An Equine Herpesvirus Type 1 (EHV-1) Ab4 Open Reading Frame 2 Deletion Mutant Provides Immunity and Protection from EHV-1 Infection and Disease.

Equine herpesvirus type 1 (EHV-1) outbreaks continue to occur despite widely used vaccination. Therefore, development of EHV-1 vaccines providing improved immunity and protection is ongoing. Here, an open reading frame 2 deletion mutant of the neuropathogenic EHV-1 strain Ab4 (Ab4ΔORF2) was tested as a vaccine candidate. Three groups of horses (n = 8 each) were infected intranasally with Ab4ΔORF2 or the parent Ab4 virus or were kept as noninfected controls. Horses infected with Ab4ΔORF2 had reduced fever and nasal virus shedding compared to those infected with Ab4 but mounted similar adaptive immunity dominated by antibody responses. Nine months after the initial infection, all horses were challenged intranasally with Ab4. Previously noninfected horses (control/Ab4) displayed clinical signs, shed large amounts of virus, and developed cell-associated viremia. In contrast, 5/8 or 3/8 horses previously infected with Ab4ΔORF2 or Ab4, respectively, were fully protected from challenge infection as indicated by the absence of fever, clinical disease, nasal virus shedding, and viremia. All of these outcomes were significantly reduced in the remaining, partially protected 3/8 (Ab4ΔORF2/Ab4) and 5/8 (Ab4/Ab4) horses. Protected horses had EHV-1-specific IgG4/7 antibodies prior to challenge infection, and intranasal antibodies increased rapidly postchallenge. Intranasal inflammatory markers were not detectable in protected horses but quickly increased in control/Ab4 horses during the first week after infection. Overall, our data suggest that preexisting nasal IgG4/7 antibodies neutralize EHV-1, prevent viral entry, and thereby protect from disease, viral shedding, and cell-associated viremia. In conclusion, improved protection from challenge infection emphasizes further evaluation of Ab4ΔORF2 as a vaccine candidate.IMPORTANCE Nasal equine herpesvirus type 1 (EHV-1) shedding is essential for virus transmission during outbreaks. Cell-associated viremia is a prerequisite for the most severe disease outcomes, abortion and equine herpesvirus myeloencephalopathy (EHM). Thus, protection from viremia is considered essential for preventing EHM. Ab4ΔORF2 vaccination prevented EHV-1 challenge virus replication in the upper respiratory tract in fully protected horses. Consequently, these neither shed virus nor developed cell-associated viremia. Protection from virus shedding and viremia during challenge infection in combination with reduced virulence at the time of vaccination emphasizes ORF2 deletion as a promising modification for generating an improved EHV-1 vaccine. During this challenge infection, full protection was linked to preexisting local and systemic EHV-1-specific antibodies combined with rapidly increasing intranasal IgG4/7 antibodies and lack of nasal type I interferon and chemokine induction. These host immune parameters may constitute markers of protection against EHV-1 and be utilized as indicators for improved vaccine development and informed vaccination strategies.

Hematologic, Biochemical, and Endocrine Parameters in Horses With Tooth Resorption and Hypercementosis.

BACKGROUND: Equine odontoclastic tooth resorption and hypercementosis (EOTRH) is a frequently diagnosed condition in adult horses. The underlying etiology is still unknown. Hematologic, biochemical, and endocrine values have not been reported in EOTRH-affected horses. OBJECTIVES: The main objective of the study was to describe the hematologic, biochemical, and endocrine parameters in horses with EOTRH. STUDY DESIGN: Descriptive cross-sectional study of client-owned animals with EOTRH. METHODS: A complete blood count, biochemistry panel, and endocrine profile were performed in horses diagnosed with EOTRH. Diagnosis was based on oral and radiographic examination findings and confirmed with histopathology. RESULTS: Eighteen horses with EOTRH aged 10 to 32 years from various regions of the United States were sampled. The only consistent abnormality on the complete blood cell count and chemistry panel was hypoalbuminemia (88%). Endocrine parameters demonstrated no major abnormalities in the functioning of the thyroid and pituitary pars intermedia. The parathyroid hormone concentration was increased in 7 (47%) of 15 horses with an elevated 25-hydroxy vitamin D in 3 (17%) of 17 horses. Main Limitations: The main limitations of this study are the small sample size and lack of age-matched and management-matched control horses. CONCLUSIONS: The relevance of elevated parathyroid hormone in this study cannot be determined due to the lack of age-based controls and large population studies. With the small population evaluated in this study, there are no obvious hematological, biochemical, and endocrine changes evident. Further evaluation with signalment-matched controls will be necessary to evaluate some trends noted in the laboratory values.

