Objective—To determine prevalences of various hemoplasma species among cats in the United States with possible hemoplasmosis and identify risk factors for and clinicopathologic abnormalities associated with infection with each species.
Animals—310 cats with cytologic evidence of hemoplasmosis (n = 9) or acute or regenerative anemia (309).
Procedures—Blood samples were tested by means of a broad-spectrum conventional PCR assay for hemoplasma DNA and by means of 3 separate species-specific real-time PCR assays for DNA from “Candidatus Mycoplasma haemominutum” (Mhm), Mycoplasma haemofelis (Mhf), and “Candidatus Mycoplasma turicensis” (Mtc).
Results—Overall prevalences of Mhm, Mhf, and Mtc infection were 23.2% (72/310), 4.8% (15/310), and 6.5% (20/310), respectively. Mixed infections were detected in 20 (6.5%) cats. Cats infected with hemoplasmas were more likely to be male than were uninfected cats. Infection with FeLV or FIV was significantly associated with infection with Mhf. Compared with uninfected cats, cats infected with Mhf had higher reticulocyte counts, nucleated RBC counts, and mean corpuscular volume; cats infected with Mhm had higher mean corpuscular volume; and cats infected with Mtc had higher monocyte counts.
Conclusions and Clinical Relevance—Results supported the suggestion that these 3 hemoplasma species commonly occur among cats in the United States and that pathogenicity of the 3 species varies.
Case Description—An 8-month-old spayed female domestic ferret (Mustela putorius furo) was referred for examination to determine the cause of lethargy and severe anemia.
Clinical Findings—Initial examination revealed that the ferret was lethargic but with appropriate mentation. The only other abnormal findings were severe pallor of the mucous membranes, nasal planum, and skin and a PCV of 8%. Pure red cell aplasia (PRCA) was diagnosed on the basis of cytologic evaluation of a bone marrow biopsy specimen.
Treatment and Outcome—Medical treatment included blood transfusions, IM administration of iron dextran, oral administration of antimicrobials and gastrointestinal tract protectants, and SC administration of erythropoietin. Once PRCA was diagnosed, the ferret was orally administered prednisone, cyclosporine, and azathioprine. Nine months after onset of treatment, the PRCA was in remission and the ferret was doing well. Immunosuppressive treatment was discontinued at 14 months after onset of treatment, and 36 months after initial examination, the ferret appeared to be healthy.
Clinical Relevance—It is important that PRCA be considered as a differential diagnosis for a ferret with severe anemia. Prolonged immunosuppressive treatment was successful in the ferret described here. (J Am Vet Med Assoc 2010;237:695-700)
Objective—To determine the effects of an external
nasal dilator strip on cytologic characteristics of bronchoalveolar
lavage (BAL) fluid in racing Thoroughbreds.
Animals—23 Thoroughbred racehorses in active
Procedure—Each horse raced on 2 occasions: once
while wearing an external nasal dilator strip and once
while not. Bronchoalveolar lavage was performed 12
to 18 hours after each race, and BAL fluid was analyzed
for RBC and leukocyte counts and hemosiderin
Results—Mean ± SEM count of RBCs in BAL fluid
when horses raced without the nasal dilator strip (84.6 ±
27.5 cells/µL) was not significantly different from count
when they raced with it (41.7 ± 12.2 cells/µL). Horses
were grouped as having mild or severe bleeding on the
basis of RBC count in BAL fluid after horses raced without
the nasal dilator strip. Mean count when horses with
severe bleeding raced without the nasal dilator strip
(271.0 ± 63.7 cells/µL) was significantly higher than
mean count when these horses raced with the strip
(93.8 ± 37.6 cells/µL). Mean count of lymphocytes in
BAL fluid was significantly lower after horses raced with
the external nasal dilator strip.
Conclusions and Clinical Relevance—Results suggest
that use of an external nasal dilator strip in
Thoroughbred racehorses may decrease pulmonary
bleeding, particularly in horses with severe exerciseinduced
pulmonary hemorrhage. ( J Am Vet Med Assoc
Objective—To determine the effects of meloxicam on values of hematologic and plasma biochemical analysis variables and results of histologic examination of tissue specimens of Japanese quail (Coturnix japonica).
Animals—30 adult Japanese quail.
Procedures—15 quail underwent laparoscopic examination of the left kidneys, and 15 quail underwent laparoscopic examination and biopsy of the left kidneys. Quail in each of these groups received meloxicam (2.0 mg/kg, IM, q 12 h; n = 10) or a saline (0.9% NaCl) solution (0.05 mL, IM, q 12 h; control birds; 5) for 14 days. A CBC and plasma biochemical analyses were performed at the start of the study and within 3 hours after the last treatment. Birds were euthanized and necropsies were performed.
