less important in managing and clearing P. multocida infections. / Ph. D.
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Epidemiology of bovine respiratory disease and mortality in commercial feedlotsBabcock, Abram Heath January 1900 (has links)
Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / David G. Renter / Bradley J. White / The objective of my research was to quantify epidemiologic parameters associated with feedlot mortality and bovine respiratory disease complex (BRDC); the most significant cause of morbidity in U.S. feeder cattle. We conducted four retrospective studies utilizing individual health and cumulative cohort-level feedlot data. We developed a database that represented 33 U.S. feedlots from six states over ten years. Each project used a subset of these data.
We found that the timing of BRDC was associated with important performance and health indices. In the first study, we evaluated the effect of the timing of individual BRDC treatments on standardized net returns. We found important performance and health measures (e.g. hot carcass weight and number treatments) driving net return differences associated with the timing of BRDC. For the second cohort-level study, we classified temporal patterns of BRDC, and evaluated associations among temporal patterns and performance and health. Temporal patterns were significantly associated with mean daily weight gain, days on feed, carcass weight, yield grade, quality grade, cumulative mortality, and retreatment risk.
We also evaluated combined mortality and culling risks and quantified the effects of risk factors using count models. All risk factors (arrival weight, gender, and arrival month) were significant and the effects were modified by one another; effects of these covariate patterns have been impossible to quantify in smaller studies.
Finally, we assessed the ability of regression models to predict cumulative BRDC morbidity based on arrival risk factors; then assessed the additional value of incorporating daily BRDC morbidity and mortality information. The percent of correctly classified cohorts did increase across days, but the effect of day was modified by weight, month, and feedlot. Information on daily morbidity was beneficial in predicting cumulative morbidity, but daily mortality provided little benefit.
Our database containing animal health and cohort-level data allowed us to generate novel information on the effects of the timing of BRDC in feedlot populations. We also demonstrated effects of covariate patterns on adverse health outcomes that heretofore had been difficult to quantify. Finally, we showed that a predictive model for BRDC may be useful for the feedlot industry; this model should be further developed with future research.
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Epidemiology, diagnosis, and prevention of bovine respiratory disease complexHanzlicek, Gregg Alan January 1900 (has links)
Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / David G. Renter / Bradley J. White / The objective of my research was to generate novel information concerning the epidemiology, diagnosis and prevention of bovine respiratory disease complex (BRDC), a common pre-weaning and post-weaning beef calf disease. To reach my objective, I conducted three prospective field trials within post-weaned calf populations, and one retrospective study of pre-weaned calves utilizing survey data.
I evaluated differences in behavior, health and performance in calves receiving multiple component health programs. Calves in a minimally invasive program, which included primarily non-injectable products, displayed less aversion to initial product administration but experienced higher BRDC morbidity (P = 0.02) and poorer performance (P = 0.04) compared to calves in a more invasive (all injectable products) program.
Secondly, in a study of Mannheimia haemolytica inoculated calves, I found that no parameter included in physical examinations, or common blood component evaluations could discern health from disease. However, disease recognition was aided by the measurement of the number of steps taken by a calf in a 24 hour period. None of the parameters that were evaluated predicted the severity of lung pathology.
Thirdly, I conducted a study in post-weaned feeder calves that determined prevalence estimates for Mollicutes in general, and Mycoplasma bovis specifically, and their respective associations with health and performance. Nasal Mollicutes prevalence was high on arrival, and differences in calf performance were associated with (P < 0.01) nasal prevalence. More than half of the calves seroconverted to M. bovis; calves not seroconverting gained more weight (0.49 kg/head/day) during the study than those calves that did seroconvert (0.35 kg/head/day).
Finally, I conducted a retrospective analysis of national U. S. cow-calf survey data to identify herd level management practices associated with pre-weaned calf BRDC. I found feeding antibiotics to pre-weaned calves, importing cattle, the number of outside visitors, economic purpose of the cow-calf operation, and breeding management of the herd were associated with herd-level pre-weaning BRDC rates.
