OBJECTIVE To determine whether plasma ACTH concentrations vary following administration of a thyrotropin-releasing hormone (TRH) solution prepared for research purposes and stored at −20°C (rTRH) or prepared by a compounding pharmacy and stored at room temperature (approx 22°C; cTRH). ANIMALS 34 adult horses. PROCEDURES The study consisted of 2 experiments. In experiment 1, each horse underwent 2 TRH stimulation tests separated by 24 hours; 10 horses were administered cTRH for the first test and rTRH for the second test (group 1), 10 horses were administered rTRH for the first test and cTRH for the second test (group 2), and 10 horses were administered rTRH for both tests (group 3). Plasma ACTH concentrations were measured at 0 (baseline) and 30 minutes after TRH administration and the delta ACTH responses (change in ACTH concentration after TRH administration) were calculated. In experiment 2, the design was the same as that for experiment 1 except there were 14 days between tests, ACTH was measured at 0 and 10 minutes after TRH administration, and 11, 9, and 10 horses were assigned to groups 1, 2, and 3, respectively. RESULTS Adverse effects associated with TRH administration included transient coughing and yawning. In experiment 1, the median delta ACTH response for the second test was significantly lower than that for the first test for all groups. In experiment 2, the median delta ACTH response did not differ significantly between the first and second tests for any group, ACTH concentrations after rTRH administration were positively correlated (rs = 0.95) with those after cTRH administration, and the mean ± SD bias in post-TRH ACTH concentration between rTRH and cTRH was 2.9 ± 12.4 pg/mL. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that the TRH stimulation test should not be repeated within 24 hours, and cTRH solution stored at room temperature could be used to effectively perform TRH stimulation testing in horses.

The aim of this study was to evaluate the antimicrobial activity and determine the minimum bactericidal concentration (MBC) of the essential oils derived from Origanum vulgare (oregano), Melaleuca alternifolia (tea tree), Cinnamomum cassia (cassia), and Thymus vulgaris (white thyme) against Salmonella Typhimurium, Salmonella Enteritidis, Escherichia coli, Staphylococcus aureus and Enterococcus faecalis. The study also investigated the ability of these different bacterial strains to develop adaptation after repetitive exposure to sub-lethal concentrations of these essential oils. The MBC of the essential oils studied was determined by disc diffusion and broth dilution methods. All essential oils showed antimicrobial effect against all bacterial strains. In general, the development of adaptation varied according to the bacterial strain and the essential oil (tea tree > white thyme > oregano). Therefore, it is important to use essential oils at efficient bactericidal doses in animal feed, food, and sanitizers, since bacteria can rapidly develop adaptation when exposed to sub-lethal concentrations of these oils.

During the United States Department of Agriculture (USDA) National Animal Health Monitoring System’s (NAHMS) 2007‐2008 beef study, producers from 24 states were offered the opportunity to evaluate their animals for internal parasites and for overall responses to treatment with anthelmintics. A lapse of 45 d was required between initial sampling and any previous treatments. Choice of anthelmintic (oral benzimidazoles, and both injectable and pour-on endectocides) was at the discretion of the producer so as not to alter the local control programs. Fresh fecal samples were collected from 20 animals, or from the entire group if less than 20, then randomly assigned to 1 of 3 participating laboratories for examination. Analyses consisted of double centrifugation flotation followed by enumeration of strongyle, Nematodirus, and Trichuris eggs (the presence of coccidian oocysts and tapeworm eggs was also noted). Where strongyle eggs per gram (epg) exceeded 30, aliquots from 2 to 6 animals were pooled for egg isolation and polymerase chain reaction (PCR) analysis for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. Results from 72 producers (19 States) indicated that fecal egg count reductions were < 90% in 1/3 of the operations. All operations exhibiting less than a 90% reduction had used pour-on macrocyclic lactones as the anthelmintic treatment. While some of these less than expected reductions could have been the result of improper drug application, PCR analyses of the parasite populations surviving treatment, coupled with follow-up studies at a limited number of sites, indicated that less than expected reductions were most likely due to anthelmintic resistance in Cooperia spp. and possibly Haemonchus spp.

