Differentiation of lymphocytic-plasmacytic enteropathy and small cell lymphoma in cats using histology-guided mass spectrometry.

BACKGROUND: Differentiation of lymphocytic-plasmacytic enteropathy (LPE) from small cell lymphoma (SCL) in cats can be challenging. HYPOTHESIS/OBJECTIVE: Histology-guided mass spectrometry (HGMS) is a suitable method for the differentiation of LPE from SCL in cats. ANIMALS: Forty-one cats with LPE and 52 cats with SCL. METHODS: This is a retrospective clinicopathologic study. Duodenal tissue samples of 17 cats with LPE and 22 cats with SCL were subjected to HGMS, and the acquired data were used to develop a linear discriminate analysis (LDA) machine learning algorithm. The algorithm was subsequently validated using a separate set of 24 cats with LPE and 30 cats with SCL. Cases were classified as LPE or SCL based on a consensus by an expert panel consisting of 5-7 board-certified veterinary specialists. Histopathology, immunohistochemistry, and clonality testing were available for all cats. The panel consensus classification served as a reference for the calculation of test performance parameters. RESULTS: Relative sensitivity, specificity, and accuracy of HGMS were 86.7% (95% confidence interval [CI]: 74.5%-98.8%), 91.7% (95% CI: 80.6%-100%), and 88.9% (95% CI: 80.5%-97.3%), respectively. Comparatively, the clonality testing had a sensitivity, specificity, and accuracy of 85.7% (95% CI: 72.8%-98.7%), 33.3% (95% CI: 14.5%-52.2%), and 61.5% (95% CI: 48.3%-74.8%) relative to the panel decision. CONCLUSIONS AND CLINICAL IMPORTANCE: Histology-guided mass spectrometry was a reliable technique for the differentiation of LPE from SCL in duodenal formalin-fixed paraffin-embedded samples of cats and might have advantages over tests currently considered state of the art.

Pneumonic Plague in a Dog and Widespread Potential Human Exposure in a Veterinary Hospital, United States.

In December 2017, a dog that had pneumonic plague was brought to a veterinary teaching hospital in northern Colorado, USA. Several factors, including signalment, season, imaging, and laboratory findings, contributed to delayed diagnosis and resulted in potential exposure of >116 persons and 46 concurrently hospitalized animals to Yersinia pestis.

Associations of environment, health history, T-zone lymphoma, and T-zone-like cells of undetermined significance: A case-control study of aged Golden Retrievers.

BACKGROUND: T-zone lymphoma (TZL), an indolent disease in older dogs, comprises approximately 12% of lymphomas in dogs. TZL cells exhibit an activated phenotype, indicating the disease may be antigen-driven. Prior research found that asymptomatic aged Golden Retrievers (GLDRs) commonly have populations of T-zone-like cells (phenotypically identical to TZL) of undetermined significance (TZUS). OBJECTIVE: To evaluate associations of inflammatory conditions, TZL and TZUS, using a case-control study of GLDRs. ANIMALS: TZL cases (n = 140), flow cytometrically diagnosed, were identified through Colorado State University's Clinical Immunology Laboratory. Non-TZL dogs, recruited through either a database of owners interested in research participation or the submitting clinics of TZL cases, were subsequently flow cytometrically classified as TZUS (n = 221) or control (n = 147). METHODS: Health history, signalment, environmental, and lifestyle factors were obtained from owner-completed questionnaires. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were estimated using multivariable logistic regression, obtaining separate estimates for TZL and TZUS (versus controls). RESULTS: Hypothyroidism (OR, 0.3; 95% CI, 0.1-0.7), omega-3 supplementation (OR, 0.3; 95% CI, 0.1-0.6), and mange (OR, 5.5; 95% CI, 1.4-21.1) were significantly associated with TZL. Gastrointestinal disease (OR, 2.4; 95% CI, 0.98-5.8) had nonsignificantly increased TZL odds. Two shared associations for TZL and TZUS were identified: bladder infection or calculi (TZL OR, 3.5; 95% CI, 0.96-12.7; TZUS OR, 5.1; 95% CI, 1.9-13.7) and eye disease (TZL OR, 2.3; 95% CI, 0.97-5.2; TZUS OR, 1.9; 95% CI, 0.99-3.8). CONCLUSIONS AND CLINICAL IMPORTANCE: These findings may elucidate pathways involved in TZUS risk and progression from TZUS to TZL. Further investigation into the protective association of omega-3 supplements is warranted.