Equine gamma herpesvirus presence and viral load are not associated with equine glandular gastric disease.

OBJECTIVE: To investigate the role of equine herpesvirus-2 (EHV-2) and equine herpesvirus-5 (EHV-5) in equine glandular gastric disease (EGGD) by visualizing and quantifying these gamma herpesviruses in EGGD-affected and normal glandular gastric mucosa of horses. A secondary objective was to describe the histopathological abnormalities in the equine gastric glandular mucosa in horses with EGGD. ANIMALS: 29 horses (n = 21 postmortem and 8 gastroscopy) categorized as normal (11), EGGD (12), or both EGGD and equine squamous gastric disease (6). METHODS: Glandular gastric mucosal samples were collected from horses by gastroscopy or postmortem. Histopathology and in situ hybridization targeting EHV-2 and EHV-5 were performed on grossly normal and abnormal glandular gastric mucosa. The number of in situ hybridization-positive cells per millimeter squared of tissue was calculated. Evaluators were blinded to groups. RESULTS: Glandular gastric tissues from horses without EGGD had higher viral loads in the mucosa than normal or abnormal tissues from EGGD horses. There was no difference in viral loads for EHV-2 or EHV-5 between grossly or endoscopically normal to abnormal gastric tissues within horses with EGGD. Lymphocytic plasmacytic gastritis was the most common histopathological abnormality, with only 3 horses having mucosal disruption (glandular ulcer or erosion). CLINICAL RELEVANCE: Equine gamma herpesviruses are unlikely to play a role in the pathophysiology of EGGD. EGGD is frequently inflammatory with occasional mucosal disruption (ulcer or erosion).

Antimicrobial resistance trends among Salmonella isolates obtained from dairy cattle in the northeastern United States, 2004-2011.

Monitoring antimicrobial resistance trends among bacteria isolated from food animals and people is necessary to inform public policy regarding appropriate antimicrobial use. Our objectives were to describe the antimicrobial resistance status of Salmonella isolates from dairy cattle in the northeastern United States and to identify trends in resistance to various antimicrobial agents over time. Data were collected retrospectively for all bovine Salmonella isolates that were obtained from samples submitted to Cornell University's Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. Temporal trends in the prevalence of resistant Salmonella were investigated for each antimicrobial agent using the Cochran-Armitage trend test. Antimicrobial susceptibility testing was performed on 2745 bovine Salmonella isolates from clinical samples submitted during the study period. Overall resistance to each antimicrobial agent ranged from 0% (amikacin, ciprofloxacin, and nalidixic acid) to 72.0% (sulfadimethoxine). There was evidence of a significantly decreasing trend in prevalence of resistance to most agents: amoxicillin/clavulanic acid (AUG), ampicillin (AMP), cefoxitin (FOX), ceftiofur (TIO), ceftriaxone (AXO), chloramphenicol (CHL), chlortetracycline (CTET), florfenicol (FFN), kanamycin (KAN), neomycin (NEO), oxytetracycline (OXY), spectinomycin (SPE), streptomycin (STR), sulfadimethoxine (SDM), sulfisoxazole (FIS), and tetracycline (TET). Among the 265 isolates that were tested using the National Antimicrobial Resistance Monitoring System (NARMS) panel, the most common resistance patterns were pansusceptible (54.0%), AUG-AMP-FOX-TIO-AXO-CHL-KAN-STR-FIS-TET (18.1%), and AUG-AMP-FOX-TIO-AXO-CHL-STR-FIS-TET (12.1%). Increasing prevalence of S. enterica serovar Cerro over the course of the study period presumably had an impact on the observed resistance trends. Nevertheless, these results do not support the notion that the current level of antimicrobial use in dairy cattle is driving an increase in the emergence and dissemination of drug-resistant Salmonella in the region served by the laboratory.

Agreement of stall-side and laboratory major crossmatch tests with the reference standard method in horses.

