Frequencies of feline blood types in the Rio de Janeiro area of Brazil

Background: The distribution and frequency of blood types in cat populations vary according to geographic region and breed. Frequencies of feline blood types in Rio de Janeiro city, as well as in other Brazilian areas, are unknown, and the risk of unmatched transfusions and neonatal isoerythrolysis has not been estimated. Objectives: The purpose of this study was to determine the frequency of feline blood types in the area of Rio de Janeiro, Brazil. Methods: EDTA blood samples were obtained from 172 nonpedigreed domestic shorthair (DSH) cats (92 female, 80 male, 3 months-20 years old) in different sites of Rio de Janeiro city. Blood typing was performed by agglutination assays using Triticum vulgaris lectin and feline anti-A serum. The hemagglutination results for type B and AB cats were confirmed by high-performance thin-layer chromatography (HPTLC) of erythrocyte membrane gangliosides. Results: The majority (163/172, 94.8%) of cats were type A, 2.9% were type B, and 2.3% were type AB. High-titer anti-A serum agglutinated RBCs from all cats in type A and type AB blood groups, with 3+ to 4+ agglutination. The probability that a type A cat would receive type B or AB blood in a first random transfusion was calculated as 2.25% and 2.20%, respectively. HPTLC analysis of glycolipids yielded a chromatographic profile characteristic of feline gangliosides for all blood groups. Conclusions: These results indicate a high prevalence of type A cats in Rio de Janeiro, Brazil, and a low frequency of type B and AB cats, consistent with what has been observed for DSH cats in other regions of the world.     

Design,optimization, and application of a conventional PCR assay with an internal control for detection of ‘Candidatus Mycoplasma turicensis’ 16S rDNA in domestic cats from Brazil

Background: ‘Candidatus Mycoplasma turicensis’ (CMtc) is a hemotrophic bacterial species that can, alone or in combination, induce anemia in cats. The diagnostic test of choice for hemoplasma infections is PCR. Conventional PCR assays have been developed for the detection of Mycoplasma haemofelis (Mhf) and ‘Candidatus M. haemominutum’ (CMhm) but not for CMtc. Although real‐time PCR assays have been reported for all of the feline hemoplasmas, the expense of necessary instrumentation precludes its use in Brazil and many other countries. Objectives: The goals of this study were to develop and optimize a conventional PCR assay to diagnose CMtc using an internal control to detect false‐negative results, and to evaluate the occurrence of CMtc infection in domestic cats from Brazil. Methods: Species‐specific primers were designed and a PCR assay was developed for the detection of CMtc 16S rDNA in cat blood. Sensitivity was determined by serial 10‐fold dilutions of plasmid and DNA extracted from blood from an experimentally infected cat. EDTA blood samples from 373 cats were collected. DNA was extracted using a silica‐based protocol and tested using the PCR assay. Results: Primer concentration, annealing temperature, and MgCl₂ concentration were optimized in the presence and absence of the internal control. Two samples negative for the internal control were excluded. Of the remaining 371 samples (117 healthy and 254 unhealthy cats), 17 (4.6%) were positive for CMtc. Conclusion: These results demonstrate the utility of an optimized PCR assay to detect CMtc in feline blood samples. We also report for the first time the prevalence of CMtc infection in domestic cats in Brazil.     

Reduction of non-specific reactions to the Brucella abortus serum agglutination test by the addition of EDTA

Serum samples taken from cattle known to be infected with Brucella abortus, and samples routinely collected as part of the brucellosis eradication scheme were tested by the serum agglutination test (SAT), complement fixation test (CFT) and the SAT modified by the addition of ethylene diamine tetra-acetic acid (EDTA). In 64 per cent of the samples giving an SAT titre greater than 100 iu, but a CFT titre 8.3 icftu or less, the agglutination reaction was sufficiently affected by the action of EDTA for the titre to drop to below 100 iu. Only 5 per cent of samples giving an SAT titre greater than 100 iu and a CFT titre of 20 icftu or greater were affected in a similar manner by EDTA, and none of the 29 sera taken from known infected animals showed a drop in titre to below 100 iu, although some with titres greater than 500 iu did show significant EDTA sensitivity.     

