Treatment success in cats with chronic enteropathy is associated with a decrease in fecal calprotectin concentrations.

Feline chronic enteropathies (FCE) are challenging to diagnose and monitor for progression and response to treatment. Fecal calprotectin might be a useful non-invasive marker to evaluate clinical endpoints of therapeutic monitoring in FCE. We evaluated fecal calprotectin concentrations in cats with FCE before and after initiation of treatment comprised of immunomodulation and/or dietary intervention. Included were 17 cats with FCE and 18 healthy controls. Clinical investigation of FCE cases included clinical severity grading (feline chronic enteropathy activity index, FCEAI) in all cats, abdominal ultrasonography in 15 cats, and gastrointestinal biopsies in 6 cats. Fecal calprotectin was measured in samples from 12 cats with FCE before treatment, all 17 FCE cats ≥6 weeks after treatment initiation, and all healthy controls. Fecal calprotectin concentrations in FCE cases before treatment (median: 61 µg/g) were significantly higher than after treatment initiation (median: 15 µg/g; p = 0.0098) and compared to controls (median: 6 µg/g; p = 0.0235) and correlated with the FCEAI scores (ρ = 0.54, p = 0.0316). Fecal calprotectin concentrations after treatment initiation were higher with more severe duodenal/proximal jejunal pathology (ρ = 0.83, p = 0.0427) and shorter intervals between sampling time points (ρ = -0.54, p = 0.0250). Relevant decreases in initially increased fecal calprotectin concentrations are seen in cats with FCE on varying treatment strategies that significantly improve or have remission of clinical signs. This supports the utility of fecal calprotectin as a surrogate biomarker to assess disease severity in FCE cases. Further studies need to evaluate fecal calprotectin concentrations longitudinally in relation to mucosal healing vs. clinical response.

Fecal Calprotectin Concentrations in Cats with Chronic Enteropathies.

Diagnosis of feline chronic inflammatory enteropathies (CIE) and the differentiation from small cell intestinal lymphoma (SCL) can be challenging. Intestinally expressed calprotectin (S100A8/A9 protein complex) appears to be part of the complex pathogenesis of feline chronic enteropathies (FCE). Fecal calprotectin is a non-invasive biomarker for intestinal inflammation in humans and dogs but has not yet been evaluated in cats. We hypothesized that fecal calprotectin (fCal) concentrations are increased in FCE, correlate with clinical and/or histologic disease severity, and distinguish cases of CIE from SCL. This case-control study included fecal samples and patient data from cats with CIE (n = 34), SCL (n = 17), other gastrointestinal (GI) diseases (n = 16), and cats with no clinical signs of GI disease (n = 32). fCal concentrations were measured using the immunoturbidimetric fCal turbo assay (Bühlmann Laboratories). Compared to healthy cats, fCal concentrations were significantly increased in CIE, SCL, and other diseases (all p < 0.0001), but were not different between these three groups (all p > 0.05), or between cats with extra-GI diseases and healthy controls. These findings suggest that fCal may have utility as a clinical biomarker for FCE but not for intestinal disease differentiation. It further supports the role of calprotectin in the pathogenesis of the spectrum of FCE, which includes CIE and SCL.

Verification of the fCAL turbo immunoturbidimetric assay for measurement of the fecal calprotectin concentration in dogs and cats.

The concentration of calprotectin in feces (fCal) is a clinically useful marker of chronic gastrointestinal inflammation in humans and dogs. No commercial assay is widely available to measure fCal in small animal medicine, to date. Thus, we verified the immunoturbidimetric fCAL turbo assay (Bühlmann) of fCal for canine and feline fecal extracts by determining linearity, spiking and recovery, and intra-assay and inter-assay variability. We determined RIs, temporal variation over 3 mo, and effect of vaccination and NSAID treatment. Observed:expected (O:E) ratios (x̄ ± SD) for serial dilutions of feces were 89-131% (106 ± 9%) in dogs and 77-122% (100 ± 12%) in cats. For spiking and recovery, the O:E ratios were 90-118% (102 ± 11%) in dogs and 83-235% (129 ± 42%) in cats. Intra- and inter-assay CVs for canine samples were ≤19% and ≤7%, and for feline samples ≤22% and ≤21%. Single-sample RIs were <41 µg/g for dogs and <64 µg/g for cats. With low reciprocal individuality indices, using population-based fCal RIs is appropriate, and moderate fCal changes between measurements (dogs 44.0%; cats: 43.2%) are considered relevant. Cats had significant (but unlikely relevant) fCal increases post-vaccination. Despite individual fCal spikes, no differences were seen during NSAID treatment. The fCAL turbidimetric assay is linear, precise, reproducible, and sufficiently accurate for measuring fCal in dogs and cats. Careful interpretation of fCal concentrations is warranted in both species during the peri-vaccination period and for some patients receiving NSAID treatment.

