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J. Anim. Sci. 2003. 81:3088-3094
© 2003 American Society of Animal Science

The role of acidogenic diets and ß-hydroxybutyate on lymphocyte proliferation and serum antibody response against bovine respiratory viruses in Holstein steers1,2,3

D. C. Donovan*, A. R. Hippen*,4, D. J. Hurley{dagger} and C. C. L. Chase*

* South Dakota State University, Brookings 57007 and and {dagger} University of Georgia, College of Veterinary Medicine, Athens 30605


   Abstract
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 Abstract
Introduction
 Experimental Procedures
 Results
 Discussion
 Implications
 Literature Cited
 
Acidogenic diets were evaluated for their effects on lymphocyte proliferation in response to Staphylococcus aureus exotoxin B (SEB), and specific lymphocyte proliferation and serum-neutralizing antibody titers to four bovine respiratory viruses in vitro. Four Holstein steer calves, with an average weight of 213 ± 42 kg, were fed a basal (control) diet consisting of 49% forage and 51% concentrate (DM basis), with 15% CP (on a DM basis). Three additional treatment diets were used: 1) the basal diet supplemented with 700 mL/d of butylene glycol (BG) to induce ketoacidosis by increasing blood ß-hydroxybutyate (BHBA); 2) the basal diet supplemented with 1.2 ± 0.1 kg/d of anionic salts (AS; Soychor 16.7, West Central Soy, Ralston, IA) to induce a metabolic acidosis; and 3) the basal diet with all forage replaced by finely ground corn and soybean meal blended to provide 15% CP (HG), to induce lactic acidosis. The calves were fed each diet for 21 d in a 4 x 4 Latin square design. Blood samples were collected on d 18, 19, and 20 of each 21-d period and analyzed for pH; concentrations of BHBA; in vitro lymphocyte proliferation to SEB, bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), parainfluenza-3 (PI-3), and bovine herpesvirus-1 (BHV-1); and titers of serum-neutralizing antibodies against the four viruses. Following treatment, the average pH of the serum samples was 7.38 for calves fed the control diet, 7.37 for the BG treatment, and 7.36 for the HG treatment, and was decreased (P < 0.05) to 7.33 for the AS treatment. All acidogenic diets decreased lymphocyte response to SEB (P < 0.05). The lymphocyte proliferative response, however, of each virus showed a different pattern of interaction with the three acidogenic diets tested. The AS diet was associated with increased lymphocyte proliferative response to BVDV and BRSV (P < 0.01) and increased serum neutralization titers to BHV-1 (P < 0.05). In calves fed the BHBA-inducing diet (BG), an increase in lymphocyte proliferation to BRSV was observed (P < 0.05). A similar relationship to blood BHBA concentration was not observed with the lymphocyte proliferation to BVDV, PI-3, or BHV-1. Titers of serum-neutralizing antibody against PI3 (P < 0.05) and BHV-1 (P < 0.01) were negatively correlated with blood pH, and titers of serum neutralizing antibodies to BHV-1 were negatively correlated to elevated circulating concentrations of BHBA (P < 0.05).

Key Words: Acidosis • Bovine • ß-Hydroxybutyrate • Ketosis • Lymphocyte • Virus


   Introduction
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 Abstract
 Introduction
Experimental Procedures
 Results
 Discussion
 Implications
 Literature Cited
 
The interaction of physiological measurements influenced by nutrition and the immune response are not clearly defined. Reviews have outlined interactions between nutrition and viral disease (Beck, 1996Go) and the effects of extracellular pH on viral infection (Lardner, 2001Go). Lofgreen (1983)Go noted an increased rate and severity of bovine respiratory disease complex when the amount of concentrate in the diet fed to calves was increased from 55% to more than 72%.

Likewise, ketosis is a risk factor for multiple disorders during lactation that make the periparturient dairy cow more susceptible to disease (Erb and Grohn, 1988Go). Increased blood concentration of ketone bodies, especially ß-hydroxybutyrate (BHBA), may contribute to increased susceptibility to disease. Increasing concentrations of ketone bodies have been observed to depress the response of bovine lymphocytes to plant mitogens (Targowski and Klucinski, 1983Go; Klucinski et al., 1988; and Franklin et al., 1991Go). Nonnecke et al. (1992)Go found inhibition of pokeweed mitogen-induced immunoglobin M secretion in vitro following the addition of BHBA at concentrations found in the plasma of severely ketotic cows.

