Development of an ELISA to detect circulating anti‐asparaginase antibodies in dogs with lymphoid neoplasia treated with Escherichia coli l‐asparaginase

Resistance to Escherichia coli l ‐asparaginase in canine lymphoma occurs frequently with repeated administration, a phenomenon often attributed, without substantiation, to the induction of neutralizing antibodies. To test the hypothesis that treated dogs develop antibodies against the drug, we created an enzyme‐linked immunosorbent assay (ELISA) to measure plasma anti‐asparaginase immunoglobulin G responses. Using samples from dogs that had received multiple doses, specific reactivity against l ‐asparaginase was demonstrated, while naïve patients' samples were negative. The optimized ELISA appeared sensitive, with endpoint titers >1 600 000 in positive control dogs. Intra‐ and inter‐assay coefficients of variation were 3.6 and 14.5%. The assay was supported by the observation that ELISA‐positive plasma could immunoprecipitate asparaginase activity. When clinical patients were evaluated, 3/10 dogs developed titers after a single injection; with repeated administration, 4/7 dogs were positive. l ‐asparaginase antibodies showed reduced binding to the PEGylated drug formulation. The ELISA should prove useful in investigating the potential correlation of antibody responses with resistance.     

Comparison of the efficacy of two anti-uric acid diets in dalmatian dogs.

Commercial anti-uric acid diets for dogs may contain insufficient protein to sustain growth and lactation. In order to investigate the efficacy of an experimental purine-free diet moderately low in protein, its effect on urinary uric acid excretion was compared with that of a commercial dog food and a commercial low-protein anti-uric acid diet. The experimental diet, commercial dog food and commercial anti-uric acid diet contained 10.0, 12.8 and 5.0 g crude protein/MJ metabolizable energy, respectively. Twelve Dalmatian dogs were subjected to a 3 x 3 Latin square study. Although the plasma uric acid concentration was significantly lower when the dogs were fed either the commercial anti-uric diet (18.7 +/- 6.0 mumol/l, mean +/- SD, n = 12) or the experimental diet (19.2 +/- 8.3 mumol/l), when compared to the commercial dog food (29.2 +/- 11.1 mumol/l), no significant decrease of uric acid concentration in urine collected before the morning meal was seen. The average concentration of urinary uric acid was 60 mumol/l. There was a significant increase in the urea:creatinine ratio in urine when the dogs were fed the experimental diet compared to the commercial anti-uric acid diet, confirming that the experimental diet contained more protein. The experimental diet lowered plasma uric acid and was relatively high in protein and thus may be suitable for use in growing and lactating dogs to prevent ammonium urate urolithiasis.     

Analysis of serum proteins in clinically healthy goats (Capra hircus) using agarose gel electrophoresis

Background: Electrophoretic patterns of serum proteins provide useful information on pathological conditions in ruminants. Their reference values, however, are dissimilar to those of other species. Reference values for goats using agarose gel as the supporting matrix have not been reported. Objective: The aim of this study was to evaluate serum concentrations of total protein and protein fractions (albumin and globulins) by means of agarose gel electrophoresis (AGE) in goats in order to establish electrophoretic reference intervals and to evaluate potential changes associated with aging. Methods: Blood was collected from 105 clinically healthy Girgentana goats by means of jugular venipuncture. Serum protein concentrations were assessed by AGE. Three age groups were compared: 1–1.5 years, 2–4 years, and 5–12 years. Results: Values (mean ± SD) were determined for concentrations of total protein (72.26 ± 6.40 g/L), albumin (31.80 ± 4.00 g/L), α‐globulins (6.40 ± 1.23 g/L), β₁‐globulins (10.50 ± 2.58 g/L), β₂‐globulins (5.18 ± 1.60 g/L), and γ‐globulins (18.65 ± 5.90 g/L) and for albumin/globulin (A/G) ratio (0.82 ± 0.20). One‐way ANOVA showed statistically significant age‐related differences for total protein and α‐globulin concentrations and A/G ratios. Age influenced protein concentrations with the 5–12‐year‐old group having higher total protein and α‐globulin concentrations and lower albumin concentration and A/G ratios than the 2–4‐year‐old group. Conclusions: This study provides reference values for total protein concentrations and protein fractions obtained by AGE in goats. Some values vary with age. Age‐specific reference intervals are reported in order to provide clinicians with an additional diagnostic aid.     

