Transfusion of compatible blood types ensures the vitality of transfused erythrocytes and avoids transfusion reactions. Cats with types A, B, and AB blood should receive transfusions of the same blood type. In a feline blood donor program, it is therefore essential to have blood donors of all blood types available.
The objectives of this study were the identification of the 3 feline blood types in Ragdoll cats, the comparison of their frequencies with those of Domestic Shorthair (DSH) cats, and the determination of whether Ragdolls are suitable donors in a feline blood donor program.
The blood type was determined by gel column agglutination from Ragdoll cats. The relationships between phenotypic traits, the origin of the cats, and the different blood types were examined. The frequencies for potential transfusion reactions and the risk for neonatal isoerythrolysis (NI) were estimated.
Of 61 typed Ragdolls, 77.1% had type A, 4.9% type B, and 18% type AB blood. The frequency of blood type A in Ragdolls was lower than in DSH cats (P = .02), while the frequency of blood type AB in Ragdolls was higher than in DSH cats (P = .0002). No relationship was found between blood type and origin of the cat or phenotypic traits. The estimated frequencies of major and minor transfusion reactions following an unmatched transfusion between Ragdolls (donors and recipients), Ragdoll donors and DSH recipients, and DSH donors and Ragdoll recipients were 4.7%, 6.7%, 4.6%, and 18.5%, 20.8%, 7.6%, respectively. The frequency of kittens at risk for NI was 5%.
The presence of all 3 feline blood types and a relatively high incidence of AB type cats make Ragdolls an ideal donor breed to include in feline blood transfusion programs.
A 6-year-old Wirehair Dachshund had a meningioma around the optic nerve that caused exophthalmos. A benign mesenchymal tumor was suspected based on the cytologic pattern of a fine-needle aspirate, and a meningioma was diagnosed by histopathologic examination. In addition to the meningioma cells, the cytologic smears included groups of cells from apparently 4 layers of normal retina. In particular, uniform rod-shaped structures in the cytologic sample could suggest rod-shaped bacteria, but these structures were identified as cylindrical outer segments of photoreceptor rod cells. Other retinal structures recognized included pigmented epithelial layer cells with their uniquely formed pigment granules, the characteristic bi-lobed, cleaved nuclei from the outer nuclear layer, and nerve tissue likely from the outer plexiform layer of the retina.
Dynamic adhesion assays allow the examination of platelet dysfunction and drug effects on platelet function.
The purpose of the study was to optimize several parameters such as type and concentration of collagen, wall shear stress, and the concentration of the platelet-activating agonist in a new biochip perfusion chamber for the study of canine platelets.
After fluorescent staining of platelets, citrated blood of 10 healthy dogs was perfused through the flow chamber coated with different concentrations of canine or bovine skin collagen. Wall shear stress ranged from 14 to 60 dynes/cm2. Protease-activating receptor 4 (PAR 4) agonist was used for platelet activation. After perfusion, platelet attachment to the collagen matrix was quantified based on fluorescent imaging. Total platelet covered area and average size of platelet covered areas were measured by planimetry.
Canine platelet adhesion was supported by ≥ 200 μg/mL canine collagen, but not bovine skin collagen. Consistent results were obtained with a wall shear stress of 14 dynes/cm2, whereas higher wall shear stress resulted in increased variability. Platelet activation with PAR 4 agonist increased the total platelet covered area and the average size of platelet covered areas.
This study indicates the need to carefully select collagen type and concentration to assess canine thrombus formation in a dynamic flow chamber. The established method should be a useful tool to determine changes in platelet–matrix interactions as an indicator of platelet activation or platelet dysfunction in dogs.
There is no agreement in current publications regarding the reliability of serum concentrations of natriuretic peptides (NPs) to detect dogs with subclinical myxomatous mitral valve disease (MMVD) and to differentiate between asymptomatic stages.
We sought to compare N-terminal pro-B-type natriuretic peptide (NT-proBNP) and pro-atrial natriuretic peptide 31-67 (proANP) concentrations between various stages of canine MMVD and to investigate the influence of age, weight, and sex.
In this prospective study, dogs were classified in different disease stages using the modified Canine Heart failure International Expert Forum (CHIEF) system. Serum NP concentrations were compared between groups.
A total of 559 samples from 116 healthy dogs and 236 dogs with MMVD were analyzed. Using cut-off values (1207 pmol/L for NT-proBNP, 1578 fmol/mL for proANP), dogs with MMVD with and without congestive heart failure (CHF) could be differentiated with a sensitivity of 83% for both and specificities of 85% and 86%, respectively. Dogs staged in CHIEF B1 and B2 could not be distinguished based on NP concentrations due to wide variation within the groups. Intact females (means 598 pmol/L and 1036 fmol/mL, respectively) had significantly higher values of both NPs than intact males (315 pmol/L and 836 fmol/mL).
NPs in canine MMVD are useful to discriminate between asymptomatic dogs and dogs with CHF. Due to a large overlap of NP-concentrations between the groups, NPs do not seem to be useful to differentiate between dogs in stages B1 and B2. Interpretation of NT-proBNP and proANP values should include consideration of sex-specific differences.