For these experiments, a decrease in blood pH resulting from ketoacidosis was hypothesized to result in decreased nonspecific and viral antigen-specific lymphocyte proliferation and in decreased titers of serum-neutralizing antibodies. The objective of this experiment was to evaluate the role of BHBA compared with other acidogenic agents in viral antigen-specific and super-antigen-driven lymphocyte proliferation and on the titers of circulating neutralizing antibodies against bovine viruses commonly affecting dairy and feedlot cattle.


   Experimental Procedures
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 Abstract
 Introduction
 Experimental Procedures
Results
 Discussion
 Implications
 Literature Cited
 
Experimental Design
Four Holstein steer calves with an average age of 120 ± 10 d and an average weight of 213 ± 42 kg were used in a 4 x 4 Latin square design with four 21-d periods. Thirty-nine days before the start of the experiment, the calves were vaccinated intramuscularly with an inactivated viral vaccine against bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), parainfluenza-3 (PI-3), and bovine herpesvirus-1 (BHV-1; Triangle 9, Fort Dodge Labs, Fort Dodge, IA; Triangle 9+ PH-K [killed virus]: bovine herpesvirus 1 [bovine rhinotracheitis virus], bovine viral diarrhea virus, parainfluenza-3, respiratory syncytial virus vaccine, Leptospira canicola, L. grippotyphosa, L. hardjo, L. icterohaemorrhagiae, L. Pomona, and Mannheimia (Pasturella) haemolytica). Twenty-one days before the experiment, the calves received an intramuscular booster dose of the same inactivated vaccine to maximize the numbers of circulating memory cells and optimize the titers of circulating antibodies.

The calves were individually fed a total mixed diet once daily at 0600. The fresh mixed diet was weighed and fed daily. Before each feeding, the refusals were weighed and removed and the amount of feed offered daily was adjusted to allow for 5% refusals.

Four dietary treatments (Table 1Go) were randomly assigned to the calves. The experimental dietary treatments were: 1) a basal diet (control, CON) containing 49% forage and 51% concentrate; 2) the basal diet supplemented with 700 mL of butylene glycol (BG; Van Walters and Rogers, Sioux City, IA) to induce ketoacidosis (Mills et al., 1984Go); 3) the basal diet supplemented with anionic salts (AS; Soychlor 16.7; West Central Soy, Ralston, IA) at 1.2 ± 0.1 kg, depending on DMI; and 4) a high-grain diet (HG) to simulate feedlot acidosis by replacing the forage portion of the basal diet with corn and soybean meal. The AS diet provided a dietary cation–anion balance of -5.0 ± 4.5 mEq/100 g to create a metabolic acidosis (West, 1992Go; Goff and Horst, 1998Go). Nutrient content of the basal diet was as follows: NEm = 1.45 Mcal/kg, NEg = 0.902 Mcal/kg, 15% CP, 19% ADF, 29% NDF, 47.51% NFC, and 3.4% ether extract (DM basis, calculated based on wet chemistry by Dairyland Labs, Arcadia, WI).


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  		Table 1. Composition of basal and treatment diets
 
All calves were fed the control diet for the first 7 d of each 21-d period. Treatments were introduced to the diets of the calves assigned to acidogenic treatments on d 8 and increased in a step-wise fashion until d 15. From d 15 to 21, calves were fed the maximum concentrations of each treatment. After completion of each period, all calves were placed on the basal diet for 7 d to normalize blood pH. Calves that received the HG diet in the preceding period were drenched with 227 g of a yeast, magnesium, calcium, and phosphorus mixture (Techmix, Inc., Stewart, MN) on d 21, and their diets were supplemented with 227 g/d of sodium bicarbonate for 7 d to restore ruminal pH.

Samples
On d 18 and 20 of each period, four blood samples were collected as follows: 10 mL of blood into Vacutainer tubes (Becton Dickinson, Franklin Lakes, NJ) containing 143 units of sodium heparin for whole-blood pH and blood gas analysis; 10 mL into Vacutainer tubes containing 0.1 mL of 15% K3-EDTA for BHBA analysis; 10 mL into Vacutainer tubes containing no additives for analysis of serum neutralizing antibodies; and 20 mL into heparinized syringes containing 400 units of porcine heparin diluted in 0.4 mL of calcium- and magnesium-free PBS for lymphocyte proliferation assays.

