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Abstract

OBJECTIVE

To determine anti-bovine respiratory syncytial virus (BRSV) antibody titers for nasal secretions and serum from beef calves following administration of a modified-live (MLV) BRSV vaccine.

ANIMALS

60 healthy newborn purebred beef calves.

PROCEDURES

Calves were randomly assigned to 1 of 3 groups: intranasal (IN)-SC (IN MLV BRSV vaccine within 24 hours of birth and SC MLV BRSV vaccine at 2 months of age), SC-IN (SC MLV BRSV vaccine within 24 hours of birth and IN MLV BRSV vaccine at 2 months of age), or NO-IN (no vaccine within 24 hours of birth and IN MLV BRSV vaccine at 2 months of age). Nasal secretion and serum samples were collected for determination of anti-BRSV antibodies within 24 hours of birth and 2 and 6 months of age.

RESULTS

Titers of anti-BRSV IgA antibodies in nasal secretions and BRSV neutralizing antibodies in serum were similar among groups at each sampling time. Within 24 hours of birth, nasal anti-BRSV IgA titers were negligible. At 2 months, mean nasal anti-BRSV IgA titers for calves in IN-SC, SC-IN, and NO-IN groups were 192.84, 224.49, and 114.71, respectively.

CONCLUSIONS AND CLINICAL RELEVANCE

Concentrations of anti-BRSV IgA antibodies in the nasal secretions and BRSV neutralizing antibodies in the serum of young beef calves following an MLV BRSV vaccine protocol that consisted of IN or SC vaccine within 24 hours of birth and vice versa at 2 months of age were not different from that following only an IN MLV BRSV vaccine at 2 months of age. However, the lack of any differences may have been attributed to other factors. (Am J Vet Res 2021;82:746–751)

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in American Journal of Veterinary Research

Abstract

OBJECTIVE

To investigate indicators of neutrophil activation in the blood of healthy and asthma-affected horses and assess associations between corticosteroid treatment and these variables.

ANIMALS

48 horses (14 with severe equine asthma [SEA], 21 with mild to moderate equine asthma [MEA], and 13 healthy controls).

PROCEDURES

In a 3-part retrospective study, hematology analyzer data for horses included in previous studies were reviewed. Neutrophil size, neutrophil light absorbance (NLA), and myeloperoxidase (MPO) index were recorded. Data for each variable were compared among groups for the entire study sample (part 1). Changes in each variable were assessed for one subset of horses (5 SEA-affected and 6 controls) after treatment for 2 weeks with dexamethasone (0.06 mg/kg, PO, q 24 h; part 2) and for another subset (8 SEA-affected horses) after the same treatment and after a 1-week post-treatment washout period (part 3).

RESULTS

All 3 variables were significantly greater for the SEA group, compared with the MEA and control groups in part 1. Following dexamethasone treatment, the control- and SEA-group NLA and MPO index significantly decreased and SEA-group neutrophil size significantly decreased in part 2; immediate posttreatment results for SEA-affected horses were similar in part 3, with significantly increased neutrophil size and nonsignificant increases in NLA and MPO index following washout.

CONCLUSIONS AND CLINICAL RELEVANCE

Results suggested horses with exacerbated SEA have larger neutrophils that contain more MPO, compared with neutrophils of MEA-affected and healthy control horses. The clinical value of these variables for the diagnosis of equine asthma was deemed limited owing to data overlap among groups. (Am J Vet Res 2021;82:737–745)

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in American Journal of Veterinary Research

Abstract

OBJECTIVE

To determine the in vitro effects of epinephrine, norepinephrine, and dobutamine on lipopolysaccharide (LPS)-stimulated production of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10) in blood from healthy dogs.

SAMPLES

Blood samples from 9 healthy dogs.

PROCEDURES

Blood samples were incubated with LPS from Escherichia coli O127:B8 or PBSS (control) for 1 hour. Afterward, the samples were incubated with 10μM epinephrine, norepinephrine, or dobutamine or with saline (0.9% NaCl) solution (control) for 23 hours. Leukocyte viability was assessed by use of trypan-blue exclusion in blood from 2 dogs to ensure cell viability was not altered by the catecholamines. Tumor necrosis factor-α, IL-6, and IL-10 concentrations were measured in the supernatant in duplicate with a canine-specific multiplex bead-based assay. Blood samples from 2 dogs were used to create dose-response curves to evaluate whether the observed cytokine modulation was dependent on catecholamine concentration.