BACKGROUND: Crossmatching is used to prevent life-threatening transfusion reactions in horses. Laboratory methods are laborious and technically challenging, which is impractical during emergencies. HYPOTHESIS/OBJECTIVES: Evaluate agreement between a stall-side crossmatch kit (KIT) and a laboratory method (LAB) in horses with known and unknown blood types. ANIMALS: Twenty-four blood-typed and alloantibody-screened healthy adult horses (Aim 1) and 156 adult horses of unknown blood type (Aim 2). METHODS: Prospective, blinded study. Expected positive (n = 35) and negative (n = 36) crossmatches among 24 antibody and blood-typed horses were used to determine sensitivity and specificity of KIT and LAB against the reference method. Agreement in 156 untyped horses was evaluated by reciprocal crossmatch (n = 156). RESULTS: Sensitivity (95% confidence interval [CI]) for LAB and KIT compared with expected reactions was 77.1% (59.9%-90.0%) and 91.4% (77.0%-98.2%), and specificity 77.8% (60.9%-89.9%) and 73.5% (55.6%-87.1%), respectively. The KIT was 100% sensitive for Aa reactions; LAB was 100% sensitive for Qab; and both were 100% sensitive for Ca. Cohen's κ agreement for LAB and KIT with expected positive and negative reactions (n = 71) was moderate (0.55 [0.36-0.74]) and substantial (0.65 [0.47-0.82]), respectively. Agreement was fair comparing LAB with KIT in Aim 1 (0.30 [0.08-0.52]) and in untyped horses in Aim 2 (0.26 [0.11-0.41]). CONCLUSIONS AND CLINICAL IMPORTANCE: Agreement between KIT and LAB with expected reactions was blood type dependent. Performance of both methods depends on blood type prevalence.

A point mutation in a herpesvirus polymerase determines neuropathogenicity.

Infection with equid herpesvirus type 1 (EHV-1) leads to respiratory disease, abortion, and neurologic disorders in horses. Molecular epidemiology studies have demonstrated that a single nucleotide polymorphism resulting in an amino acid variation of the EHV-1 DNA polymerase (N752/D752) is significantly associated with the neuropathogenic potential of naturally occurring strains. To test the hypothesis that this single amino acid exchange by itself influences neuropathogenicity, we generated recombinant viruses with differing polymerase sequences. Here we show that the N752 mutant virus caused no neurologic signs in the natural host, while the D752 virus was able to cause inflammation of the central nervous system and ataxia. Neurologic disease induced by the D752 virus was concomitant with significantly increased levels of viremia (p = 0.01), but the magnitude of virus shedding from the nasal mucosa was similar between the N752 and D752 viruses. Both viruses replicated with similar kinetics in fibroblasts and epithelial cells, but exhibited differences in leukocyte tropism. Last, we observed a significant increase (p < 0.001) in sensitivity of the N752 mutant to aphidicolin, a drug targeting the viral polymerase. Our results demonstrate that a single amino acid variation in a herpesvirus enzyme can influence neuropathogenic potential without having a major effect on virus shedding from infected animals, which is important for horizontal spread in a population. This observation is very interesting from an evolutionary standpoint and is consistent with data indicating that the N752 DNA pol genotype is predominant in the EHV-1 population, suggesting that decreased viral pathogenicity in the natural host might not be at the expense of less efficient inter-individual transmission.

Live-attenuated recombinant equine herpesvirus type 1 (EHV-1) induces a neutralizing antibody response against West Nile virus (WNV).

The immunogenicity in horses of a recombinant equine herpesvirus type 1 (EHV-1) vaccine expressing West Nile virus (WNV) prM and E proteins was studied. To construct the recombinant EHV-1, two-step en passant mutagenesis was employed for manipulation of a bacterial artificial chromosome (BAC) of vaccine strain RacH. Recombinant EHV-1 stably expressed the WNV prM and E proteins as demonstrated by indirect immunofluorescence and Western blotting. In addition, growth properties in vitro of the EHV-1/WNV recombinant were found to not be significantly different from those of the parental virus. To determine if vaccination of horses induces an antibody response, 10 horses were allocated in two groups. Group A consisted of six horses that were vaccinated three times with the recombinant EHV-1/WNV virus in 28- to 31-day intervals. Group B consisted of four horses that were sham-vaccinated using the same regimen. Serum was collected on days 0, 31, 45 and 66. Plaque reduction neutralization test and IgG(T)- and IgGb-specific WNV E antibody-capture ELISAs were used. After a single vaccination (day 31), at least four of the six horses from group A had detectable levels of serum neutralizing antibodies against WNV, and three horses retained SN titers until the end of the study. None of the horses in the control group B sero-converted. On days 31 and 45, five of the six horses in group A had a marked increase of WNV-specific IgG(T), and at least four exhibited modestly elevated WNV-specific IgGb titers. From the results, we concluded that the EHV-1 vectored virus is able to express the WNV structural proteins and that vaccination of horses results in the induction of WNV E-protein-specific IgG(T), IgGb, and neutralizing antibodies.

