Endotoxin-induced changes in the hemostatic system in neonatal calves: the effect of antiserum to a mutant Escherichia coli (J-5)

Changes in the hemostatic system were studied in 22 neonatal calves given a small dosage of Escherichia coli endotoxin (0.5 microgram/kg) by slow (5-hour) IV infusion. The effect of pretreatment with an antiserum to mutant of E coli O111:B4 (J-5) was evaluated. The platelet count, plasma fibrinogen concentration, prothrombin time, and activated partial thromboplastin time changed significantly from base line during and after endotoxin infusions in all calves. The mean platelet count was significantly decreased from 1 through 24 hours after endotoxin infusion was started. Mean plasma fibrinogen was decreased 2 through 12 hours after endotoxin infusion was started. The mean prothrombin time and activated partial thromboplastin time were significantly greater than base line at 3 to 6 hours and 3 to 12 hours, respectively, after endotoxin infusion was started. Serum concentration of fibrinolytic degradation products remained less than 10 micrograms/ml. Bovine J-5 antiserum did not prevent the endotoxin-induced changes in the hemostatic system of these neonatal calves.     

Clotting profiles and selected hematology of captive Speke's gazelles (Gazella spekei).

Manual restraint and jugular venipuncture were used to obtain blood for hematology and coagulation tests for 18 captive Speke's gazelles (Gazella spekei). The hematocrit and hemoglobin values were slightly higher in Speke's gazelles than in domestic ruminants. The Speke's gazelles had a mean prothrombin time of 15.1 sec and a mean activated partial thromboplastin time of 24.2 sec. The pregnant female Speke's gazelles had shorter activated partial thromboplastin times than the males, but the difference was not significant. Ideally, prothrombin times and activated partial thromboplastin times would be compared to a healthy conspecific during a suspected bleeding crisis. Baseline prothrombin and activated partial thromboplastin times are presented here for Speke's gazelles because clotting times for exotic hoofstock are quite limited.     

Effects of Escherichia coli endotoxin on leukocyte and platelet counts, fibrinogen concentrations, and blood clotting in colostrum-fed and colostrum-deficient neonatal calves

Effects of a single IV injection of Escherichia coli endotoxin on hemogram and clotting function were compared in colostrum-fed and colostrum-deficient neonatal calves. Before endotoxin administration, the 2 groups of calves only differed in their prothrombin times. After endotoxin administration, there were significant differences (P less than 0.005) between colostrum-fed and colostrum-deficient calves in total leukocyte, segmented neutrophil, nonsegmented neutrophil, and lymphocyte (P less than 0.05) counts and partial thromboplastin time. Significant time dependent changes were observed in the aforementioned parameters and in platelet count and fibrinogen concentration. Seemingly, colostrum feeding improved the calf's ability to respond to endotoxin challenge exposure probably because of improved granulopoietic activity.     

Hematology of experimental acute Sarcocystis bovicanis infection in calves. II. Serum biochemistry and hemostasis studies

Of four Holstein-Friesian calves infected with 200,000 sporocysts of Sarcocystis bovicanis, three become ill and died on days 35, 55, and 59 of a 63-day experiment. No control calves became ill or died. Serum biochemicals and hematologic indicators of hemostasis from both groups were measured throughout the experiment. Creatine phosphokinase values for both groups increased markedly during acute infection. Lactic dehydrogenase and aspartate aminotransferase values were high in infected calves on days 25 to 35 and days 24 to 63, respectively, indicating injury of muscle, liver, or other tissues. Sorbitol dehydrogenase values were significantly higher for infected than for control calves on days 25 and 35, indicating liver injury. Serum bilirubin and blood urea nitrogen values were significantly increased in three anemic infected calves from day 25 or 26 to day 35, probably reflecting destruction of erythrocytes. The fourth infected calf was not anemic and had no hyperbilirubinemia and only minimal azotemia. Serum protein and albumin values decreased in infected calves on days 21 to 30 or 35, when, although hypoalbuminemia persisted, total protein concentration increased. Glucose, calcium, sodium, and chloride values decreased in infected calves slightly before onset of illness and remained low throughout the experiment. Potassium, magnesium, and phosphorus values did not differ between infected and control calves. Activated partial thromboplastin time and Russell's viper venom time were normal; prothrombin time was significantly higher from day 27 to day 49 in infected calves. This pattern was interpreted as evidence for acquired factor VII deficiency. Abnormal retraction of blood clots and enlarged platelets in blood smears, which indicate platelet dysfunction and increased platelet turnover, respectively, were seen on days 27 through 35 in anemic infected calves. Values for thrombin time (three calves) and fibrin degradation product concentration (one calf) increased just before death of the infected calves.     

