Paraneoplastic hypereosinophilia in a dog with intestinal T-cell lymphoma

A 9-year-old, intact male Doberman Pinscher was examined because of anorexia and weakness. Results of a CBC showed severe, microcytic, hypochromic anemia with mild eosinophilia (2944 cells/microL, reference interval 100-1250/microL) and thrombocytosis. Hypoferremia, hypoferritinemia, and a positive fecal occult blood test supported a diagnosis of iron deficiency anemia secondary to chronic intestinal hemorrhage. Abdominal ultrasound evaluation showed a thickened small intestinal loop, of which representative specimens were obtained during exploratory laparotomy. Histologically, the intestinal wall was infiltrated by a neoplastic population of large, round, lymphoid cells with vesicular chromatin, 1 or more prominent nucleoli, and a high number of mitotic figures. The cells were closely admixed with mature eosinophils, but were negative for metachromatic granules with toluidine blue. Immunohistochemically, tumor cells were positive for CD3, and negative for CD21, Pan B, and CD79a. A diagnosis of intestinal T-cell lymphoma was made. Chemotherapy was begun, with 30 mg/m;2 of doxorubicin administered intravenously every 3 weeks. Eosinophil concentration was 880/microL 2 weeks after surgery (on day 15 after presentation) but increased markedly to 62,914/microL on day 30, 62,400/microL on day 37, and 39,444/microL on day 58 after presentation. An association between hypereosinophilia and T-cell lymphoma is well established in human patients, in whom production of IL-5 by neoplastic T cells has been demonstrated. Hypereosinophilia has been reported only rarely with intestinal lymphoma in cats and horses, and with T-cell lymphoma in dogs.     

Assessment of biomarkers influencing treatment success on small intestinal lymphoma in dogs

This study aimed to determine a reliable therapeutic biomarker for localized small intestinal lymphoma (SIL) in dogs based on clinical and histopathological features. We retrospectively investigated 84 dogs with localized SIL, including 36 dogs receiving surgery and 48 dogs receiving chemotherapy. The dogs receiving surgery were divided into two subgroups: 18 dogs (group 1) with overall survival (OS) <120 days (median OS) and 18 dogs (group 2) with OS ≥120 days. Correspondingly, the dogs receiving chemotherapy were divided into 24 dogs (group 3) with OS <98 days (median OS) and 24 dogs (group 4) with OS ≥98 days. Clinical, haematological, histopathological and immunohistochemical analyses were comparatively evaluated among the four subgroups. There was no significant difference in OS between the surgery and chemotherapy groups. In dogs receiving surgery, the rate of Ki67‐positive cells was significantly increased in group 1 compared to group 2 and showed no significant difference between groups 3 and 4. In dogs receiving chemotherapy, the rate of O6‐methylguanine‐DNA methyltransferase (MGMT) was significantly higher in group 3 than in group 4 and showed no significant difference between groups 1 and 2. Additionally, our data showed that OS in dogs with higher Ki67 expression might be significantly increased by chemotherapy than by surgery, that of those with higher MGMT expression might be significantly increased by surgery than by chemotherapy, and Ki67 and MGMT were independent of each other. Indices of Ki67 and MGMT are suggested therapeutic biomarkers to determine the optimal first‐line treatment for localized SIL in dogs.     

Detection of monoclonality in intestinal lymphoma with polymerase chain reaction for antigen receptor gene rearrangement analysis to differentiate from enteritis in dogs

The diagnosis of canine intestinal lymphoma by morphological examination is challenging, especially when endoscopic tissue specimens are used. The utility of detection of antigen receptor gene rearrangement by polymerase chain reaction (PARR) in canine lymphoma has been well established, but its usefulness to distinguish enteritis and intestinal lymphoma remains unclear. In this retrospective study we assessed clonality of 29 primary canine intestinal lymphoma, 14 enteritis and 15 healthy control cases by PARR analysis, using formalin‐fixed, paraffin‐embedded full‐thickness tissue specimens. We could detect monoclonal rearrangements in 22 of 29 canine intestinal lymphomas [76%; 95% confidence interval (CI) 56–90%] and polyclonal rearrangements in all of the enteritis and healthy control cases (100%; CI 88–100%). We revealed a predominance of T‐cell phenotype compared to B‐cell phenotype (85%; CI 65–96% and 15%; CI 4–35%, respectively). We showed that PARR analysis contributes to differentiation of canine intestinal lymphoma from enteritis and to phenotyping of lymphomas.     