Cytokine profiles of peripheral blood mononuclear cells isolated from septic and healthy neonatal foals.

BACKGROUND: Septicemia initiates the production of pro-inflammatory (interleukin [IL] 1-beta [IL-1beta], interferon-gamma [IFN-gamma], IL-6), and anti-inflammatory (IL-4) cytokines. The transcription of some of these proteins (IL-8, IL-6) is linked to endotoxin-induced activation of the toll-like receptor 4 (TLR4) on peripheral blood mononuclear cells (PBMC). HYPOTHESES: Septic foals fail to increase gene expression of IFN-gamma. Nonsurviving septic foals exhibit distinctive cytokine profiles. ANIMALS: Twenty-one septic and 20 healthy neonatal foals. METHODS: Using real-time polymerase chain reaction, gene expression of IFN-gamma, IL-1beta, IL-6, IL-4, IL-8, TLR4, and beta-actin in PBMC were measured in samples obtained from septic foals at 0, 24, and 72 hours (T = 0, 24, and 72 hours) after admission to the Cornell University Hospital for Animals. Control foals were sampled at comparable times. RESULTS: At T=0 hours, septic foals exhibited a 6-fold decrease in gene expression of IL-4 and a 5-fold increase in gene expression of TLR4. Gene expression of IFN-gamma, IL-6, IL-8, or of IL-1beta did not differ between the 2 groups of foals at T = 0 hours. In septic foals that died (n = 3), there was a 15-fold increase in IL-6 at T = 0 hours compared to survivors. CONCLUSIONS AND CLINICAL IMPORTANCE: Septic foals, unlike septic human infants, up-regulate TLR4 gene expression, which may enhance pro-inflammatory cytokine production. Despite the presence of sepsis, IFN-gamma was not up-regulated. Additional studies are needed to verify that increased IL-6 expression is associated with a poor prognosis in septic foals.

First demonstration of equid gammaherpesviruses within the gastric mucosal epithelium of horses.

Horses commonly develop gastric mucosal ulcers, similar to humans, a condition known as equine gastric ulcer syndrome (EGUS) that can lead to poor performance and lost training time and care expenses. Unlike humans, however, an infectious bacterial cause of ulcers has not been conclusively identified. Herpesviruses, while well-established causative agents of diseases such as cold sores, genital lesions, and certain types of cancer, have also been implicated in the development of a subset of gastric ulcers in humans. The presence of equid herpesviruses in the gastrointestinal tract and their potential contribution to EGUS has not been evaluated. Here, we provide the first evidence of equid gammaherpesviruses 2 and 5 (EHV-2 and -5) within the epithelium of the gastric mucosa of horses. These viruses were initially detected by a nested PCR screen of gastric tissue samples obtained from client- and university-owned horses with and without ulcers; however, no association with EGUS was found in this limited sample set. We then validated a highly sensitive in situ hybridization (ISH) assay and used this assay to characterize the distribution of these viruses in necropsy gastric tissue samples from five racehorses. Analyses revealed frequent EHV-2 and EHV-5 co-infections within the gastric mucosal epithelium, regardless of the ulcer status. These results are the first to demonstrate the presence of equid gammaherpesviruses in the gastric mucosa of horses and warrants further investigation to determine the contribution of these viruses to the development of EGUS and/or other gastrointestinal diseases.

Detection of Salmonella spp. in veterinary samples by combining selective enrichment and real-time PCR.