My research projects generated unique information concerning the epidemiology of important pathogens, differences among preventive health programs, objective BRDC diagnostic parameters, and pre-weaning BRDC risk factors. These research studies reinforce the complexity of BRDC and demonstrate the pathogen, animal and management factors affecting BRDC risk in pre- and post-weaned beef calves.
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Qualification and quantification of bacterial pathogen load in acute bovine respiratory disease cases.Roof, Clinton January 1900 (has links)
Master of Science / Department of Clinical Sciences / Michael D. Apley / One hundred ninety four steers, bulls, and heifers weighing 182-318 kg were purchased at an Arkansas sale barn and shipped 12 hours to a northern Kansas feedlot. There was no previous history of treatment and the cattle had been delivered to the sale barn within the 24 hour period prior to the sale. The objectives of the study were to evaluate (1) bacterial pathogen isolates in different locations in the respiratory tract, (2) pathogen load in clinically ill and clinically normal calves, and (3) compare histological damage that may be a result of clinical disease. Fifteen calves were identified with signs of acute bovine respiratory disease (BRD) based on clinical score and a minimum rectal temperature of 40° C. An additional 5 calves with no clinical signs and rectal temperatures < 40° C were selected as controls. Cattle were humanely euthanized following recording of antemortem clinical observations. At postmortem, samples for microbiologic and histologic (hematoxylin and eosin stain) analysis were collected from grossly normal and/or consolidated tissue in each lung lobe. Samples were also collected from the tonsils and trachea. Quantification of the BRD pathogens per gram were determined for each positive site and then converted to total counts for each animal. Total colony forming units (CFU) of pathogens in the entire lung for cattle with identified pathogens ranged from 2x10[superscript]7 – 2x10[superscript]8 CFU for Pasturella multocida and 9x10[superscript]6 – 9x10[superscript]8 CFU for Mannheimmia haemolytica. Total visual estimated percent consolidation ranged from 0.0% to 45.0% of the total lung. Isolated pathogens from the upper and lower respiratory tract were compared and showed to have no significant agreement. Histology scores of 0-4 were assigned to the tissue samples and compared to the quantified BRD pathogens to test a possible association between the pathologic process and the total agents in that tissue sample. A significant difference in bacterial counts between histology scores of zero or 1 and a histology score of 4 was observed.
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Near infrared spectroscopy: a potential method to detect undifferentiated bovine respiratory diseaseFox, Jeffrie Thomas January 1900 (has links)
Master of Science / Department of Diagnostic Medicine/Pathobiology / Larry C. Hollis / Mark F. Spire / Two studies were undertaken to evaluate the use of Near Infrared Spectroscopy (NIRS) to determine arterial oxygen saturation (StO[subscript]2) in cattle with naturally-occurring Undifferentiated Bovine Respiratory Disease (UBRD) and experimentally-induced UBRD utilizing Mannheimia haemolytica.
The first study was a natural infection model utilizing 679 beef heifers weighing approximately 227 kg (500 pounds) originating from a southeastern U.S. salebarn. Heifers were evaluated for UBRD upon feedlot arrival, at revaccination, at day 35 on feed, at re-implant time, and two weeks prior to shipment for slaughter. Animals deemed to have UBRD were treated for UBRD and data was collected for 5 days following treatment, while a comparable healthy cohort was also evaluated at the time of treatment. There was a trend for NIRS to be able to predict the incidence of subsequent UBRD when cattle were evaluated on arrival (p=0.0552). However, the ability to detect UBRD in clinically ill cattle was not significantly different (p>0.1690) when compared to healthy cohorts in this model.
When carcass characteristics were evaluated at each time point, NIRS StO[subscript]2 values were able to differentiate between yield grades of animals with UBRD and healthy cohorts when evaluated at revaccination, day 35, re-implant, and pre-shipping (p<0.0199). NIRS tended to be able to differentiate yield grades at initial processing (p=0.0513). StO[subscript]2 was not a predictor of quality grade at any time point (p>0.1023), nor was there any correlation between lung lesions at slaughter and StO[subscript]2 (p>0.2292).