A cross‐sectional study of intestinal parasitoses in dogs and children of the periurban area of La Plata (Buenos Aires,Argentina): Zoonotic importance and implications in public health

A cross‐sectional study was performed between school term dates 2014 and 2015 to diagnose intestinal parasites in dogs and children living with them. The socio‐environmental characteristics and hygiene practices of the children were also evaluated in terms of risk factors for parasitic infection of periurban neighbourhoods of La Plata (Buenos Aires, Argentina). Serial coproparasitological samples of 78 dogs and 211 children were analysed by means of concentration and flotation techniques. Socio‐environmental variables and hygiene practices of children were evaluated through semi‐structured questionnaires which were answered by every family. The study showed that 82.1% of dogs were parasitized. The specific richness was of 11 species; Ancylostoma caninum (69.2%), Uncinaria stenocephala (41.0%), Trichuris vulpis (28.2%) and Toxocara canis (21.8%) were the most prevalent. The study also revealed that 67.8% of children were positive. Also, 11 species were identified and the most prevalent were Blastocystis sp. (36.0%), Enterobius vermicularis (27.5%) and Giardia lamblia (21.3%). The risk for parasitosis was higher in 6‐year‐old children and older (OR = 1.9, 95% IC: 1.0–3.7) and in those who did not wash their hands or did it occasionally after playing with their pets (OR = 2.8, 95% IC: 1.4–5.5). Blastocystis sp. and Entamoeba coli infection risks were greater in children whose parents had a basic level of education (OR = 3.4, 95% IC: 1.3–8.7 and OR = 3.6, 95% IC: 0.8–15.9, respectively). In addition, the risk of infection for E. coli was higher in children who lived in floodable houses (OR = 4.4, 95% IC: 0.9–16.6). Likewise, the risk of infection for E. vermicularis was greater in children with onychophagia (OR = 1.6, 95% IC: 0.7–3.7) and in 6 year olds and older whose parents completed only primary studies (OR = 3.6, 95% IC: 1.4–9.1). The results obtained show the existence of a worrying epidemiological scenario that stresses the importance of zoonotic parasitosis as a serious problem of public health.     

Salivary cortisol and behavior in therapy dogs during animal-assisted interventions: A pilot study

Animal-assisted interventions (AAIs) have been associated with positive effects on human psychological and physiological health. Although the perception of quality standards in AAIs is high, only few investigations have focused on potential welfare implications for therapy dogs linked to their performance in AAIs. The standardized program “multiprofessional animal-assisted intervention (MTI)” has been carried out in adult mental health care, significantly improving patients' prosocial behaviors. In the present study, we monitored salivary cortisol and behavioral measures in therapy dogs that participated in MTI group therapy sessions in an in-patient substance abuse treatment facility. Work-related activity (lay, sit, stand, walk, and run), behavior (lip licking, yawning, paw lifting, body shake, tail wagging, and panting), response to human action (taking food treats and obeying commands), and salivary cortisol levels were analyzed over the course of 5 subsequent MTI working sessions in experienced therapy dogs (N = 5), aged 5.4 ± 2.8 years (mean ± standard deviation). Salivary cortisol levels decreased from presession to postsession in sessions 1, 2, and 3. However, only in session 4 and 5, postsession cortisol levels were significantly lower than presession levels (P = 0.043). There was no difference between salivary cortisol levels sampled on a nonworking day at home and work-related levels sampled at the therapy site. None of the behavioral parameters varied significantly over the course of the 5 MTI sessions. Both lip licking (P = 0.038) and body shake (P = 0.021) were positively correlated with the decline in cortisol during session 5. The study results suggest that trained dogs are not being stressed by repeated participation in in-patient substance abuse therapy sessions. Further investigation into the effects of animal-assisted therapy on dogs' physiological markers and behavior is warranted.     

Laryngeal silicone stent as a treatment option for laryngeal paralysis in dogs: a preliminary study of 6 cases

BackgroundLaryngeal paralysis is a common idiopathic degenerative neurological disease in older medium-to-large breed dogs, with surgical correction of the obstruction being the treatment of choice.ObjectivesThis study evaluated the use of laryngeal silicone stents to treat canine laryngeal paralysis in dogs where classic surgical treatment was not accepted by the owners.MethodsDogs diagnosed with laryngeal paralysis, for which the owners refused arytenoid lateralization surgery as a first-line treatment, were treated with laryngeal silicone stents.ResultsSix dogs with bilateral laryngeal paralysis were included in the study. All dogs showed improvement in clinical signs immediately after the procedure. No clinical signs or radiographic changes were noted in four out of six dogs in the follow-up visit performed 1 wk later. One dog was suspected of aspirating water while drinking, but the signs disappeared after repositioning the stent. Another dog had a relapse of stridor due to caudal migration of the stent. This dog underwent arytenoid lateralization surgery because larger stents are not commercially available. At the time of writing, between seven and 13 mon after stent placement, no significant incidents have occurred in four dogs, and all owners report a satisfactory quality of life.ConclusionsLaryngeal silicone stenting is an interesting alternative for treating dogs with acquired laryngeal paralysis when the owners refuse classic arytenoid lateralization surgery. Furthermore, stent placement can be a temporary solution to stabilize these dogs until a permanent surgical treatment can be performed.     

