Hematological and coagulation profiles in dogs experimentally infected with Angiostrongylus vasorum (Baillet, 1866).

Hematological and coagulation profiles were studied in crossbred dogs experimentally infected with Angiostrongylus vasorum. Two groups of five dogs were experimentally inoculated with 50 and 100 third stage infective larvae (L(3)) of A. vasorum per kilogram of body weight. A third group of five uninfected animals was used as control. One sample of 10 ml of blood was collected from each animal on the 10, 20, 30, and 45 days after inoculation (dai) and at 30-day intervals thereafter for the remainder of the 210-day experimental period. The blood sample was used for the complete hemogram and platelet count, as well as measurements of prothrombin time, partial thromboplastin time and factors V and VIII. Anemia was observed in infected dogs, 6 weeks after the infection. The eosinophils presented peaks in four periods after infection. Thrombocytopenia became accentuated on the 72 dai. Decreased prothrombin time activity and increased partial thromboplastin time were observed at the 6 and 9 weeks after infection and decreased of factors VIII and V activities occurred from 4 to 6 weeks after infection. It may be conclude that infection by A. vasorum in dogs may cause a discrete anemia during the acute phase which is probably regenerative. In addition, important hemostatic alterations due to the infection suggest a chronic intravascular consumption coagulopathy.     

Blood biochemical profiles of thai indigenous and Simmental x Brahman crossbred cattle in the Central Thailand

Plasma biochemical profiles were studied in 112 mature (3 to 5-year-old) healthy cattle comprised of 61 Thai indigenous and 51 Simmental x Brahman crossbred male and cyclic female cattle at Nongkwang (Central Thailand) Livestock Research and Breeding Center, Thailand. Data were analysed for the effect of breed and sex. The results showed that the plasma glucose and gamma-glutamyl transferase (GGT) in the two breeds were significantly (P < 0.05) different. Furthermore, the urea, creatinine, albumin, total protein, aspartate amino transferase (AST), alanine amino transferase (ALT) and alkaline phosphatase (ALP) levels in Thai indigenous were significantly (P < 0.01) higher than in crossbred cattle. However, creatine kinase did not significantly differ in crossbred and indigenous animals. A sex difference was found in glucose level with male Thai indigenous having significantly higher levels (P < 0.05) than the other three groups. Plasma urea concentration in male crossbred cattle was lower than in the other groups (P < 0.05). Female crossbred cattle had significantly (P < 0.05) lower plasma creatinine levels than the other animals. Furthermore, levels of albumin in male and total protein in female crossbred were the lowest (P < 0.05) among the groups. The AST, ALT, ALP and GGT levels were significantly (P < 0.05) different between male and female. Female crossbred cattle had the lowest (P < 0.05) AST and GGT levels, whereas lowest (P < 0.05) ALT and ALP concentration was determined in male individuals of these breeds.     

Biochemical abnormalities observed in serum of dogs infected with large Babesia in Warsaw (Poland)

Biochemical abnormalities observed in canine babesiosis are related to the severity of the disease. The primary biochemical abnormalities found in affected dogs are: increase of the serum activity of transaminases and alkaline phosphatase, azotemia, and hypoglycemia. The purposes of this study were: 1) to estimate biochemical abnormalities in dogs infected with large Babesia in Warsaw and 2) to evaluate statistically changes observed during canine babesiosis in dogs from Warsaw. Samples of serum were collected from dogs naturally infected with large Babesia. Among 2023 positive samples, 202 were randomly selected. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatinine, blood urea nitrogen (BUN), total serum protein (TSP), albumin and blood glucose concentration were determined with a clinical chemistry analyser. Elevated activity of ALT, AST and ALP was detected accordingly in: 64.9, 92.6 and 31.7% of dogs. Elevated creatinine concentration and BUN were detected accordingly in 30.7 and 62.4% of dogs. Decrease of TSP, albumin, BUN, and hypoglycemia was detected accordingly in: 19.8, 32.7, 1.5 and 18.3% of dogs. The most common biochemical abnormalities found in affected dogs were: increase of activity of transaminases and ALP, elevated creatinine concentration, hypoalbuminemia and hypoglycemia. These abnormalities resulted from hepatopathy, renal failure and fasting.     