Results—No adverse effects of treatments were observed, and no significant changes in values of hematologic variables were detected during the study. Plasma uric acid concentrations and creatine kinase or aspartate aminotransferase activities were significantly different before versus after treatment for some groups of birds. Gross lesions identified during necropsy included lesions at renal biopsy sites and adjacent air sacs (attributed to the biopsy procedure) and pectoral muscle hemorrhage and discoloration (at sites of injection). Substantial histopathologic lesions were limited to pectoral muscle necrosis, and severity was greater for meloxicam-treated versus control birds.
Conclusions and Clinical Relevance—Meloxicam (2.0 mg/kg, IM, q 12 h for 14 days) did not cause substantial alterations in function of or histopathologic findings for the kidneys of Japanese quail but did induce muscle necrosis; repeated IM administration of meloxicam to quail may be contraindicated.
Objective—To determine the optimal osteogenic source of equine mesenchymal stem cells (eMSCs) and optimize collection of and expansion conditions for those cells.
Animals—10 adult Quarter Horses and 8 newborn Thoroughbred foals.
Procedures—eMSCs were isolated from bone marrow (BM), adipose tissue, and umbilical cord blood and tissue, and the osteogenic potential of each type was assessed. Effects of anatomic site, aspiration volume, and serum type on eMSC yield from BM were investigated.
Results—BM-eMSCs had the highest overall expression of the osteogenic genes Cbfa1, Osx, and Omd and staining for ALP activity and calcium deposition. There was no significant difference in BM-eMSC yield from the tuber coxae or sternum, but yield was significantly greater from the first 60-mL aspirate than from subsequent aspirates. The BM-eMSC expansion rate was significantly higher when cells were cultured in fetal bovine serum instead of autologous serum (AS).
Conclusions and Clinical Relevance—eMSCs from BM possessed the highest in vitro osteogenic potential; eMSCs from adipose tissue also had robust osteogenic potential. The tuber coxae and the sternum were viable sources of BM-eMSCs in yearlings, and 60 mL of BM aspirate was sufficient for culture and expansion. Expanding BM-eMSCs in AS to avoid potential immunologic reactions decreased the total yield because BM-eMSCs grew significantly slower in AS than in fetal bovine serum. Additional studies are needed to determine optimal ex vivo eMSC culture and expansion conditions, including the timing and use of growth factor—supplemented AS. (Am J Vet Res 2010;71:1237-1245)
Objective—To evaluate N-hydroxysuccinimide (NHS)-biotin labeling of equine RBCs and determine posttransfusion survival of autologous equine RBCs stored in citrate phosphate dextrose adenine-1 (CPDA-1) for 0, 1, 14, and 28 days.
Animals—13 healthy adult Thoroughbreds.
Procedures—Serial dilutions of biotin and streptavidin-phycoerythrin (PE) were evaluated in vitro in blood collected from 3 horses. One horse was used to determine RBC distribution and recovery. Twelve horses were allocated to 4 groups for in vivo experiments in which blood was collected into CPDA-1. Blood was labeled with biotin and reinfused or stored at 4°C for 1, 14, or 28 days prior to labeling with NHS-biotin and reinfusion. Posttransfusion blood samples were collected 15 minutes and 1, 2, 3, 5, 7, 14, 21, 28, and 35 days after reinfusion. Biotin-labeled RBCs were detected via flow cytometry by use of streptavidin-PE. Posttransfusion lifespan of RBCs and RBC half-life were determined.
Results—Optimal biotin concentration was 0.04 pg of biotin/RBC, and the optimal streptavidin-PE ratio was 1.2 μg of streptavidin-PE/1 × 106 RBCs. Posttransfusion lifespan of autologous RBCs was 99, 89, 66, and 59 days after storage for 0, 1, 14, and 28 days, respectively. Storage did not result in significant alterations in RBC lifespan. Mean posttransfusion RBC half-life was 50, 45, 33, and 29 days for 0, 1, 14, and 28 days of storage, respectively.
Conclusions and Clinical Relevance—Biotin can be used to label equine RBCs for RBC survival studies. Posttransfusion survival of equine autologous RBCs was greater than previously reported.
OBJECTIVE To investigate renal, gastrointestinal, and hemostatic effects associated with oral administration of multiple doses of meloxicam to healthy Hispaniolan Amazon parrots (Amazona ventralis).
