Mannheimia Haemolytica Challenge Research Articles

Zinc Source and Concentration Altered Physiological Responses of Beef Heifers during a Combined Viral-Bacterial Respiratory Challenge.

Simple SummaryBovine respiratory disease is one of the greatest health challenges cattle producers face and is most commonly treated with antibiotics. With the current push to reduce the use of antibiotics in livestock production, producers are looking at non-pharmaceutical alternatives such as nutritional supplements. This study aimed to determine if different forms of zinc supplementation could reduce some of the negative health effects associated with bovine respiratory disease. Overall, cattle supplemented with ZinMet (zinc methionine/organic zinc) responded better during the disease as evidenced by blood parameters, decreased lesion severity, and decreased fever. Conversely, cattle fed a large dose of zinc sulfate (inorganic zinc) displayed a higher fever and blood parameters that indicated a greater sickness response. Findings from this study suggest that the type and amount of zinc fed to cattle may influence their response to bovine respiratory disease.To determine the effects of zinc supplementation on the immune response to a combined viral-bacterial respiratory disease challenge, thirty-two beef heifers (255 ± 15 kg) were subjected to a 30-d period of Zn depletion, then randomly assigned to one of three treatment diets fed for 30 d before the challenge: (1) supplementation with 100 mg of Zn from Zn sulfate/kg of DM (Zn100), (2) supplementation with 200 mg of Zn from Zn sulfate/kg of DM (Zn200), and (3) supplementation with 80 mg of Zn/kg of DM from zinc methionine and 20 mg of Zn from Zn sulfate/kg of DM (ZinMet). After the 30-d supplementation period, all heifers were fitted with indwelling vaginal temperature (VT) devices and intra-nasally challenged with 1 × 108 PFU bovine herpesvirus-1 on d -3, and then allowed to rest in outdoor pens for 3 d. On d 0, each heifer was challenged intra-tracheally with an average dose of 2.38 × 107 CFU Mannheimia haemolytica (MH), fitted with an indwelling jugular catheter, and then moved into individual stalls in an environmentally-controlled enclosed barn. Whole blood samples were collected at 1-h (serum) and 2-h (complete blood counts) intervals from 0 to 8 h, and at 12, 24, 36, 48, 60, 72, 168, and 360 h relative to MH challenge. Data were analyzed using the MIXED procedure of SAS specific for repeated measures with fixed effects of treatment, time, and their interaction. There was a treatment effect (p < 0.01) for VT such that Zn200 heifers had greater VT than Zn100 and ZinMet heifers. There was a trend (p = 0.10) for a serum cortisol treatment effect with Zn100 heifers having greater cortisol than ZinMet heifers. Total leukocytes and lymphocytes were greater (p ≤ 0.01) in Zn100 heifers than Zn200 and ZinMet heifers, whereas monocytes were less (p = 0.05) in ZinMet heifers than Zn100 and Zn200 heifers. Concentrations of IL-6 were greater (p = 0.02) in ZinMet heifers than Zn100 and Zn200 heifers. Concentrations of IFN-γ were greater in Zn200 heifers than ZinMet heifers at 0 h, and Zn100 heifers from 0 to 12 h post-MH challenge (treatment x time p = 0.02). Serum haptoglobin was not affected by treatment or treatment x time (p ≥ 0.36) but increased over time (p < 0.01) in all groups. There was a trend (p = 0.11) for ZinMet heifers to have less severe nasal lesion scores than Zn100 heifers. The observed differential physiological responses in this study indicate that zinc source and concentration may alter the response to a bovine respiratory challenge in heifers.

Differential haptoglobin responsiveness to a Mannheimia haemolytica challenge altered immunologic, physiologic, and behavior responses in beef steers.