Determination of MDR1 gene expression for prediction of chemotherapy tolerance and treatment outcome in dogs with lymphoma

The multidrug resistance gene 1(MDR1) expression levels were analysed in 27 dogs with different types of malignant lymphomas receiving a standard chemotherapy protocol. Blood samples were used for MDR1 real‐time PCR expression analysis. Treatment tolerance and outcome were evaluated on a regular basis by clinical examination and client questioning. Dogs developing severe adverse effects under treatment showed significantly lower basal MDR1 gene expression levels when compared with those who tolerated the drugs well. In the longitudinal MDR1 gene expression analysis during treatment, four dogs showed a greater than two‐fold MDR1 up‐regulation, compared to baseline expression. All four of these dogs, but none of the others, showed disease progression. In conclusion, basal and follow‐up MDR1 gene expression levels could be of predictive value for the occurrence of severe adverse drug reactions and/or the development of MDR during chemotherapy for lymphoma in dogs.     

Dietary protein concentration affects intestinal microbiota of adult cats: a study using DGGE and qPCR to evaluate differences in microbial populations in the feline gastrointestinal tract

The objective of this study was to identify qualitative and quantitative differences in microbial populations of adult cats fed diets containing different protein concentrations. Following a 4 week baseline period, eight healthy adult domestic short‐hair queens (>1‐year‐old) were randomly allotted to a moderate‐protein (MP; n = 4) or high‐protein (HP; n = 4) diet for 8 weeks. Fresh faecal samples were collected after baseline and 8 weeks on treatment and stored at ?80 °C. Following DNA extraction, samples were analyzed using denaturing gradient gel electrophoresis to distinguish qualitative changes between diets. Quantitative polymerase chain reaction was used to measure E. coli, Bifidobacterium, Clostridium perfringens, and Lactobacillus populations. Compared to baseline, cats fed MP had a bacterial similarity index of 66.7% as opposed to 40.6% similarity for those fed HP, exhibiting marked changes in intestinal bacteria of cats fed HP. Bifidobacterium populations were greater (p < 0.05) in cats fed MP versus HP (9.44 vs. 5.63 CFU/g). Clostridium perfringens populations were greater (p < 0.05) in cats fed HP than MP (12.39 vs. 10.83 CFU/g). In this experiment, a high‐protein diet resulted in a dramatic shift in microbial populations. Decreased Bifidobacterium population in cats fed HP may justify prebiotic supplementation for such diets.     

Risk factors for development of sterile haemorrhagic cystitis in canine lymphoma patients receiving oral cyclophosphamide: a case–control study

Sterile haemorrhagic cystitis (SHC) is a known risk of cyclophosphamide treatment; however, most canine reports are case series. This case–control study examined risk factors for SHC in dogs with lymphoma receiving oral cyclophosphamide. Twenty‐two dogs with SHC and 66 control dogs were identified. On univariate analysis, SHC risk factors included age (P = 0.041), induction protocol (P = 0.021) and cumulative cyclophosphamide dose (P = 0.002). On multivariate analysis, increasing cumulative cyclophosphamide dose was associated with increased risk of SHC and the ‘short’ induction protocol (protocol 1) was associated with decreased risk. Controlling for age and induction protocol, odds of SHC increased by 2.21 per 750 mg m^?2 increase in cyclophosphamide dose (P = 0.001). SHC from oral cyclophosphamide is a predominately delayed toxicity resulting from high cumulative doses.     

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.     

Effects of α2-adrenergic drugs on small intestinal motility in the horse: An in vitro study

The effects of selective α₂-agonists (xylazine, detomidine and medetomidine) and antagonists (yohimbine and atipamezole) on in vitro small intestine motility in the horse were evaluated. Samples of equine jejunum were placed in isolated organ baths and drug-induced modifications of motility were measured by means of an isotonic transducer. All tested α₂-agonists dose-dependently reduced both spontaneous and electrically-evoked phasic contractions. Conversely, α₂-antagonists were ineffective when tested alone, and showed a heterogeneous and dose-independent ability to inhibit agonist activity. In particular, the antagonism exerted by higher concentrations of both yohimbine and atipamezole against α₂-agonists was weaker than when lower concentrations were used. The data are indicative of the presence of both pre- and post-synaptic α₂-adrenoceptors with inhibitory activity on equine jejunum motility, and support a possible therapeutic utility of these drugs in horse intestinal disorders associated with hypermotility.     