Unusual IgM-secreting multiple myeloma in a dog

A 4-year-old castrated male dog was evaluated because of multiple-limb lameness. Signs of pain were elicited during palpation of the regions of the proximal tibial metaphyses and distal left radial diaphysis. Radiography revealed osteolytic lesions of the long bones. Blood analyses revealed hypercalcemia and transient cytopenias. Serum protein electrophoresis did not reveal a monoclonal gammopathy; however, urine protein electrophoresis revealed Bence Jones proteinuria. Serial diagnostic sampling of bone lesions, immunohistochemical staining methods, and serum and urine protein immunoelectrophoresis were required to establish a diagnosis of multiple myeloma. Two IgM components were identified via serum protein immunoelectrofixation. The dog improved clinically after initiation of chemotherapy with melphalan and prednisone; however, the dog ultimately was euthanatized because of pathologic fracture. The case was unique because there was lack of vertebral involvement, an unusual gammopathy, and difficulty in identifying myeloma cells via serial sampling.     

New biomarkers for increased intestinal permeability induced by dextran sodium sulphate and fasting in chickens

Increased intestinal permeability (IP) can lead to compromised health in chickens. As there is limited literature on in vivo biomarkers to assess increased IP in chickens, the objective of this study was to identify a reliable biomarker of IP using DSS ingestion and fasting models. Male Ross chickens (n = 48) were reared until day 14 on the floor pen in an animal care facility, randomized into the following groups: control, DSS and fasting (each with n = 16), and then placed in metabolism cages. DSS was administered in drinking water at 0.75% from days 16 to 21, while controls and fasted groups received water. All birds had free access to feed and water except the birds in the fasting group that were denied feed for 19.5 h on day 20. On day 21, all chickens were given two separate oral gavages comprising fluorescein isothiocyanate dextran (FITC‐d, 2.2 mg in 1 ml/bird) at time zero and lactulose, mannitol and rhamnose (LMR) sugars (0.25 g L, 0.05 g M and 0.05 g R in 2 ml/bird) at 60 min. Whole blood was collected from the brachial vein in a syringe 90 min post‐LMR sugar gavage. Serum FITC‐d and plasma LMR sugar concentrations were measured by spectrophotometry and high‐performance ion chromatography respectively. Plasma concentrations of intestinal fatty acid binding protein, diamine oxidase, tight junction protein (TJP), d ‐lactate and faecal α‐antitrypsin inhibitor concentration were also analysed by ELISA. FITC‐d increased significantly (p < 0.05) after fasting compared with control. L/M and L/R ratios for fasting and L/M ratio for DSS increased compared with control chickens (p < 0.05). TJP in plasma was significantly increased due to fasting but not DSS treatment, compared with controls. Other tests did not indicate changes in IP (p > 0.05). We concluded that FITC‐d and LMR sugar tests can be used in chickens to assess changes in IP.     

Effect of dosage and application mode of l‐carnitine on plasma lipid and egg‐yolk cholesterol of turkeys,hatchability of eggs and post‐hatch growth of their offsprings