Indicator traits associated with disease resiliency would be useful to improve the health and welfare of feedlot cattle. A post hoc analysis of data collected previously (Kayser et al., 2019a) was conducted to investigate differences in immunologic, physiologic, and behavioral responses of steers (N = 36, initial BW = 386 ± 24 kg) that had differential haptoglobin (HPT) responses to an experimentally induced challenge with Mannheimia haemolytica (MH). Rumen temperature, DMI, and feeding behavior data were collected continuously, and serial blood samples were collected following the MH challenge. Retrospectively, it was determined that 9 of the 18 MH-challenged steers mounted a minimal HPT response, despite having similar leukocyte and temperature responses to other MH-challenged steers with a greater HPT response. Our objective was to examine differences in behavioral and physiological responses between MH-challenged HPT responsive (RES; n = 9), MH-challenged HPT nonresponsive (NON; n = 9), and phosphate-buffered saline-inoculated controls (CON; n = 18). Additionally, 1H NMR analysis was conducted to determine whether the HPT-responsive phenotype affected serum metabolite profiles. The RES steers had lesser (P < 0.05) cortisol concentrations than NON and CON steers. The magnitude of the increases in neutrophil concentrations and rumen temperature, and the reduction in DMI following the MH challenge were greatest (P < 0.05) in RES steers. Univariate analysis of serum metabolites indicated differences between RES, NON, and CON steers following the MH challenge; however, multivariate analysis revealed no difference between HPT-responsive phenotypes. Prior to the MH challenge, RES steers had longer (P < 0.05) head down and bunk visit durations, slower eating rates (P < 0.01) and greater (P < 0.05) daily variances in bunk visit frequency and head down duration compared with NON steers, suggesting that feeding behavior patterns were associated with the HPT-responsive phenotype. During the 28-d postchallenge period, RES steers had decreased (P < 0.05) final BW, tended (P = 0.06) to have lesser DMI, and had greater (P < 0.05) daily variances in head down and bunk visit durations compared with NON steers, which may have been attributed to their greater acute-phase protein response to the MH challenge. These results indicate that the HPT-responsive phenotype affected feeding behavior patterns and may be associated with disease resiliency in beef cattle.

Efficacy of statistical process control procedures to identify deviations in continuously measured physiologic and behavioral variables in beef steers experimentally challenged with Mannheimia haemolytica.

The objective of this experiment was to determine if statistical process control (SPC) procedures coupled with the remote continuous collection of feeding behavior patterns, accelerometer-based behaviors, and rumen temperature can accurately differentiate between animals experimentally inoculated with Mannheimia haemolytica (MH) or PBS. Thirty-six crossbred steers (BW = 352 ± 23 kg) seronegative for MH were randomly assigned to bronchoselective endoscopic inoculation with MH (n = 18) or PBS (n = 18). Electronic feed bunks were used to measure DMI and feeding behavior traits, accelerometer-based neck collars measured feeding- and activity-behavior traits, and ruminal thermo-boluses measured rumen temperature. Data were collected for 28 d prior to and following inoculation. Steers inoculated with MH exhibited elevated (P < 0.02) levels of neutrophils and rumen temperature indicating that MH challenge effectively stimulated immunologic responses. However, only nine of the MH steers exhibited increased serum haptoglobin concentrations indicative of an acute-phase protein response and one displayed clinical signs of disease. Shewhart charts (SPC procedure) were used for two analyses, and sensitivity was computed using all MH-challenged steers (n = 18), and a subset that included only MH-challenged haptoglobin-responsive steers (n = 9). Specificity was calculated using all PBS steers in both analyses. In the haptoglobin-responsive only analysis, DMI and bunk visit (BV) duration had the greatest accuracy (89%), with accuracies for head-down (HD) duration, BV frequency, time to bunk, and eating rate being less (83%, 69%, 53%, and 61%, respectively). To address the diurnal nature of rumen temperature, data were averaged over 6-h intervals, and quarterly temperature models were evaluated separately. Accuracy for the fourth quarter rumen temperature was higher (78%) than the other quarterly temperature periods (first = 56%, second = 50%, and third = 67%). In general, the accelerometer-based behavior traits were highly specific ranging from 82% for ingestion to 100% for rest, rumination, and standing. However, the sensitivity of these traits was low (0% to 50%), such that the accuracies were moderate compared with feeding behavior and rumen temperature response variables. These results indicate that Shewhart procedures can effectively identify deviations in feeding behavior and rumen temperature patterns to enable subclinical detection of BRD in beef cattle.

Pre-weaning plane of nutrition and Mannheimia haemolytica dose influence inflammatory responses to a bovine herpesvirus-1 and Mannheimia haemolytica challenge in post-weaning Holstein calves