Addition of Mesenchymal Stem Cells to Autologous Platelet-Enhanced Fibrin Scaffolds in Chondral Defects: Does It Enhance Repair?

BACKGROUND: The chondrogenic potential of culture-expanded bone-marrow-derived mesenchymal stem cells (BMDMSCs) is well described. Numerous studies have also shown enhanced repair when BMDMSCs, scaffolds, and growth factors are placed into chondral defects. Platelets provide a rich milieu of growth factors and, along with fibrin, are readily available for clinical use. The objective of this study was to determine if the addition of BMDMSCs to an autologous platelet-enriched fibrin (APEF) scaffold enhances chondral repair compared with APEF alone. METHODS: A 15-mm-diameter full-thickness chondral defect was created on the lateral trochlear ridge of both stifle joints of twelve adult horses. In each animal, one defect was randomly assigned to receive APEF+BMDMSCs and the contralateral defect received APEF alone. Repair tissues were evaluated one year later with arthroscopy, histological examination, magnetic resonance imaging (MRI), micro-computed tomography (micro-CT), and biomechanical testing. RESULTS: The arthroscopic findings, MRI T2 map, histological scores, structural stiffness, and material stiffness were similar (p > 0.05) between the APEF and APEF+BMDMSC-treated repairs at one year. Ectopic bone was observed within the repair tissue in four of twelve APEF+BMDMSC-treated defects. Defects repaired with APEF alone had less trabecular bone edema (as seen on MRI) compared with defects repaired with APEF+BMDMSCs. Micro-CT analysis showed thinner repair tissue in defects repaired with APEF+BMDMSCs than in those treated with APEF alone (p < 0.05). CONCLUSIONS: APEF alone resulted in thicker repair tissue than was seen with APEF+BMDMSCs. The addition of BMDMSCs to APEF did not enhance cartilage repair and stimulated bone formation in some cartilage defects. CLINICAL RELEVANCE: APEF supported repair of critical-size full-thickness chondral defects in horses, which was not improved by the addition of BMDMSCs. This work supports further investigation to determine whether APEF enhances cartilage repair in humans.

Concentrations of serum amyloid A and plasma fibrinogen in horses undergoing emergency abdominal surgery.

OBJECTIVE: To compare the perioperative response of serum amyloid A (SAA) to fibrinogen in horses requiring exploratory celiotomy for colic and to determine if SAA could be used to predict complications and outcome. DESIGN: Prospective observational clinical study. SETTING: University teaching hospital. ANIMALS: Eighteen horses undergoing exploratory celiotomy for colic. Inclusion criteria for the study included survival and anesthetic recovery from exploratory celiotomy, no history of surgery within the past year. INTERVENTIONS: Blood was obtained via jugular venipuncture before surgery (time 0) and at 24, 48, 72, and 96 hours after recovery from anesthesia. MEASUREMENTS AND MAIN RESULTS: Quantitative and semiquantitative fibrinogen, SAA, total nucleated cell counts, and total protein were evaluated at each time point. Multivariable linear regression was used to assess differences at each time point and after grouping horses according to duration of colic prior to surgery, strangulating surgical lesion or not, presence of systemic inflammatory response syndrome (SIRS) on admission, and postsurgical complications. Significant (P < 0.05) increases in SAA concentrations occurred in all cases after surgery compared to fibrinogen concentration, which only demonstrated a mild, clinically insignificant increase postsurgery. SAA concentrations were also significantly increased (P < 0.05) in cases identified with SIRS prior to surgery and postoperatively at 48 (P = 0.05) and 72 hours (P = 0.02) in horses that developed complications. CONCLUSIONS: Measurement of SAA is a more sensitive indicator of inflammation than fibrinogen in the perioperative period of horses requiring exploratory celiotomy for colic. Serial measurement of SAA at 48, 72, and 96 hours after surgery may be helpful to determine risk of complications and guide postoperative management. Measurement of SAA on admission also allows for quantification of SIRS when it is detected clinically.