Hematologic evaluation of normal and anemic lambs with the Technicon H*1 using EDTA or heparin as anticoagulants

Abstract: A study was performed to evaluate blood from young lambs using the Technicon H*1 hematology analyzer, with emphasis on RBC parameters, comparison of tripotassium EDTA and heparin, and the effects of storage on heparinized blood. Blood samples from lambs 2 days to 18 weeks of age were analyzed within 6 hours, revealing a high precision, except for WBC counts in heparinized blood. The HCT values estimated by the H*1 correlated well (r²= .90) with those obtained by the microhematocrit method. Mean hematologic values obtained for heparinized blood differed by up to 4% from values obtained for blood collected into EDTA. WBC counts decreased 8.4% in heparinized blood stored at 4°C for 1 day, but differences observed in RBC counts were ≤ 2%. Problems occurred when analyzing blood from young lambs with low hemoglobin values because the H*1 incorrectly counted microcytes with volumes of < 10 fL as platelets. When the necessary corrections were performed, the H*1 was useful for analyzing RBC parameters in lamb blood collected both into EDTA and into heparin.     

Effect of venipuncture site and anticoagulant on selected hematologic values in black rhinoceros (Diceros bicornis).

Paired blood samples were collected from the ear and radial vein of four captive healthy adult black rhinoceroses (Diceros bicornis). Samples were collected using heparin or ethylenediaminetetraacetic acid (EDTA) as an anticoagulant. Packed cell volume (PCV) and total protein (TP) values were compared between samples drawn from the two venipuncture sites and treated with the two anticoagulants to determine whether statistically significant variation occurred. No significant difference in the grouped values was observed when venipuncture sites (ear and radial vein) were compared using the same anticoagulant (heparin). However, when comparing different anticoagulants (EDTA and heparin) used to collect blood from the radial vein, the grouped-heparinized samples had higher mean PCV and TP values than did the EDTA-treated samples. These differences may be important when performing serial sampling in a sick rhinoceros and suggest that the choice of anticoagulant should be consistent, although selection of venipuncture site may be less important when monitoring selected hematologic values in black rhinoceroses.     

The influence of anticoagulants on the measurement of total protein concentration in equine peritoneal fluid

The aim of this study was to evaluate the influence of two commonly used anticoagulants (K3EDTA and lithium heparin) on refractometric and spectrophotometric measurement of total protein (TP) concentration in equine peritoneal fluid samples. The influence of a commercial solution of K3EDTA, a solution of K3EDTA in distilled water and lithium heparin on the refractometric and spectrophotometric (biuret) quantification of TP content in peritoneal fluid samples was assessed. Total protein concentration measured by refractometry was consistently overestimated in samples with commercial K3EDTA. The solution of K3EDTA in distilled water only caused TP overestimation at high K3EDTA concentrations (>5 micromol/ml). By contrast, lithium heparin did not influence the refractometric values of TP. Neither anticoagulant modified TP values when measured by the biuret method. In conclusion, the use of K3EDTA as anticoagulant may result in a significant overestimation of TP values of peritoneal fluid samples measured by refractometry.     

The comparison of thiobarbituric acid reactive substances (TBARS) concentrations in plasma and serum from dairy cattle

The aim of this study was to compare thiobarbituric acid reactive substances (TBARS) concentrations in serum, plasma with heparin (heparin plasma), and plasma with ethylenediaminetetraacetic acid disodium salt (EDTA plasma) as anticoagulants from dairy cattle. Serum, heparin plasma, and EDTA plasma TBARS were not sufficiently strongly correlated to allow accurate prediction of one set of values from the other. Heparin plasma TBARS concentrations were found to be lower, and were affected by the duration of mixing during the assay process. The results suggest that it is necessary to differentiate TBARS concentrations between different sample types such as serum, heparin plasma, and EDTA plasma. For measurements of TBARS concentrations in cattle, EDTA plasma samples may be more suitable than the other samples.     

The hemagglutinating activity of Pseudomonas aeruginosa strains isolated from humans and animals

The presence and type of adhesins occurring in Pseudomonas aeruginosa strains were determined by hemagglutination test with a 3% suspension of normal and trypsin-treated human group A erythrocytes, with or without the addition of sugar inhibitors (D-mannose, D-glucose, L-fucose, D-galactose, D-fructose, lactose, N-acetylneuraminic acid, N-acetylglucosamine and N-acetylgalactosamine). This study showed that a low percentage of Pseudomonas aeruginosa strains caused the agglutination of normal erythrocytes. Trypsin treatment of erythrocytes did not affect the hemagglutinating properties, indicating that hemagglutination was not dependent upon a trypsin-sensitive protein on the erythrocytes surface. Most of the studied strains agglutinated RBCs at 37 degrees C. A great variability in the inhibiting activity on studied strains was observed among the carbohydrates tested. These results demonstrated the predominant role of N-acetylglucosamine, N-acetylgalactosamine and N-acetylneuraminic acid for Pseudomonas aeruginosa adhesion to RBCs.     

