Effects of pituitary pars intermedia dysfunction and Prascend (pergolide tablets) treatment on endocrine and immune function in horses.

It remains unclear how pituitary pars intermedia dysfunction (PPID) and pergolide treatment (Prascend [pergolide tablets]) affect endocrine and immune function in horses. To evaluate these effects, blood was collected regularly from 28 university-owned horses (10 Non-PPID, 9 PPID control [PC], and 9 PPID treatment [PT]) over approximately 15 mo. Pergolide treatment was initiated after Day 0 collections. Analyses included ACTH, insulin, total cortisol, free cortisol, complete blood counts, plasma myeloperoxidase, and cytokine/receptor gene expression in basal whole blood and in vitro stimulations (PMA/ionomycin, heat-inactivated Rhodococcus equi, and heat-inactivated Escherichia coli) of whole blood and peripheral blood mononuclear cells (PBMCs). The results were analyzed using a linear mixed model (SAS 9.4) with significance set at P < 0.05. Significant group (P = 0.0014) and group-by-time (P = 0.0004) effects were observed in resting ACTH such that PT horses differed from Non-PPID horses only at Day 0. PT horses had significantly lower changes in ACTH responses to thyrotropin-releasing hormone stimulation tests than PC horses at non-fall time points only, mid-late February 2018 (P = 0.016) and early April 2018 (P = 0.0172). When PT and PC horses did not differ, they were combined before comparison to Non-PPID horses. No significant group or group-by-time effects were seen in resting insulin, total cortisol, or free cortisol; however, significant time effects were observed in these measures. PPID horses had lower absolute lymphocyte (P = 0.028) and red blood cell (P = 0.0203) counts than Non-PPID horses. In unstimulated whole blood, PPID horses had increased IL-8 expression compared with Non-PPID horses (P = 0.0102). In addition, PPID horses had decreased interferon γ production from PBMCs after stimulation with R. equi (P = 0.0063) and E. coli (P = 0.0057) and showed increased transforming growth factor ß expression after E. coli stimulation (P = 0.0399). The main limitations of this study were a limited sample size and an inability to truly randomize the PPID horses into treatment groups. Resting ACTH is likely the best choice for determining successful responses to pergolide. Neither PPID nor pergolide appears to influence insulin, total cortisol, and free cortisol. As measured, systemic immune function was altered in PPID horses, and it is likely that these horses are indeed at increased risk of opportunistic infection. Despite reducing ACTH, pergolide treatment did not appear to influence immune function.

Effects of subcutaneous injections of a long acting moxidectin formulation in grazing beef cattle on parasite fecal egg reduction and animal weight gain.

Trials were conducted in Arkansas, Idaho, Illinois and Wisconsin using a common protocol to evaluate effectiveness and safety of a long acting (LA), oil-based injectable formulation of moxidectin in beef cattle grazing spring and/or summer pastures. At each site, 150 cattle (steers and/or heifers) were blocked based on pretreatment fecal strongyle egg counts (EPG) and then randomly assigned to treatments within blocks. Presence of naturally acquired parasitic infections, confirmed by presence of parasite eggs in feces, was a prerequisite for study enrollment. Within each block of three animals, two received moxidectin LA injectable on day 0 at a dosing rate of 1.0 mg moxidectin/kg b.w. into the dorsal aspect of the proximal third of the ear, and one received a placebo control treatment. Cattle were weighed before treatment and on day 55 or 56 (55/56) after treatment. Fecal samples were also collected from 10 randomly selected blocks of animals at each site on days 14, 28 and 55/56 for EPG quantification. Average daily gain (ADG) was computed over the posttreatment period. Data pertaining to ADG and EPG were combined across sites and analyzed by mixed model analysis of variance to assess the fixed effect of treatment and random effects of site, block within site and the treatment by site interaction. Compared to placebo-treated controls, the geometric means of fecal EPG counts from cattle treated with moxidectin LA injectable were reduced 99.8% 14 days after treatment, 99.1% 28 days after treatment and 96.7% 55/56 days after treatment. Rate of weight gain by cattle treated with moxidectin LA injectable was 0.59 kg/day, or 23% (0.11 kg/day) more than placebo-treated controls (P<0.05). None of the cattle treated with moxidectin LA injectable exhibited signs of macrocyclic lactone toxicosis. Summarized across all study sites, proportions of cattle that received concurrent therapeutic treatments were similar among treatment groups. Study results demonstrate that moxidectin cattle LA injectable administered at a dosing rate of 1.0 mg moxidectin/kg b.w. to grazing beef cattle was effective and safe.