OBJECTIVE To investigate serum and plasma serotonin concentrations, percentage of serotonin-positive platelets, level of surface-bound platelet serotonin expression (mean fluorescence intensity [MFI]), and platelet activation (CD62 expression) in platelet-rich plasma from Cavalier King Charles Spaniels with myxomatous mitral valve disease (MMVD). ANIMALS Healthy dogs (n = 15) and dogs with mild MMVD (18), moderate-severe MMVD (19), or severe MMVD with congestive heart failure (CHF; 10). PROCEDURES Blood samples were collected from each dog. Serum and plasma serotonin concentrations were measured with an ELISA, and surface-bound platelet serotonin expression and platelet activation were determined by flow cytometry. RESULTS Dogs with mild MMVD had higher median serum (746 ng/mL) and plasma (33.3 ng/mL) serotonin concentrations, compared with MMVD-affected dogs with CHF (388 ng/mL and 9.9 ng/mL, respectively), but no other group differences were found. Among disease groups, no differences in surface-bound serotonin expression or platelet activation were found. Thrombocytopenic dogs had lower serum serotonin concentration (482 ng/mL) than nonthrombocytopenic dogs (731 ng/mL). In 26 dogs, a flow cytometry scatterplot subpopulation (FSSP) of platelets was identified; dogs with an FSSP had a higher percentage of serotonin-positive platelets (11.0%), higher level of surface-bound serotonin expression (MFI, 32,068), and higher platelet activation (MFI, 2,363) than did dogs without an FSSP (5.7%, 1,230, and 1,165, respectively). An FSSP was present in 93.8% of thrombocytopenic dogs and in 29.5% of nonthrombocytopenic dogs. CONCLUSIONS AND CLINICAL RELEVANCE A substantive influence of circulating serotonin on MMVD stages prior to CHF development in Cavalier King Charles Spaniels was not supported by the study findings. An FSSP of highly activated platelets with pronounced serotonin binding was strongly associated with thrombocytopenia but not MMVD.

OBJECTIVE To develop a model of hip joint synovitis on the basis of intra-articular injection of a sodium urate suspension in dogs and to characterize associated gait changes. ANIMALS 6 healthy adult dogs. PROCEDURES Each dog was sedated, and synovitis was induced by injection of 1 mL of a sodium urate suspension (20 mg/mL) into the right hip joint under ultrasonographic guidance. Observational and instrumented gait analyses to determine temporospatial, kinetic, and kinematic variables were performed prior to and 4, 8, and 24 hours after sedation and synovitis induction. RESULTS Injection of a sodium urate suspension into the hip joint of healthy dogs resulted in lameness of the ipsilateral pelvic limb as determined by observational and instrumented gait analyses. For all dogs, lameness was clinically detectable within 1.5 to 2 hours after injection, reached its maximum intensity at 4 hours after injection, and had subsided by 24 hours after injection. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that injection of a sodium urate suspension into the hip joint of healthy dogs reliably induced synovitis and signs of pain and lameness in the ipsilateral pelvic limb that lasted 24 hours. This model can be used in conjunction with instrumented gait analysis to provide information on gait changes associated with hip joint disease and might be useful for evaluating the efficacy of analgesics or other interventions for the treatment of hip joint disease in dogs.

OBJECTIVE To investigate the association of bovine respiratory disease (BRD) or vaccination with serologic response in calves. ANIMALS 94 Holstein calves. PROCEDURES To assess the association between BRD and antibody titers, 38 calves < 3 months old that were treated for BRD were matched with 38 untreated calves. To investigate the effect of vaccination on antibody titers, 24 calves were randomly assigned to be vaccinated against bovine respiratory syncytial virus (BRSV), bovine viral diarrhea virus types 1 and 2, bovine herpesvirus type 1 (BHV1), and parainfluenza virus type 3 at 2 weeks of age (n = 6), 5 weeks of age (6), and both 2 and 5 weeks of age (6) or were assigned to be unvaccinated controls (6). Blood samples were obtained at I, 2, 5, and 12 weeks for determination of serum neutralization antibody titers against the vaccine viruses, bovine coronavirus, and Mannheimia haemolytica. Antibody rates of decay were calculated. RESULTS Calves with initial antibody titers against BRSV < 1:64 that were treated for BRD had a slower rate of anti-BRSV antibody decay than did similar calves that were not treated for BRD. Calves with high initial antibody titers against BRSV and BHV1 had lower odds of BRD than did calves with low initial antibody titers against those 2 pathogens. Vaccination at 2 or 5 weeks of age had no effect on the rate of antibody decay. CONCLUSIONS AND CLINICAL RELEVANCE Clinical BRD and the serologic response of dairy calves were associated with initial antibody titers against BRSV and BHV1. Serologic or clinical responses to viral exposure may differ in calves with low passive immunity.

OBJECTIVE To compare antibody responses of horses naturally infected with West Nile virus (WNV) and those vaccinated against WNV, to identify whether vaccination interferes with the ability to diagnose WNV infection, and to determine the duration of antibody responses after vaccination. SAMPLE Sera from horses naturally infected with WNV (n = 10) and adult WNV-naïve horses before and after vaccination with a live canarypox virus–vectored vaccine (7) or a killed virus vaccine (8). PROCEDURES An established WNV IgM capture ELISA was used to measure IgM responses. Newly developed capture ELISAs were used to measure responses of 8 other WNV-specific immunoglobulin isotypes. A serum neutralization assay was used to determine anti-WNV antibody titers. RESULTS WNV-specific IgM responses were typically detected in the sera of WNV-infected horses but not in sera of horses vaccinated against WNV. Natural infection with and vaccination against WNV induced an immunoglobulin response that was primarily composed of IgG1. West Nile virus–specific IgG1 was detected in the sera of most horses 14 days after vaccination. Serum anti-WNV IgG1 and neutralizing antibody responses induced by the killed-virus vaccines were higher and lasted longer than did those induced by the live canarypox virus–vectored vaccine. CONCLUSIONS AND CLINICAL RELEVANCE On the basis of these findings, we recommend that horses be vaccinated against WNV annually near the beginning of mosquito season, that both IgM and IgG1 responses against WNV be measured to distinguish between natural infection and vaccination, and that a WNV IgG1 ELISA be used to monitor anti-WNV antibodies titers in vaccinated horses.