Improving understanding of early behavioral indicators of lumbosacral disease in working dogs using 3D visualization of skeletal movements during working tasks: Feasibility study

An improved understanding of the early behavioral indicators of lumbosacral disease in working dogs may allow earlier interventions and help reduce premature retirement because of disability. However, recognition of early behavioral indicators can be challenging in stoic, high-drive working dogs because they often mask clinical signs. The purpose of this feasibility study was to develop a technique for visualizing canine skeletal movements during working tasks and to describe veterinary clinical specialist opinions on the utility of the visualization technique. Three detection-trained police dogs with a recent history of working task deficits and suspected lumbosacral disease were recruited for the study. Conventional and motion capture video recordings were acquired as dogs performed walking and search high working tasks. Whole-body computed tomography (CT) scans were acquired using clinical multislice CT scanners. Image data from motion capture recordings and whole-body CT scans were analyzed and merged. Three-dimensional (3D) computer animation video clips of skeletal movements were created for each dog and each task, using multiple viewing angle perspectives. Interactive meetings with veterinary clinical specialist reviewers were used to refine point placements for the final renderings. Veterinary clinical specialists reviewed final 3D animation movie clips and recorded their opinions on the utility for the visualization technique. Veterinary clinical specialists reported that the computer animations helped them recognize behavioral characteristics that they had not initially noticed in physical examinations. Potential applications for this visualization technique include creating educational training aids for veterinary students, owners, and handlers; assisting veterinarians in planning rehabilitative treatments; and assisting researchers in developing computer models for biomechanical analyses. Future controlled prospective studies are needed in a large number of normal and affected working dogs to improve accuracy of the visualization technique and test the effect of the technique on observer performance.     

MOPP chemotherapy for treatment of resistant lymphoma in dogs: a retrospective study of 117 cases (1989-2000)

The purpose of this retrospective study was to evaluate the efficacy and toxicity of the MOPP chemotherapy protocol (mechlorethamine, vincristine, procarbazine, and prednisone) as a rescue regimen in dogs with lymphoma. One hundred seventeen dogs that had resistance to previously administered chemotherapy were evaluated. Before treatment with MOPP, all dogs received a median of 6 chemotherapy drugs for a median duration of 213 days. Thirty-one percent (36 of 117) had a complete response (CR) to MOPP for a median of 63 days, and 34% (40 of 117) had a partial response (PR) for a median of 47 days. Sixteen percent (19 of 117) had stable disease (SD) for a median of 33 days. Predictors for response to MOPP were not identified. Gastrointestinal (GI) toxicity occurred in 28% (33 of 117) of the dogs, and 13% (15 dogs) required hospitalization. Five dogs developed septicemia, and 2 died as a result. MOPP was an effective treatment for dogs with resistant lymphoma and was well tolerated by the majority of affected dogs.     

A newly designed radiation therapy protocol in combination with prednisolone as treatment for meningoencephalitis of unknown origin in dogs: a prospective pilot study introducing magnetic resonance spectroscopy as monitor tool

Background

A plethora of treatment options have been described for canine meningoencephalitis of unknown origin (MUO), yet a gold standard has not been established. The aim of this prospective pilot study was to document the effect of a newly designed 30 Gray (Gy) radiation therapy (RT) protocol plus corticosteroids as treatment for focal and multifocal MUO, to monitor clinical and imaging changes during the course of the disease with conventional magnetic resonance imaging (MRI) and proton MR Spectroscopy (H-1 MRS) and to detect the occurrence of radiation related side effects.

Results

Six dogs (3 with focal and 3 with multifocal lesions) were included in the study. The RT protocol used consisted of 30 Gy in 10 fractions. The neurological status of all six dogs improved during RT, with 3 of 6 cases returning to a normal condition. One dog was euthanized early during follow-up (<3 weeks after end of RT). Three month follow up MRI was normal in one dog and improved in 3 dogs and H-1 MRS normalized in 4. In the dog without improvement of the MRI lesions, the N-acetyl aspartate continued to decrease, while choline and creatine concentrations remained stable during that time. This dog was euthanized 18 month after the end of RT due to relapse. One dog was lost to follow up 12 month after completion of RT. The other 3 dogs are still alive at the time of writing.