An 8-year-old castrated male hound mix was referred to the Purdue University Veterinary Teaching Hospital for severe lameness, pollakiuria, and dyschezia. On presentation, the dog was nonweight bearing on the right rear limb and the right carpus was diffusely swollen. Synovial fluid analysis from the right carpus revealed a population of epithelial cells displaying marked anisocytosis, anisokaryosis, multinucleation, and prominent, variably sized nucleoli. A metastatic carcinoma with presumed prostatic or urothelial origin was diagnosed based on cytomorphology. Subsequent cytologic evaluation of peripheral lymph nodes revealed the presence of a similar neoplastic population. The dog was euthanized and synovial fluid from both stifle joints, as well as impression smears of the prostate gland, were collected. Carcinoma cells were identified in each stifle joint and in the prostate gland. Immunocytochemistry was performed on synovial fluid smears from 2 of the joints (right stifle and right carpus) and on impression smears of the prostate gland. The neoplastic population in the joints and prostate gland showed strong immunoreactivity to uroplakin III, a urothelial marker, indicating metastasis of a transitional cell carcinoma to multiple joints. In addition, evidence for epithelial to mesenchymal transition was identified using cytokeratin, an epithelial marker, and vimentin, a mesenchymal marker. A necropsy was performed and histopathology confirmed the presence of metastatic transitional cell carcinoma in various tissues. This case illustrates the importance of considering metastatic disease when a patient is presented with severe lameness and joint pain, and the clinical utility of synovial fluid cytology for diagnosis of metastasis in these cases.
An EDTA-anticoagulated blood sample collected from a 1.5-year-old, intact male, English Bulldog was submitted for a CBC. The CBC data and blood smear evaluation revealed borderline high hematocrit (54%, reference interval 37–55%), inappropriate rubricytosis, moderate leukopenia due to both mature neutropenia and lymphopenia, and mild thrombocytopenia. Numerous leukocytes showed evidence of karyolysis, pyknosis, and karyorhexis, and apoptotic bodies were frequent in the background. Many neutrophils had botryoid nuclei characterized by increased numbers of nuclear segments radially arranged with spoke-like, delicate chromatin filaments connecting the segments centrally. The finding of botryoid nuclei and inappropriate rubricytosis was indicative of severe hyperthermia, such as heatstroke. The dog had been exercised a long time during conditions of high temperature and humidity until he collapsed. The dog was diagnosed with severe heatstroke, hypovolemic shock, disseminated intravascular coagulation, and multiorgan dysfunction syndrome. Despite aggressive treatment, the patient died of cardiopulmonary arrest. Botryoid nuclei are frequent in people with heatstroke. In the authors' experience, botryoid nuclei are seen commonly in dogs with heatstroke, but they have never been reported in veterinary medicine. The presence of petechiation with only mild thrombocytopenia and inappropriate rubricytosis also is suggestive of heatstroke and manifests ongoing life-threatening vascular derangement.
Rickettsial agents, including those in the genera Anaplasma, Ehrlichia, Neorickettsia, and Rickettsia, are important and common vector-borne pathogens of dogs and cats. Disease induced by these organisms ranges from clinically inapparent to severe and potentially fatal. However, laboratory confirmation of a rickettsial etiology can be complicated by a number of factors, including the wide spectrum of disease induced by these organisms, an often low and widely fluctuating level of organism present in infected animals, cross-reactions on serologic and molecular assays, and the presence of co-infections. Correct diagnosis is most likely to be reached when multiple diagnostic strategies, including careful microscopic examination of stained blood films or tissues, both specific and broad serologic tests, and a suite of molecular detection assays, are used in concert. Accurate interpretation of diagnostic tests requires awareness of the likelihood for multiple agents, including novel organisms, to be responsible for the results seen in a given patient. This review provides an overview of current strategies used to diagnose rickettsial infections in dogs and cats.
A chronic loss of canine α1-proteinase inhibitor (cα1-PI) into the gastrointestinal (GI) tract could change the systemic proteinase-proteinase inhibitor balance. Serum cα1-PI concentrations have not been studied in dogs with well-defined GI diseases.
To further evaluate serum cα1-PI concentrations in dogs with GI diseases, the objectives of this study were to (1) analytically validate a previously developed fecal cα1-PI immunoassay to determine serum concentrations, (2) determine a population-based reference interval (RI) and assess the clinical utility, (3) determine stability of serum cα1-PI, (4) determine the intra-individual variation in healthy dogs, and (5) determine the clinically relevant magnitude of change of serum cα1-PI.
Prestudy validation of the 125I-cα1-PI immunoassay included linearity, spiking recovery, and intra- and inter-assay precision. A RI was calculated with samples of healthy dogs. Stability at −20°C was tested on 36 samples. Intra-individual variation was assessed using samples collected from 11 healthy dogs over a 12-week period.
The cα1-PI radioimmunoassay (RIA) was linear, accurate, precise, and reproducible. Serum cα1-PI decreased by 11% after one year at −20°C. Analytical, intra-individual, inter-individual, and total variation were 6.4, 9.9, 9.0, and 25.3%, respectively. The RI for serum cα1-PI was 732–1802 mg/L (n = 87); there were no differences between sex and age groups. The index of individuality was 1.31. As analytical variation was > ½ inter-individual variation, the minimum critical difference was not determined.
The results of this study provide the basis for further evaluating serum cα1-PI in dogs with GI disease. Using a population-based RI for serum cα1-PI appears to be appropriate.