Hematological Measures
The heparinized blood was collected by 0700 h and immediately transported on ice to the Brookings Hospital Respiratory Care Unit (Brookings, SD) for analysis. The temperature of the samples was adjusted to 38.6°C, and samples were analyzed for pH, partial pressures of CO2 and O2, HCO3-, and base excess by blood gas analyzer within 1 h of sampling (Instrumentation Laboratory, IL1620, Lexington, MA). If samples are stored at 4°C, blood pH will decrease at less than 0.01 pH unit/h, and pCO2 increases by only approximately 0.5 mm Hg/h at 2 to 4°C (Scott et al., 1999Go).

The EDTA-treated blood was centrifuged at 1,171 x g for 20 min at 4°C for collection of plasma. A 1-mL aliquot was retained and stored at -20°C for analysis of BHBA (Williamson and Mellanby, 1974Go) by the Clinical Chemistry Laboratory of the South Dakota State University Animal Disease Research and Diagnostic Laboratory.

Lymphocyte Proliferation
Proliferation assays were performed by methods described previously (Rutten et al., 1990Go). Peripheral blood mononuclear cells (PBMC) were isolated by single-step density gradient centrifugation on Histopaque 1083 (Sigma Chemical Co., St. Louis, MO). The heparizined blood collected in syringes was diluted 1:4 with calcium- and magnesium-free PBS, and 40 mL of the diluted blood was layered over 10 mL of Histopaque 1083. The tubes were centrifuged at 800 x g for 45 min at 10°C. Four gradient layers were present in the 50-mL conical tube (from bottom to top, respectively): red blood cells, histopaque, mononuclear cells, and diluted plasma. The mononuclear cell layer was collected and transferred to a 15-mL conical tube. Cells were rinsed with 10 mL of PBS by centrifugation at 400 x g for 15 min. The PBMC pellet was suspended in 10 mL of PBS, and the cells were counted using a hemocytometer in the presence of trypan blue to determine cell viability (Sigma Chemical Co.). The PBMC cell density was adjusted to 3 x 106 viable cells/mL in Roswell Park Memorial Institute-1640 media (Sigma Chemical Co.) containing 10% horse serum (vol/vol; Hyclone Laboratories, Logan, UT), 2 mM pyruvate (Sigma Chemical Co.), 50 µg/mL of gentamicin sulfate (Sigma Chemical Co.), and an additional 2 mM L-glutamine (Sigma Chemical Co.). Horse serum, rather than fetal bovine serum, was used because bovine lymphocyte proliferation is consistently greater in horse serum than in fetal bovine serum, and horse serum also avoids interference with antiviral bovine antibodies found in fetal bovine serum.

Super-antigen-induced proliferation was assessed by addition of Staphylococcus aureus exotoxin B (SEB) to mononuclear cultures. The SEB is a broad-spectrum mitogen used to monitor the functional capacity of the mononuclear cells and to normalize small day-to-day differences in culture conditions. The SEB was used at 0.1 µg/mL, a concentration found in preliminary studies to be optimal for the culture conditions used in the experiment.

The NY-1 isolate of BVDV type 1 (American Type Culture Collection [ATCC], Rockville, MD), PI-3 (National Veterinary Services Laboratory [NVSL], Ames, IA), and the Cooper (Colorado-1) isolate of BHV-1 (ATCC) were propagated and titered in Madin-Darby bovine kidney cells (ATCC). The BRSV (NVSL) was propagated and titered in bovine turbinate cells isolated from fetal calves. The cells were obtained from the South Dakota State University Animal Disease Research and Diagnostics Laboratory. The BVDV, BRSV, PI-3, and BHV-1 were stored at -80°C in minimum Eagle’s essential medium (MEM; Sigma Chemical Co.) until added to culture.