The effect of dosage and application mode of l ‐carnitine on plasma lipid and egg‐yolk cholesterol of breeder turkeys, hatchability of eggs and post‐hatch growth response was investigated using 180 breeder hens. The hens were assigned to six dietary treatments in a 2 × 3 factorial arrangements of two application modes of l ‐carnitine (diet and drinking water) supplemented at 0, 50 and 100 ppm (mg/kg or mg/l) levels, respectively. Each treatment was replicated five times with six hens per replicate. Dietary inclusion of 50 ppm l ‐carnitine showed the lowest (p < 0.01) plasma total cholesterol (TC) and low‐density lipoprotein concentration (LDL). Breeder hens offered 50 ppm l ‐carnitine with no regard to application mode recorded the highest (p < 0.01) plasma high‐density lipoprotein (HDL). Hens offered 50 and 100 ppm l ‐carnitine irrespective of application mode also showed reduced (p < 0.01) egg‐yolk TC concentration at 32 weeks of age. Dietary supplementation of 50 ppm l ‐carnitine for breeder turkeys recorded the lowest (p < 0.01) egg‐yolk triglyceride (TG) at 40 weeks of age. Hens offered 50 ppm l ‐carnitine irrespective of application mode recorded the highest (p < 0.05) hen‐day egg production. Incidence of dead‐in‐shell also reduced (p < 0.05) with increasing dosage of l ‐carnitine. Dietary supplementation of 50 ppm and oral application in drinking water of 100 ppm l ‐carnitine for breeder turkeys resulted in highest (p < 0.05) egg fertility. Offsprings from breeder hens fed diets supplemented with l ‐carnitine recorded no post‐hatch mortality. Highest (p < 0.05) post‐hatch final live weight and weight gain was obtained with poults obtained from hens fed diet supplemented with 50 ppm l ‐carnitine. In conclusion, dietary supplementation of 50 ppm l ‐carnitine for turkey hens showed improved serum lipid profile, egg fertility, reduced dead‐in‐shell, egg‐yolk cholesterol and resulted in improved post‐hatch growth performance.     

l‐carnitine and fat type in the maternal diet during gestation and lactation modify the fatty acid composition and expression of lipid metabolism‐related genes in piglets

This study examined the influence of adding different amounts of maternal dietary l ‐carnitine and two fat types on fatty acid (FA) composition and the expression of lipid metabolism‐related genes in piglets. The experiment was designed as a 2 × 2 factorial with two fat types (3.5% soyabean oil, SO, and 3.5% fish oil, FO) and two levels of l ‐carnitine (0 and 100 mg/kg) added to the sows' diets. A higher proportion of n‐3 polyunsaturated fatty acids (PUFA) and a lower ratio of n‐6/n‐3 PUFA in sow milk and piglet tissues were observed in the FO groups than in the SO groups. Adding l ‐carnitine increased the proportion of C16:1 in sow milk and decreased n‐3 PUFA in piglet subcutaneous fat. Hepatic peroxisome proliferator‐activated receptor α (PPAR‐α) was more abundantly expressed in piglets from the FO groups than from the SO groups (p < 0.05), whereas stearoyl‐CoA‐desaturase (SCD), sterol regulatory element binding protein‐1 (SREBP1) and ?6‐desaturase (D6D) genes were less expressed in the FO groups compared with piglets from the SO groups. The expression of fatty acid synthase (FAS) genes was decreased in the SO groups with l ‐carnitine compared to that of the other dietary treatments. No differences among dietary treatments were observed with regard to the expression of acetyl‐CoA carboxylase (ACC). In conclusion, FO and l ‐carnitine supplementation in sows affect FA composition and hepatic gene expression in piglets.     

Predictive value of hypoglycin A and methylencyclopropylacetic acid conjugates in a horse with atypical myopathy in comparison to its cograzing partners

Hypoglycin A (HGA) was detected in blood and urine of a horse suffering from atypical myopathy (AM; Day 2, serum, 8290 μg/l; urine: Day 1, 574, Day 2, 742 μg/l) and in its cograzing partners with a high variability (46–1570 μg/l serum). Over the period of disease, the level of the toxic metabolites (methylencyclopropylacetic acid [MCPA]‐conjugates) increased in body fluids of the AM horse (MCPA‐carnitine: Day 2, 0.246, Day 3, 0.581 μmol/l serum; MCPA‐carnitine: Day 2, 0.621, Day 3, 0.884 μmol/mmol creatinine in urine) and HGA decreased rapidly (Day 3, 2430 μg/l serum). In cograzing horses MCPA‐conjugates were not detected. HGA in seeds ranged from 268 to 367 μg/g. Although HGA was present in body fluids of healthy cograzing horses, MCPA‐conjugates were not detectable, in contrast to the AM horse. Therefore, increasing concentrations of MCPA‐conjugates are supposed to be linked with the onset of AM and both parameters seem to indicate the clinical stage of disease. However, detection of HGA in body fluids of cograzing horses might be a promising step in preventing the disease.     