Temporal effects of 3 commonly used anticoagulants on hematologic and biochemical variables in blood samples from macaws and Burmese pythons

BACKGROUND: Few studies have been done to evaluate anticoagulants for use with blood samples from birds and reptiles. Heparin currently is the most commonly used anticoagulant in practice, but may adversely affect blood cell staining and quantitation. OBJECTIVE: The purpose of this study was to evaluate the effects of lithium heparin, K3-EDTA, and sodium citrate, with and without the addition of albumin, on hematologic variables in macaw (Ara sp) and python (Python molurus bivittatus) blood samples. METHODS: Blood samples from 10 macaws and 10 Burmese pythons were collected in heparin-coated syringes and placed into tubes containing either lithium heparin, K3-EDTA, or sodium citrate with and without the addition of 0.25 mL of a 22% bovine serum albumin solution. Cell lysis was determined by counting the number of lysed cells/200 WBCs in Wright's-Giemsa-stained blood smears and by qualitative evaluation of pink plasma in microhematocrit tubes. A CBC was done after 3, 12, and 24 hours of storage at 4 degrees C in anticoagulant-containing tubes and results were compared with those obtained at 0 hour for the heparin-coated syringe sample. A biochemical panel also was done at each time point in similarly stored lithium-heparin samples. RESULTS: Hemolysis was significantly increased in citrated samples from both macaws and pythons beginning at 12 hours. At 24 hours, 19 of 30 (63%) macaw samples in all anticoagulants had >100 lysed cells/200 WBCs. There were no significant differences in hematologic values in samples from pythons collected in heparin or EDTA at any time point. No significant differences were found in the number of lysed cells or in other hematologic data in samples with albumin. Glucose concentration decreased and potassium concentration increased significantly over time in heparinized blood samples. CONCLUSIONS: Based on the results of this study, whole blood samples anticoagulated with lithium heparin or EDTA should be evaluated within 12 hours (macaws) or 24 hours (pythons) of collection and stored at 4 degrees C for best results. Citrate should be avoided as it may result in increased cell lysis. The addition of albumin does not prevent cell lysis.     

Myelofibrosis in a cat (abstract)

A 2-year-old female domestic shorthair cat was presented with ill-health and marked ascites. Clinically, there was marked pallor of mucous membranes. Two haematological examinations and transudate-exudate analyses of peritoneal fluid were conducted 1 week apart, as well as a coagulation screen and routine serum biochemistry. The haematological examinations revealed a severe persistent non-regenerative anaemia. There were no abnormalities within the neutrophil and lymphocyte cell lines. Very occasional nucleated red blood cells (RBCs) in peripheral circulation and some RBC agglutination were seen. Analysis of peritoneal fluid disclosed a modified transudate, and small numbers of erythroid and myeloid precursors were observed on the second occasion. The cat was FIV/FeLV-negative, and there were no coagulation abnormalities. Following necropsy and histopathological examination, myelofibrosis and extramedullary haematopoiesis were seen. Thoracic masses were hyperplastic lymph nodes in which there was evidence of extramedullary haematopoiesis. The exact cause of the myelofibrosis was not obvious but it may have been a case of myeloid metaplasia, however not all the characteristics of this entity were present.     

Identification of Pentatrichomonas hominis in feline fecal samples by polymerase chain reaction assay

Pentatrichomonas hominis is considered to be a commensal protozoan of the vertebrate digestive tract. On the basis of light microscopic examination of feces, some investigators presumptively identified P. hominis as a causative agent of feline diarrhea. However, molecular identification of P. hominis infection in the cat has not been reported. Another trichomonad, Tritrichomonas foetus, is recognized as an intestinal pathogen in cats and often presumptively diagnosed on the basis of the presence of trichomonads in diarrheic feces. It is of importance to determine if cats are natural hosts for P. hominis, as the presence of this organism could result in inaccurate assumption of T. foetus infection. In this study, we used a species-specific PCR assay to identify P. hominis 18S rRNA genes in fecal samples collected from a convenience population of cats in which a high prevalence of T. foetus infection had been previously identified (cat show) or suspected (submitted for T. foetus diagnostic testing). The prevalence of T. foetus infection in these samples was 31% and 28.6%, respectively. P. hominis infection was identified by PCR of DNA extracted from feces of five cats (1.9% and 2.1% of fecal samples, respectively). All cats in which P. hominis was identified were also infected with T. foetus. PCR identification of P. hominis infection in the cat should facilitate future studies to determine the pathogenicity of this species and enable differentiation of P. hominis from other known or as-yet unidentified species of trichomonads that may infect cats.     