The second study involved 12 head of 181 kg (400 pound) heifers which were subjected to an experimental challenge model of Mannheimia haemolytica. Animals were evaluated daily and StO[subscript]2 readings recorded 12 hours pre-inoculation, at inoculation, 6, 12 and 24 hours post inoculation and daily for the next 12 days. While NIRS could not definitively differentiate healthy cohort cattle from challenge cattle (p>0.0713), there were trends toward challenge cattle having lower StO[subscript]2 values than healthy controls.
The authors conclude that while these studies did not provide conclusive evidence of the ability of NIRS to detect UBRD, further studies with a machine that is specifically calibrated and designed for use with cattle should be performed.
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Various factors involved in control, treatment, and investigation of bovine respiratory disease in high risk feedlot cattleTorres, Siddartha January 1900 (has links)
Doctor of Philosophy / Department of Animal Science and Industry / Chris Reinhardt / Dan Thomson / Bovine Respiratory Disease Complex (BRDC) is the most common and costly disease in feedlot cattle in North America. Annual economic losses are estimated to be US$1 billion due to mortality, reduced performance, and treatment costs. The disease is a multifactorial syndrome caused by a combination of environmental factors, management practices, animal susceptibility, and viral and bacterial pathogens. The objectives of this dissertation were to evaluate two injectable antimicrobials for the treatment and control of BRDC in feedlot cattle, investigation of factors associated with BRDC mortality and morbidity, and to develop control charts based on statistical process control (SPC) principles to monitor cattle mortality rates.
Two multi-site prospective studies were conducted to evaluate the comparative efficacy of the administration of gamithromycin and tulathromycin for the treatment and control of BRDC. A total of 2,529 animals were enrolled at two commercial feedlot locations to evaluate the efficacy of the antimicrobials to control BRDC. Morbidity due to BRDC was higher (P = 0.03) among calves receiving gamithromycin compared with those receiving tulathromycin; however, treatments were considered bioequivalent (P < 0.05) for BRDC mortality, case fatality rate and re-treatment rate. Final BW, ADG, DMI and F:G, were similar (P<0.05) between the groups of calves receiving gamithromycin and tulathromycin. For the evaluation of treatment efficacy, a total of 1,049 calves were enrolled in the study. Re-treatment rate was higher among animals treated with gamithromycin compared with those treated with tulathromycin. Treatments were bioequivalent (P < 0.05) for case fatality rate, final BW, and ADG.
To evaluate factors associated with BRDC, a retrospective study was conducted to analyze BRDC mortality and morbidity associated with initial body weight, rectal temperature, and castration and dehorning (tipping) at processing. Calves with lighter weights and fever at processing were at greater risk of mortality and morbidity due to BRDC. Also, bulls castrated at processing were at higher risk of developing BRDC. Finally, we developed control charts based on SPC principles to monitor and identify “normal” and special cases of variation of mortality rate. In feedlot cattle, monitoring lots of cattle through SPC principles can be used as a powerful tool for continuous improvement.
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Comparison of gamithromycin, tilmicosin and tulathromycin: metaphylactic treatments in high risk calves for bovine respiratory diseaseMiller, Tanner J. January 1900 (has links)
Master of Science / Department of Clinical Sciences / Daniel U. Thomson / Bovine Respiratory Disease (BRD) continues to be one of the largest animal health concerns in the cattle industry. BRD is a multifaceted group of pathogens, both viral and bacterial, that take advantage of an immune compromised calf to cause disease. This study took aim at comparing metaphylactic treatments for BRD in both the feedlot and pasture setting.