Serum protein concentrations from clinically healthy horses determined by agarose gel electrophoresis

Background: Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and α‐, β‐, and γ‐globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported. Objectives: The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm‐blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined. Methods: Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi‐automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within‐run and within‐assay precision. Data from warm‐blooded and draught horses were compared using the Mann–Whitney U test. Results: Within‐run and within‐assay CVs were <5% for all protein fractions. No significant difference was found between warm‐blooded and heavy draught horses and so combined reference intervals (2.5–97.5%) were calculated for total protein (51.0–72.0 g/L), albumin (29.6–38.5 g/L), α₁‐globulin (1.9–3.1 g/L), α₂‐globulin (5.3–8.7 g/L), β₁‐globulin (2.8–7.3g/L), β₂‐globulin (2.2–6.0 g/L), and γ‐globulin (5.8–12.7 g/L) concentrations, and albumin/globulin ratio (0.93–1.65). Conclusion: Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions.     

Application of the slope‐ratio growth assay technique to estimate tryptophan availability in soybean meal fed to young rats

The slope‐ratio assay for rat was used to determine whether tryptophan (Trp) availability in soybean meal (SBM) is affected by the presence of other amino acids (AAs). In a preliminary study, rats were fed graded levels of Trp‐supplemented diets to establish the Trp concentration range over which the weight gain response was linear. This range was found to be from 0.04% to 0.12% Trp. Subsequently, rats were fed basal (0.045% Trp) or Trp‐supplemented diets from three different sources: l ‐Trp alone, SBM, or l ‐Trp mixed with other AAs to reflect AA levels in the test SBM (AA‐mix). Weight gain in rats increased linearly with supplemental Trp intake (p < .05) for all Trp sources. Compared to the slope achieved with l ‐Trp alone, the estimated availability of Trp in SBM was 84.4%, while for the AA‐mix it was 93.4%. It is evident that the 6.6% reduction in l ‐Trp availability in AA‐mix is due to metabolic costs derived from excess levels of other AAs beside Trp, given that the absorption of crystalline l ‐Trp in the small intestine is 100%. In conclusion, the Trp availability of SBM was estimated to be around 90.4% (i.e., 84.4/93.4 × 100) after correcting for the effects of the other AAs in SBM.     

Plasma concentration‐dependent suppression of endogenous hydrocortisone in the horse after intramuscular administration of dexamethasone‐21‐isonicotinate

Detection times and screening limits (SL) are methods used to ensure that the performance of horses in equestrian sports is not altered by drugs. Drug concentration–response relationship and knowledge of concentration–time profiles in both plasma and urine are required. In this study, dexamethasone plasma and urine concentration–time profiles were investigated. Endogenous hydrocortisone plasma concentrations and their relationship to dexamethasone plasma concentrations were also explored. A single dose of dexamethasone‐21‐isonicotinate suspension (0.03 mg/kg) was administered intramuscularly to six horses. Plasma was analysed for dexamethasone and hydrocortisone and urine for dexamethasone, using UPLC‐MS/MS. Dexamethasone was quantifiable in plasma for 8.3 ± 2.9 days (LLOQ: 0.025 μg/L) and in urine for 9.8 ± 3.1 days (LLOQ: 0.15 μg/L). Maximum observed dexamethasone concentration in plasma was 0.61 ± 0.12 μg/L and in urine 4.2 ± 0.9 μg/L. Terminal plasma half‐life was 38.7 ± 19 h. Hydrocortisone was significantly suppressed for 140 h. The plasma half‐life of hydrocortisone was 2.7 ± 1.3 h. Dexamethasone potency, efficacy and sigmoidicity factor for hydrocortisone suppression were 0.06 ± 0.04 μg/L, 0.95 ± 0.04 and 6.2 ± 4.6, respectively. Hydrocortisone suppression relates to the plasma concentration of dexamethasone. Thus, determination of irrelevant plasma concentrations and SL is possible. Future research will determine whether hydrocortisone suppression can be used as a biomarker of the clinical effect of dexamethasone.     