The BHV-1 lymphocyte stimulation was done with UV-inactivated BHV-1 based on studies in our laboratory and those by Griebel et al. (1990)Go that indicated that UV-inactivated BHV-1 stimulation yielded a more consistent and reproducible lymphocyte response than did live virus. A General Electric ultraviolet light bulb (GEG64T6-germicidal) was used to inactivate BHV-1. One milliliter of virus was placed into a 10 x 100 mm sterile dish (Becton Dickinson), 8 cm from a UV bulb. The virus was exposed to UV light for 20 min to crosslink viral DNA. Inactivation of BHV-1 was verified by placing serially diluted samples into tissue culture. The virus was determined to be properly inactivated when no cytopathic effect was observed in BT cells after 120 h of incubation. Inactivated BHV-1 was added to the cultures based on an original multiplicity of infection of 2.0 tissue culture infective dose at a 50% infective rate (TCID50) per cell. For the other three viruses (BVDV, BRSV, PI-3), lymphocyte stimulation was done with live virus using a multiplicity of infection of 0.1 viral particles per cell. For each virus, 100 µL of virus preparation (BHV-1, BVDV, BRSV, or PI-3) was added to 100 µL of PBMC (3 x 105 cells), and cultures were prepared in quadruplicate. Samples were incubated for 120 h in a humidified atmosphere of 5% CO2 at 37°C to allow for multiple rounds of cell replication. Six hours before termination of the assay, cells were pulsed with 10 µL of 3H thymidine (20 µCi/mL, 6.7 Ci/mM). Cells were harvested onto a glass fiber filter with a semiautomated cell harvester (model PHD, Cambridge Technology, Watertown, MA). The incorporation of the radioactivity into the DNA of actively dividing cells was determined using a liquid scintillation spectrophotometer (LS1801, Beckman Instruments, Inc., Fullerton, CA), and was expressed as the specific incorporation (i.e., counts/min in antigen-stimulated cells minus counts/min in medium control cells).

Virus Neutralization Assay
Blood collected in tubes containing no additive was allowed to clot and then centrifuged at 1,171 x g for 20 min at 4°C. The serum was then removed from the tube and stored at -20°C in 4-mL 12 x 75 mm polypropylene tubes for analysis of neutralizing antibodies. All of the serum was stored at 20°C until it was tested.

The serum was thawed and heat inactivated at 56°C for 30 min to remove complement before conducting the virus neutralization assays (OIE, 1992Go). A microtiter plate dilution series was set up by placing 50 µL of MEM containing 0.4% penicillin and streptomycin solution (125 µg/mL of penicillin and 250 µg/mL of streptomycin; Sigma Chemical Co.) and 0.4% fungizone (1 µg/mL, Gibco BRL, Grand Island, NY) into each well of a 96-well flat bottom plate (Cellstar, Greiner Labortechnik, Charleston, SC). The test serum was diluted 1:4 with MEM; 50 µL of the diluted test serum from each calf was placed into the first row of wells in duplicate and a twofold serial dilution series was made across the plate.

Each of the stock viruses (BHV-1, BVDV type 1, Singers [NVSL], BRSV, or PI-3) was diluted in MEM to create a working stock of 4,000 to 6,000 TCID50 per milliliter (200 to 300 TCID50/50 µL). Fifty microliters of virus working stock was dispensed into each well of the plate containing diluted serum or medium as a control. To prepare back titrations, 200 µL of working stock of each virus was diluted into 1.8 mL of the prepared MEM and tested at 10-1, 10-2, and 10-3 dilutions. Back titrations were conducted with each assay and used to assess cytopathic effect of the viruses. The plates were incubated in a 5% CO2 incubator for 1 h at 37°C.

The bovine turbinate cells (passages 18 and 19, ATCC, Rockville, MD) were grown to approximately 90 to 95% confluence in 75-cm2 flasks (Corning, Corning, NY) containing the described MEM solution with 10% irradiated fetal bovine serum (Hyclone Laboratories). During the incubation of the plates containing serum dilutions and virus, the bovine turbinate cells were trypsinized with 1x trypsin (Sigma Chemical Co.) and suspended in 80 mL of MEM per flask. Fifty microliters of cells (1 x 104) was added to each well and the plates were returned to the 5% CO2 incubator. The cytopathic effect endpoint for BHV-1-, BRSV-, or PI-3-infected cells was determined at 120 h. The endpoint for BVDV-infected cells was determined at 72 h (OIE, 1992Go).