A comparison of peritoneal fluid values in mares following bilateral laparoscopic ovariectomy using a vessel sealing and dividing device versus placement of two ligating loops.

The objective of this study was to determine the effect of bilateral laparoscopic ovariectomy on peritoneal fluid values in mares and compare how this effect was modified by the method of ovarian vessel hemostasis used. Ten mares undergoing standing bilateral laparoscopic ovariectomy were used in a randomized clinical study. During surgery, blood vessels within the mesovarium were either: (1) sealed and transected with a vessel sealing and dividing device (VSDD), or (2) ligated using two loops placed proximal to each ovary and then the mesovarium transected using laparoscopic scissors. The ovaries were removed through the ipsilateral body wall. Abdominocentesis was performed before surgery and 24 h and 72 h after surgery. Markers of peritoneal inflammation, as measured by total nucleated cell count, total protein (TP) and red blood cell count via abdominocentesis, were consistently increased for all groups compared to pre-operative values. The mean (range) of TP for the VSDD group was 4.14 (3.9-4.5) g/dL, and that for the ligating loop group was 3.18 (2.7-3.5) g/dL. Use of the VSDD resulted in significantly greater TP concentrations in the abdominal fluid at 24 h and 72 h post-operatively when compared to a ligating loop (P <0.001 and 0.04, respectively).

Comparison of routine urinalysis and urine Gram stain for detection of bacteriuria in dogs.

OBJECTIVE: To determine the utility of performing urine Gram stain for detection of bacteriuria compared to routine urine sediment examination and bacterial aerobic urine culture. DESIGN: Prospective, observational study. SETTING: University teaching hospital. ANIMALS: Urine samples acquired via cystocentesis through convenience sampling from 103 dogs presenting to a tertiary referral institution. INTERVENTIONS: All samples underwent routine urinalysis, including sediment examination, as well as urine Gram stain and quantitative bacterial aerobic urine culture. MEASUREMENTS AND MAIN RESULTS: The urine Gram stain demonstrated improved sensitivity (96% versus 76%), specificity (100% versus 77%), positive predictive value (100% versus 83%), and negative predictive value (93% versus 69%) when identifying bacteriuria, compared to routine urine sediment examination. CONCLUSIONS: The urine Gram stain is highly sensitive and specific when detecting the presence of bacteria in canine urine samples. Gram staining should be considered when bacteriuria is highly suspected and requires rapid identification while bacterial culture is pending.

Comparison of peritoneal fluid values after laparoscopic cryptorchidectomy using a vessel-sealing device (Ligasure™) versus a ligating loop and removal of the descended testis.

OBJECTIVE: To determine the effect of unilateral laparoscopic cryptorchidectomy and removal of the descended testis on peritoneal fluid values, and to compare effect between 2 methods for cryptorchid testis vessel hemostasis. STUDY DESIGN: Randomized clinical study. ANIMALS: Stallions (n = 10) with unilateral abdominal cryptorchid testis. METHODS: During standing laparoscopic cryptorchidectomy, blood vessels within the mesorchium of the cryptorchid testis were either sealed and transected with the LigaSure Atlas™ or 2 ligating loops were placed proximal to the testis and the tissue transected with laparoscopic scissors. The testis was removed through the body wall. After laparoscopic cryptorchidectomy, stallions were anesthetized and the descended testis was removed using a closed technique leaving the scrotal incision open. Abdominocenteses were performed before surgery, and 24, and 72 hours after surgery. RESULTS: Values for peritoneal total nucleated cell count (TNCC), total protein concentration (TP), and red blood cell count (RBCC) were all elevated at 24 and 72 hours when compared with baseline. Median TNCC for LigaSure™ (59,780 cells/µL) was nearly twice that of the ligating loop (32,880 cells/µL) at 24 hours postoperatively. There was no statistically significant difference in TNCC, TP, or RBCC between groups. CONCLUSIONS: TNCC, TP, and RBCC increase appreciably from baseline 24 hours after laparoscopic cryptorchidectomy and closed castration but are markedly reduced by 72 hours.