Insulin-like growth factor (IGF-1) is an important mediator of tissue repair in horses.
The aim of the study was to evaluate whether IGF-1 could be measured reliably in equine serum and tendon tissue extracts, using an IGF-1 ELISA kit developed for human serum and plasma.
A glycyl-glycine pretreatment protocol of samples was compared with the pretreatment procedure recommended by the manufacturer. Intra- and inter-assay imprecision were evaluated by repeated measurements of equine serum pools. Assay inaccuracy was determined based on the linearity of serially diluted equine serum samples and tendon tissue extracts. The recovery of IGF-1 was evaluated in serum and tendon tissue extracts spiked with known amounts of IGF-1.
The range of IGF-1 released using the manufacturer's pretreatment was between 23% and 56% of the amount released using the gly-gly pretreatment in different equine samples. In serum pools with low, intermediate, and high IGF-1 concentrations, intra-assay imprecision was 4.0%, 4.0%, and 3.1%, respectively, and inter-assay imprecision was 13.9%, 7.3%, and 12.8%, respectively. The recovery of serially diluted serum was 96 ± 3% when diluted with serum, and 72 ± 15% when diluted with PBS. The recovery after dilution was 108 ± 17% in tendon tissue extracts. Recovery from serum spiked with a fixed amount of IGF-1 was 101 ± 5% and 99 ± 7% from tendon tissue samples.
The IDS Octeia IGF-1 ELISA kit can be used for measuring IGF-1 in equine serum and tendon tissue extract after pretreatment with glycyl-glycine.
A 7-year-old spayed female German Wirehaired Pointer was presented with difficulty breathing after being found seizing in a water-filled drainage ditch while out hunting. Aspirates from a tracheal wash contained numerous degenerate neutrophils, fewer macrophages, some of which contained basophilic debris, low numbers of extracellular diatoms, and a single intracellular short bacterial rod. As the dog continued to clinically decline and could not be weaned from oxygen support, the owners chose euthanasia. The major necropsy finding was a severe granulomatous bronchopneumonia that was likely due to aspiration of foreign material based on the microscopic presence of plant-like material, bi-refringent crystalline material, non-cellular debris, and occasional fungal structures. Diatoms are a class of algae that live primarily in water. Diatom analysis has been used, with some controversy, in human forensics to assist in documenting drowning as the cause of death. In this case, given the clinical history, the presence of diatoms and inflammation in the tracheal wash were interpreted as a likely result of the aspiration of surface water. To our knowledge, this is the first reported case of diatoms observed in a cytologic specimen in a nonhuman mammal with aspiration pneumonia.
Sternal bone marrow aspiration in dogs is not commonly performed as it is considered technically challenging in smaller dogs. However, the sternum is readily accessible and associated with less pain from aspiration compared with other sites.
The aim of the study was to investigate feasibility, ease, number of attempts, safety, and sample quality of sternal bone marrow aspirates in small dogs.
Bone marrow aspirates were obtained in a randomized order from 3 sites in 26 clinically healthy Beagles under general anesthesia. Samples were obtained from the sternum using one-inch 20- or 22-gauge hypodermic needles, from the right greater tubercle of the humerus, and the right iliac crest using 18-gauge Illinois needles. The difficulty of each procedure was scored. Two types of bone marrow smears were prepared and reviewed by a pathologist unaware of site of aspiration or dog. The number of particles per slide and overall slide quality were scored. The site of aspiration and the cranial thoracic wall were evaluated at autopsy for evidence of trauma or pneumothorax.
The number of attempts and time for bone marrow aspiration were greater for ilium than for sternum or humerus, but the sternum was the easiest to aspirate. Smear quality and particle number were similar for all sites. Neither trauma at the site of aspiration nor pneumothorax were identified.
Aspiration of sternal bone marrow with hypodermic needles is feasible and safe in Beagle dogs. Samples equivalent in quality to those from the humerus or ilium can be obtained from clinically normal dogs.
Avian tuberculosis is an important disease affecting all species of birds and is most often caused by Mycobacterium avium or Mycobacterium genavense. Blood proteins are important diagnostic constituents in gastrointestinal, hepatic, renal, and infectious diseases.
The aim of the study was to compare serum protein profiles of domestic pigeons (Columba livia var. domestica) infected with Mycobacterium avium subsp. avium (MAA), with healthy pigeons.
Serum samples were collected from 80 pigeons with clinical signs of tuberculosis, all kept in the same loft. All birds were necropsied and cultured for mycobacteriosis; positive cultures were typed for MAA by PCR reactions targeting 16S rRNA, IS901 and IS1245. The concentration of total serum proteins was determined by the biuret method and spectrophotometry. Individual protein fractions were analyzed by cellulose acetate electrophoresis and extrapolated based on total protein concentration. For statistical analysis, the infected birds were compared with healthy pigeons.
A total of 37 pigeons with culture results positive for MAA were selected and allocated to 2 groups, a culture-positive group with macroscopic lesions of tuberculosis and another without macroscopic lesions. Six protein fractions were identified: prealbumin, albumin, alpha-1, alpha-2, and beta globulins and gamma globulins. Concentrations of total protein, beta globulins and gamma globulins were statistically significantly higher in the infected pigeons when compared with the control group. There were no significant differences between the groups of birds with or without macroscopic lesions.