Autologous conditioned serum: the comparative cytokine profiles of two commercial methods (IRAP and IRAP II) using equine blood

Reasons for performing study: Osteoarthritis (OA) is one of the most prevalent and debilitating conditions affecting the horse. Autologous conditioned serum (ACS), commercially available as IRAP and IRAP II, is a recently developed treatment for OA in which plasma is prepared from venous blood by incubation with glass beads for 24 h. This product has been shown to increase anti‐inflammatory cytokines and growth factors in human blood. However, data for equine ACS preparations are lacking. Objectives: To characterise the protein profiles produced by commercially available ACS systems in equine blood. Methods: Blood was drawn from 5 horses into 6 groups: red top vacutainer (control), IRAP and IRAP II, with and without heparin. Samples were collected 1 or 24 h post draw and analysed for IL‐1ra, IL‐10, IGF‐1, TGF‐β, TNF‐α and IL‐1β using ELISAs. Results: Twenty‐four hour IRAP and IRAP II samples contained significantly higher levels of all cytokines relative to 1 h serum controls. At 24 h, IRAP II contained significantly higher levels of IL‐1ra and IRAP contained significantly higher levels of TNF‐α, compared to 24 h controls. In addition, TGF‐β, IL‐10 and IL‐1β in IRAP and IRAP II sera were similar to 24 h serum controls. The addition of heparin significantly reduced levels of IGF‐1, TNF‐α and TGF‐β, and significantly elevated levels of IL‐1ra. Conclusions: The cytokine profile that IRAP II produced is modestly better than IRAP. Incubation of whole blood in glass tubes stimulated cytokine synthesis, although not as efficiently as IRAP II. Potential relevance: Although high levels of IL‐1ra were found in ACS, elevation of other factors suggests these cytokines play a previously understated role in clinical improvements. Because ACS has been shown to alleviate clinical symptoms of OA, the present study suggests that factors other than IL‐1ra alone might be involved in its clinical efficacy. Species‐dependent elevations of cytokines warrant further investigation and optimisation of the systems appears to be necessary based on the differences between human and equine blood.     

Immune‐mediated myasthenia gravis in a methimazole‐treated cat

A 12‐year‐old female neutered ragdoll crossbred cat was presented for investigation of generalised weakness and regurgitation. The cat was being treated with transdermal methimazole for hyper‐thyroidism, which had been diagnosed 10 weeks previously. An acetylcholine receptor antibody titre was consistent with acquired myasthenia gravis. Withdrawal of methimazole and treatment with pyridostigmine was followed by resolution of clinical signs and reduction of the acetylcholine ‐receptor antibody titre. Medical control of hyperthyroidism was subsequently achieved with carbimazole, administered in conjunction with pyridostigmine, and no recurrence of clinical signs was observed. Myasthenia gravis is an uncommon but clinically significant adverse effect of methimazole therapy in cats, and may be caused by immunomodulatory properties of this drug. An adverse drug reaction should be considered in cats receiving methimazole that develop myasthenia gravis, and potentially also other immune‐mediated disorders.     