A multiplex polymerase chain reaction assay for the identification of Mannheimia haemolytica, Mannheimia glucosida and Mannheimia ruminalis.

The objective of this study was to design a multiplex PCR assay to identify Mannheimia haemolytica, Mannheimia glucosida and Mannheimia ruminalis. The multiplex PCR included primer sets HP, amplifying a DNA region from an unknown hypothetical protein, Lkt and Lkt2, amplifying different regions of the leukotoxinD gene, and 16S to amplify universal bacterial sequences of the 16S rRNA gene. Based on positive amplification, isolates were delineated as M. haemolytica (HP, Lkt, 16S), M. glucosida (HP, Lkt, Lkt2, 16S), or M. ruminalis (HP, 16S). The validity of the assay was examined against 22 reference strains within the family Pasteurellaceae and 17 field isolates (nasal) that had been collected previously from feedlot cattle and tentatively identified as M. haemolytica based on morphology and substrate utilization. Additionally, 200 feedlot cattle were screened for M. haemolytica using multiplex PCR. Forty-four isolates from 25 animals were identified as M. haemolytica. The PCR assay positively identified all M. haemolytica, as confirmed by phenotypic tests and clustering based upon cellular fatty acid methyl ester (FAME) profiles. Selected nasal isolates that exhibited evidence of haemolysis, but were M. haemolytica-negative based on PCR, were also confirmed negative by phenotypic and FAME analyses. The multiplex PCR assay required no additional phenotypic tests for confirmation of M. haemolytica, within the group of bacteria tested.

Assessment of the likelihood of the introduction of foot-and-mouth disease through importation of live animals into the Malaysia-Thailand-Myanmar peninsula.

OBJECTIVE: To assess the likelihood of an introduction of foot-and-mouth disease (FMD) into the Malaysia-Thailand-Myanmar (MTM) peninsula through terrestrial movement of livestock. ANIMALS: 89,294 cattle and buffalo legally moved into the MTM peninsula. PROCEDURES: A quantitative risk assessment was conducted by use of a stochastic simulation. Patterns of livestock movement were ascertained through review of relevant governmental records and regulations and by interviewing farmers, traders, and local officers when the records did not exist. Parameters identified in the process were the probabilities of livestock having FMD and of FMD infection going undetected during import processes. The probability of an animal accepted for import having FMD was also assessed. Sensitivity analysis was performed to determine the effects that each parameter had on the model. RESULTS: The simulation yielded an average consignment prevalence of 10.95%. Typically, each animal in a quarantine facility had a 2.7% chance of having an inapparent form of FMD infection; hence, it was likely an animal would not be identified as infected. Findings revealed that the mean probability of an animal accepted for import having FMD was 2.9%, and the risk was as high as 11%. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the model allowed for the evaluation of movement regulations currently imposed in the MTM peninsula. Evidence from the study suggested that current practices in animal movement were far from efficient in preventing introduction of FMD-infected animals into the MTM region, and additional measures will be necessary.

Utility of polymerase chain reaction for analysis of antigen receptor rearrangement in staging and predicting prognosis in dogs with lymphoma.