The pathology of acute Nolletia gariepina poisoning of cattle

Toxicity in cattle by the shrub Nolletia gariepina was induced experimentally by intraruminal administration of 3 g/kg dried, milled plant material as a single dose. The animals had to be starved for 24 hours before dosing, as dosing on a full rumen did not induce any signs of toxicity during 5 days of observation and clinical pathology monitoring. Clinical signs were not specific and varied according to the duration (acute versus subacute) of the toxicological process. Clinical pathological parameters indicated renal and to a lesser extent hepatic damage, with raised serum concentrations of urea, creatinine, aspartate aminotransferase (AST) and gamma glutamyl transferase (GGT). Increased urinary sodium and potassium concentration and GGT activity, as well as proteinuria, were evident. Histological and electron microscopic examinations revealed acute renal tubular epithelial cell degeneration and necrosis, especially of the proximal convoluted tubules. Mild hepatocellular degeneration was also noticeable.     

Infection of dogs by experimental inoculation with human isolates of Giardia duodenalis: clinical and laboratory manifestations

The susceptibility of dogs to experimental inoculation with trophozoites and cysts of human isolates of Giardia duodenalis and the clinical and laboratory profiles of infection of these animals were studied. Two groups (A and B), each comprising three dogs, were inoculated with G. duodenalis trophozoites and cysts, respectively. A third group of two dogs was not inoculated and remained as control. After inoculation feces were collected daily to determine the pre-patent period, by flotation in 33% zinc sulfate solution. Blood samples (5mL) were collected from animals at 15-day intervals during the 165 days of the experimental period and were used to carry out the hemogram and biochemical evaluation of the levels of total protein, albumin, alanine aminotransferase, gamma glutamyltransferase, urea and creatinine. A prepatent period was observed at 5-6 days post-inoculation (p.i.) in the inoculated dogs, with cysts eliminated for approximately 3 months. No alterations were seen in the clinical parameters evaluated. Anemia was observed at 15 p.i. in the inoculated dogs. The mean eosinophil count of inoculated groups was higher than that of the control (p< or =0.05) but none of the biochemical parameters analyzed presented significant differences. The results of this study show that G. duodenalis from human isolates is able to infect dogs with minimal systemic manifestations without producing clinical signs of giardiasis.     

Effects of storage time and freezing temperature on clinical chemical parameters from canine serum and heparinized plasma

To assess changes in 24 blood constituents in frozen serum and heparinized plasma, blood samples were drawn from 10 clinically normal German Shepherd army dogs. The storage characteristics of nine enzymes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, lipase), and 15 metabolites and minerals (albumin, bile acids, bilirubin, calcium, cholesterol, creatinine, fructosamine, glucose, magnesium, phosphate, potassium, protein, sodium, triglycerides, urea) were studied. Parallel samples of serum and heparinized plasma were stored for 90 and 240 days at two different storage temperatures, -200 degrees C and -700 degrees C. Sixteen of the 24 analytes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, bile acids, calcium, cholesterol, creatinine, fructosamine, magnesium, phosphate, urea) showed statistically significant (p < 0.05) changes during the storage period related to storage time, storage temperature, and sample type. Seven of the analytes (amylase, GGT, GLDH, LDH, bile acids, fructosamine, magnesium) showed changes of possible clinical importance with mean differences from baseline larger than 20% for the enzymes and 10% for the metabolites and minerals during the storage periods.     

Plasma biochemistry of one-humped camel (Camelus dromedarius): Effects of anticoagulants and comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals but limited informations are existing for camel plasma biochemistry. Eleven clinically healthy one-humped camels were blood sampled in different tubes containing different anticoagulants and plain tubes for harvesting plasma and serum. The concentrations of glucose, cholesterol, triglyceride, total bilirubin, urea, creatinine, total protein, albumin, calcium, inorganic phosphorus, magnesium, and chloride and the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatine kinase (CK) and gamma glutamyl transferase (GGT) were measured. Except for the amounts of AST, ALT, CK, total bilirubin, and inorganic phosphorus, other measured parameters were significantly lower in citrated plasma than in serum. Most parameters did not show any difference, but significant increase for CK activity and significant decrease for GGT, cholesterol, creatinine and chloride were seen when heparin was used as anticoagulant. Using EDTA as an anticoagulant caused a significant difference in the amounts of some measured parameters in plasma except glucose, AST, ALT, GGT, cholesterol, albumin, total protein, bilirubin, and triglyceride in comparison with serum.     