Conclusions

RT with 30 Gy in 10 fractions can provide an additional option for anti-inflammatory treatment of focal and multifocal MUO. The protocol used for treatment monitoring was feasible while no side effects of RT could be observed during the follow up period. Moreover, H-1 MRS could represent a new and non-invasive tool to control the progression of the disease during the treatment course.     

Use of spray-cooling technology for development of microencapsulated capsicum oleoresin for the growing pig as an alternative to in-feed antibiotics: a study of release using in vitro models

The aim of this study was to develop sustained release microspheres of capsicum oleoresin as an alternative to in-feed additives. Two spray-cooling technologies, a fluidized air bed using a spray nozzle system and a vibrating nozzle system placed on top of a cooling tower, were used to microencapsulate 20% of capsicum oleoresin in a hydrogenated, rapeseed oil matrix. Microencapsulation was intended to reduce the irritating effect of capsicum oleoresin and to control its release kinetics during consumption by the animal. Particles produced by the fluidized air bed process (batch F1) ranged from 180 to 1,000 microm in size. The impact of particle size on release of capsaicin, the main active compound of capsicum oleoresin, was studied after sieving batch F1 to obtain 4 formulations: F1a (180 to 250 microm), F1b (250 to 500 microm), F1c (500 to 710 microm), and F1d (710 to 1,000 microm). The vibrating nozzle system can produce a monodispersive particle size distribution. In this study, particles of 500 to 710 microm were made (batch F2). The release kinetics of the formulations was estimated in a flow-through cell dissolution apparatus (CFC). The time to achieve a 90% dissolution value (T90%) of capsaicin for subbatches of F1 increased with the increase in particle size (P < 0.05), with the greatest value of 165.5 +/- 13.2 min for F1d. The kinetics of dissolution of F2 was slower than all F1 subbatches, with a T90% of 422.7 +/- 30.0 min. Nevertheless, because CFC systems are ill suited for experiments with solid feed and thus limit their predictive values, follow-up studies were performed on F1c and F2 using an in vitro dynamic model that simulated more closely the digestive environment. For both formulations a lower quantity of capsaicin dialyzed was recorded under fed condition vs. fasting condition with 46.9% +/- 1.0 vs. 74.7% +/- 2.7 for F1c and 32.4% +/- 1.4 vs. 44.2% +/- 2.6 for F2, respectively. This suggests a possible interaction between capsaicin and the feed matrix. Moreover, 40.4 +/- 3.9% of the total capsaicin intake in F2 form was dialyzed after 8 h of digestion when feed had been granulated vs. 32.4 +/- 1.4% when feed had not been granulated, which suggests that the feed granulation process could lead to a partial degradation of the microspheres and to a limitation of the sustained release effect. This study demonstrates the potential and the limitations of spray-cooling technology to encapsulate feed additives.     

Effect of gentamicin on CD3+ T-lymphocyte proliferation for treatment of equine recurrent uveitis: An in vitro study

Objective

The objective of the study was to determine the effect of gentamicin on CD3+ T-lymphocyte proliferation and cell viability using an in vitro cell culture model as a means of investigating the mechanism of action of low-dose intravitreal gentamicin injection.

Animals Studied

Three adult horses with no evidence of ophthalmic or systemic disease.

Procedure

Peripheral blood lymphocytes were treated with gentamicin at concentrations 37.5 μg/mL, 112.5 μg/mL, 187 μg/mL, 375 μg/mL, or 750 μg/mL then stimulated to proliferate with concanavalin A (ConA). 4′,6-diamidino-2-phenylindole (DAPI) and carboxyfluoroscein succinimidyl ester (CSFE) were used as markers of cell viability and cell proliferation, respectively. Following 5-day culture, live cell counts and CSFE fluorescent intensity data were collected via automated cell count and flow cytometry. The experimental design was duplicated using preservative-free gentamicin and a proprietary brand formulation. Statistical analysis was performed using two-way ANOVA with Tukey's multiple comparison test.

Results

No statistically significant comparisons in CD3+ T-lymphocyte live cell counts and geometric mean fluorescent intensity of CSFE were identified between gentamicin concentrations or formulations.

Conclusions

Gentamicin had no effect on equine peripheral blood CD3+ T-lymphocyte cell viability and proliferation in concentrations ranging from “safe” to “retinotoxic” in relation to intravitreal injection volumes. Low-dose intravitreal gentamicin may not suppress the Th1- and Th17-mediated immune response.