Data Analysis
Repeated measures were reduced to a single mean for each collection period (i.e., d 18 to 20 of each feeding period). The lymphocyte proliferation data was evaluated for internal clustering to determine outliners using the Q-test, and values were rejected when the Q value was greater than 0.76 (Skoog and West, 1969Go). All data were analyzed by the MIXED model procedures of SAS (SAS Inst. Inc., Cary, NC), with diet and feeding period as main effects. The residual sums of squares were used as the error term. Preplanned comparisons were made for the effects of: 1) control vs. acidogenic diets (CON vs. AS + BG + HG); 2) ketogenic diets vs. other acidogenic agents (BG vs. AS + HG); and 3) inorganic vs. organic acids from diets (AS vs. HG + BG). Regression procedures of SAS were used to determine: 1) lymphocyte proliferation of each virus vs. blood pH and BHBA, and 2) serum neutralization vs. blood pH and BHBA. Significance was declared at P < 0.05 and trends at P < 0.10.


   Results
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 Abstract
 Introduction
 Experimental Procedures
 Results
Discussion
 Implications
 Literature Cited
 
Dry Matter Intake and Body Weight
The average DMI for the 3-d sampling period (d 18, 19, and 20) was greatest for calves fed the control diet and least for those fed HG (8.19, 7.23, 7.11, and 6.78 kg/d for CON, AS, BG, and HG, respectively; P < 0.05).

Hematological Measures
The blood pH tended to be decreased (P < 0.10) under all acidosis-inducing treatments compared with the control diet (Figure 1Go). Blood pH was decreased (P < 0.05) in calves fed AS to a greater degree than for the other treatment diets. The induction of metabolic acidosis by AS was associated with a decrease (P < 0.01) in base excess. Blood base excess was also tended to be lower (P < 0.01) for calves fed all acidogenic diets compared with calves fed the control diet (5.88, 1.06, 5.55, and 4.50 for CON, AS, BG, and HG, respectively). Blood HCO3 concentrations also tended to be increased (P < 0.10) for calves fed the control diet compared with acidogenic diets averaging 30.6, 27.0, 30.2, and 30.0 mg/dL for CON, AS, BG, and HG, respectively, and was decreased (P < 0.05) in calves fed AS vs. BG + HG.



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  		Figure 1. Blood pH via jugular venipuncture in calves fed a control diet (CON) or diets containing anionic salts (AS), butylene glycol (BG), or high grain (HG). Effects in model include: CON vs. AS + BG + HG, P < 0.10; and AS vs. HG + BG, P < 0.05.

 
The concentration of BHBA in plasma was increased (P < 0.01) in calves fed BG compared with each of the other dietary treatments (Figure 2Go). As anticipated, the BG diet induced a mild ketonemic condition.



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  		Figure 2. Plasma ß-hydroxybutyrate (BHBA) via jugular venipuncture in calves fed a control diet (CON), or diets containing anionic salt (AS), butylene glycol (BG), or high grain (HG). Effects in model include: BG vs. AS + HG, P < 0.01.

 
Lymphocyte Proliferation
Super-antigen-induced proliferation was assessed using SEB (Figure 3aGo). The ketosis inducing diet, and each of the other acidosis inducing diets resulted in decreased (P < 0.05) lymphocyte response to SEB stimulation in vitro.



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  		Figure 3. Proliferation response of lymphocytes of: a) Staphylococcus aureus exotoxin B (SEB); b) bovine viral diarrhea virus (BVDV); c) bovine respiratory syncytial virus (BRSV); d) parainfluenza-3 (PI-3); and e) bovine herpes virus-1 (BHV-1) in calves fed a control diet (CON) or diets containing anionic salts (AS), butylene glycol (BG), or high grain (HG). Data are expressed as counts per minute (cpm). Effects in model include: a) SEB, CON vs. AS + BG + HG, P < 0.05; b) BVDV, CON vs. AS + BG + HG, P < 0.01; c) BRSV, BG vs. AS + HG, P < 0.10; d) PI-3, AS vs. HG + BG, P < 0.10; e) BHV-1, CON vs. AS + BG + HG, P < 0.05.

 
A positive association (P < 0.01) existed between blood pH and lymphocyte proliferation after in vitro stimulation with BVDV (Table 2Go). Proliferative response to BVDV was less (P < 0.01) for all acidogenic diets than for lymphocytes from calves fed the control diet (Figure 3bGo).


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  		Table 2. Correlations between lymphocyte proliferation and blood metabolites
 
The proliferative response of lymphocytes to BRSV (Figure 3cGo) isolated from serum of calves fed BG tended to be greater (P < 0.10) than for CON, AS, and HG. In contrast to the expected results, increases in serum BHBA were associated (P < 0.05) with enhanced proliferation of cells stimulated with BRSV in vitro (Table 2Go).