The objectives of this study were to determine whether plane of nutrition (PON) of milk replacer previously provided to calves, and dosage level of Mannheimia haemolytica (MH), influenced inflammatory responses to a combined viral-bacterial respiratory challenge. Holstein calves (1 d of age; n = 30) were assigned to treatments in a 2 × 3 factorial with pre-weaning PON and MH dose as main effects (n = 5 per treatment). Calves were fed either a low (LPN; n = 15) or a high PON (HPN; n = 15) from birth through weaning. Calves fed LPN were fed 436 g of dry matter (DM) per day of milk replacer until weaning, and HPN calves were fed 797 g of DM per day of milk replacer from d 1 to 10 and 1,080 g of DM per day from d 11 until weaning. Calf starter and water were offered ad libitum. Calves were step-down weaned beginning at d 54 and moved into an enclosed barn at d 70. Indwelling rectal temperature (RT) recording devices and jugular catheters were inserted at d 80. Calves were challenged with 1.5 × 108 plaque-forming units (pfu) per mL of bovine herpesvirus-1 (BHV-1) in each nostril at d 81 and with either 106, 107, or 108 cfu of MH at d 84. Blood samples were collected at varying intervals relative to BHV-1 and MH challenges. Four LPN calves either died or were euthanized soon after the 144-h observation period, whereas all HPN calves survived the entire observation period. As dosage of MH administered increased, acute and systemic inflammatory responses increased. Higher doses of MH resulted in increased leukocyte, neutrophil, and haptoglobin concentrations in infected calves. Data from the current study suggest that the highest dose, 108 cfu, triggered weaned calves' acute disease response, whereas the lower doses, 106 and 107 cfu, caused more moderate inflammation and disease. The effects of PON on inflammation responses to the disease challenge indicated that calves previously fed the LPN diet had more severe pathophysiological responses. Calves fed LPN showed higher peripheral neutrophil and leukocyte counts and serum haptoglobin concentrations following the BHV-1 challenge. Additionally, following the MH challenge, LPN calves had higher peripheral neutrophil counts, neutrophil-to-lymphocyte ratios, and serum tumor necrosis factor-α concentrations. These data demonstrate that higher doses of MH increase the acute inflammatory response and prolong inflammation, and that calves previously fed LPN responded more severely to the combined viral-bacterial respiratory challenge.

Pathogenesis of co-infections of influenza D virus and Mannheimia haemolytica in cattle

Bovine respiratory disease (BRD) is economically significant, and influenza D virus (IDV) is commonly identified in cattle with BRD. Mannheimia haemolytica (MHA) is an opportunistic bacterial contributor to BRD; surveillance data suggest that MHA and IDV co-infection occurs in cattle. The objective of this study was to evaluate the synergistic pathogenesis in cattle co-infected with IDV and MHA. Sixteen dairy calves were randomly assigned to four groups of four calves. The IDV + MHA + group received D/bovine/C00046 N/Mississippi/2014 (D/46 N) intranasally at 0 days post-inoculation (DPI) and Mannheimia haemolytica D153 (MHA D153) intratracheally at 5 DPI. The IDV + MHA- group received only D/46 N at 0 DPI; the IDV-MHA + group received only MHA D153 at 5 DPI; and the IDV-MHA- group received neither agent. Clinical scores were calculated twice daily. At 10 DPI, IDV + MHA+, IDV-MHA+, and IDV-MHA- calves were euthanized and evaluated for pathologic lesions. The IDV + groups seroconverted to IDV by 10 DPI. Clinical scores were higher in IDV + groups than IDV- groups on 2–5 DPI (p = 0.001). After MHA challenge on 5 DPI, clinical scores (6–10 DPI) were slightly lower in IDV+MHA+ group than IDV-MHA+ group (p < 0.05) but not significantly different between MHA+ groups and MHA- groups. The average gross pathology score was higher for IDV-MHA+ group than groups IDV-MHA- and IDV+MHA+; however, no significant differences were identified among groups. Under the conditions of this study, infection with IDV before MHA enhance neither clinical disease nor lung pathology, relative to calves infected with MHA alone.

Effects of Mannheimia haemolytica challenge with or without supplementation of Saccharomyces cerevisiae boulardii strain CNCM I-1079 on immune upregulation and behavior in beef steers.