Synovial Fluid Analysis and Microscopic Assessment of Macrophage Quantities and Morphology in Equine Septic Arthritis.

OBJECTIVE: Research and provision of data on macrophages by cytological synovial fluid analysis and light microscopy in horses with septic arthritis MATERIAL AND METHODS: Records of 167 synovial fluid samples were evaluated and subdivided into different groups: (1) non-septic, (2) haematogenous septic arthritis in foals and (3) traumatic/iatrogenic septic arthritis. The effect of joint lavage on synovial fluid cytology and on the occurrence of macrophage phenotypes was investigated. RESULTS: Regardless of aetiology and age of the horse, macrophage concentrations in synovial sepsis are decreased to a median of 5-6 % (unaffected joints: 23.5 %) and further diminished by joint lavage. Microscopic assessment led to the identification of 4 phenotypes. Morphological characteristics of type 1 showed similarities to monocytes and predominated in unaffected and in septic joints after lavage. CONCLUSION AND CLINICAL RELEVANCE: Macrophages are highly versatile by altering their phenotype. A morphological assessment by light microscopy is easily applicable. Type 1 presumably contributes to joint homeostasis.

Synbiotic-IgY Therapy Modulates the Mucosal Microbiome and Inflammatory Indices in Dogs with Chronic Inflammatory Enteropathy: A Randomized, Double-Blind, Placebo-Controlled Study.

Chronic inflammatory enteropathy (CE) is a common cause of persistent gastrointestinal signs and intestinal inflammation in dogs. Since evidence links dysbiosis to mucosal inflammation, probiotics, prebiotics, or their combination (synbiotics) may reduce intestinal inflammation and ameliorate dysbiosis in affected dogs. This study's aim was to investigate the effects of the synbiotic-IgY supplement on clinical signs, inflammatory indices, and mucosal microbiota in dogs with CE. Dogs with CE were enrolled in a randomized prospective trial. Twenty-four client-owned dogs were fed a hydrolyzed diet and administered supplement or placebo (diet) for 6 weeks. Dogs were evaluated at diagnosis and 2- and 6-week post-treatment. Outcome measures included clinical activity, endoscopic and histologic scores, inflammatory markers (fecal calprotectin, C-reactive protein), and composition of the mucosal microbiota via FISH. Eleven supplement- and nine placebo-treated dogs completed the trial. After 6 weeks of therapy, clinical activity and endoscopic scores decreased in both groups. Compared to placebo-treated dogs, dogs administered supplement showed decreased calprotectin at 2-week post-treatment, decreased CRP at 2- and 6-week post-treatment increased mucosal Clostridia and Bacteroides and decreased Enterobacteriaceae in colonic biopsies at trial completion. Results suggest a beneficial effect of diet and supplements on host responses and mucosal microbiota in dogs with CE.

Malnutrition as the cause of recumbency in suckler cows associated with Trypanosoma theileri infection.