Effect of l‐glutamic acid supplementation on performance and nitrogen balance of broilers fed low protein diets

The aim of this study was to evaluate the effect of protein reduction and supplementation of l ‐glutamic acid in male broiler diets. A total of 648 chicks of the Cobb 500 strain were distributed in a completely randomized design with six treatments and six replications with eighteen birds per experimental unit. The study comprised pre‐starter (1–7 days), starter (8–21 days), growth (22–35 days) and final (36–45 days) phases. The first treatment consisted of a control diet formulated according to the requirements of essential amino acids for each rearing phase. The second and third treatments had crude protein (CP) reduced by 1.8 and 3.6 percentage points (pp) in relation to the control diet respectively. In the fourth treatment, l ‐glutamic acid was added to provide the same glutamate level as the control diet, and in the last two treatments, the broilers were supplemented with 1 and 2 pp of glutamate above that of the control diet respectively. The reduction in CP decreased the performance of broilers and the supplementation of l ‐glutamic acid did not influence performance when supplied in the diets with excess of glutamate. The lowest excreted nitrogen values were observed in the control diet, and treatments 2 and 3, respectively, in comparison with treatments with the use of l ‐glutamic acid (5 and 6). Retention efficiency of nitrogen was better in the control diet and in the treatment with a reduction of 1.8 pp of CP. It was verified that the serum uric acid level decreased with the CP reduction. A reduction in CP levels of up to 21.3%, 18.8%, 18.32% and 17.57% is recommended in phases from 1 to 7, 8 to 21, 22 to 35 and at 36 to 42 days, respectively, with a level of glutamate at 5.32%, 4.73%, 4.57%, 4.38%, also in these phases.     

Effect of L‐theanine on meat quality,muscle amino acid profiles,and antioxidant status of broilers

This study investigated the effect of L ‐theanine on carcass traits, meat quality, muscle antioxidant capacity, and amino acid (AA) profiles of broilers. Three hundred 1‐day‐old Ross 308 male broilers were randomly allotted to five groups with six replicates. Birds were fed the basal diet or basal diet with 300, 600, 900, or 1,500 mg/kg L ‐theanine for 42 consecutive days. The results showed that L ‐theanine quadratically increased dressing percentage, eviscerated percentage, and leg muscle yield (p < .05). Meanwhile, drip loss, cooking loss, shear force, L*_24h, and muscle lactate content decreased quadratically in response to dietary L ‐theanine supplementation (p < .05), while pH_24h and muscle glycogen content were quadratically improved by L ‐theanine (p < .05). Notably, the contents of muscle malondialdehyde and protein carbonyl, and the activities of muscle total antioxidant capacity, catalase, and glutathione peroxidase decreased quadratically in response to dietary L ‐theanine supplementation (p < .05), suggesting that the oxidative stress level of muscle was decreased quadratically. Moreover, L ‐theanine quadratically increased the concentrations of most of muscle essential AA, nonessential AA, and flavor AA (p < .05). In conclusion, L ‐theanine can be used as a valuable feed additive to modulate carcass traits, meat quality, muscle antioxidant status, and AA profiles of boilers, and its optimum addition level is 600 mg/kg based on the present study.     