Objectives of this experiment were to examine the effects of live yeast (LY) supplementation on immunological, physiological, and behavioral responses in steers experimentally challenged with Mannheimia haemolytica (MH). Thirty-six crossbred Angus steers (BW = 352 ± 23 kg) seronegative for MH were allocated within a 2 × 2 factorial arrangement: Factor 1 = roughage-based diet with LY (Saccharomyces cerevisiae boulardii CNCM I-1079, 25 g·per steer daily) or negative control (CON). Factor 2 = bronchoselective endoscopic inoculation with MH or phosphate buffer solution (PBS). Steers were fed their respective diets for 28 d prior to MH challenge on day 0. Reticulo-rumen temperature (RUT; ThermoBolus, Medria) was measured continuously at 5-min intervals and rectal temperature on days -4, 0, 1, 2, 3, 5, 7, 10, and 14 relative to MH inoculation. Compared with PBS-treated steers, the steers inoculated with MH had increased (P < 0.05) RUT from 2 to 24 h, reaching a zenith (>41 °C) 9 to 11 h post-MH challenge, whereas rectal temperature was increased (P < 0.04) in MH-inoculated steers on day 1 post-MH challenge. Supplementation with LY increased (P < 0.05) rectal temperature on days 0, 7, and 10, relative to CON steers. There were inoculation x day interactions (P < 0.01) for lymphocyte, neutrophil, leukocyte, and haptoglobin concentrations. Steers challenged with MH had increased (P < 0.05) neutrophil concentration from days 1 to 3, leukocyte concentration on days 1 and 2 and haptoglobin concentration on days 1 to 5 post-MH challenge compared with PBS-treated steers. Steers supplemented with LY exhibited increased (P < 0.02) cortisol throughout the study compared with the CON treatment. Following inoculation, MH-challenged steers exhibited reduced (P < 0.05) DMI, eating rate, frequency, and duration of bunk visit (BV) events compared with PBS-treated steers. Results from this study demonstrate that the experimental challenge model effectively stimulated acute-immune responses and behavioral changes that are synonymous with naturally occurring bovine respiratory disease (BRD). However, supplementation with LY minimally altered the impact of the MH challenge on physiological and behavioral responses in this study. Continuously measured RUT was more sensitive at detecting febrile responses to MH challenge than rectal temperature. These results serve to guide future research on behavioral and physiological alterations exhibited during a BRD infection.

Duration of immunity of a four-valent vaccine against bovine respiratory diseases

This study demonstrated the duration of immunity over 6 months of a vaccine against key bovine respiratory disease pathogens: Parainfluenza 3, Bovine Respiratory Syncytial Virus, Bovine Viral Diarrhoea and Mannheimia haemolytica. This was performed by challenge on colostrum-deprived calves at the age of 2 weeks. Recent European field isolates were used as challenge strains. Clinical signs and pathogen excretion or presence were monitored. Field relevance of the viral challenge strains was analysed using phylogenic analysis. Significant reduction of excretion of the 3 viruses in vaccinated animals was a consistent finding, demonstrating the efficacy of the vaccine. Reducing shedding is indeed key to interrupting the infection transmission chain and helping to achieve the protective effects of immunisation that extend beyond the individual. A significant reduction of clinical signs and lung lesions following the Mannheimia haemolytica challenge was also observed in vaccinated animals versus controls. Comparison of the challenge strains to an array of global and European strains, including recent ones, demonstrated a high genetic proximity, supporting the potential for the vaccine to maintain similar levels of efficacy in the field over a 6-month period post vaccination.

0101 Influence of dietary supplementation with a Saccharomyces cerevisiae fermentation product prototype on the pathophysiological response to a combined intranasal bovine herpesvirus-1 and intratracheal Mannheimia haemolytica challenge in Holstein steers

0101 Influence of dietary supplementation with a Saccharomyces cerevisiae fermentation product prototype on the pathophysiological response to a combined intranasal bovine herpesvirus-1 and intratracheal Mannheimia haemolytica challenge in Holstein steers

Efficacy and safety of a novel DNA immunostimulant in cattle

Four studies were performed to evaluate efficacy (Studies 1 and 2) and safety (Studies 3 and 4) of a novel DNA immunostimulant (Zelnate, Bayer Healthcare) in cattle. Zelnate (ZEL) was administered concurrently with (Study 1) or 24 hours after (Study 2) a Mannheimia haemolytica (Mh) challenge. Holstein steers (3 to 4 months of age; n=32/treatment, Study 1; n=40/treatment, Study 2) received either ZEL or a negative control (CON) on day 0 (Study 1) or day 1 (Study 2). Calves were challenged intratracheally with 60 mL of Mh (10 to the 7th power CFUs/mL, Study 1; 10 to the 8th power CFUs/mL, Study 2) on day 0 and clinically observed to day 5. Lung scores were obtained at the time of premature death or after necropsy on day 5. ZEL mitigated lung lesions in this Mh model, compared to the CON (Studies 1 and 2), and significantly reduced mortality compared to the CON (P &lt; 0.05; Study 2). ZEL was shown to be safe at the injection site (Study 3) and among large populations in different field scenarios (Study 4). ZEL demonstrated efficacy by mitigating lung lesions (Studies 1 and 2) and decreasing mortality (Study 2) in this Mh disease model. Studies 3 and 4 confirmed that ZEL is safe for use in cattle.