BACKGROUND: Recumbent cows are a diagnostic challenge because of a wide range of differential diagnoses, which include trauma, neurological and metabolic disorders, malnutrition and mineral deficiencies. This case report describes recumbent suckler cows that presented as a herd problem. In addition to weakness due to inanition, Cu and Se deficiencies were considered as possible aetiologies of the recumbency. Furthermore, Trypanosoma (T.) theileri, a blood parasite of unknown importance in Germany, was detected in the blood of some cows. CASE PRESENTATION: Three recumbent cows were referred to the Clinic for Ruminants and Swine, Faculty of Veterinary Medicine of the University of Leipzig. They were unable to rise and had low body condition scores and rough hair coats. Haematological and serum biochemical analyses showed neutrophilia, electrolyte imbalances, increased activities of muscle and liver enzymes and decreased concentrations of trace elements, especially Copper (Cu) and Selenium (Se). T. theileri was detected in a routine blood smear from one cow. The cows did not respond to an intensive care protocol, which included intravenous fluids and electrolytes, mineral substitution, non-steroidal anti-inflammatories and antibiotics, and were therefore euthanized or died. Postmortem examination showed cachexia, subcutaneous and scleral oedema and muscular dystrophy, especially in the hind limbs. Follow-up examination of the herd of origin produced similar findings including the detection of T. theileri in a large proportion of the herd. Ration analysis revealed considerable undersupply of several nutrients. CONCLUSIONS: Based on all findings, an aetiological diagnosis of trace mineral and nutrient deficiency with possible involvement of T. theileri was made.

Proline-rich antimicrobial peptide Api137 is bactericidal in porcine blood infected ex vivo with a porcine or human Klebsiella pneumoniae strain.

OBJECTIVES: Klebsiella pneumoniae is an emerging invasive pathogen in humans and pigs. Resistance against multiple antibiotics in this species is a major health concern and the development of new antibiotics is urgently needed. The objective of this study was to investigate the effects of proline-rich antimicrobial peptides (PrAMPs) on the survival of K. pneumoniae strains in porcine blood. METHODS: We established a bactericidal assay with K. pneumoniae in fresh blood drawn from 4-week-old piglets. PrAMPs, namely the apidaecins Api137 and Api802 as well as the oncocin Onc112, were added to ex vivo-infected whole blood samples in order to study their bactericidal effects and, in the case of Api137, also immune responses. RESULTS: A porcine invasive and a human iucA+rmpA+ K. pneumoniae strain showed prominent proliferation in porcine blood. Application of Api137 resulted in a dose-dependent prominent bactericidal effect killing the invasive porcine K. pneumoniae strain. Addition of 8 µg/mL Api137 also resulted in complete killing of the human iucA+rmpA+ strain. Cytotoxicity, haemolysis and induction of the pro-inflammatory cytokine tumour necrosis factor-alpha (TNFα) in K. pneumoniae-infected porcine blood treated with Api137 was comparable with values obtained after application of 10 µg/mL cefquinome. CONCLUSION: We describe a new non-rodent model for invasive K. pneumoniae bacteraemia and present promising data for the PrAMP Api137 for the control of infection with hypervirulent K. pneumoniae strains.

[Validation of a cow-side blood test for beta-hydroxybutyrate concentration in dairy cows]. / Überprüfung eines Schnelltestsystems zur Erfassung der Betahydroxybutyrat-Konzentration im Blut von Milchkühen.

OBJECTIVE: Clinical ketosis is common during the dairy cows' transition period and is responsible for considerable economic loss. Early identification of cows with subclinical ketosis is the first step for maintaining the health and productivity of dairy cows. The goals of the study were two-fold: The first was to examine the usefulness of a mobile test device as a cow-side test; and the second was to compare BHB concentrations measured by the ketometer using capillary blood and blood collected from the coccygeal vessels with values determined by a reference method in the laboratory using jugular blood. MATERIAL AND METHODS: Blood samples were collected from a jugular vein or the coccygeal vessels in 81 dairy cows at 7 time points (14 and 7 days pre-partum and 7, 14, 21, 28, and 42 days post-partum) for kinetic enzymatic measurement of BHB concentration in the laboratory. Blood samples were concurrently collected from the coccygeal vessels or by pricking the vulvar lip at the transition of the skin to the mucosa (capillary blood) to determine BHB concentration using the WellionVet BELUA ketometer (MED TRUST GmbH, Marz, Austria). RESULTS: Initial errors in operating the ketometer were quickly eliminated with experience. BHB concentrations of jugular blood measured in the laboratory were 0.07 mmol/l lower than those measured in coccygeal blood. The mean BHB concentration measured in coccygeal and capillary blood using the WellionVet BELUA ketometer did not significantly differ but were 0.13 and 0.12 mmol/l respectively, lower than the mean jugular vein concentrations measured in the laboratory. CONCLUSION: The WellionVet BELUA ketometer is useful for determination of BHB concentration in cows provided that the manufacturer's specifications are followed. Capillary blood is best collected at the transition from the vulvar skin to its mucosa. The device generates rapid results that correlate well with BHB concentrations determined in the laboratory and with the results obtained from different blood collection sites. It is ideally suited for monitoring dairy cows for subclinical ketosis using capillary or coccygeal blood.