Platelet concentrations and platelet-associated IgG in greyhounds

BACKGROUND: Greyhounds have lower platelet concentrations (PC) than dogs of other breeds have. No underlying cause has been investigated. HYPOTHESIS: We hypothesized that Greyhounds have lower mean PC because of breed variation, not immune-mediated causes. Our secondary hypothesis was that PC is dependent on the method of analysis. ANIMALS: Sixty privately owned Greyhounds in Kansas. METHODS: Blood samples were collected into evacuated glass tubes containing ethylenediamine tetraacetic acid (EDTA). Blood smears were evaluated for platelet clumps. All 60 samples had PC determined by manual, impedance, and buffy coat analyzer methods. Results of the 60 samples were compared with results of samples with (n = 25) and without (n = 35) clumps, and with control dogs. Platelets were assayed for the presence of surface-associated antigen (PSAIgG) by direct immunofluorescence. RESULTS: The mean PC was below that of the control dogs for the impedance method (P < .001). No significant difference in PC was detected between analysis methods or between samples with or without platelet clumps. Three of 60 (5%) of the Greyhounds had PC between 50,000 and 100,000/microL with impedance analysis; no samples had < 100,000/microL via buffy coat analysis. PSAIgG was not identified in any samples. CONCLUSIONS AND CLINICAL IMPORTANCE: The mean Greyhound PC for the impedance method was below the reference interval for control dogs but was not significantly different from PC determined by other methods. An immune-mediated cause for the lower PC was unlikely because no samples had PSAIgG. The decreased PC is most consistent with breed variation. As only 0-5% of samples, depending on analysis method, had PC < 100,000/microL, a Greyhound with a PC < 100,000/microL is not necessarily consistent with breed variation, thus diagnostic testing is indicated.     

Cold agglutinin activity in 2 dogs

A 5‐year‐old neutered male Mastiff and an 8‐year‐old spayed female Labrador Retriever were presented to the University of Minnesota Veterinary Medical Center. The Mastiff was presented for evaluation of lameness and pyoderma one month prior in Missouri, where he tested positive for Ehrlichia canis by serum ELISA test, treated with doxycycline. PCR for Ehrlichia sp, Anaplasma sp, Babesia sp, and Bartonella sp, and PCR for antigen receptor rearrangement were negative, serum protein electrophoresis (SPE) revealed polyclonal gammopathy, and mildly reactive lymphoid cells were seen cytologically. The Labrador presented with a proliferative rostral mandibular gingival mass and lipomas for further presurgical evaluation of cold agglutinin activity documented by a commercial laboratory 2 years earlier prior to removal of a grade II mast cell tumor. This dog had a negative SNAP4Dx, normal SPE, and persistently increased serum ALP activity and polyuria/polydipsia suggestive for hyperadrenocorticism. Both dogs had markedly agglutinated RBC in the EDTA samples that dispersed with warming, and normal plasma color. Cold agglutinin activity was demonstrated by direct saline agglutination testing using whole blood and washed erythrocytes demonstrating agglutination at 30°C, 25°C, 15°C, and 4°C, but not at 37°C . CBC results (ADVIA 2120i) from the Mastiff revealed no significant differences in the RBC results obtained at room temperature (RT) and at 37°C; however, the RT run demonstrated negative bias in neutrophil and platelet concentrations attributed to rapid RBC settling. This uncommon hematologic condition may cause artifacts on the automated leukogram and platelet count, and may be subclinical for long periods.     

Heparin-induced agglutination of erythrocytes in horses

Heparin was administered subcutaneously 2 times a day for 4 days to 5 horses. An additional group of 5 horses was used as time-matched controls. Significant decreases in PCV, erythrocyte count, and hemoglobin concentration were observed during heparin therapy. The mean corpuscular volume (MCV) of the heparin-treated horses increased to a peak value of 66.1 fl on the last day of treatment. Erythrocyte creatine concentration and glucose 6-phosphate dehydrogenase activity increased moderately during the treatment. These data indicated that the rapid, profound increase in MCV during heparin therapy was not primarily a result of release of large immature erythrocytes from the bone marrow. A second experiment was subsequently performed, using 3 horses. These horses were given heparin 2 times a day, as was done in the first experiment. Saline wet mounts of erythrocyte suspensions were examined once a day for the presence of agglutination. Cell suspensions were examined with or without exposure to a dilute trypsin solution, and erythrocyte counts were done on each suspension, using an electronic cell counter. Agglutination of erythrocytes was evident on the first day of treatment and became more pronounced as treatment progressed. Exposure to trypsin solution reversed the agglutination. The apparent erythrocyte count decreased and MCV increased sharply in the samples processed normally, but there was little change in those suspensions exposed to trypsin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of a rapid agglutination method for detection of equine red cell surface antigens (Ca and Aa) as part of pretransfusion testing