Bighorn sheep × domestic sheep hybrids survive Mannheimia haemolytica challenge in the absence of vaccination

Bighorn sheep (BHS, Ovis canadensis) are much more susceptible than domestic sheep (DS, Ovis aries) to pneumonia caused by leukotoxin (Lkt)-producing members of the Family Pasteurellaceae, particularly Mannheimia haemolytica and Bibersteinia trehalosi. Leukotoxin is widely accepted as the critical virulence factor of these bacteria since Lkt-negative mutants do not cause death of BHS. Typically, DS carry Lkt-positive M. haemolytica and/or B. trehalosi as commensal bacteria in their nasopharynx. In contrast, most BHS do not carry Lkt-positive M. haemolytica or B. trehalosi, or carry Lkt-negative strains in their nasopharynx. In previous studies, we demonstrated that unimmunized DS resist M. haemolytica challenge while BHS succumb to it. We hypothesized that Lkt-neutralizing antibodies, induced by Lkt-positive M. haemolytica and/or B. trehalosi innately carried by DS in their nasopharynx, render them less susceptible to infection by these bacteria. In this study we developed BHS×DS F1 hybrids by artificial insemination of domestic ewes with BHS semen. F1 hybrids were fertile, and produced F2 hybrids and back-crosses. The F1, F2, and back-crosses were raised together with domestic ewes. All these animals acquired Lkt-positive M. haemolytica and/or B. trehalosi, and developed high titers of Lkt-neutralizing antibodies in the absence of vaccination. Furthermore, all of these animals resisted challenge with lethal dose of M. haemolytica. These results suggest that lack of previous exposure to Lkt is at least partially responsible for fatal pneumonia in BHS when they acquire Lkt-positive M. haemolytica and/or B. trehalosi from DS when the two species commingle.

Pulmonary lesions and clinical disease response to Mannheimia haemolytica challenge 10 days following administration of tildipirosin or tulathromycin1,2

This clinical trial evaluated the impact of metaphylactic antimicrobial administration 10 d before experimental inoculation with Mannheimia haemolytica (MH) to mitigate pulmonary lesions. Thirty-three crossbreed heifers were procured as a single group and were randomly allocated to 1 of 3 blocks and to treatment, tildipirosin (ZUP; 4 mg/kg) or tulathromycin (DRX; 2.5 mg/kg) or saline (SAL; 1 mL/45.5 kg), within block on arrival at Kansas State University. All trial procedures were staggered by 7-d intervals for each block, resulting in all animals within a block receiving treatment, challenge, and necropsy on the same dates. Heifers within each block received an endoscopic MH challenge 10 d following treatment administration (d 0) and were housed in individual indoor stalls for 3 d postchallenge. Clinical illness scores (CIS), respiration quality scores, appetite scores, and injection site reactions were recorded on all animals from d 0 through d 13. Rectal temperatures were measured once daily on all animals from d 8 through d 13. Heifers were necropsied, and lung lesions were evaluated on d 13. Lung lesion data were evaluated using nonparametric methods (Kruskall-Wallis), and standard least squares models were used to evaluate the remaining variables. The pulmonary lesion scores (percentage of affected lung) ranged from 3.3% to 39.8% for all heifers with 92% (11/12) of ZUP-treated heifers having <10% lesions. Tildipirosin-treated heifers had lower (P < 0.05) lung lesion scores when compared with DRX- and SAL-treated heifers. Lung weight expressed as a percentage of BW was lower (P < 0.05) in ZUP heifers compared to DRX- and SAL-treated heifers. The probability of receiving abnormal CIS, appetite scores, and respiratory scores was lower (P < 0.05) in ZUP-treated heifers compared to DRX- and SAL-treated animals. This study showed that heifers treated with tildipirosin 10 d before MH challenge have less pulmonary damage and fewer clinical signs of illness compared to heifers treated with DRX or SAL.

LKTA and PlpE small fragments fusion protein protect against Mannheimia haemolytica challenge

Bovine respiratory disease (BRD) complex is a major cause of economic losses for the cattle backgrounding and feedlot industries. Mannheimia haemolytica is considered the most important pathogen associated with this disease. Vaccines against M. haemolytica have been prepared and used for many decades, but traditional bacterins have failed to demonstrate effective protection and their use has often exacerbated disease in vaccinated animals. Thus, the BRD complex continues to exert a strong adverse effect on the health and wellbeing of stocker and feeder cattle. Therefore, generation of recombinant proteins has been helpful in formulating enhanced vaccines against M. haemolytica, which could confer better protection against BRD. In the present study, we formulated a vaccine preparation enriched with recombinant small fragments of leukotoxin A (LKTA) and outer-membrane lipoprotein (PlpE) proteins, and demonstrated its ability to generate high antibody titers in rabbits and sheep, which protected against M. haemolytica bacterial challenge in mice.