Plasma biochemistry of one-humped camel (Camelus dromedarius): effects of anticoagulants and comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals but limited informations are existing for camel plasma biochemistry. Eleven clinically healthy one-humped camels were blood sampled in different tubes containing different anticoagulants and plain tubes for harvesting plasma and serum. The concentrations of glucose, cholesterol, triglyceride, total bilirubin, urea, creatinine, total protein, albumin, calcium, inorganic phosphorus, magnesium, and chloride and the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatine kinase (CK) and gamma glutamyl transferase (GGT) were measured. Except for the amounts of AST, ALT, CK, total bilirubin, and inorganic phosphorus, other measured parameters were significantly lower in citrated plasma than in serum. Most parameters did not show any difference, but significant increase for CK activity and significant decrease for GGT, cholesterol, creatinine and chloride were seen when heparin was used as anticoagulant. Using EDTA as an anticoagulant caused a significant difference in the amounts of some measured parameters in plasma except glucose, AST, ALT, GGT, cholesterol, albumin, total protein, bilirubin, and triglyceride in comparison with serum.     

Haematological and biochemical changes in dogs naturally infected with Angiostrongylus vasorum before and after treatment

Haematological and biochemical parameters were studied prospectively in 48 dogs naturally infected with Angiostrongylus vasorum in a primary care setting. Samples for analysis were obtained when treatment was started and 42days afterwards. Prior to treatment, 21% of affected dogs exhibited eosinophilia, whereas increased total white blood cell (WBC) counts and neutrophilia were observed in only 4.2%. WBC counts and concentrations of neutrophils, eosinophils and monocytes decreased significantly from days 0 to 42, indicating that, even in dogs without elevated absolute blood values, a low grade inflammatory response may be present in dogs with A. vasorum infection. Biochemical changes (especially an increase in serum globulins and a decrease in serum fructosamine) were in agreement with the findings of other studies. The results show that the diagnosis of canine angiostrongylosis should not be excluded based on unremarkable haematological and blood biochemical parameters. They also support our recent finding that a low serum fructosamine concentration may be associated with infection with A. vasorum.     

Cytological and parasitological analysis of bronchoalveolar lavage fluid for the diagnosis of Angiostrongylus vasorum infection in dogs

Bronchoalveolar lavage (BAL) is a procedure that retrieves cells and other elements from the lungs for evaluation, which helps in the diagnosis of many pulmonary diseases. The aims of this work were to perform this procedure in dogs in the acute and chronic phases of an Angiostrongylus vasorum infection for cytological analysis and to evaluate the potential of this technique as a diagnostic method for this lung-heart worm. The BAL procedure was performed through the use of an endotracheal tube on seven A. vasorum infected dogs and on five non-infected dogs lined as a control group. Sixty days post-infection (dpi) active and live larvae were retrieved from the bronchoalveolar fluid (BALF) of all infected dogs. Furthermore, in one animal it was possible to retrieve larvae in its BALF before the pre-patent period. This work reports that the A. vasorum infection resulted in an increase of relative neutrophils and eosinophils counts. In contrast, there was a significant decrease in the alveolar macrophage relative count in infected animals from 60 to 330 dpi. This study shows that the BAL is an accurate technique for the diagnosis of canine angiostrongylosis. Moreover, the technique allows us to retrieve cells and other elements that line the lung surface for cytological evaluation, which provides information about inflammatory diseases, and the diagnosis and prognosis of pulmonary parasites such as A. vasorum.     