In lymphoid neoplasia, molecular assays to confirm clonality rely on the fact that lymphoid cells normally contain DNA regions with unique sequences, resulting from recombination of the V, D, and J genes. The purpose of this study was to determine the utility of polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) for molecular staging and predicting prognosis in canine lymphoma. We hypothesized that the PARR assay would offer a sensitive method for detecting neoplastic cells in blood, and that the presence of such cells would be a negative prognostic finding compared with dogs with no detectable circulating tumor cells. Eighty-six patients with histologically or cytologically confirmed lymphoma were studied from initial diagnosis until death or euthanasia. All patients had PARR assays of a representative tumor-infiltrated lymph node and peripheral whole blood. Sixty-two patients had clonal rearrangements detected in the lymph node and were able to be staged by PARR. Seventeen patients (27%) had no detectable tumor in their blood and 45 (73%) were blood positive. Our findings showed that (1) PARR correlated with clinical stage in that the PARR assay was more likely to detect tumor cells in blood in stage 5 lymphomas, (2) PARR was more sensitive for detecting circulating tumor cells than visual assessment of blood or bone marrow because 80% of stage 3 lymphomas were blood-PARR-positive, and (3) PCR stage was not prognostic for disease-free interval (DFI) or survival.

Association between exercise-induced pulmonary hemorrhage and performance in Thoroughbred racehorses.

OBJECTIVE: To determine whether exercise-induced pulmonary hemorrhage (EIPH) was associated with racing performance inThoroughbred horses not medicated with furosemide and not using nasal dilator strips. DESIGN: Observational cross-sectional study. ANIMALS: 744 two- to 10-year-old Thoroughbred horses racing in Melbourne, Australia. PROCEDURE: Horses were enrolled prior to racing, and a tracheobronchoscopic examination was performed after 1 race. Examinations were recorded on videotape, and presence and severity (grade 0 to 4) of EIPH were subsequently determined by 3 observers blinded to the horses' identity. Race records were abstracted for each horse examined. RESULTS: Overall, 52.1% of horses eligible for participation in the study were examined, and horses that were examined did not differ from horses that were not examined in regard to age, sex distribution, or proportion of horses that won or finished in the first 3 positions. Horses with EIPH grades < 1 were 4.0 times as likely to win, 1.8 times as likely to finish in the first 3 positions, and 3.03 times as likely to be in the 90th percentile or higher for race earnings as were horses with grades > 2. Horses with EIPH grades > 1 finished significantly farther behind the winner than did horses without EIPH. However, odds that horses with grade 1 EIPH would win or finish in the first 3 positions were not significantly different from odds for horses without EIPH. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that EIPH is associated with impaired performance in Thoroughbred racehorses not medicated with furosemide and not using nasal dilator strips.

Comparison of the odds of isolation, genotypes, and in vivo production of major toxins by Clostridium perfringens obtained from the gastrointestinal tract of dairy cows with hemorrhagic bowel syndrome or left-displaced abomasum.

OBJECTIVE: To compare the frequency of isolation, genotypes, and in vivo production of major lethal toxins of Clostridium perfringens in adult dairy cows affected with hemorrhagic bowel syndrome (HBS) versus left-displaced abomasum (LDA). DESIGN: Case-control study. ANIMALS: 10 adult dairy cattle with HBS (cases) and 10 adult dairy cattle with LDA matched with cases by herd of origin (controls). PROCEDURE: Samples of gastrointestinal contents were obtained from multiple sites during surgery or necropsy examination. Each sample underwent testing for anaerobic bacteria by use of 3 culture methods. The genotype of isolates of C. perfringens was determined via multiplex polymerase chain reaction assay. Major lethal toxins were detected by use of an ELISA. Data were analyzed with multivariable logistic regression and chi2 analysis. RESULTS: C. perfringens type A and type A with the beta2 gene (A + beta2) were the only genotypes isolated. Isolation of C. perfringens type A and type A + beta2 was 6.56 and 3.3 times as likely, respectively, to occur in samples from cattle with HBS than in cattle with LDA. Alpha toxin was detected in 7 of 36 samples from cases and in 0 of 32 samples from controls. Beta2 toxin was detected in 9 of 36 samples from cases and 0 of 36 samples from controls. CONCLUSIONS AND CLINICAL RELEVANCE: C. perfringens type A and type A + beta2 can be isolated from the gastrointestinal tract with significantly greater odds in cattle with HBS than in herdmates with LDA. Alpha and beta2 toxins were detected in samples from cows with HBS but not from cows with LDA.

Tracheobronchoscopic assessment of exercise-induced pulmonary hemorrhage in horses.