Clinical Chemistry Investigations in Recumbent and Healthy German Holstein Cows After the Fifth Day in Milk.

INTRODUCTION: Recumbency is a frequent symptom occurring throughout lactation. Its cause can be related to the energy or mineral metabolism, or to trauma or infectious diseases. We compared various clinical chemistry parameters between healthy and recumbent cows and between cows with different causes of recumbency and determined if hypocalcaemia manifests in later lactation. MATERIAL AND METHODS: Recumbent (n = 32) and healthy (n = 32) German Holstein cows were studied. After clinical examination, a serum sample was taken to measure the concentrations of Mg, Ca, Fe, Na, K, Pi, ß-hydroxybutyrate, total bilirubin, non-esterified fatty acids (NEFA), urea, and creatinine as well as activities of alkaline phosphatase, aspartate aminotransferase (AST), creatine kinase (CK), and γ-glutamyl transferase in recumbent cows > 5 d in milk and control cows matched for age, lactation number, and pregnancy stage. RESULTS: In recumbent cows, mean serum concentrations of NEFA, bilirubin, and CK were statistically higher, while those of Fe, K, and Pi were significantly lower. Parameters compared between different recumbency diagnoses showed some descriptive Fe, K, urea, and AST differences, but these were not statistically significant. CONCLUSION: The results show that only a limited number of parameters have diagnostic besides therapeutic value. Although of minor importance in our study, hypocalcaemia should be considered a cause of recumbency, even outside the typical risk period of parturient paresis.

Comparison of ELISA and capillary electrophoresis with laser-induced fluorescence detection in the analysis of Ochratoxin A in low volumes of human blood serum.

In this paper the determination of Ochratoxin A (OTA) in low volumes of human blood serum by enzyme-linked immunosorbent assay (ELISA) is compared with an appropriate capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) method. In order to use ELISA for high-throughput analysis in epidemiological studies no sample cleanup was performed. Both methods showed a limit of detection (LOD) of 0.5 ng/mL. Comparing the precisions of both methods, the data show that the quantified concentrations in ELISA are higher than the corresponding concentrations in the CE-LIF method. Using a matrix calibration curve instead of a standard calibration curve the reproducibilities of both methods are comparable. No additional matrix effect could be observed by adding phenylalanine as probable matrix compound to the serum.

[Pituitary syndrome caused by neoplasia in a 17-month-old Holstein Friesian heifer]. / Hirnbasissyndrom infolge eines Tumors bei einer 17 Monate alten Deutsch-Holstein-Färse.

A 17-month-old Holstein Friesian heifer was presented after one day with dysphagia, slight paralysis of the tongue and swelling of the eyelids. Clinical examination of the animal revealed an extended posture of the head and neck, severely increased salivation, reduced lingual tone and mandibular paralysis with complete absence of the swallowing reflex. The right eye showed a drooping eyelid, mucopurulent discharge, exposure keratitis, corneal opacity and miosis. On the left side, a moderate exophthalmos and slight mucous discharge from the nostril were observed. Neurological examination revealed the absence of multiple cranial nerve reflexes suggesting a pituitary syndrome. On X-rays, a soft-tissue opacity with sharp margins and a diameter of approximately 5 cm was seen. It was located ventral to the ethmoid bone with possible intraneurocranial origin. Rhinoscopically, a mass located distal to the ethmoid bone with an uneven, slightly reddish surface partly covered by purulent exudate was visualised. Post-mortem examination of the euthanized animal confirmed neoplasia, which ranged from the fossa hypophysialis of the corpus ossis basisphenoidalis to the ethmoid bone. Histopathologic findings matched a small, round, blue cell tumour.