Larval output of infected and re-infected dogs with Angiostrongylus vasorum (Baillet, 1866) Kamensky, 1905

Canine angiostrongylosis is a nematode infection in domestic dogs and wild canids. A natural infection in a domestic dog frequently leads to pneumonia, loss of physical performance, coughing, anemia, cardiac insufficiency, pulmonary fibrosis and death. The main diagnostic method is based on the finding of Angiostrongylus vasorum first-stage larvae (L1) in infected dog feces. With this objective, 11 experimentally exposed to 100 third-stage larvae (L3) per kilogram of body weight (mean = 885.45 L3/animal; S.E. = 77.7). The animals were monitored for 300 days post-single-infection (PI) and the quantity of L1 output measured. Our results showed an irregular excretion of L1 and a variation in the pre-patent period (33-76 days) and the number of L1 excreted by individual animals (1-1261 L1/g). After 300 days PI, five dogs were exposed a second time and monitored for 300 days post-re-infection (PRI) (=600 days PI). The quantity of L1 output demonstrated that double exposed dogs also presented an irregular excretion of L1 but a smaller variation in the number of L1 excreted by individual animals (4-550 L1/g).     

一起群发性奶牛尿素中毒的临床研究

2004年南京某奶牛公寓由于饲养户在奶牛饲料中添加尿素方法不当,引起群发性尿素中毒。该牛群共有奶牛87头,发病64头,药物治疗56头,治愈54头,死亡10头,均为成母牛。作者调查了发病牛群的病史、临床症状、病理变化、治疗情况,对中毒奶牛的血液进行生化分析,并与正常奶牛比较,结果血清谷草转氨酶(AST)、γ-谷氨酰转移酶(GGT)、碱性磷酸酶(ALP)、磷酸肌酸激酶(CK)、磷酸肌酸激酶同工酶(CK-MB)、肌酐(CRE)、尿素氮(UN)、葡萄糖(GLU)、乳酸脱氢酶(LDH-L)等差异极显著(P<0·01),尿素(UREA)、血清K等差异显著(P<0·05),这些指标活性升高说明病牛心肌、胃肠平滑肌、肝肾等存在与病死牛的病理变化相一致的损伤。     

Detection of specific antibodies in dogs infected with Angiostrongylus vasorum

Canine angiostrongylosis, caused by the nematode Angiostrongylus vasorum, is an emerging cardiopulmonary disease in Europe which can be fatal if left untreated. We determined the diagnostic value of the specific detection of antibodies against A. vasorum adult somatic antigen, adult excretory/secretory (E/S) antigen and first stage larvae (L1) somatic antigen in ELISAs. Also, A. vasorum adult somatic antigen purified by monoclonal antibodies (mAb) was evaluated in a sandwich-ELISA. Among the crude antigens, the best sensitivities when testing 21 naturally infected dogs were obtained using adult E/S and somatic antigen (85.7% and 76.2%, respectively), which were comparable with the results of the sandwich-ELISA based on mAb-purified antigens (81%). The ELISA performed with L1 antigen had the lowest sensitivity (42.9%). In experimentally inoculated dogs, the sensitivities ranged from 97.7% to 100% with all test settings. The specificity was 98.8% (92.5-99.9%, 95% CI) with all ELISAs using sera of 82 randomly selected dogs. Cross-reactions using adult somatic, adult E/S and L1 somatic antigen were observed in sera of dogs infected with Crenosoma vulpis, Dirofilaria immitis, Dirofilaria repens, and Eucoleus aerophilus. In contrast, using the mAb-purified antigens, the cross-reactions were minimal. Depending on the antigens used, specific antibodies were detected starting between 13 and 21 days post experimental inoculation (dpi), and at latest between 35 and 48 dpi, thus before or around the onset of patency. The serological follow-up of four A. vasorum-infected dogs after anthelmintic treatment at 88 dpi showed a decrease of antibody levels after drug administration, and the animals became seronegative 2-9 weeks later. Two untreated dogs remained seropositive. In four dogs treated 4 dpi, virtually no antibody-reaction was detectable, with the exception of the ELISA performed with L1 antigen. The early detection of specific antibodies against A. vasorum by ELISA represents a valid alternative for a reliable diagnosis and for follow-up investigations after anthelmintic treatment.     