Rapid screening for enteric bacterial pathogens in clinical environments is essential for biosecurity. Salmonella found in veterinary hospitals, particularly Salmonella enterica serovar Dublin, can pose unique challenges for culture and testing because of its poor growth. Multiple Salmonella serovars including Dublin are emerging threats to public health given increasing prevalence and antimicrobial resistance. We adapted an automated food testing method to veterinary samples and evaluated the performance of the method in a variety of matrices including environmental samples ( n = 81), tissues ( n = 52), feces ( n = 148), and feed ( n = 29). A commercial kit was chosen as the basis for this approach in view of extensive performance characterizations published by multiple independent organizations. A workflow was established for efficiently and accurately testing veterinary matrices and environmental samples by use of real-time PCR after selective enrichment in Rappaport-Vassiliadis soya (RVS) medium. Using this method, the detection limit for S. Dublin improved by 100-fold over subculture on selective agars (eosin-methylene blue, brilliant green, and xylose-lysine-deoxycholate). Overall, the procedure was effective in detecting Salmonella spp. and provided next-day results.

Prognostic value of plasma L-lactate concentration measured cow-side with a portable clinical analyzer in Holstein dairy cattle with abomasal disorders.

BACKGROUND: L-Lactate has been used as a prognostic indicator for ill humans and animals. A portable analyzer that measures L-lactate could help veterinarians decide to proceed with correction of a displaced abomasum. HYPOTHESES: The likelihood of a dairy cow with a displaced abomasum remaining in the herd can be predicted by lactate concentration and other variables. ANIMALS: Thirty-four healthy early-lactation dairy cows, and 131 cows with abomasal displacements (DA) presented to Cornell University, Ithaca, NY. METHODS: Plasma L-lactate was measured using a commercial analyzer (i-STAT). A cow had a positive outcome (PO) if she remained in the herd 30 days after surgical correction of the displaced abomasum and a negative outcome (NO) if she was culled or died in that time. A multivariable model with physical examination and clinicopathologic variables for predicting NO for cows with right-sided abomasal displacements was constructed. RESULTS: The median plasma L-lactate was 0.54 mM/L (interquartile range, 0.42-0.74) in healthy lactating Holstein cows. In cows with right-sided displaced abomasa, median plasma L-lactate concentrations were higher in cows with NO (5.88 mM/L) versus PO (3.23 mM/L) (P = .002). In a multivariable model, which identified chloride, heart rate, and L-lactate as the best fitting variables for cows with right-sided displacements, the probability of NO increased as L-lactate increased. CONCLUSIONS AND CLINICAL IMPORTANCE: Plasma L-lactate concentration might be a useful predictor of productive outcomes in cows with right-sided abomasal disorders.

Antimicrobial resistance trends among Salmonella isolates obtained from horses in the northeastern United States (2001-2013).

OBJECTIVE To describe the antimicrobial resistance patterns of Salmonella isolates obtained from horses in the northeastern United States and to identify trends in resistance to select antimicrobials over time. SAMPLE 462 Salmonella isolates from horses. PROCEDURES Retrospective data were collected for all Salmonella isolates obtained from equine specimens that were submitted to the Cornell University Animal Health Diagnostic Center between January 1, 2001, and December 31, 2013. Temporal trends in the prevalence of resistant Salmonella isolates were investigated for each of 13 antimicrobials by use of the Cochran-Armitage trend test. RESULTS The prevalence of resistant isolates varied among antimicrobials and ranged from 0% (imipenem) to 51.5% (chloramphenicol). During the observation period, the prevalence of resistant isolates decreased significantly for amoxicillin-clavulanic acid, ampicillin, cefazolin, cefoxitin, ceftiofur, chloramphenicol, and tetracycline and remained negligible for amikacin and enrofloxacin. Of the 337 isolates for which the susceptibility to all 13 antimicrobials was determined, 138 (40.9%) were pansusceptible and 192 (57.0%) were multidrug resistant (resistant to ≥ 3 antimicrobial classes). The most common serovar isolated was Salmonella Newport, and although the annual prevalence of that serovar decreased significantly over time, that decrease had only a minimal effect on the observed antimicrobial resistance trends. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that current antimicrobial use in horses is not promoting the emergence and dissemination of antimicrobial-resistant Salmonella strains in the region served by the laboratory.

West Nile virus-specific immunoglobulin isotype responses in vaccinated and infected horses.

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.

Survival of foals with experimentally induced Rhodococcus equi infection given either hyperimmune plasma containing R. equi antibody or normal equine plasma.