Rumen temperature change monitored with remote rumen temperature boluses after challenges with bovine viral diarrhea virus and Mannheimia haemolytica12

Remote rumen temperature monitoring is a potential method for early disease detection in beef cattle. This experiment was conducted to determine if remotely monitored rumen temperature boluses could detect a temperature change in steers exposed to bovine viral diarrhea virus (BVDV) and challenged with a common bovine respiratory disease pathogen, Mannheimia haemolytica (MH). Twenty-four Angus crossbred steers (BW = 313 ± 31 kg) were allotted to 1 of 4 treatments: 1) no challenge (control); 2) challenge by a 72-h exposure to 2 steers persistently infected with BVDV; 3) bacterial challenge with MH; and 4) viral challenge by a 72-h exposure to 2 steers persistently infected with BVDV followed by bacterial challenge with MH (BVDV + MH). Remotely monitored rumen temperature boluses programmed to transmit temperature every minute were placed in the rumen before the time of exposure to steers persistently infected with BVDV. Rectal temperatures were taken before MH challenge (0) and at 2, 4, 6, 12, 18, 24, 36, 48, 72, and 96 h after MH challenge. Rumen temperatures were recorded 3 d before (-72 h; period of BVDV exposure) through 14 d after (336 h) MH challenge. Rumen temperatures were analyzed as a randomized complete block design with a 2 × 2 factorial arrangement of treatments and a first-order autoregressive covariance structure for repeated measures. A treatment × day interaction was observed for average daily rumen temperature (P < 0.01). A treatment difference (P < 0.01) was observed on d 0, when MH-challenged steers had greater rumen temperatures than steers not challenged with MH. There was no BVDV × day interaction (P > 0.01). Rumen temperatures averaged every 2 h resulted in a BVDV × hour interaction (P < 0.01) and an MH × hour interaction (P < 0.01). The BVDV × hour differences occurred at h -18 to -14, 40 to 46, 110, 122, and 144 to 146 (P < 0.01). The MH × hour difference occurred at h 4 to 24 (P < 0.01). Maximum rumen temperature was increased (P < 0.01) for BVDV (0.8 °C), MH (1.2 °C), and BVDV + MH (1.3 °C) compared with the control. On average, rumen temperatures measured by the boluses at the same time points as the rectal temperatures were 0.13 °C less than rectal temperatures, and the 2 body temperatures were highly correlated (r = 0.89). Rumen temperature boluses appear to have potential as a tool for detecting temperature changes associated with adverse health events such as exposure to bovine respiratory disease and BVDV.

Effects of exposure to calves persistently infected with bovine viral diarrhea virus type 1b and Mannheimia haemolytica challenge on animal performance, nitrogen balance, and visceral organ mass in beef steers1,2

Bovine viral diarrhea viruses (BVDV) have been isolated alone or in combination with other viral and bacterial pathogens in animals diagnosed with bovine respiratory disease (BRD), a disease causing major economic loss to the feedlot industry. The objective of this experiment was to determine the effects of Mannheimia haemolytica challenge after short-term exposure (72 h) to bovine viral diarrhea virus type 1b (BVDV1b) persistently infected (PI) calves on performance, N balance, and organ mass in finishing cattle. Treatments (6 steers/treatment; initial BW = 314 +/- 31 kg) were 1) steers not exposed to steers PI with BVDV nor challenged with M. haemolytica (control; CON); 2) steers exposed to 2 steers PI with BVDV1b (BVD) for 72 h; 3) steers intratracheally challenged with M. haemolytica (MH); or 4) steers exposed to 2 steers PI with BVDV1b for 72 h and challenged with M. haemolytica (BVD+MH). There were 12 h between exposure to PI steers and challenge with M. haemolytica. Steers were housed in metabolism stanchions during the first 5 d after the M. haemolytica challenge and on d 7 to 11, 28 to 32, and for 5 d before slaughter (average 119 d on feed) to determine N balance and were weighed every 28 d. At slaughter, carcass and organ mass data were collected. Data were analyzed as a randomized complete block design with a 2 x 2 factorial arrangement of treatments, and steer was used as the experimental unit. From d -3 (beginning of PI steer exposure) to 4, steers challenged with M. haemolytica had less (P = 0.04) ADG than steers not challenged with M. haemolytica. In addition, steers exposed to steers PI with BVDV tended (P = 0.09) to have less ADG and G:F across the entire finishing period than steers not exposed to BVDV. Before slaughter, retained N expressed as grams per day (P = 0.03) and as a percentage of N intake (P = 0.04) was less in BVD steers compared with steers not exposed to BVDV. There were no effects (P > 0.10) of BVDV exposure or M. haemolytica challenge on empty BW (EBW) or carcass characteristics. Expressed as a percentage of EBW, HCW was less (P = 0.02) and total offal weight was greater (P = 0.02) for steers challenged with M. haemolytica compared with steers not challenged. Results are in agreement with those reported in larger scale finishing studies and suggest that acute exposure to BRD-related pathogens can have long-term effects on animal performance.