RESULTS

Incubation of blood with epinephrine and norepinephrine significantly increased LPS-stimulated production of IL-10, compared with the control. Epinephrine and norepinephrine significantly decreased LPS-stimulated production of TNF-α, compared with the control. Epinephrine and norepinephrine did not significantly alter LPS-stimulated production of IL-6. Dobutamine did not alter catecholamine production.

CONCLUSIONS AND CLINICAL RELEVANCE

Epinephrine and norepinephrine, but not dobutamine, had immunomodulatory effects on LPS-stimulated TNF-α and IL-10 production in blood from healthy dogs in this in vitro model of sepsis. Data suggested that dobutamine may have immune system-sparing effects in dogs with sepsis.

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in American Journal of Veterinary Research

Abstract

OBJECTIVE

To use a biopolymer delivery system to investigate the ability of interleukin (IL)-4 to recruit neutrophils into subcutaneous tissues of equids.

ANIMALS

16 horses and 2 ponies.

PROCEDURES

Animals were assigned to 3 experiments (6/experiment). Effects of recombinant equine (Req) IL-4 (100, 250, or 500 ng/site) versus a positive control (ReqIL-8; 100 ng, 250 ng, or 1 μg/site) and a negative control (Dulbecco PBSS or culture medium) on neutrophil chemotaxis were assessed after SC injection into the neck with an injectable biopolymer used as the vehicle. Tissue samples including the biopolymer plug were collected by biopsy at various time points from 3 hours to 7 days after injection. Neutrophil infiltration was evaluated by histologic scoring (experiments 1, 2, and 3) or flow cytometry (experiment 3).

RESULTS

Histologic neutrophil infiltration scores did not differ significantly among treatments at most evaluated time points. On flow cytometric analysis, log-transformed neutrophil counts in biopsy specimens were significantly greater for the ReqIL-8 treatment (1 μg/site) than the negative control treatment at 3 but not 6 hours after injection; results did not differ between ReqIL-4 and control treatments at either time point. Negative control treatments induced an inflammatory response in most equids in all experiments.

CONCLUSIONS AND CLINICAL RELEVANCE

Flow cytometry was a more reliable method to estimate neutrophil migration than histologic score analysis. The ReqIL-4 treatment did not induce a detectable neutrophil response, compared with the negative control treatment in this study. Evidence of inflammation in negative control samples suggested the biopolymer is not a suitable vehicle for use in equids.

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in American Journal of Veterinary Research

Abstract

OBJECTIVE

To quantify acute immunologic and metabolic responses of beef heifers following topical administration of transdermal flunixin meglumine (TDFM) at various times relative to bovine herpesvirus 1 (BHV1) and Mannheimia haemolytica challenges.

ANIMALS

32 beef heifers (mean body weight, 170 kg).

PROCEDURES

Heifers were assigned to 1 of 4 groups. Heifers in the control group did not receive TDFM, whereas 1 dose of TDFM (3.3 mg/kg) was topically applied to heifers of groups A, V, and B at −144, −72, and 0 hours. All heifers were inoculated with 1 × 108 plaque-forming units of BHV1 in each nostril at −72 hours and with 1.18 × 106 CFUs of M haemolytica intratracheally at 0 hours. Vaginal temperature was recorded and blood samples were collected for quantification of select immunologic and metabolic biomarkers at predetermined times from −144 to 360 hours.

RESULTS

Mean vaginal temperature was similar between group A and the control group. Mean vaginal temperatures for groups V and B were generally lower than that for the control group following BHV1 and M haemolytica challenges, respectively. Mean neutrophil oxidative burst capacity and L-selectin expression at 0 hours were significantly decreased for group V relative to the other groups. Other biomarkers did not differ among the groups at any time.

CONCLUSIONS AND CLINICAL RELEVANCE

Results suggested that topical administration of TDFM to beef cattle effectively alleviated pyrexia without adverse effects on acute immunologic or metabolic responses when TDFM was administered at the same time as, but not before, respiratory pathogen challenge.