BACKGROUND: Blood typing before transfusion minimizes the risk of transfusion reactions and prevents immunization of the recipient against incompatible RBC antigens. The major RBC antigens that warrant identification before packed RBC or whole blood transfusions in horses are Ca and Aa. Standard blood-typing protocols are time-consuming (2.5-3.0 hours) and impractical in emergency settings. OBJECTIVES: The purpose of this study was to determine whether equine RBCs could be typed for Ca and Aa antigens using sera from horses with RBC antibodies in a modified rapid (15 minute) blood-typing protocol. METHODS: Serum was obtained from a horse with anti-Ca antibodies and from another horse with anti-Aa antibodies. The presence of agglutinating antibodies was confirmed with antibody screening. Venous blood samples, collected in citrate-phosphate-dextrose, were obtained from 21 horses of various breeds. Samples were blood typed in the Veterinary Medical Teaching Hospital Hematology Laboratory using standard methodology. Washed RBCs from each of the 21 horses were incubated individually with anti-Ca and anti-Aa sera at dilutions of 1:4, 1:8, and 1:16 for 15 and 30 minutes at room temperature and 37 degrees C. RESULTS: Of the 21 horses, 13 were identified as Aa+/Ca+, four were Aa+/Ca-, two were Aa-/Ca+, and two were Aa-/Ca-. All 17 Aa-positive horses had a positive agglutination reaction at all dilutions of anti-Aa serum, incubation times, and temperatures, while all Aa-negative horses were negative. Each Ca-positive horse had a positive agglutination reaction at all incubation time points and temperatures up to the 1:16 dilution of the anti-Ca serum. All Ca-negative horses were negative at all times, temperatures, and dilutions of anti-Ca serum. Use of the modified protocol on 26 hospitalized horses resulted in accurate typing, based on complete antibody screens. CONCLUSIONS: These results support the hypothesis that equine RBCs can be blood typed using a rapid (15 minute) protocol, at room temperature, for the presence of Ca and Aa antigens using equine-derived antisera. This technique may be beneficial for pretransfusion testing of equine patients in an emergency setting.     

A serologic survey of Oklahoma cats for antibodies to feline immunodeficiency virus, coronavirus, and Toxoplasma gondii and for antigen to feline leukemia virus

A serologic survey was done on 618 cat sera submitted to the Oklahoma Animal Disease Diagnostic Laboratory between July 1, 1987 and June 30, 1988. The samples were collected from clinically normal and sick cats. The sera were tested for the presence of antibodies to feline immunodeficiency virus by a commercial immunoassay, to a coronavirus by an indirect fluorescent antibody test, and to Toxoplasma gondii by a commercial latex agglutination test and for the presence of feline leukemia virus antigen with one of 3 different commercial assay kits. Ten percent of the sera had antibodies to feline immunodeficiency virus, 35% had antibodies to a coronavirus, and 22% had antibodies to Toxoplasma gondii. Feline leukemia virus antigen was detected in 15% of the sera. Thirty-two percent of the sera had evidence of exposure to 2 or more of the agents.     

Effect of storage on bovine erythrocyte cholinesterase activity.

Bovine whole blood and washed erythrocyte samples with either sodium heparin or EDTA as the anticoagulant were stored at -24, 2 to 3, 21, or 37 C for up to 8 weeks to determine the stability of erythrocyte cholinesterase (ChE) activity. An automated colorimetric method was used to measure ChE activity. Regression analysis was used to estimate the rate of decline of ChE activity of samples to 95% of normal. Samples stored at -24 and 2 to 3 C were very stable, and ChE activity was maintained for several weeks or longer. Samples stored at 21 C maintained an activity level not less than 95% of normal for at least 9 days and those stored at 37 C for at least 2 days. The type of anticoagulant used did not appear to affect significantly the stability of activity.     

Comparison of the effect of dipotassium ethylenediaminetetraacetic acid and lithium heparin on hematologic values in the green iguana (Iguana iguana).

We compared the effects of dipotassium ethylenediaminetetraacetic acid (EDTA) and lithium heparin on hematologic values of green iguanas (Iguana iguana). Thirty-two privately owned sibling iguanas had blood drawn, and the sample was divided into three components: an EDTA tube, a heparin tube, and a nonanticoagulated blood smear. A full reptilian complete blood count was performed on each anticoagulated sample, and white blood cell (WBC) and leukocyte differential counts were performed on the whole-blood smears. Heparin and EDTA samples differed significantly in absolute values of thrombocytes, WBC, heterophils, and monocytes. The EDTA had no significant effect on the packed cell volume or plasma protein values, and the white blood count and differential counts produced with EDTA were more similar to those of the nonanticoagulated blood smear than were the counts produced with heparin.