Lymphocyte proliferation in response to PI-3 tended to be increased (P < 0.10) in serum from calves fed AS compared with BG and HG (Figure 3dGo). Accordingly, a negative association (P < 0.01) existed between blood pH and lymphocyte proliferation after in vitro stimulation with PI-3 (Table 2Go).

The lymphocyte proliferative responses to BHV-1 were decreased (P < 0.05) after feeding all acidogenic diets (Figure 3eGo), and this decrease was positively associated (P < 0.01) with pH (Table 2Go).

Serum-Neutralizing Antibodies
Serum-neutralizing antibody titers to BVDV and BRSV were not affected in calves fed acidogenic diets and were not correlated with blood pH or BHBA concentration (data not shown). Serum neutralizing titers to PI-3 were not affected by dietary treatments; however, the serum-neutralizing antibody to PI-3 was negatively correlated (r = -0.37, P < 0.05) with blood pH. Antibody titers to BHV-1 tended to be greater (P < 0.10) when calves were fed the AS diet compared with the control diet (3.8 vs. 2.2 log2). Also, comparison of the AS diets vs. the two diets containing organic acids (HG and BG) demonstrated that the calves fed organic acids had greater (P < 0.05) titers to BHV-1 (3.8, 2.4 log2). Negative correlations were also observed for the serum-neutralizing antibody to BHV-1 respective to serum pH (r = -0.45, P < 0.01) and BHBA concentrations (r = -0.37, P < 0.05).


   Discussion
Top
 Abstract
 Introduction
 Experimental Procedures
 Results
 Discussion
Implications
 Literature Cited
 
Correlations between the physiological state, serum concentrations of ketone bodies, and blood pH with lymphoproliferative response against bovine viruses and the titers of serum-neutralizing antibodies against bovine virus showed different patterns for each of the viruses tested in our experiment. Ketone bodies have been associated with immunosuppressive activity (Franklin et al., 1991Go). The function of polymorphonuclear leukocytes was generally inhibited with observations of reduced chemotaxis, respiratory activity, and bactericidal capacity at decreased pH (Lardner, 2001Go).

An analysis of correlation between BHBA on lymphocyte proliferation and serum-neutralizing antibody titers failed to uncover an effect on immune response to virus. The association between blood pH and circulating antibody titer in this study was more dependent on blood pH than on the circulating concentration of BHBA. Nonnecke et al. (1992)Go demonstrated that a mixture of ketone bodies, approximating plasma levels of severely ketotic cows, inhibited mitogen-induced immunoglobin M secretion in vitro. In contrast, in this study, lymphocyte proliferation to BRSV by lymphocytes isolated from calves with elevated BHBA was greater than for lymphocytes isolated from calves fed the other acidosis inducing diets.

Lymphocyte proliferation in response to the addition of BVDV or PI-3 was increased when low blood pH was measured. In contrast, proliferation in response to the addition of BHV-1 to the cultures was decreased when low blood pH was measured.

An increase in the number and frequency (or fraction) of antigen-specific cells present is an essential step in adaptive immunity. This allows effective response against pathogens by the body (Janeway et al., 1999Go).

In conclusion, the proliferative responses of each virus tested showed a different pattern of interaction with the three acidogenic diets tested. In particular, ketoacidosis- and mineral acidosis-inducing diets seem to have different effects on lymphoproliferation in response to the four viruses tested. Titers of serum-neutralizing antibody against BVDV and PI-3 were negatively correlated with blood pH and titers to only BHV-1 were negatively correlated with elevated circulating concentrations of BHBA.


   Implications
Top
 Abstract
 Introduction
 Experimental Procedures
 Results
 Discussion
 Implications
Literature Cited
 
A model for evaluation of immune response in cattle under the production stressors of ketosis and acidosis has been described. Lymphocytes from calves that were ketotic or acidotic exhibited a generally decreased responsiveness to super-antigen stimulus. Responsiveness to viral stimulation, however, was not predictable. These results imply that the responsiveness of cattle, when exposed to specific viruses, may depend on nutritional and metabolic status, and that consideration of these factors may improve the efficacy of vaccination programs.