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in American Journal of Veterinary Research

Abstract

OBJECTIVE

To evaluate effects of poly(ADP-ribose) polymerase-1 (PARP1) inhibitors on the production of tumor necrosis factor-α (TNF-α) by interferon-γ (IFN-γ)– and lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs) of horses as an in vitro model of inflammation in horses.

SAMPLE

1,440 samples of PBMCs from 6 healthy research horses.

PROCEDURES

From heparinized whole blood samples, PBMC cultures were obtained. An initial dose-response trial on 48 PBMC samples from 2 horses (24 samples each) was used to determine concentrations of IFN-γ and LPS for use as low- and high-level stimulation concentrations. Seventy-two PBMC samples from 6 horses were assigned equally to 1 of 4 PARP1 inhibition categories: no PARP1 inhibitor (PARP1 inhibition control); 2-((R)-2-methylpyrrolidin-2-yl)-1H-benzimidazole-4-carbozamide dihydrochloride (ABT888);4-(3-(1-(cyclopropanecarbonyl)piperazine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one (AZD2281); or N-(6-oxo-5,6-dihydrophenanthridin-2-yl) -N,N-dimethylacetamide hydrochloride (PJ34). Samples of PBMCs from each horse and each PARP1 inhibition category were then assigned to 1 of 3 levels of IFN-γ and LPS stimulation: none (control), low stimulation, or high stimulation. After a 24-hour incubation period, a TNF-α ELISA was used to measure TNF-α concentration in the supernatant. Results were compared across treatments and for each horse. Data were analyzed with repeated-measures ANOVA.

RESULTS

Median TNF-α concentration was significantly lower for PJ34-treated, high-level stimulated PBMCs than for PARP1 inhibition control, high-level stimulated PBMCs; however, no other meaningful differences in TNF-α concentration were detected among the inhibition and stimulation combinations.

CONCLUSIONS AND CLINICAL RELEVANCE

Findings suggested that PJ34 PARP1 inhibition may reduce TNF-α production in horses, a potential benefit in reducing inflammation and endotoxin-induced damage in horses.

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in American Journal of Veterinary Research

Abstract

OBJECTIVE To characterize activation and expression of immune genes of chicken macrophages after in vitro stimulation with lipopolysaccharide (LPS) and mouse erythrocytes.

ANIMALS Five 15-day-old chickens and 2 BALB/c mice.

PROCEDURES Macrophages were extracted from chicken bone marrow or peripheral blood and then stimulated with cytokines secreted from cell lines L929 and HD11. Stimulated chicken macrophages were further cocultured with LPS or mouse erythrocytes, and gene transcription of some distinctive cytokines was detected by use of a real-time PCR assay.

RESULTS Morphological features and phagocytic function of macrophages were characterized. Activated macrophages had an elongated shape with a large cell nucleus, and they had phagocytic function. Distinctive genes encoding the surface marker gene CD11b were identified; high quantities of CD11b were transcribed. Relative transcription of chicken genes BF and BL in mature cells cocultured with both stimuli was lower than for control cells. However, the quantity of genes encoding M1- or M2-distinctive cytokines (interleukin [IL]-1β, IL-10, IL-12, inducible nitric oxide synthase, tumor necrosis factor-α, and transforming growth factor-β) that were transcribed differed significantly between stimulation with LPS and mouse erythrocytes.

CONCLUSIONS AND CLINICAL RELEVANCE Chicken macrophages were differentially stimulated by LPS and mouse erythrocytes, which suggested that in vitro stimulation can distinctly influence the transcription and expression of immune genes of chicken macrophages.

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in American Journal of Veterinary Research

Abstract

OBJECTIVE To evaluate cell-mediated and humoral immune responses of calves receiving 2 doses of a dual-adjuvanted vaccine containing inactivated bovine herpesvirus type 1 (BHV1) and bovine viral diarrhea virus types 1 (BVDV1) and 2 (BVDV2) before and after exposure to BHV1.

ANIMALS 24 Holstein steers negative for anti-BHV1 antibodies and proliferative cell-mediated immune responses against BHV1 and BVDV.