In the feedlot study, heifers (n=579, 403.7 ± 27.4 lbs) from Southwest Texas were identified as being high risk for BRD and shipped to the Clayton Livestock Research Center in Clayton, NM. Cattle were randomly allocated within truck load lots into 18 to 20 head treatment pens (30 pens; 3 treatments; 10 reps). Cattle were given one of three metaphylactic treatments based on the randomly assigned treatment for their pen within a replicate. The three antibiotic treatments administered at initial processing were: 1) Tulathromycin (2.5 mg/kg), 2) Tilmicosin (13.3 mg/kg), and 3) Gamithromycin (6.0 mg/kg). Cattle were fed a typical commercial starter diet for the first 56-60 d with a step-up ration change at day 28. At the end of the feeding period, pens were weighed and body weights recorded. Dry Matter Intake, morbidity, and mortality were recorded by CLRC personnel daily. Cattle administered tulathromycin had higher daily gains than cattle administered gamithromycin by 0.29 lbs/d (P<.01) and tended (P=0.09) have higher daily gains than cattle that received tilmicosin by 0.18 lbs/d. Tulathromycin treated cattle tended (P = 0.12) to have improved feed efficiency compared to gamithromycin treated cattle. Cattle that received tulathromycin (5.2%) had lower morbidity rates (P < .02) than tilmicosin (14.6%) and gamithromycin (12.79%) treated cattle. There were no treatment differences in dry matter intake or mortality in cattle.
For the wheat pasture study, heifers (n=120, 393.2 ± 28.6 lbs) from the same origin and risk were shipped to the CLRC and processed before being trailed to a nearby wheat pasture. Cattle were randomly assigned into three treatment groups (3 treatments, 40 reps), and were given one of three metaphylactic treatments. The three antibiotic treatments administered at initial processing were: 1) Tulathromycin (2.5 mg/kg), 2) Tilmicosin (13.3 mg/kg), and 3) Gamithromycin (6.0 mg/kg). Cattle were allowed to graze on wheat for 54 days with free-choice Hi-Pro mineral mixed with Lasalocid, an ionophore. After 54 days on wheat pasture, the cattle were trailed back to the CLRC facilities and final individual weights were recorded. Morbidity and mortality were recorded daily by CLRC personnel. No differences were identified for ADG (P=0.98), morbidity (P=0.46) or mortality (P=0.36) among the three treatment groups.
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Utilizing Lactipro (Megasphaera elsdenii NCIMB 41125) to accelerate adaptation of cattle to high-concentrate diets and improve the health of high-risk calvesMiller, Kevin Alan January 1900 (has links)
Doctor of Philosophy / Department of Animal Sciences and Industry / J. S. Drouillard / Four experiments were conducted to evaluate the potential for accelerating adaption to high-concentrate diets and improving the health of high-risk calves, by administering an oral dose of Megasphaera elsdenii. Lactipro, a probiotic containing a live culture of Megasphaera elsdenii (10[superscript]9 CFU/mL), was orally dosed to cattle (100 mL) at initial processing. In experiment 1, heifers were adapted to a high-concentrate diet in 21 d without Lactipro or dosed with Lactipro and adapted in 16 d, 11 d, 6 d, or fed the high-concentrate diet d 1. Accelerating adaptation decreased DMI (P = 0.09), ADG, and efficiency (P < 0.05) during the first 30 d compared to cattle adapted over 21 d. Over 129 d, DMI and ADG were not different (P ≥ 0.12), but efficiency tended (P = 0.08) to be better for cattle adapted over 21 d. Liver abscesses and carcass traits were not different between treatments, with the exceptions of LM area (quadratic, P < 0.01) and marbling (linear, P = 0.07), which tended to increase with accelerated adaptation protocols. In experiment 2, cattle dosed with Lactipro and placed directly onto the finishing diet required less roughage (P < 0.01), tended to have lower DMI (P = 0.07), better quality grade (P ≤ 0.07), and similar ADG and efficiency (P ≥ 0.14) compared to cattle adapted over 18 d without Lactipro. Over the first 24 d DMI, DM fecal output, and apparent total tract NDF digestibility were greater (P < 0.01) for cattle adapted over 18 d. In Experiment 3, dosing calves, of Mexican origin, with Lactipro did not affect performance (P ≥ 0.16). Second antibiotic therapies for undifferentiated bovine respiratory disease (UBRD) tended (P = 0.06) to be lower in calves dosed Lactipro. In Experiment 4, dosing calves from Texas with Lactipro increased DMI, ADG, and efficiency (P ≤ 0.05). The incidence of 1st and 2nd antibiotic therapies for UBRD (P < 0.01) and medical cost were decreased for calves dosed Lactipro (P < 0.05). Dosing cattle with Lactipro allows for accelerated adaption to high-concentrate diets, decreases reliance on roughages, and improves health during the receiving period.