Clinical evaluation of serum alcohol dehydrogenase activity in horses with acute intestinal obstruction.

OBJECTIVES: To measure serum alcohol dehydrogenase (ADH) activity in horses with acute intestinal obstruction and to determine the diagnostic and prognostic utility of this analyte. DESIGN: Prospective observational study. SETTING: University Veterinary Hospital. ANIMALS: Thirty healthy horses (control group) and 77 horses with acute intestinal obstruction, including 36 horses with nonstrangulating obstruction (23 with left ventral colon impaction and 13 with left dorsal displacement [G1], 22 with small intestinal strangulation [G2], and 19 with colon torsion [G3]). INTERVENTIONS: Serum ADH activity was assayed spectrophotometerically in all horses. Serum lactate concentration and hepatic enzyme (aspartate aminotransferase, gamma-glutamyl transferase, glutamate dehydrogenase) activities were measured using an automatic analyzer. MEASUREMENTS AND MAIN RESULTS: The median [interquartile range] serum ADH activity in healthy horses was 10.5 [8.7-11 U/L]. ADH activity was significantly increased (P<0.05) in G1=16.5 [13.8-18 U/L], G2=40 [20-74.9 U/L], and G3=63.2 [40-78 U/L] compared with healthy controls. Aspartate aminotransferase and glutamate dehydrogenase activities were also significantly increased in G3 in comparison with controls. ADH activity was correlated with serum lactate concentration in G1 and G3, respectively (P<0.01, r=0.55 and 0.8). Other liver enzymes did not show any significant correlation with lactate. ADH activity was directly related to the probability of strangulation; odds ratio=1.11. ADH activity >20 U/L had 80.6% specificity and 80.5% sensitivity for discriminating horses with strangulating obstruction. Twelve horses euthanized before surgery were excluded from the outcome analysis. Increasing ADH activity was associated with nonsurvival; odds ratio=1.03. ADH activity <80 U/L had 94.44% specificity and 66.67% sensitivity for survival. CONCLUSION: Serum ADH activity may be a useful clinical parameter in detecting intestinal strangulation in horses and may provide some prognostic value in horses with acute intestinal obstruction.

[Free hemoglobin and tumor necrosis factor-alpha in the blood of horses with colic or acute colitis]. / Freies Hämoglobin und Tumor-Nekrose-Faktor-alpha im Blut von Pferden mit Kolik oder akuter Kolitis.

A total of 50 adult horses were classified into 4 groups: healthy horses (group 1, n = 14), horses with non-strangulation obstruction (group 2, n = 13), horses with strangulation obstruction (group 3, n = 11) and horses with acute colitis and severe inflammation (peritonitis or thrombophlebitis) (group 4, n = 12). Eleven euthanized horses recreated from group 3 (4 horses with small intestinal strangulation obstruction and hemolytic peritoneal fluid) and group 4 (7 horses with septic peritonitis) were taken as group 5. Free hemoglobin (fr. Hb) and tumor necrosis factor-alpha (TNFa) were measured by method of Kahn and by Duo Set ELISA Development Kit. The concentrations of fr. Hb and TNF-a were significantly higher (p <0.01) in group 3 (medians: 8.0 micromol/l; 14665 pg/ml) and group 4 (9.1; 15081) in comparison to healthy horses (0.61; 573) and group 2 (0.5; 821), whereas group 2 showed no significant difference with group 1.There was a positive correlation between fr. Hb and TNF-alpha (r = 0.82; P = 0.01) in group 4. Concentrations of both fr. Hb and TNF-alpha were significantly (p <0,01) higher in euthanized horses of group 5 than in medically treated horses of group 2.The high concentration of fr. Hb could be a proinflammatory mediator to stimulate high production ofTNF-alpha known as a proinflammatory cytokine. Both of these parameters could contribute to the pathogenesis of systemic inflammatory response syndrome: fever, tachycardia, tachypnoe, leukopenia, thrombocytopenia, and lactic acidosis. Based on this, increased concentrations of both fr. Hb and TNF-alpha could be a guarded prognostic indicator in horses with strangulation obstruction, acute colitis or septic peritonitis.