Statistically significant differences in total protein, and beta and gamma globulin concentrations in all infected pigeons suggest that serum protein electrophoresis represents a nonspecific but valuable indicator for tuberculosis in pigeons.
Peritoneal fluid (PF) analysis is a valuable diagnostic tool in equine medicine. Markers such as serum amyloid A (SAA) and haptoglobin (Hp) could facilitate the diagnosis of inflammatory abdominal conditions.
The objectives were to (1) establish reference intervals (RI) for SAA and Hp in serum and PF in healthy horses, (2) compare SAA and Hp concentrations between healthy horses and horses with colic, and (3) to assess the correlation between serum and PF concentrations.
Serum amyloid A and Hp concentrations were determined by automated assays in prospectively enrolled healthy reference horses and horses with colic. RIs were calculated, group concentrations were compared by Student's t-test, and Pearson′s correlation for serum and PF concentrations were determined.
In healthy horses (n = 62) the measurements for SAA were below the detection limit (0.5 mg/L) in 94% of serum samples and 98% of PF samples. Horses with colic (n = 61) had statistically significantly increased SAA concentrations in serum (P < .0001) and PF (P = .0013). While PF Hp concentrations were increased in horses with colic the serum concentrations of Hp were decreased (P < .0001). There was a strong correlation between paired serum and PF SAA concentrations (n = 94, R = .72, P < .0001), whereas the correlation between paired serum and PF Hp was weak (n = 94, R = .22, P = .0382). Finally, horses with colic tended to have serum SAA and PF Hp concentrations above the RIs.
With the apparent difference between healthy horses and horses with colic and the presently established RIs, serum SAA and PF Hp concentrations represent potential valuable diagnostic markers for inflammatory abdominal conditions in that species.
Transfusion of RBC concentrates may cause adverse effects in the recipient, particularly when stored > 2 weeks. Prestorage removal of WBCs and platelets (leukoreduction, LR) improves clinical outcome in the human recipient. As blood ages during storage, progressive alterations in the structure and function of the cells occur. Changes in cell membranes may lead to formation of microparticles (MPs) in stored blood.
The aim of the study was to quantify MP concentration in supernatants from canine RBC concentrates from 11 clinically healthy dogs.
Whole blood units (n = 11) were collected and randomized either to be stored without LR (n = 5), or to be subject to prestorage LR (n = 6). Whole blood was processed for the generation of RBC concentrates, from which aliquots were aseptically collected weekly for 5 weeks. Supernatants from the concentrates were evaluated for phosphatidylserine-expressing MPs by flow cytometry using staining with Annexin-V-phycoerythrin.
Microparticle counts were similar between non-LR and LR units on storage days 0 and 7, but were significantly higher in non-LR units on days 14, 21, 28, and 35. MPs increased during the 35-day storage by a mean (SD) of 1.8 (1.4)-fold in LR units and 5.5 (3.1)-fold in non-LR units.
There was marked formation of phosphatidylserine-expressing MPs during storage beyond 7 days in canine RBC concentrates. Prestorage LR attenuated the generation of MPs.
Obesity is the most common nutritional disorder in today's dog population and the major risk factor for a number of related diseases. However, the exact pathogenesis of obesity-related complications is not always clear. In people, butyrylcholinesterase (BChE) is suspected to be involved in lipoprotein metabolism and has also been associated with the pathogenesis of inflammatory disease, one of the potential complications related to obesity.
The aim of the study was to evaluate the effect of experimentally induced weight loss on BChE and acetylcholinesterase (AChE) in obese dogs to elucidate the possible relationship between these 2 enzymes and obesity.
Six obese intact female Beagle dogs were allocated to a weight loss program for 3 months. BChE was measured in serum samples using butyrylcholine as substrate, whereas AChE was measured in whole blood after inhibition of BChE with ethopropazine and using acetylcholine as a substrate.
After rapid weight loss serum BChE activities were statistically significantly lower (P < .05), whereas AChE activities were higher (P < .01). There was a positive correlation between serum BChE activity and concentrations of total cholesterol (TCHOL, P < .001), low-density lipoprotein-cholesterol (LDL-C, P < .001), and triglycerides (P < .05). A negative correlation was detected between serum BChE and AChE activities (P < .0001), and between AChE activity and serum levels of TCHOL (P < .01), LDL-C (P < .01) and high-density lipoprotein-cholesterol (P < .05).
Short-term weight loss in obese intact female Beagle dogs resulted in opposite effects in 2 cholinesterase isoenzyme activities, namely lower BChE and higher AChE activities.
In cats, mastocytemia is considered to be confined to animals with mast cell tumors (MCT), whereas in dogs it is associated with diverse diseases.
The objective of this retrospective study was to investigate the diagnostic and prognostic significance of mastocytemia in cats.
All blood smears and buffy coat (BC) smears on which mast cells were identified over a 6-year period were retrospectively reviewed and mast cells counted. Mastocytemic cats were classified based on their clinical diagnosis.