The purpose of this study was to determine if colostrum-deprived foals with experimentally induced Rhodococcus equi pneumonia have a decreased severity of the disease and decreased mortality rate when given hyperimmune (HI) R. equi antibody plasma (R. equi titer at least 100 % and virulence-associated protein A [VapA] at least 10000) prophylactically versus when given normal equine plasma (R. equi titer less than 20 % and VapA less than 160). Sixteen colostrum-deprived foals (R. equi titer less than 5 %) each received normal equine plasma in the first 24 hours of life (R. equi titer less than 20 %). At 14 days of age, six foals were given normal equine plasma and 10 foals were given HI plasma. All foals were subsequently infected intrabronchially with a pathogenic strain of R. equi (2.5 x 10 sup 8; organisms) at 21 days of age. Repeated physical examinations, weight measurements, complete blood cell counts, fibrinogen measurements, and thoracic radiographs (ventrodorsal and lateral) were performed to help determine the severity of the disease. Foals given HI plasma had significantly higher R. equi ELISA titers (42.4 %) than those given normal plasma (20.9 %) on the day of experimental infection. Mortality rates and severity of disease were statistically similar (P >.05) for the groups. Although none of the foals was treated with antibiotics, several with severe R. equi pneumonia recovered. Either HI or normal equine plasma administered to foals in the first few weeks of life caused no adverse effects and may be protective against R. equi, although the exact constituent responsible for protection is undetermined and requires further investigation.

Maternal T-lymphocytes in equine colostrum express a primarily inflammatory phenotype.

The purpose of this study was to characterize maternal immune cells in colostrum of mares. Cell phenotypes and cytokine secretion from mare peripheral blood mononuclear cells (PBMC) and cells from colostrum were analyzed by flow cytometry and by multiplex cytokine analysis. Equine colostral leukocytes were composed of mainly CD8(+) and CD4(+) lymphocytes. CD8(+) cells were significantly enriched in colostrum compared to PBMC (n=35). Colostral T-cells (n=13) responded to stimulation with PMA/ionomycin with a significantly higher magnitude of IL-17 (p=0.037) and similar IFN-γ concentrations (p=0.305), while IL-4 (p=0.0002) and IL-10 (p=0.0002) production was decreased compared to PBMC. CD4(+) and CD8(+) T-cells in colostrum produced IFN-γ (n=4). The findings show that colostrum T-cells can produce all four cytokines investigated here but most cells are polarized toward IL-17 and IFN-γ production and an inflammatory phenotype. Maternal T-cells likely migrate to the colostrum in a selective manner and may have specific roles in neonatal immune development.

Salmonella enterica serovar Oranienburg outbreak in a veterinary medical teaching hospital with evidence of nosocomial and on-farm transmission.

Nosocomial salmonellosis continues to pose an important threat to veterinary medical teaching hospitals. The objectives of this study were to describe an outbreak of salmonellosis caused by Salmonella enterica serovar Oranienburg within our hospital and to highlight its unique features, which can be used to help mitigate or prevent nosocomial outbreaks in the future. We retrospectively analyzed data from patients that were fecal culture-positive for Salmonella Oranienburg between January 1, 2006, and June 1, 2011, including historical, clinical, and pulsed-field gel electrophoresis (PFGE) data. Salmonella Oranienburg was identified in 20 horses, five alpacas, and three cows during this time frame, with dates of admission spanning the period from August, 2006, through January, 2008. We consider most of these patients to have become infected through either nosocomial or on-farm transmission, as evidenced by molecular subtyping results and supportive epidemiologic data. Interpretation of PFGE results in this outbreak was challenging because of the identification of several closely related Salmonella Oranienburg subtypes. Furthermore, a high percentage of cases were fecal culture-positive for Salmonella Oranienburg within 24 h of admission. These patients initially appeared to represent new introductions of Salmonella into the hospital, but closer inspection of their medical records revealed epidemiologic links to the hospital following the index case. Cessation of this outbreak was observed following efforts to further heighten biosecurity efforts, with no known cases or positive environmental samples after January, 2008. This study demonstrates that a Salmonella-positive culture result within 24 h of admission does not exclude the hospital as the source of infection, and it underscores the important role played by veterinary medical teaching hospitals as nodes of Salmonella infection that can promote transmission outside of the hospital setting.

Evaluation of metaphylactic RNA interference to prevent equine herpesvirus type 1 infection in experimental herpesvirus myeloencephalopathy in horses.