Serum protein electrophoresis values for free-ranging and zoo-based koalas (Phascolarctos cinereus)

In a clinical setting, especially with species of special interest, it is important to use all clinical pathology testing options for general health monitoring and diagnosis. Protein electrophoresis (EPH) has previously been shown to be an important adjunct tool in veterinary medicine. Serum samples from 18 free-ranging and 12 zoo-based koalas (Phascolarctos cinereus) were subject to EPH analysis. Significant differences were found between the two groups for the following values: total protein, albumin, beta globulins, and albumin-globulin ratio (P < 0.05). By using the combined data, the minimum-maximum values for the EPH fractions were as follows: total protein 5.0-7.8 g/dl, albumin 2.8-4.7 g/dl, alpha-1 globulins 0.5-1.1 g/dl, alpha-2 globulins 0.3-0.7 g/dl, beta globulins 0.4-1.0 g/dl, gamma globulins 0.2-1.0 g/dl, and albumin-globulin ratio 1.0-2.1.     

Cutaneous T‐cell lymphoma – Sézary syndrome in a Boxer

A 7‐year‐old male Boxer with a 3.5‐year history of atopy and food hypersensitivity was presented with multiple poorly circumscribed nodules and maculae of the skin and tongue, and jaundiced mucosal membranes. Cytologic and histopathologic examination of the skin lesions revealed cutaneous epitheliotropic lymphoma. Cells were CD3⁺ and CD8⁺ in flow cytometry. The CBC showed a moderate leukocytosis with 16% atypical lymphocytes with irregularly cleaved nuclei. Flow cytometric phenotyping of peripheral blood showed an elevated proportion of the CD8⁺ T‐lymphocyte subpopulation, indicating a malignant population of T‐cell origin, and the electropherogram of the PCR antigen receptor rearrangement produced a monoclonal peak for TCRγ. Liver enzyme activities were markedly increased and abdominal ultrasound examination showed increased echogenicity of the liver and enlarged abdominal lymph nodes. Fine‐needle aspirates of the liver confirmed infiltration with lymphocytes exhibiting the same morphology as the cells detected in skin and peripheral blood. Treatment was induced with L‐asparaginase, lomustine, and prednisone. Partial clinical remission of the skin and tongue lesions was achieved within 10 days, and hematologic abnormalities resolved. Despite further treatment with L‐asparaginase and lomustine, the dog relapsed within one month and was euthanized. Presence of malignant lymphocytes in skin, peripheral blood, and liver indicate a rare variant of leukemic cutaneous T‐cell lymphoma, equivalent of Sézary syndrome in a dog. This case report describes the use of flow cytometry as a complementary tool for lymphocyte characterization of skin lesions for the first time.     

Dystrophin‐deficient muscular dystrophy in a Norfolk terrier

A six‐month‐old male entire Norfolk terrier was presented with a 3‐month history of poor development, reluctance to exercise and progressive and diffuse muscle atrophy. Serum creatine kinase concentration was markedly elevated. Magnetic resonance imaging of the epaxial muscles revealed asymmetrical streaky signal changes aligned within the muscle fibres (hyperintense on T2‐weighted images and short‐tau inversion recovery with moderate contrast enhancement on T1‐weighted images). Electromyography revealed pseudomyotonic discharges and fibrillation potentials localised at the level of the supraspinatus, epaxial muscles and tibial cranialis muscles. Muscle biopsy results were consistent with dystrophin‐deficient muscular dystrophy. The dog remained stable 7 months after diagnosis with coenzyme Q10 and l ‐carnitine; however after that time, there was a marked deterioration and the owners elected euthanasia. This case report describes the clinical presentation, magnetic resonance imaging, electrodiagnostic and histopathological findings with immunohistochemical analysis in a Norfolk terrier with confirmed dystrophin‐deficient muscular dystrophy, which has not been previously described in this breed.     