OBJECTIVE To determine whether thioredoxin (TRX)-1 can be used as a valid biomarker for oxidative stress in dogs. ANIMALS AND SAMPLES 10 Beagles and Madin-Darby canine kidney cells. PROCEDURES Madin-Darby canine kidney cells were used to verify antigen cross-reactivity between human and canine anti–TRX-antibodies. Dogs were assigned to receive 21% or 100% O2 (5 dogs/group) via an artificial respirator during a 3-hour period of isoflurane anesthesia (starting at 0 hours). Blood and urine samples were collected before (baseline) and at 6, 12, 24, and 48 hours after commencement of inhalation anesthesia. Concentrations of TRX-1 and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in plasma and urine samples were analyzed; urine concentrations were reported as ratios against urine creatinine concentration. RESULTS Canine TRX-1 was recognized by monoclonal human anti-TRX-1 antibodies (clones of adult T-cell leukemia-derived factor [ADF]-11 and ADF21) by western blot analysis. Results of an ELISA indicated that plasma TRX-1 concentration and urine TRX-1-to-creatinine concentration ratio increased rapidly after the 3-hour period of hyperoxia with maximal peaks at 12 and 6 hours, respectively. Urine 8-OHdG-to-creatinine concentration ratio also increased significantly after hyperoxia induction. However, unlike the rapid increase in urine TRX-1-to-creatinine concentration ratio, maximal urine 8-OHdG-to-creatinine concentration ratio was attained at 48 hours after hyperoxia induction. These variables remained unchanged from baseline in the control group. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that human anti-TRX monoclonal antibodies cross-reacted with canine TRX, and plasma TRX-1 concentrations were rapidly increased in dogs following an oxidative stress challenge. Thus, TRX may be a valuable clinical biomarker for detecting oxidative stress more rapidly than 8-OHdG in dogs.

The aim of the present study was morphological and histochemical analysis of the lacrimal gland (LG) in African black ostrich Struthio camelus domesticus in the embryonic and postnatal period. Studies were conducted on 50 ostriches aged between the 28th day of incubation until 7 months old. Tissue sections were stained with haematoxylin and eosin, Azan trichrome, periodic acid-Schiff, Alcian blue pH 2.5, aldehyde fuchsin and Hale's dialysed iron. The LG in ostrich was classified as a tubulo-acinar type. The primordia of the lobes were determined in the LG structure on the 28th day of incubation, whilst the weakly visible lobes with acini and tubules were observed on the 40th day of incubation. Morphometric studies of the LG showed steady growth, characterised by an increase in both length and width. Histometric measurements of lobe size showed little difference between the first, second and third age groups, whilst in the fourth age group a marked increase in size of lobes was observed. The study showed that, apart from morphological changes, during the growth of the LG the character of acid mucopolysaccharides changed. Sulphated acid mucopolysaccharides were indicated, particularly with aldehyde fuchsin (AF) staining in the fourth age group. The Hale's dialysed iron (HDI) staining showed a low concentration of carboxylated acid mucopolysaccharides in the first and second age groups and a higher concentration in the third and fourth age groups. Periodic acid-Schiff staining (PAS)-positive cells were observed in each age group, but only a small number of cells with a weakly PAS-positive reaction were demonstrated in the first age group.

A cross-sectional study was conducted from November 2011 to April 2012 on a total of 384 pigs from two privately owned intensive farms in Bishoftu, Ethiopia. The objectives of the study were to identify and determine the prevalence of common parasites of pigs. For the determination of gastrointestinal (GIT) parasites, faecal samples were collected from the study animals and subjected to standard parasitological examination techniques. Physical examination was conducted for the presence of skin parasitic lesions and skin scrapings were collected to determine prevalence of ectoparasites. The overall prevalence of GIT parasites in the pigs was 25% (96/384). Examination of faecal samples revealed the ova or oocysts of four different gastrointestinal parasites, namely Coccidia (12%), Strongyles (5.2%), Ascaris suum (4.9%) and Trichuris suis (2.9%). Mixed infection by at least two parasite species was observed in 3.65% (14/384) of the pigs. The only ectoparasite species identified was Sarcoptes scabiei var. suis, with a prevalence of 2.6%. This study indicates that pig parasites are a major problem in the study area, hence implementation of strategic control measures and appropriate hygienic management systems are recommended to reduce the prevalence of parasites.