OBJECTIVE: To determine the interobserver variability of assessment of exercise-induced pulmonary hemorrhage (EIPH) during tracheobronchoscopic examination in horses. ANIMALS: 747 Thoroughbred racehorses. Procedures-850 tracheobronchoscopic examinations were performed within 2 hours of racing for the horses. Examinations were recorded on videotape, and EIPH and its severity were assessed independently by 3 veterinarians. Concordance was determined by calculation of the Cohen weighted kappa statistic and tabulation of scores assigned by each observer. RESULTS: Weighted kappa statistics ranged from 0.75 to 0.80. In 99.4% of observations, all observers agreed or 2 of 3 agreed and the third differed by < or = 1 grade. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that interobserver reliability of tracheobronchoscopic assessment of EIPH in Thoroughbred racehorses is high when the examination is conducted by experienced veterinarians. Concordance among investigators is sufficient to justify use of this grading system for further studies and clinical descriptions of EIPH.

Pulmonary ultrasonographic abnormalities associated with naturally occurring equine influenza virus infection in standardbred racehorses.

The purpose of this investigation was to determine if naturally occurring acute infectious upper respiratory disease (IRD) caused by equine influenza virus is associated with ultrasonographically detectable pleural and pulmonary abnormalities in horses. Standardbred racehorses were evaluated for signs of IRD, defined as acute coughing or mucopurulent nasal discharge. For every horse with IRD (n = 16), 1 or 2 horses with no signs of IRD and the same owner or trainer (n = 30) were included. Thoracic ultrasonography was performed within 5-10 days of the onset of clinical disease in horses with IRD. Horses without IRD were examined at the same time as the horses with IRD with which they were enrolled. The rank of the ultrasound scores of horses with IRD was compared to that of horses without IRD. Equine influenza virus was identified as the primary etiologic agent associated with IRD in this study. Mild lung consolidation and peripheral pulmonary irregularities were found in 11 (69%) of 16 of the horses with IRD and 11 (37%) of 30 of control horses. Lung consolidation (median score = 1) and peripheral irregularities scores (median score = 1) were greater in horses with IRD compared to horses without IRD (median score = 0; P < .05). Pleural effusion was not observed. Equine influenza virus infection can result in abnormalities of the equine lower respiratory tract. Despite the mild nature of IRD observed in this study, lung consolidation and peripheral pulmonary irregularities were more commonly observed in horses with clinical signs of IRD. Further work is needed to determine the clinical significance of these ultrasonographic abnormalities.

Survey of complications and antimicrobial use in equine patients at veterinary teaching hospitals that underwent surgery because of colic.

OBJECTIVE: To determine current practices regarding use of antimicrobials in equine patients undergoing surgery because of colic at veterinary teaching hospitals. DESIGN: Survey. SAMPLE POPULATION: Diplomates of the American College of Veterinary Surgeons performing equine surgery at veterinary teaching hospitals in the United States. PROCEDURE: A Web-based questionnaire was developed, and 85 surgeons were asked to participate. The first part of the survey requested demographic information and information about total number of colic surgeries performed at the hospital, number of colic surgeries performed by the respondent, and whether the hospital had written guidelines for antimicrobial drug use. The second part pertained to nosocomial infections. The third part provided several case scenarios and asked respondents whether they would use antimicrobial drugs in these instances. RESULTS: Thirty-four (40%) surgeons responded to the questionnaire. Respondents indicated that most equine patients undergoing surgery because of colic at veterinary teaching hospitals in the United States received antimicrobial drugs. Drugs that were used were similar for the various hospitals that were represented, and for the most part, the drugs that were used were fairly uniform irrespective of the type of colic, whereas the duration of treatment varied with the type of colic and the surgical findings. The combination of potassium penicillin and gentamicin was the most commonly used treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study document the implementation of recommendations by several authors in veterinary texts that antimicrobial drugs be administered perioperatively in equine patients with colic that are undergoing surgery. However, the need for long-term antimicrobial drug treatment in equine patients with colic is unknown.