   Footnotes
 
1 This research was partly funded by the South Dakota State University Research Support Fund. Back

2 Published with the approval of the director of the South Dakota Agric. Exp. Stn. as publication No. 3304 of the Journal Series. Back

3 The authors express their gratitude to L. Braun for his assistance with laboratory methodology. Back

4 Correspondence: Department of Dairy Science, DM 109, Box 2104 (phone: 605-688-5490; fax: 605-688-6276; E-mail: arnoldhippen{at}sdstate.edu).

Received for publication July 2, 2002. Accepted for publication August 27, 2003.


   Literature Cited
Top
 Abstract
 Introduction
 Experimental Procedures
 Results
 Discussion
 Implications
 Literature Cited
 


Beck, M. A. 1996. The role of nutrition in viral disease. Nutr. Biochem. 7:683–690.

Erb, H. N., and Y. T. Grohn. 1988. Epidemiology of metabolic disorders in periparturient dairy cows. J. Dairy Sci. 71:2557–2571.

Franklin, S. T., J. W. Young, and B. J. Nonnecke. 1991. Effects of ketones, acetate, butyrate, and glucose on bovine lymphocyte proliferation. J. Dairy Sci. 74:2507–2514.[Abstract]

Goff, J. P., and R. L. Horst. 1998. Use of hydrochloric acid as a source of anions for prevention of milk fever. J. Dairy Sci. 81:2874–2880.[Abstract]

Griebel, P. J., H. B. Ohmann, M. J. P. Lawman, and L. A. Babiuk. 1990. The interaction between bovine herpesvirus-1 and activated bovine T lymphocytes. J. Gen. Virol. 71:369–377.[Abstract/Free Full Text]

Janeway, C. A., P. Travers, and M. Walport. 1999. Immunologic memory. Page 402 in Immunobiology: The Immune System in Health and Disease. 4th ed. Garland Publishing, New York, NY.

Lardner, A. 2001. The effects of extracellular pH on immune function. J. Leukocyte Biol. 69:522–530.[Abstract/Free Full Text]

Lofgreen, G. P. 1983. Nutrition and management of stressed beef calves. Vet. Clin. N. Am. Large Anim. Pract. 5:87–101.

Mills, S. E., R. R. Lyle, D. C. Beitz, and J. W. Young. 1984. In vitro hepatic gluconeogenesis during experimental ketosis produced in calves by 1,3-butanediol and phlorizin.J. Dairy Sci. 67:2265–2275.

Nonnecke, B. J., S. T. Franklin, and J. W. Young. 1992. Effects of ketones, acetate, and glucose on in vitro immunoglobulin secretion by bovine lymphocytes. J. Dairy Sci. 75:982–990.[Abstract]

OIE. 1992. Manual of Standards for Diagnostic Tests and Vaccines. Office International des Epizootics, Paris, France.

Oster, T., W. Davidson, and S. Ehl. 2002. Virus clearance and immunopathology by CD8+ T cells during infection with respiratory syncytial virus are mediated by IFN-{gamma}. Eur. J. Immunol. 32:2117–2123.[Medline]

Rutten, V. P. M., G. H. Wentick, W. A. C. deJong, A. C. A. van Exsel, and E. J. Hensen. 1990. Determination of BHV-1 specific immune reactivity in naturally infected and vaccinated animals by lymphocyte proliferation assays. Vet. Immunol. Immunopathol. 15:259–267.

Scott, M. G., J. W. Heusel, V. A. Lehyrs, and Ole Siggaard-Anderson. 1999. Electrolytes and Blood Gases. Page 1082 in Textbook of Clinical Chemistry. 3rd ed. C. A. Birtes and E. Ashwood, ed. W. B. Saunders, Philadelphia, PA.

Skoog, D. A., and D. M. West. 1969. Rejection of data. Page 45 in Fundamentals of Analytical Chemistry. 2nd ed. Holt, Rinehart, and Winston, Inc., New York, NY.

Targowski, S. P., and W. Klucinski. 1983. Reduction in mitogenic response of bovine lymphocytes by ketone bodies. Am. J. Vet. Res. 44:828–830.[Medline]

West, J. W. 1992. Cation-anion balance: Its role in lactating cow nutrition. Page 15 in Feedstuffs. Miller Publishing Co., Carol Stream, IL.

Williamson, D. H., and J. Mellanby. 1974. D-(-)-3-Hydroxybutyrate. Page 1836 in Methods of Enzymatic Analysis. Vol. 4. 2nd ed. H. U. Bergmeyer, ed. Acad. Press, London, England.



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