Chemical composition of alfalfa silage with waste date and its feeding effect on ruminal fermentation characteristics and microbial protein synthesis in sheep

This study was conducted to evaluate the effect of feeding ensiled alfalfa with waste date on ruminal fermentation characteristics, microbial protein synthesis, protozoa population and blood parameters in sheep. Eight rams were used in a 2 × 2 change over design. Each experimental period consisted of 21 days including 16 days for adaptation and 5 days for sampling. For ensiling, fresh alfalfa (Medicago sativa L.) with different levels of waste date (Phoenix dactylifera L.) were mixed together and ensiled in 100‐l containers for 45 days. Chemical composition of silages such as dry matter (DM), crude protein, NH₃‐N, organic matter, NDF, ADF and pH were determined. Then, it was used as 30% (DM basis) in diets. The experimental diets were as follows: (i) control (diet containing alfalfa silage without waste date), (ii) diet containing alfalfa silage with 5 g waste date/95 g DM, (iii) diet containing alfalfa silage with 10 g waste date/90 g DM, and (iv) diet containing alfalfa silage with 15 g waste date/85 g DM. The results of this experiment showed that adding waste date to alfalfa during ensiling, improved silage quality, DM and energy level. Total protozoa population and all of holotrich, cellulolytic and entodinia in rumen fluid were increased linearly by increasing the level of waste date. Nitrogen (N) intake, urinary N excretion and nitrogen retention were affected by dietary treatments. Also, allantoin, uric acid, total purine derivatives, microbial protein synthesis, cholesterol level and blood urea nitrogen were significantly different. In conclusion, direct ensilage of alfalfa can be attained by mixing 15 g waste date/85 g DM and positive associative effects such as increased metabolizable energy (ME) and silage quality occurred.     

Successful Treatment of Disseminated Nocardiosis Caused by Nocardia veterana in a Dog

A 5‐year‐old male castrated Lhasa Apso cross was evaluated for a 1‐month history of inappetence, lethargy, gagging, and progressive right thoracic limb lameness. Synovial fluid analysis revealed nonseptic suppurative inflammation, and a diagnosis of immune‐mediated polyarthritis (IMPA) was made. After 3 months of treatment with prednisone and later cyclosporine, the dog developed multiple firm cutaneous and subcutaneous masses and a focal mass within the jejunum. Cultures of blood, urine, skin lesions, and the jejunal mass identified Nocardia veterana by matrix‐absorption laser desorption ionization‐time‐of‐flight mass spectrometry (MALDI‐TOF MS) and allowed for earlier identification of the organism compared to more traditional secA1 gene sequencing. Immunosuppressive drug treatment was discontinued, and the dog was treated for 3 months by administration of trimethoprim‐sulfamethoxazole (TMS). No recurrence of clinical signs was reported 1 year later. This case report highlights the clinical utility of MALDI‐TOF MS, particularly for the rapid identification of slow‐growing, fastidious organisms.     

Proof-of-concept study: Evaluation of plasma and urinary electrolytes as markers of response to L-asparaginase therapy in dogs with high-grade lymphoma

Response to chemotherapy is one of the most important prognostic factors in dogs with lymphoma. The objective of this feasibility study was to evaluate if clinical responses to a specific cytotoxic agent (L-asparaginase) could be anticipated by measuring analyte concentrations in plasma and urine concentrations of lymphoma-bearing dogs. We hypothesized that potassium and phosphate concentrations in plasma and urine would be higher in dogs that completely responded to therapy. Plasma and urine samples of dogs with lymphoma were obtained before 12 and 24 hours after intramuscular L-asparaginase injections. Peripheral lymph node volumes were evaluated according to the Veterinary Cooperative Oncology Group standardized criteria. Plasma and urine electrolyte, calcium, phosphate, creatinine, urea, total protein, and albumin concentrations were measured, and the fractional excretions of each electrolyte were calculated. Statistical analyses compared complete vs partial responders using a linear regression model. Contrast analyses were also performed to differentiate the mean of each group, with adjustments made with the Benjamini-Hochberg procedure. Fourteen dogs were included, eight with complete responses, and six with partial responses. Plasma phosphate concentrations were significantly higher at 12 hours (P = .0003) and 24 hours (P = .009) after complete responses to therapy. This study demonstrates the potential use of plasma and urine analyte monitoring after chemotherapy induction. Plasma phosphate measurements represent a potential indicator of early responses to L-asparaginase therapy. Larger population studies are warranted to confirm these preliminary results.