PROCEDURES Calves were randomly assigned to 3 groups. The vaccinated group (n = 10) received 2 doses of vaccine on days 0 and 21. Control (n = 10) and seeder (4) groups remained unvaccinated. Calves were commingled during the study except for the 3-day period (days 53 to 55) when seeders were inoculated with BHV1 (1.04 × 107 TCID50, IV) to serve as a source of virus for challenge (days 56 through 84). Rectal temperature and clinical illness scores were monitored, and blood and nasal specimens were obtained for determination of clinicopathologic and immunologic variables.

RESULTS After BHV1 challenge, mean rectal temperature and clinical illness scores were lower for vaccinates than controls. In vaccinates, antibody titers against BHV1 and BVDV2, but not BVDV1, increased after challenge as did extracellular and intracellular interferon-γ expression, indicating a T helper 1 memory response. Additional results of cell marker expression were variable, with no significant increase or decrease associated with treatment.

CONCLUSIONS AND CLINICAL RELEVANCE Calves administered 2 doses of a killed-virus vaccine developed cell-mediated and humoral immune responses to BHV1 and BVDV, which were protective against disease when those calves were subsequently exposed to BHV1.

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in American Journal of Veterinary Research

Abstract

OBJECTIVE To evaluate the effect of an immunotherapeutic product on concentrations of anti–Pythium insidiosum antibodies in dogs.

ANIMALS 7 healthy hound-crossbreds.

PROCEDURES Antibody concentrations were evaluated before (day 0) and after administration of the immunotherapeutic product. The immunotherapeutic product was administered on days 0, 7, and 21. Serum was obtained on days 0, 7, 14, 21, 28, 35, 42, 49, and 56. Anti–P insidiosum antibody concentrations were measured and reported as the percentage positivity relative to results for a strongly positive control serum.

RESULTS Mean ± SD percentage positivity before administration of the immunotherapeutic product was 7.45 ± 3.02%. There was no significant change in anti–P insidiosum antibody concentrations after administration of the product, with percentage positivity values in all dogs remaining within the range expected for healthy dogs (3% to 15%).

CONCLUSIONS AND CLINICAL RELEVANCE Administration of the immunotherapeutic product to healthy dogs in accordance with the manufacturer's suggested protocol did not induce a significant change in anti–P insidiosum antibody concentrations. These results suggested that administration of the immunotherapeutic product may not interfere with postadministration serologic monitoring. However, further investigations will be required to determine whether there is a similar effect in naturally infected dogs.

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in American Journal of Veterinary Research

Abstract

OBJECTIVE To determine the in vitro effect of calcitriol on indicators of immune system function in endotoxin-primed blood samples from healthy dogs.

SAMPLE Blood samples from 6 healthy adult dogs.

PROCEDURES Leukocytes were primed by incubation of blood samples with lipopolysaccharide (LPS; endotoxin) or PBS solution (unprimed control group) for 1 hour. Following priming, blood samples were incubated with calcitriol (2 × 10−7M) or ethanol (control substance) for 24 hours. After sample incubation, LPS-stimulated leukocyte production of tumor necrosis factor (TNF) and interleukin-10 (IL10) was measured with a canine-specific multiplex assay, and apoptosis and toll-like receptor 4 (TLR4) expression were evaluated via flow cytometry.

RESULTS LPS stimulation of unprimed leukocytes but not endotoxin-primed leukocytes resulted in a significant increase in TNF and IL10 production, confirming the presence of endotoxin tolerance in dogs in vitro. Endotoxin priming significantly increased neutrophil viability with no effect on lymphocyte viability or TLR4 expression by neutrophils and monocytes. Calcitriol exposure significantly decreased LPS-stimulated production of TNF by unprimed and endotoxin-primed leukocytes. Conversely, calcitriol exposure had no effect on IL10 production by unprimed leukocytes but did significantly increase IL10 production by endotoxin-primed leukocytes. Calcitriol had no significant effect on the degree of neutrophil or lymphocyte apoptosis, nor was neutrophil and monocyte TLR4 expression affected in unprimed or endotoxin-primed leukocytes.

CONCLUSIONS AND CLINICAL RELEVANCE These data indicated that calcitriol induced an anti-inflammatory shift in unprimed and endotoxin-primed canine leukocytes in vitro, without compromising neutrophil and monocyte TLR4 expression or altering the viability of neutrophils and lymphocytes in canine blood samples.

Restricted access
in American Journal of Veterinary Research