Mastocytemia was identified on 40 blood smears and 13 BC smears from 33 cats. The incidence of mastocytemia detected in cats during routine CBCs was 0.33% (40/12,116 CBCs). Twenty-two of 33 mastocytemic cats (67%) had visceral (n = 17) or cutaneous MCT (n = 7), including 2 that had concurrent visceral and cutaneous involvement. In 3 additional cases (9%), visceral MCT was clinically suspected, but no cytologic or histopathologic evaluation of visceral organs was performed. MCT was excluded in 3 of 33 mastocytemic cats (9%) with a final diagnosis of lymphoid neoplasia (n = 2) and multiorgan hemangiosarcoma (n = 1). Five additional animals (15%) had a diagnosis other than MCT, including lymphoma (n = 2) and chronic renal failure (n = 3), but no cytologic or histopathologic evaluation of the spleen was performed. Blood smears from cats with confirmed MCT had 1–113 mast cells per smear, whereas cats in which MCT was excluded had 1–2 mast cells per smear.
Data confirm that mastocytemia is rare and most commonly found in cats with visceral MCT; however, rare circulating mast cells may also be seen with neoplasms other than MCT.
Small hematology analyzers for veterinary practices improve point-of-care diagnostic testing for companion animals. Validation of these instruments is needed to ensure the accuracy of results.
The objective was to validate the Celltac alpha hematology analyzer for feline and canine blood samples.
Canine and feline blood samples were analyzed on a Celltac alpha and a Sysmex XT-2000iV. Manual methods were used for the WBC differential count, PCV, and feline platelet (PLT) count. Flagging and precision of the new instrument were analyzed. Correlation and Bland–Altman analyses were used to compare the methods.
A total of 623 blood samples (363 canine, 260 feline) were analyzed. Within-batch precision of the Celltac showed acceptable coefficients of variation (CV) for WBC count (< 4%), PLT count (< 8%), hemoglobin (HGB) concentration (< 3%), and HCT (< 3%), while precision was poor for leukocyte subpopulations. HGB concentration and WBC count had good agreement between the methods. CV for the granulocyte (GRAN) count was 2–9% in cats and 6–29% in dogs. CV for the lymphocyte (LYM) count was 8–20% in cats and 13–51% in dogs. Negative bias and a proportional systematic error were apparent for PLT count, feline HCT, and eosinophil count. Analytical error flags and incomplete results were reported for 11.8% of canine and 25.4% of feline samples.
Leukocytosis and leukopenia were reliably detected by the Celltac alpha. The instrument provided acceptable results for total WBC count, GRAN count, HGB concentration, and HCT in canine blood samples, but PLT count was systematically overestimated. In feline blood samples, both low and high PLT counts were inaccurate and a proportional systematic error for HCT led to overestimation of this variable. Imprecision for WBC differential counts was high.
Lake Sturgeon (Acipenser fulvescens) is a long-lived freshwater fish distributed throughout the Great Lakes region and is a threatened species in New York State. The species' unique life cycle makes it useful as an indicator of ecosystem health, and efforts to repatriate Lake Sturgeon to their historic range are underway. However, comprehensive hematologic and biochemical reference intervals that would be valuable for assessing the health status of individual animals have not been reported.
The objective of this study was to determine normal hematologic and plasma biochemical reference intervals for healthy wild Lake Sturgeon.
Blood samples were collected from 52 wild Lake Sturgeon caught in gill nets in the St. Lawrence River. Heparinized whole blood and plasma samples were analyzed using standard techniques. Reference intervals were calculated using the robust method following elimination of outliers and Box–Cox transformation of data.
Hematologic reference intervals were as follows: PCV 17–38%, estimated WBC count 2740–23,150/μL, neutrophils 193–6121/μL, eosinophils 0–558/μL, other granulocytes/heterophils 0–488/μL, lymphocytes 1447–14,044/μL, and monocytes 55–1684/μL. Plasma biochemical reference intervals were as follows: aspartate aminotransferase 333–1746 U/L, calcium 1.85–2.80 mmol/L, chloride 95–123 mmol/L, creatine kinase 776–35,536 U/L, glucose 2.94–14.76 mmol/L, glutamate dehydrogenase 6–30 U/L, phosphate 2.03–5.81 mmol/L, potassium 2.34–4.24 mmol/L, sodium 122.9–151.1 mmol/L, total protein 2.0–4.4 g/dL, triglycerides 1.07–5.12 mmol/L, and uric acid 1–251 μmol/L.
Reference intervals reported here will be useful for health assessment of wild and repatriated Lake Sturgeon.
The economic consequences from mortality of sheep after blood loss can be considerable. To date there are no reports addressing hematologic, blood gas, and biochemical changes in ovine blood stored in CPDA-1 bags.
The aim of this study was to investigate hematologic, blood gas, and biochemical alterations resulting from storage of ovine blood in CPDA-1 bags to establish transfusion protocols in sheep.
From each of 7 healthy 8-month-old sheep 450 mL of blood were collected into CPDA-1 bags and stored for 35 days in at 3–6°C. Samples were taken from the bags at days 0, 7, 14, 21, and 35. Whole blood was used to assess PCV, MCV, RBC count, pH, pO2, pCO2, and concentrations of bicarbonate, sodium, and lactate. Plasma was used to measure potassium, hemoglobin, and glucose concentrations.
The PCV remained stable throughout the storage period, while plasma hemoglobin and MCV began to increase on days 7 and 21, respectively. The RBC count began to decrease on day 21. Blood pH decreased and pCO2 increased steadily throughout the storage period. Potassium concentration increased from 3.8 to 18.3 mmol/L on day 7 and remained high thereafter. In contrast, sodium concentration began to decrease on day 7.