Effects of exposure to calves persistently infected with bovine viral diarrhea virus type 1b and subsequent infection with Mannheima haemolytica on clinical signs and immune variables: Model for bovine respiratory disease via viral and bacterial interaction1,2

The objective was to determine effects of an intratracheal Mannheimia haemolytica challenge after 72-h exposure to bovine viral diarrhea virus type 1b (BVDV1b) persistently infected (PI) calves on serum antibody production, white blood cell count (WBC), cytokine concentrations, and blood gases in feedlot steers. Twenty-four steers (initial BW = 314 +/- 31 kg) were randomly allocated to 1 of 4 treatments (6 steers/treatment) arranged as a 2 x 2 factorial. Treatments were 1) steers not exposed to steers PI with BVDV nor challenged with M. haemolytica (control; CON); 2) steers exposed to 2 steers PI with BVDV for 72 h (BVD); 3) steers intratracheally challenged with M. haemolytica (MH); and 4) steers exposed to 2 steers PI with BVDV for 72 h and challenged with M. haemolytica (BVD+MH). There were 12 h between exposure to PI steers and challenge with M. haemolytica. Rectal temperature was increased (P < 0.001) for MH and BVD+MH during the initial 24 h after the M. haemolytica challenge. For MH and BVD+MH, total WBC count was increased (P < 0.01) at 36 h post M. haemolytica challenge compared with CON, whereas in BVD steers, WBC count was decreased (P < 0.01). Total lymphocyte count was increased (P = 0.004) during the initial 72 h post BVDV exposure for the BVD and BVD+MH groups compared with MH and CON, and this difference remained at 96 h post M. haemolytica challenge. An increased (P < 0.001) total neutrophil count was observed during the initial 36 h for the MH group and at 72 h for the BVD+MH challenge group. Interleukin 1beta, IL-6, and tumor necrosis factor alpha (TNFalpha) concentrations were greater (P <or= 0.04) in BVD steers than steers not exposed to BVDV, and interferon gamma, IL-1beta, and TNFalpha were greater (P <or= 0.02) in MH steers than in steers not challenged with M. haemolytica. There were BVD x MH interactions (P <or= 0.04) for plasma carbon dioxide, carbonic acid, and sodium bicarbonate concentrations. Challenge with M. haemolytica decreased carbon dioxide, carbonic acid, and sodium bicarbonate concentrations to a greater extent when steers were not exposed to PI steers compared with steers exposed to BVDV. We conclude that the challenge model was successful at inducing bovine respiratory disease (BRD) associated with BVDV and M. haemolytica. Understanding the physiological changes in morbid animals will lead to improved strategies for decreasing severity and economic losses associated with BRD.

Effects of melengestrol acetate on the inflammatory response in heifers challenged with Mannheimia haemolytica1

Previous research from our laboratory has indicated that melengestrol acetate (MGA) added to the diet during the first 35 d after arrival in the feedlot improves growth rates and tends to reduce chronic respiratory disease in heifers naturally challenged with bovine respiratory disease. The current study was conducted to provide further insight into the possible immunomodulatory effects of MGA. Crossbred heifers (n = 48; 232 +/- 5.5 kg of BW) were used in a randomized complete block design to determine the effects of MGA on lung pathology and markers of inflammation after Mannheimia haemolytica challenge. On d 0, cattle were blocked by BW and randomly assigned, within block, to diets (54% concentrate) that provided 0 or 0.5 mg of MGA per heifer daily for the duration of the experiment. Inoculum containing from 1.3 x 10(9) to 1.7 x 10(9) cfu of M. haemolytica (20 mL) was instilled at the bifurcation of the trachea on d 14. Blood samples were collected, clinical observations were made, and rectal temperatures were recorded for each animal at 0, 12, 24, 48, 72, 96, 120, and 138 h after inoculation. Heifers fed MGA had greater circulating concentrations of eosinophils and postchallenge concentrations of segmented neutrophils and white blood cells (P < 0.01) than controls, as well as elevated plasma protein, serum haptoglobin, and fibrinogen after M. haemolytica challenge (P < 0.01). Heifers fed MGA had lower plasma glucose (P < 0.01), greater plasma urea N (P = 0.02), and elevated respiratory indices (P < 0.01) compared with controls. Necropsies performed on d 6 after inoculation suggested that M. haemolytica challenge was relatively mild, because lesions were confined to a small portion of the lungs. On a 0 to 100 scale, average lung lesion scores were 3 and 1 for MGA-fed and control groups, respectively (P < 0.06). Heifers fed MGA before mild M. haemolytica challenge were more susceptible to infection, as evidenced by a greater number of heifers fed MGA exhibiting pulmonary lesions 138 h after inoculation than controls (14 out of 23 vs. 6 out of 24 for MGA and controls, respectively; P < 0.02).