Persistent activity of moxidectin long-acting injectable formulations against natural and experimentally enhanced populations of lice infesting cattle.

A study was conducted under a common protocol in Wisconsin and Wyoming, USA, to evaluate therapeutic and persistent efficacy of two long-acting injectable formulations of moxidectin against lice populations infesting cattle. At each site, 30 beef calves were blocked into groups of three based on naturally acquired Linognathus vituli populations, then randomly assigned to treatments within blocks. Treatments, injected subcutaneously into the proximal third of the ear on Day 0, included saline, a long-acting oil-based formulation containing 10% moxidectin given at the rate of 1 mg moxidectin/kg body weight (M10/1.0), or a long-acting oil-based formulation containing 15% moxidectin given at the rate of 0.75 mg moxidectin/kg b.w. (M15/0.75). Species of sucking and chewing lice were quantified on nine predilection sites before treatment, then 28, 63, 98, 133 and 168 days after treatment. During intervals between lice counts after Day 28, study animals from the three treatment groups were commingled for 32 days with two lice-free sentinels plus four to six seeder calves with infestations of both sucking and chewing lice. Following each 32-day commingling interval, seeder and sentinel animals were removed, and principal animals were sorted into pens by treatment. Lice were quantified on sentinel animals on the day of removal, and lice were quantified on principal study animals 3 days after removal of sentinel and seeders. Moxidectin was generally not efficacious against Bovicola bovis in the injectable formulations tested, whereas Haematopinus eurysternus infestations were inadequate to judge product effectiveness. Based on geometric means, both M15/0.75 and M10/1.0 provided statistically significant therapeutic efficacy against existing infestations of L. vituli and Solenopotes capillatus (100% efficacy on Day 28), and provided persistent protection against reinfestation with L. vituli and S. capillatus (efficacy >97%) for at least 133 days following treatment.

Detection of shared magnetic antigenic determinants on whole Moraxella bovis pili by use of antisera to cyanogen bromide-cleaved M. bovis pilus protein.

OBJECTIVE: To determine the ability of antisera against cyanogen bromide-cleaved pili from 4 strains of Moraxella bovis to react with whole or nondenatured pili. SAMPLE POPULATION: Antisera to 4 strains of M. bovis produced by New Zealand White rabbits. PROCEDURE: Pili from 4 strains of M. bovis were collected and purified. Pilus proteins (pilin) were cleaved, using cyanogen bromide. Whole pilus and cyanogen bromide-cleaved pilin were injected into rabbits. Antisera were serially diluted, reacted with 4 strains of M. bovis, and examined by immunoelectron microscopy and indirect immunofluorescence. RESULTS: Antisera to whole pili aggregated and distorted pili from homologous strains, but pili from heterologous strains were unaffected. Antisera to cleaved pilin fragments resulted in partial aggregation and thickening of homologous and heterologous pili, suggestive of heterospecific antibodies. Attachment of antibodies to pili was detected by indirect immunofluorescence, indicating a strong reaction of antisera to whole pili with homologous pili. Weak cross-reactions were evident with certain heterologous strains. In contrast, antisera to cleaved pilin fragments reacted strongly with pili from homologous and heterologous strains. CONCLUSIONS AND CLINICAL RELEVANCE: We detected shared antigenic determinants on pili from various strains of M. bovis that were not immunogenic in intact pili. These sites were immunogenic after cleavage of pilus protein with cyanogen bromide, and antisera produced to protein fragments reacted with whole pili from heterologous strains of the organism. Vaccines produced from cyanogen bromide-treated pili may induce broader immunity against infectious bovine keratoconjuctivitis than that provided by currently available vaccines.

Immunoblot analysis of cyanogen bromide-cleaved Moraxella bovis pilin reveals presence of shared antigenic determinants on pili from heterologous strains.