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The association between calfhood BRDC and subsequent departure from the herd, milk production, and reproduction: an observational, retrospective studySchaffer, Aaron P. January 1900 (has links)
Master of Science / Department of Clinical Sciences / Daniel U. Thomson / Bovine respiratory disease (BRDC) is a multifaceted disease that causes considerable economic loss in both the beef and dairy industry. However, because there may be lingering effects of BRDC in the dairy industry, the full economic costs of the disease may not be realized. These lingering effects may include an increased departure from the herd, decreased milk production, and decreased reproductive performance. All of these outcomes are affected by multiple variables, but proper statistical modeling can control for many of the different influences. Few studies have attempted to look at the long term effects of calfhood BRDC but consistent associations have not been identified. The objective of this observational, retrospective study was to determine the association between calfhood BRDC prior to 120 d of age and departure from the herd, milk production, and reproductive performance. The association between the occurrence of BRDC with departure from the herd prior to first calving depended on the birth year, but for 5 of the 6 years, animals that contracted BRDC were 1.62 to 4.98 times more likely to leave the herd than animals that did not contract BRDC. In addition, animals that contracted BRDC were also 1.28 times more likely to leave the herd between first and second calving than animals that did not contract BRDC. Furthermore, the age at BRDC occurrence was associated with departure from the herd. Animals that contracted BRDC pre-weaning were 2.62 times more likely to leave the herd prior to first calving than animals that contracted BRDC post-weaning. However, age of BRDC occurrence was not significantly associated with departure from the herd between first and second calving. The occurrence of BRDC during the first 120 d of life was associated with a 233 kg decrease in lactation-one production, but was not significantly associated with production in subsequent lactations. Finally, calfhood BRDC was not significantly associated with a decreased reproductive performance measured by calving interval. In conclusion, calfhood BRDC negatively impacts productivity both prior to and after first calving.
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The application of systems thinking in cattle productionShane, Douglas January 1900 (has links)
Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / Robert L. Larson / Bradley J. White / Applying systems methods to cattle production requires investigators to think about whole systems when addressing study objectives. The research conducted for this dissertation emphasized studying whole systems using different methods. We studied cattle production systems through mathematical simulation and new indirect monitoring technologies. While the methods used for the research in this dissertation may be very different, all utilized systems methods to address the study objectives.
Firstly, we applied systems thinking methods and developed a dynamic, deterministic systems simulation of cow-calf production over a 10-year horizon. This model was used to investigate the effects the duration of postpartum anestrus (dPPA) has on reproductive performance. A large range of dPPA have been reported, so various primiparous cow and multiparous cow dPPA were simulated. We found that increasing the dPPA for primiparous and multiparous cows had a negative impact on herd performance and that the dPPA is an important factor in determining cow-calf performance success. We then used the cow-calf simulation to explore the effects of breeding nulliparous cows prior to the rest of the herd, known as providing Heifer Lead Time (tHL). We found that increasing tHL improved herd performance, especially with longer dPPA for primiparous cows.
Secondly, real-time location systems (RTLS) were used to indirectly monitor cattle behavior. These systems have been used to determine the amount of time cattle spend at eating and drinking locations. We modeled the probability of cattle participating in eating and drinking behavior when determined to be at these locations by RTLS and found that significant differences exist between individual calves and period of the day.
Finally, we explored associations between bovine respiratory disease (BRD) and animal-to-animal contacts as determined by RTLS in beef cattle. We found that the probability of BRD diagnosis was associated with the amount of time 4 days’ ago that a calf was in calf-contact with calves assumed to be shedding BRD pathogens.
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