The results show that ovine blood undergoes hematologic, blood gas, and biochemical changes during storage. Further studies are required to establish RBC viability in CPDA-1 bags after a storage period of 35 days.
Rangelia vitalii is a tick-transmitted piroplasm that causes both hemolytic and hemorrhagic disease in dogs in Brazil.
The aim of this study was to evaluate the response of the bone marrow in dogs experimentally infected with R vitalii during the acute stage of the disease.
For this study, 2 groups of a total of 12 young dogs were used. Group A was composed of healthy dogs (n = 5), and group B consisted of animals infected with R vitalii (n = 7). Blood samples were collected on days 0, 10, 20, and 30 post-inoculation and stored in EDTA tubes for a full hematology profile, including a reticulocyte count. On days 10 and 20, bone marrow samples were collected, stained, and examined.
In infected dogs anemia was identified on days 10 and 20 post-inoculation (P < .01), and on day 20 reticulocytosis was present. Infected dogs had leukopenia due to neutropenia and eosinopenia, along with lymphocytosis and monocytosis, when compared with control animals. In bone marrow, the myeloid:erythroid ratio was significantly decreased (P < .05) in infected dogs due to increased erythroid precursors.
Dogs experimentally infected with R vitalii develop regenerative extravascular hemolytic anemia accompanied by erythroid hyperplasia in the bone marrow. During the acute phase of the disease, leukopenia due to neutropenia and eosinopenia suggests intense tissue recruitment of these cells in response to the endothelial damage caused by this parasite.
A 7-year-old male castrated Jack Russell Terrier was presented to the oncology service at the University of California–Davis Veterinary Medical Teaching Hospital for evaluation of suspected lymphoma. The dog had several enlarged lymph nodes and moderate lymphocytosis. Aspirates of an enlarged inguinal lymph node contained a bimorphic population of large immature lymphocytes and smaller cells with plasmacytoid features. Both cell types often contained a single large cytoplasmic inclusion that varied from clear to pale pink to sky blue. Cytologic changes were interpreted as most consistent with lymphoid neoplasia. Based on the predominantly mature cell morphology and some morphologic heterogeneity, the peripheral lymphocytosis was interpreted as most likely reactive in nature. However, the immunophenotype of the cells (CD20+, CD21+, CD79a+, MUM-1+, and MHCII+) and clonality assays showed that tissue and blood lymphocytes were neoplastic B cells with clonal identity despite their different morphologic appearances. The cytoplasmic inclusions were positive with periodic acid-Schiff and were immunoreactive for IgM and IgG. By transmission electron microscopy, inclusions consisted of aberrant rough endoplasmic reticulum; a few small Russell bodies were also noted. A final diagnosis of high-grade B-cell lymphoma with plasmacytoid differentiation, atypical cytoplasmic inclusions, and secondary leukemia was made. Chemotherapy was initiated, but the dog was euthanized due to severe and uncontrolled seizures 9 months after the initial diagnosis. This case extends the morphologic repertoire of canine plasmacytoid neoplasms and emphasizes their continuum with multicentric lymphoma. This case also demonstrates the need for advanced diagnostic techniques in establishing blood involvement in lymphoma in some instances.
A 12-year-old, spayed female, mixed-breed dog was presented for acute hematuria, stranguria, polyuria, and polydipsia, as well as lameness for 8 days. Previous medical history included treatment for infection with Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, and Dirofilaria immitis 6.5 years prior to presentation. Besides persistently increased antibody titers to E canis and A phagocytophilum, polyclonal gammopathy with a monoclonal spike and moderate hypercalcemia were observed. There was marked hematuria, and Staphylococcus aureus was cultured from urine. Two weeks after successful treatment of the urinary tract infection, radiographs showed an extensive destructive monostotic lesion of the right humerus. Cytologic examination of fine-needle aspirates of this lesion revealed a neoplastic round cell population suggestive of multiple myeloma. The dog was treated with melphalan and prednisolone for suspected multiple myeloma and doxycycline for suspected ehrlichiosis and anaplasmosis. Treatments lead to resolution of the clinical signs, hypercalcemia, and monoclonal gammopathy, and there was radiographic improvement of bone lesions; polyclonal gammopathy persisted. About one year after presentation the dog was still in clinical remission. This is a rare report of a dog with suspected multiple myeloma and a history of multiple chronic infectious diseases, suggesting that chronic infection and uncontrolled long-term stimulation of the immune system could contribute to the pathogenesis of multiple myeloma.
An 11-year-old, castrated male, Domestic Medium Hair cat was presented to the University of Florida Small Animal Hospital with a 2-week history of upper respiratory infection and increased serum globulins, as reported by the referring veterinarian. Physical examination was unremarkable other than melanosis of the left iris, with no evidence of ocular, nasal, or respiratory disease. Laboratory abnormalities included moderate nonregenerative anemia, mild leukopenia, mild hyperfibrinogenemia, severe hyperglobulinemia, mild hypoalbuminemia, and hypocholesterolemia. Abdominal radiographs and ultrasonographic examination revealed mild splenomegaly with no other abnormalities. Thoracic radiographs revealed no abnormalities. Cytologic evaluation of fine-needle aspirates from the spleen, liver, and bone marrow revealed numerous plasma cells and many vacuolated macrophages exhibiting marked phagocytosis of mature erythrocytes and platelets, occasionally metarubricytes and leukocytes, and rarely plasma cells. The cytologic interpretation was multiple myeloma and associated hemophagocytic syndrome (HPS). Serum protein electrophoresis revealed a monoclonal gammopathy, providing further evidence for a multiple myeloma. To the authors' knowledge, this is the first report of HPS secondary to neoplasia in a cat.