Changes in the serum urea: creatinine ratio in dogs with babesiosis, haemolytic anaemia, and experimental haemoglobinaemia

The purpose of this study was to determine serum urea and creatinine concentrations, the derived urea : creatinine (UC) ratios, haemoglobin concentrations and glomerular filtration rates (GFR) in dogs with haemolytic anaemia and those with experimentally induced anaemia and/or haemoglobinaemia. There were 25 dogs with babesiosis (group 1), 13 control animals (group 2), six dogs with induced haemoglobinaemia and anaemia (group 3), six with induced haemoglobinaemia (group 4), and 14 with immune-mediated haemolytic anaemia (IMHA) (group 5). The median serum urea concentration was 11.18 mmol/L (group 1), 4.3 mmol/L (group 2), 4.3 mmol/L (group 3), 4.35 mmol/L (group 4), and 8.5 mmol/L (group 5). Median serum creatinine was 67 μmol/L (group 1), 75 μmol/L (group 2), 78.5 μmol/L (group 3), 84 μmol/L (group 4), and 82 μmol/L (group 5). Median serum haemoglobin was 1.3g/L (group 1), 0.8 g/L (group 2), 9 g/L (group 3), 3g/L (group 4), and 1.3g/L (group 5). The median UC ratio was 41.35 (group 1), 15.36 (group 2), 14.18 (group 3), 13.6 (group 4), and 14.15 (group 5). GFR was normal in all five groups. Serum urea concentration and the UC ratio were significantly greater in dogs with babesiosis than in those with IMHA, experimentally induced anaemia and/or haemoglobinaemia.     

Autochtonous infection of dogs and slugs with Angiostrongylus vasorum in Hungary

On the course of a helminthological survey of the dogs of Baranya County, Hungary Angiostrongylus vasorum infection was detected in two asymptomatic dogs. Identification of the parasite was based on morphology of the first-stage larvae (L1) isolated from droppings, and successful experimental infection with first stage larvae to laboratory reared Discus rotundatus and Lissachatina fulica snails, in order to exclude species of the family Filaroididae that have similar larvae to A. vasorum. While angiostrongylosis is widespread among foxes, this is the first report of A. vasorum infection in housedog in Hungary. In gardens, where infected dogs were being kept 91 specimens of 6 species of limacid and arionid slugs were collected of which 5 specimens of Arion lusitanicus were found to carry larvae of A. vasorum. Dogs usually do not ingest such large slugs willingly. Frogs are known to act as paratenic hosts in the life cycle of A. vasorum. Since one of the infected dogs harboured also infection with the intestinal trematode Alaria alata, of which frogs certainly play the role of the second intermediate host, therefore it is assumed that in this case the dog became infected with A. vasorum by eating frogs.     

Effects of heparin, citrate, and EDTA on plasma biochemistry of sheep: Comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals, but limited information is existing for sheep plasma biochemistry. Ten clinically healthy Baloochi breed of sheep were blood sampled in different tubes containing each anticoagulants and plain tube for harvesting plasma and serum. The concentrations of glucose, cholesterol, total bilirubin, urea, creatinine, total protein, albumin, calcium, inorganic phosphorus, and magnesium and the activity of aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine kinase (CK) and gamma glutamyl transferase (GGT) were measured. Except for the amounts of GGT, bilirubin and inorganic phosphorus, other measured parameters were significantly lower in citrated plasma than that of serum. For corrected citrated plasma significant differences were seen for the concentrations of glucose, creatinine, calcium and the activity of ALP.Most parameters did not show any difference, but significant increase was seen for albumin concentration when heparin was used as an anticoagulant. Using EDTA as anticoagulant caused a significant difference for the concentrations of some of the measured parameters in plasma except glucose, GGT, cholesterol, albumin, bilirubin, CK, and inorganic phosphorus comparing with serum.     

Effects of general anesthesia and surgery on renal function in healthy dogs

OBJECTIVES: To evaluate renal function in healthy dogs undergoing general anesthesia and ovariohysterectomy without concurrent IV administration of fluids. ANIMALS: 35 healthy client-owned dogs. PROCEDURE: Dogs were medicated with promazine hydrochloride (0.05 mg/kg of body weight, SC) approximately 45 minutes before induction of anesthesia with thiopental sodium (10 to 15 mg/kg, IV). Anesthesia was maintained with 2% halothane in oxygen. Ovariohysterectomies were performed by senior veterinary students under the direct supervision of a veterinary surgeon. Renal function was assessed (serum urea and creatinine concentrations, fractional clearance of sodium, urine alkaline phosphatase [ALP] and gamma-glutamyltransferase [GGT] activities, urine specific gravity, and enumeration of renal tubular epithelial cells in urine sediment) prior to and 24 and 48 hours after surgery. RESULTS: Duration of general anesthesia ranged from 80 to 310 minutes. Urine specific gravity and ALP activity and serum urea and creatinine concentrations did not change over time. Fractional clearance of sodium decreased 24 and 48 hours after surgery, whereas urine GGT activity and the ratio of urine GGT activity to urine creatinine concentration increased 24 hours after surgery, compared with presurgery values. Renal tubular epithelial cells increased in number in urine sediment from 11 of 35 (31.4%) dogs and 5 of 35 (14.3%) dogs 24 and 48 hours after surgery, respectively. However, this increase was not clinically relevant. CONCLUSIONS AND CLINICAL RELEVANCE: Intravenous administration of fluids to healthy dogs undergoing general anesthesia and elective surgery may not be necessary for maintenance of renal homeostasis.     