ANIMALS 12 Hispaniolan Amazon parrots.
PROCEDURES Birds were assigned to receive meloxicam oral suspension (1.6 mg/kg, PO, q 12 h) and 2.5 mL of tap water inserted into the crop by use of a gavage tube (n = 8) or the equivalent volume of tap water only (control group; 4) for 15 days. Urine and feces were collected 2 hours after treatment administration each day. Feces were evaluated for occult blood. Results of a CBC and serum biochemical analysis and measured N-acetyl-β-d-glucosaminidase (NAG) activity and whole blood clotting time were evaluated before, during, and after completion of treatments. Results of urinalysis and measured urine NAG activity were also evaluated.
RESULTS Birds treated with meloxicam had a significant increase in number of WBCs and decrease in PCV from before to after treatment. The PCV also decreased significantly, compared with results for the control group; however, WBC count and PCV for all birds remained within reference ranges throughout the study. One parrot treated with meloxicam had a single high value for urine NAG activity.
CONCLUSIONS AND CLINICAL RELEVANCE Meloxicam administered orally at the dosage used in this study caused no apparent negative changes in several renal, gastrointestinal, or hemostatic variables in healthy Hispaniolan Amazon parrots. Additional studies to evaluate adverse effects of NSAIDs in birds will be needed.
Objective—To conduct serologic surveillance for
Leishmania spp in English Foxhounds from a kennel,
as well as recipients of blood from these dogs, and
determine whether L infantum organisms could be
transmitted via blood transfusion.
Design—Serologic prevalence survey.
Animals—120 English Foxhounds and 51 dogs of various
breeds receiving blood from these donors.
Procedure—Foxhound blood donors, foxhound nondonors,
and nonfoxhound blood recipient dogs were
evaluated serologically for Leishmania spp by indirect
fluorescent antibody testing. Dogs that received
packed RBC (PRBC) transfusions from foxhound
donors from mid-1996 through mid-2000 were identified.
Furthermore, dogs were serologically evaluated
if they had received fresh frozen plasma (FFP) transfusions
in 1999 and 2000 from seropositive foxhound
Results—Thirty percent of the English Foxhounds were
seropositive for Leishmania spp (titer ≥ 1:16), although
the degree of seropositivity varied considerably during
the period. Furthermore, 57 foxhounds had been used
as donors from 1996 to 2000, and 342 units of PRBC
had been transfused to at least 227 patients. All 25
dogs screened that received PRBC from seronegative
foxhound donors tested negative, whereas 3 of 7 dogs
that received PRBC from seropositive donors tested
positive. All 9 dogs that received FFP from seropositive
foxhound donors remained seronegative.
Conclusions and Clinical Relevance—To our knowledge,
this report documents the first transmission of
Leishmania spp by blood transfusion. The use of foxhounds
as blood donors may not be advisable in
North America. (J Am Vet Med Assoc 2001;219:1081–1088)
Objective—To optimize the isolation and culture of mesenchymal stem cells (MSCs) from umbilical-cord blood (UCB), identify variables that predicted successful MSC isolation, and determine whether shipping, processing, and cryopreservation altered MSC viability, recovery rates, and expansion kinetics.
Sample Population—UCB samples from 79 Thoroughbred and Quarter Horse mares.
Procedures—UCB samples were processed to reduce volume and remove RBCs. Nucleated cells (NCs) were cryopreserved or grown in various culture conditions to optimize MSC monolayer expansion and proliferation. Donor and UCB-sample factors were analyzed to determine their influence on the success of MSC isolation and monolayer expansion.
Results—MSCs capable of multilineage in vitro differentiation were expanded from > 80% of UCB samples. Automated UCB processing and temperature-controlled shipping facilitated sterile and standardized RBC reduction and NC enrichment from UCB samples. The number of NCs after UCB samples were processed was the sole variable that predicted successful MSC expansion. The UCB-derived MSCs and NCs were successfully cryopreserved and thawed with no decrease in cell recovery, viability, or MSC proliferation. The use of fibronectin-coated culture plates and reduction of incubator oxygen tension from 20% to 5% improved the MSC isolation rate. Some UCB-derived MSC clones proliferated for > 20 passages before senescence. Onset of senescence was associated with specific immunocytochemical changes.
Conclusions and Clinical Relevance—Equine UCB samples appeared to be a rich source of readily obtainable, highly proliferative MSCs that could be banked for therapeutic use.