A recent genome-wide scan using the canine minimal screening set 2 (MSS-2) showed that cobalamin deficiency appears to be hereditary in Chinese Shar Peis and is linked to the microsatellite markers DTR13.6 and REN13N11 on canine chromosome 13.
The goal of this study was to evaluate the MYC_CANFA gene, which is the closest known gene with a distance of approximately 0.06 megabases (Mb) to the microsatellite marker DTR13.6, for any mutations in this breed.
Microsatellite markers (Myc and G15987) for genotyping and primers for sequencing were used to evaluate the MYC_CANFA gene. The genotype and gene sequence were compared between cobalamin-deficient Shar Peis, Shar Peis with normal serum cobalamin concentrations, and the DNA sequences published as part of the Ensemble Genomic map.
Neither the microsatellite markers (Myc and G15987) nor the sequences of the MYC_CANFA gene showed a significant difference among both groups of Shar Peis and the published canine DNA sequence.
The data presented here suggest that cobalamin deficiency in Shar Peis is not related to any mutations of the MYC_CANFA gene according to the genotyping and sequencing results in this study. Further investigations are warranted to find a potential genomic locus in proximity to DTR13.6 and REN13N11 that shows mutations in cobalamin-deficient Shar Peis.
Establishment of reference intervals (RI) for serum biochemical analytes is important for monitoring the health of different breeds of horses.
The aim of this study was to measure and report RI for serum biochemical analytes of the Polish Konik horse.
Blood samples were collected from 74 clinically healthy Polish Konik horses living under controlled natural conditions. These were adult primitive horses, aged 3–15 years, including 28 males (21 stallions, 7 geldings) and 46 mares. Serum analytes were measured and analyzed using a commercial automated analyzer.
The following RI (medians) were comparable to previously published RI in horses: albumin 34.8–41.3 g/L (38.0); ALP 124–309 U/L (216); ALT 6–33 U/L (19); AST 300–566 U/L (433); calcium 2.8–3.2 mmol/L (3.0); chloride 95–102 mmol/L (99); cholesterol 2.1–3.4 mmol/L (2.7); CK 167–488 U/L (328); creatinine 101–170 μmol/L (136); glucose 2.7–5.3 mmol/L (4.0); lipase 303–1143 U/L (723); magnesium 0.8–1.0 mmol/L (0.9); phosphate 0.9–1.5 mmol/L (1.2); potassium 2.4–5.0 mmol/L (3.7); sodium 138–144 mmol/L (141); total bilirubin 8–16 μmol/L (11.7); total protein 63.5–78.9 g/L (71.2); triglycerides 0.0–0.3 mmol/L (0.1); urea 2.2–7.3 mmol/L (4.7).
Reference intervals established in this study provide a valuable preliminary baseline for assessment of serum analytes in healthy Polish Konik horses.
Milk fever, a metabolic disease of dairy cattle, is associated with perturbations of calcium homeostasis, the pathogenesis of which is not yet completely understood.
The aim of this study was to investigate plasma concentrations of calcitonin gene-related peptide and selected minerals and metabolites in periparturient cows with and without milk fever.
Plasma concentrations of calcitonin gene-related peptide, as well as calcium, phosphate, magnesium, iron, glucose, lactate, and cortisol, were determined in multiple plasma samples from Jersey cows with and without spontaneous milk fever.
Cows affected by milk fever (n = 5) had lower concentrations of calcitonin gene-related peptide (P = .038) and inorganic phosphate (P < .001) in plasma than did the controls (n = 5). Also, these cows tended to have lower calcium concentrations (P = .071). Magnesium, iron, lactate, glucose, and cortisol concentrations were comparable between both groups of cows (P > .10). Around the day of calving, plasma concentrations of lactate, glucose, and cortisol increased and the concentration of iron decreased in all cows (P ≤ .01).
Despite the limited number of cows evaluated, this report is the first to indicate lowered concentrations of calcitonin gene-related peptide as part of the metabolic changes during milk fever in cows. Further work with a larger cohort of animals is warranted to understand the precise role of calcitonin gene-related peptide and the potential associations with disturbances in plasma minerals typically observed during milk fever.
Therapeutic decisions regarding urinalysis are commonly based on the presence of white and red blood cells. Traditionally, numbers per high-power field are estimated using wet-mount microscopic examination. This technique is not standardized and counts are likely prone to inaccuracy. In addition, differentiation of leukocyte types is not possible.
The aims of this study were to (1) compare WBC and RBC estimates using wet-mount examination with counts obtained using a hemocytometer, (2) assess if a hematology automated analyzer (Sysmex ST-2000iV/XT) provides reliable WBC and RBC counts in urine comparable to hemocytometer counts, and (3) evaluate air-dried Wright–Giemsa-stained urine drop sediment preparations for the determination of differential leukocyte counts.