Biochemical and histological responses of Rattus novergicus (Wistar) infected by Echinostoma paraensei (Trematoda: Echinostomatidae)

Tests were performed to evaluate the biochemical alterations in Rattus norvegicus after infection by the intestinal trematode Echinostoma paraensei. The rodents received 150 metacercariae each, serum samples were collected and the parasite load was quantified weekly until the fifth week of infection. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALKP), gamma-glutamyl transferase (GGT), bilirubin, glucose, total proteins and fractions and hepatic glycogen were determined. All the animals exposed to the metacercariae were infected in the first week and worms were recovered up to the third week after infection. The levels of AST, ALT, GGT, bilirubin and globulin rose in the first and/or second week and declined thereafter to levels near those of the control group. In contrast, the level of total proteins in the plasma fell significantly in the first week while the ALKP activity went down only in the fourth and fifth weeks in relation to the control group. There was no significant difference in the levels of albumin, glycogen and glucose. Infection by E. paraensei in R. norvegicus causes changes in the hepatic function, possibly resulting from the cholestasis produced by the partial obstruction of the bile duct by the helminths.     

Effect of storage on some constituents of camel serum

SUMMARY Effects of storage at room temperature (23–25°C) and refrigeration (4–5°C) on various biochemical constituents of camel serum were investigated. Albumin, globulin, calcium, phosphorus, cholesterol, aspartate aminotransferase (AST), alkaline phosphatase (AP) and gamma glutamyltransferase (GGT) did not change over 9 days when stored at 4–5°C. At 4–5°C, creatinine, iron and glucose in camel sera remained stable for 6 days; total protein for 7 days; and blood urea nitrogen for 8 days. Decreased activities in creatine kinase (CK), lactate dehydrogenase (LDH) and alanine aminotransferase (ALT) were apparent after 1, 6 and 7 days, respectively. At room temperature, total protein, albumin, globulin, calcium and phosphorus were stable throughout the 9 days. Changes in glucose and iron occurred after 3 days. Stability at room temperature for LDH was 1 day; AST, 3 days; GGT and ALT, 6 days; and AP, 8 days. CK activity had already declined by 4 hours and by 9 days, only 34% activity remained.     

Effect of repeated freeze-thaw cycles on routine plasma biochemical constituents in canine plasma

BACKGROUND: The effect of repeated freeze-thaw cycles on plasma constituents has not been assessed in dogs, although such a procedure is not uncommon to use in routine laboratory practice. OBJECTIVE: To assess the effect of freeze-thaw cycles on routine plasma constituents in healthy dogs. METHODS: Six healthy adult dogs were used. Blood was sampled and placed in heparinized tubes. After centrifugation, plasma was separated into 5 aliquots. One aliquot was considered as the reference aliquot and used immediately for the assay of all of the biochemical constituents. All of the other aliquots were stored at 20 degrees C. Three aliquots underwent 1, 2, or 3 freeze-thaw cycles during a 1- to 3-day period. The last aliquot remained at 20 degrees C throughout the study and was thawed on the third day. The following biochemical constituents were assayed: glucose, urea, creatinine, total proteins, sodium, potassium, chloride, calcium, phosphates, aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), and alkaline phosphatase (ALP). RESULTS: No clinically relevant change was observed between the different aliquots for all of the constituents. CONCLUSION: Repeated freeze-thaw cycles do not cause changes in the biochemical constituents studied in canine plasma.