OBJECTIVE: To evaluate metaphylactic RNA interference to prevent equine herpesvirus type 1 (EHV-1) infection in experimental herpesvirus myeloencephalopathy in horses and to determine whether horses infected with a neuropathogenic strain of the virus that develop equine herpesvirus myeloencephalopathy (EHM) have differences in viremia. ANIMALS: 13 seronegative horses. PROCEDURES: EHV-1 strain Ab4 was administered intranasally on day 0, and small interfering RNAs (siRNAs [EHV-1 specific siRNAs {n = 7} or an irrelevant siRNA {6}]) were administered intranasally 24 hours before and 12, 24, 36, and 48 hours after infection. Physical and neurologic examinations, nasal swab specimens, and blood samples were collected for virus isolation and quantitative PCR assay. Data from the study were combined with data from a previous study of 14 horses. RESULTS: No significant difference was detected in clinical variables, viremia, or detection of EHV-1 in nasal swab specimens of horses treated with the EHV-1 targeted siRNAs (sigB3-siOri2) versus controls. No significant differences in viremia were detected between horses that developed EHM and those that did not. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of siRNAs targeted against EHV-1 around the time of EHV-1 infection was not protective with this experimental design. Horses infected with the neuropathogenic EHV-1 strain Ab4 that developed EHM did not have a more pronounced viremia.

Cerebrospinal fluid eosinophilia is a sensitive and specific test for the diagnosis of Parelaphostrongylus tenuis in camelids in the northeastern United States.

Aberrant migration of Parelaphostrongylus tenuis in camelids results in neurologic deficits, recumbency, and sometimes death. An antemortem diagnosis of P. tenuis in camelids is typically based upon the presence of characteristic asymmetric neurologic deficits, known exposure to white-tailed deer, cerebrospinal fluid (CSF) eosinophilia, and response to treatment. The diagnostic accuracy of CSF eosinophil percentage for the diagnosis of P. tenuis in camelids has not been critically examined. The objective of the current study was to determine the sensitivity (Se) and specificity (Sp) of CSF eosinophil percentage, CSF eosinophil concentration, total nucleated cell concentration, and protein concentration for the antemortem diagnosis of P. tenuis. Medical records of camelids admitted to Cornell University with clinical signs of neurologic disease, CSF analysis, and necropsy were examined from January 2000 through December 2009. Se and Sp were determined by receiver operating characteristic curves in camelids diagnosed with P. tenuis (n = 13) or other conditions (n = 24) based on postmortem examination. More than 17% of eosinophils in CSF had a Se of 85% and Sp of 92% for P. tenuis diagnosis (area under the curve [AUC]: 0.87; SE AUC: 0.07; P < 0.0001; 95% confidence interval [CI] AUC: 0.72-0.96), and >1.4 eosinophils/µl of CSF had a Se of 85% and Sp of 96% (AUC: 0.9; SE AUC: 0.06; P < 0.0001; 95% CI AUC: 0.76-0.97). Cerebrospinal fluid eosinophil percentage and concentration are sensitive and specific methods for diagnosing P. tenuis antemortem in camelids residing in regions endemic to white-tailed deer.

The effect of siRNA treatment on experimental equine herpesvirus type 1 (EHV-1) infection in horses.

Available vaccines fail to induce lasting and protective immunity to equine herpesvirus 1 (EHV-1) associated diseases. RNA interference is a novel approach showing promise for therapeutic use in outbreak situations. This study examined the effect of small interfering RNA (siRNA) on clinical signs as well as the presence of live virus and viral DNA in nasal secretions and peripheral blood mononuclear cells (PBMCs) in horses experimentally infected with EHV-1. siRNA targeting two EHV-1 genes (glycoprotein B and the origin binding protein) was administered 12h before and 12h after intranasal infection with EHV-1. Control horses received siRNA targeting firefly luciferase. A significantly smaller proportion (0/10) of horses receiving siRNA targeting viral genes required euthanasia due to intractable neurologic disease as compared to horses in the control group (3/4; p=0.01). There was no significant difference in the presence of live virus or viral DNA in the nasal secretions or PBMCs between the two groups. Future studies are necessary to define the relative contributions of host and virus factors in the development of the neurological form of the infection and to determine an optimal dosing regimen for metaphylactic or therapeutic use of siRNA for treating EHV-1 infection.