WBC and RBC counts were obtained by performing wet-mount estimates, manual hemocytometer counts, and Sysmex automated counts on 219 canine and feline urine samples. Results were correlated using Spearman rank correlation. Air-dried Wright–Giemsa stained sediment drop preparations (n = 215) were examined for differential counts of leukocytes.
A low but significant association was found between WBC estimates on wet-mount examination and hemocytometer counts (rho = 0.37, P < .01). There was a high and significant association when RBC counts were compared between wet-mount and hemocytometer evaluation (rho = 0.7, P < .01). There was very high and significant interassay correlation between Sysmex data from duplicate samples for what the analyzer classified as WBC (rho = 0.97, P < .01) and RBC (rho = 0.94, P < .01). Low correlations were found between the Sysmex RBC counts and both wet-mount estimates and hemocytometer RBC counts (rho = 0.43, P < .01 and rho = 0.39, P < .01, respectively). Cell preservation in the air-dried sediment preparations was so poor that differential counts could not be performed.
WBC and RBC estimates on wet-mount examination agreed with hemocytometer counts and are therefore considered adequate. The Sysmex ST-2000iV/XT did not provide reliable cell counts under the conditions used.
The presence of human breast carcinoma micrometastases in bone marrow is associated with poor overall survival, poor breast-cancer-specific survival, poor disease-free survival, and poor distant disease-free survival. In veterinary practice, the detection of micrometastases as a component of clinical staging is a routine practice for lymphomas and mast cell tumors, but not for carcinomas.
This prospective study evaluated whether the identification of micrometastases from various carcinomas in dogs and cats in bone marrow using cell block cytology is technically feasible and whether it correlates with routine cytologic examination.
Thirteen dogs and 4 cats with various types of carcinomas were available for analysis. Routine and cell block cytologic evaluation combined with immunocytochemical staining with antibodies to CKAE1/AE3 and CK7 were performed on all bone marrow samples.
Bone marrow micrometastasis was demonstrated by both methods in 2 dogs with advanced disease. In one case cells were immunoreactive for both CKAE1/AE3 and CK7.
This study demonstrates that cell block cytology is a practical and useful method for bone marrow evaluation and is suitable for cytokeratin immunocytochemical analysis.
Reliable enumeration of mast cells and eosinophils in equine bronchoalveolar lavage (BAL) fluid is important because small increases in the percentages of these cells support the clinical diagnosis of inflammatory airway disease (IAD). Increases in BAL neutrophils also occur with IAD but are not specific due to overlap between IAD and recurrent airway obstruction (RAO).
The objectives of this study were to evaluate the reliability of a standard 400-cell leukocyte differential count and an alternate method evaluating 5 microscopic fields at 500× magnification in equine BAL fluid cytocentrifuged preparations.
BAL samples from 60 horses with and without pulmonary inflammation were evaluated using 400-cell and 5-field leukocyte differential counting methods. Reliability of enumeration of each leukocyte type was assessed by calculating and comparing intraclass correlation coefficients (ICC). Reliability of mast cell enumeration was further evaluated by comparing ICCs of slides with different cell densities.
Reliability was higher for all cell types with the 5-field method; however, overall the difference between methods was not statistically significant. Neutrophil reliability was high (ICC > 0.90) with both methods. Adequate reliability (ICC > 0.85) for mast cells was achieved only with the 5-field method on slides with higher cell density.
Enumeration of mast cells is unreliable when the standard 400-cell differential counting method is used, whereas the 5-field method on slides with higher cell density reached acceptable reproducibility. Neutrophil percentages were highly reliable with both methods.
Mast cell tumors are the most common cutaneous tumor in the dog and are often diagnosed via fine-needle aspiration and cytology. Many veterinary practices use Diff-Quik stain for these cases because it is easy to use and provides rapid results. Anecdotal reports suggest that Diff-Quik does not stain mast cell tumor granules well and that increased duration of fixation time can improve staining quality; however, this has not been prospectively evaluated.
The aim of this study was to determine if varying fixation time would affect the staining quality of mast cell granules using the Diff-Quik stain. The null hypothesis was that there would be no difference in the staining of the granules based on duration of time in the fixation solution.
Fine-needle aspirates of cutaneous mast cell tumors were obtained from 21 dogs and distributed on multiple slides. These slides were then stained in Diff-Quik at varying fixation times (ie, 5 seconds, 30 seconds, 1 minute, 2 minutes). One slide was stained with modified Wright stain as a control. Mast cell staining quality was evaluated either by blinded clinicopathologic review (n = 12) or by computer analysis of photomicrographs (n = 6). Results were compared with histopathologic grade.
There was no difference in staining quality among groups.
Alteration in fixation time using Diff-Quik does not improve staining characteristics of mast cell tumors.
Mesocestoides cestode infections in dogs are well known for causing severe peritonitis with larvae or larval fragments (metacestodes, tetrathyridia, or calcareous corpuscles) frequently observed cytologically in peritoneal fluid samples. This case report describes the cytologic and clinical features of 2 dogs infected with cestode larvae, with one case confirmed and the other presumed to be Mesocestoides sp. In these 2 unusual cases, cestode larvae or larval fragments were found in fine-needle aspirates of the liver and a mesenteric lymph node, but no organisms were found in peritoneal fluid samples. The data presented in this report indicate that clinical pathologists should not rule out Mesocestoides sp cestodiasis based on the absence of larvae in peritoneal fluid samples from dogs.