Effects of melengesterol acetate on inflammatory response during Mannheimia haemolytica challenge (2005)

Previous trials conducted at Kansas State University demonstrated that melengesterol acetate (MGA) increased growth rates and tended to reduce chronic sickness in heifers naturally challenged with undifferentiated bovine respiratory disease. Our study was conducted to gain further insight into the mode of action of MGA. Crossbred heifers (n=47; 511 lb) were used to evaluate effects of MGA on lung pathology and markers of inflammation in cattle after an intrabronchial Mannheimia haemolytica challenge. On day 0, cattle were assigned to diets (54% concentrate) that provided 0 or 0.5 mg MGA per heifer daily. On day 14 each heifer was intrabronchially inoculated with M. haemolytica. Blood samples were collected from each heifer immediately before inoculation and 12, 24, 48, 72, 96, 120, and 138 hours after inoculation. Heifers were then euthanized for postmortem examination. After the challenge, heifers fed MGA had greater numbers of neutrophils and white blood cells, as well as greater serum haptoglobin and fibrinogen concentrations. The incidence of post-challenge lung lesions was greater in heifers fed MGA, and lung lesion scores tended to be more severe in heifers fed MGA, compared with those of controls. These data indicate that MGA does not reduce inflammation in heifers 138 hours after M. haemolytica challenge, suggesting that there are other modes of action for the beneficial effects on growth and reduction of chronicity in feedlot heifers.

Dose determination and confirmation of a long-acting formulation of ceftiofur (ceftiofur crystalline free acid) administered subcutaneously for the treatment of bovine respiratory disease.

The objective of this work was to determine and confirm an effective dose of ceftiofur crystalline free acid sterile oil suspension (CCFA-SS, 100 mg ceftiofur equivalents (CE)/mL], a long-acting single-administration ceftiofur formulation, for the treatment of the bacterial component of bovine respiratory disease (BRD). Study 1 was a dose determination study that used an intratracheal Mannheimia haemolytica (Pasteurella haemolytica) challenge model to evaluate single-administration doses of CCFA-SS at 0.0, 1.1, 2.2, 3.3, 4.4 or 5.5 mg CE/kg body weight (BW) for the treatment of BRD. Data from this study were used to select doses for field testing in three multi-location clinical studies. In Study 2, the efficacy of a single administration dose of CCFA-SS at 4.4 mg CE/kg BW was compared with a negative control for the treatment of naturally occurring BRD in feedlot cattle. Treatments were administered when uniform clinical signs of BRD were present. Study 3 used a design similar to Study 2, and compared single-administration doses of CCFA-SS at 3.0 or 4.4 mg CE/kg BW with the positive-control tilmicosin (Micotil(R) 300 Injection, Elanco Animal Health) at 10 mg/kg BW. Study 4 compared the efficacy of single doses of CCFA-SS of 1.1-8.8 mg CE/kg BW with tilmicosin at 10 mg/kg BW. A total of 1176 cattle were included in these clinical studies. In Study 1, a dose of 4.55 mg CE/kg BW was determined to be effective. This was rounded to 4.4 mg CE/kg for field testing. In Study 2, a single dose of CCFA-SS at 4.4 mg CE/kg BW had a higher treatment success rate on day 14 (61%) than negative controls (26%, P < 0.01). However, in Study 3 this dose was judged to be at the beginning of an efficacious dose range for the treatment of BRD when compared with tilmicosin. In Study 4, day 28 treatment success rates were higher for CCFA-SS at 4.4-8.8 CE/kg BW than for tilmicosin (P=0.002) or the noneffective CCFA-SS dose of 1.1 mg CE/kg BW (P < 0.001). Based on decision criteria for Study 4, the effective dose was determined to be 4.4-5.5 mg CE/kg BW. These clinical studies demonstrated that a single dose of CCFA-SS (100 mg CE/mL) administered subcutaneously (s.c.) in the neck at 4.4-5.5 mg CE/kg BW is an effective treatment for BRD in feedlot cattle. However, this route of administration is no longer being considered for this formulation because of the ceftiofur residues that are present at the injection site for extended periods of time.