Moraxella bovis pilus proteins, collected and purified from four strains of M. bovis, were cleaved with cyanogen bromide. Two major fragments were produced. Antisera were produced in rabbits to the pilin protein fragments and to whole uncleaved pili from these strains. Immunoblots of whole and cyanogen bromide-cleaved pilin were reacted with the homologous and heterologous antisera to whole pili and cleaved pilin. Antisera to whole pili reacted strongly with homologous pilin. Weaker and inconsistent reactions were detected with heterologous pilin. Antisera produced to cyanogen bromide-cleaved pilin proteins reacted strongly with homologous and heterologous pilin fragments and uncleaved pilin proteins. These findings demonstrate the presence of conserved antigenic determinants on pili from heterologous strains that are non-immunogenic in the intact pilus but are immunogenic after treatment with cyanogen bromide. Cyanogen bromide-treated pilus preparation might have potential as a vaccine because antibodies are induced against heterologous strains of M. bovis, whether these cross-reactive antibodies are protective remains to be determined.

Validation of synthetic peptide enzyme immunoassays in differentiating two subgroups of ruminant respiratory syncytial virus.

Subgroup-specific peptide-based enzyme immunoassays from each respective G-glycoprotein of the ovine and the bovine respiratory syncytial virus (RSV) were developed to detect RSV-specific IgG responses in cattle. Antigenic peptides from the respective G-glycoprotein were identified from the extracellular central hydrophobic region (amino acids 158-189) located between 2 mucin-rich regions. These antigenic peptides identified by epitope mapping from each G-glycoprotein were synthesized and used to develop the subgroup-specific enzyme immunoassays. The negative cutoff for each enzyme immunoassay was established as the mean optical density of indirect immunofluorescent antibody-negative bovine sera plus 3 SDs. The sensitivity (82.9%) and specificity (100%) of the bovine enzyme immunoassay and the specificity (95.8%) of the ovine enzyme immunoassay were determined by comparison with indirect immunofluorescence (used as the "gold standard"). The negative and positive predictive values were calculated for each assay. The presence of serum antibody to ovine RSV in cattle implies that this virus infects cattle and may contribute to the pathogenesis of bovine respiratory disease.

Prevalence of ovine and bovine respiratory syncytial virus infections in cattle determined with a synthetic peptide-based immunoassay.

Subgroup-specific peptide-based enzyme-linked immunosorbent assays from the G-protein of the ovine and bovine respiratory syncytial virus (RSV), respectively, were used to determine the prevalence of the ovine and bovine subgroup strains of RSV infections in cattle. A total of 1,102 bovine serum samples were obtained from 6 diagnostic laboratories located in the northwestern and the southeastern USA and were tested for antibody to either the bovine or ovine subgroups of RSV. Antibody to viruses from each subgroup was present in samples from each region and all states tested. The Southeast had a higher prevalence of the bovine subgroup strains (69.5%). Then did the Northwest (40.9%). The prevalence of the ovine strain was similar for the two regions (16.7% in the southeast, 14.9% in the northwest). The overall prevalence was 56.6% for the bovine strain and 15.9% for the ovine strain. These results suggest members of the ovine subgroup of RSV circulate in the cattle population but with less frequency than those viruses of the bovine subgroup.

Bleeding diathesis associated with variant von Willebrand factor in a Simmental calf.

A 10-month-old Simmental heifer was examined because of a 10-day history of epistaxis and aural hematomas. Examination of the calf also revealed hemarthrosis. Initial laboratory data indicated that platelet count, platelet size, prothrombin time, and partial thromboplastin time were not different from a clinically normal (control) cow. Mucosal bleeding time was prolonged, and platelet adhesion to glass beads was less than expected. The clinical signs, prolonged bleeding time, and platelet adhesion defect were corrected by infusion of bovine plasma. Subsequent laboratory testing revealed that the affected calf had a truncated multimeric structure of von Willebrand factor (vWF), low vWF activity, and impaired platelet aggregation in response to adenosine diphosphate, but concentration of vWF was not different from that in clinically normal control animals. These data were consistent with a diagnosis of variant von Willebrand disease. The relationship of this disease to the thrombopathy of Simmental cattle is unclear.