Ontogeny of bovine hemostasis

Ontogeny of selected hemostatic system components was studied in 120 bovine fetuses which had been divided into eight monthly gestational age groups. Fetal blood was subjected to the following tests: platelet count, partial thromboplastin time, prothrombin time, thrombin time, fibrinogen quantitation, and assays for prothrombin and factors V and VIII. Platelet numbers corresponding to adult numbers were in fetal blood at least as early as gestation day 60, and their numbers varied only slightly thereafter. Bovine blood was incapable of in vitro coagulation at gestation day 90, with all samples coagulating by gestation day 150. Fetal coagulation screening test times (partial thromboplastin time, prothrombin time, and thrombin time) shortened during gestation and were near times of adults at birth. Of the four individual coagulation factors tested, only factor VIII reached adult values in the fetus in utero. Amounts of fibrinogen, prothrombin, and factors V and VIII in the neonate exceeded that of normal adult cattle.     

Effects of single-dose L-asparaginase on coagulation values in healthy dogs and dogs with lymphoma.

Ten healthy dogs and 10 dogs with multicentric lymphoma were given a single dose of L-asparaginase at a rate of 10,000 IU/m2 of body surface. Assessment of concentrations of contributors to the coagulation process and of the ability to coagulate including antithrombin III, one-stage prothrombin time, prothrombin-proconvertin time, activated partial thromboplastin time, plasminogen, fibrinogen, and platelet number were performed prior to drug administration (day 0). These tests were repeated 24 hours (day 1), 48 hours (day 2), and 7 days after treatment with L-asparaginase. Antithrombin-III concentrations were significantly lower in the dogs with lymphoma than in healthy dogs on days 0, 1, 2, and 7; however, with the exception of day 1, mean values remained within normal limits. There was also a difference between the 2 groups in prothrombin/proconvertin values on day 7 and in platelet number on day 2, with the lymphoma group having significantly shorter prothrombin/proconvertin time than healthy dogs, and the difference in platelet numbers being associated with increased counts in the healthy dogs. Data obtained from the healthy dogs and dogs with lymphoma for each coagulation test were pooled for each treatment day (0, 1, 2, and 7), and day-0 values for each coagulation test were compared with data obtained on days 1, 2, and 7. Antithrombin-III concentration on day 7 was significantly lower than on day 0, prothrombin/proconvertin time on day 1 was significantly longer than on day 0, and fibrinogen concentrations on days 1 and 2 were significantly lower than on day 0.(ABSTRACT TRUNCATED AT 250 WORDS)     

Blood coagulation times in the European brown hare (Lepus europaeus)

Background:  Many causes of mortality in the European brown hare, such as bacterial and viral infections, anticoagulant poisoning, and trauma, may result in hemorrhage. There are, however, no reference values concerning blood clotting in this species.  Objectives:  The aim of this study was to determine reference values for blood coagulation times and related parameters in healthy European brown hares.  Methods:  Blood samples from 30 clinically healthy adult hares (15 males and 15 females) were obtained. Hares were physically restrained for blood collection from the cephalic vein into tubes containing citrate and EDTA.  Results:  Mean ± SD were obtained for thrombin time (TT) (13.97 ± 1.37 seconds), prothrombin time (PT) (13.32 ± 2.15 seconds), activated partial thromboplastin time (APTT) (16.73 ± 1.86 seconds), fibrinogen concentration (2.98 ± 1.06 g/L), and platelet count (355.28 ± 128.73 × 10⁹/L). Conclusions: Reference values for blood coagulation times and other parameters associated with blood clotting will be useful in the laboratory evaluation of hemorrhage in the European brown hare.     

Evaluation of microwave-thawed canine plasma for transfusion

Units of fresh-frozen canine plasma were thawed rapidly in a microwave oven, using a mean of 15.4 thawing periods of 10 seconds each. The activated partial thromboplastin times, the one-stage prothrombin times, concentrations of fibrinogen, factor-VIII coagulant activity, and von Willebrand factor antigen of microwave-thawed units were not significantly different from those measured in small aliquots of the same units thawed in a warm water bath (n = 5). Five dogs were given a unit of their own microwave-thawed plasma. During the 24 hours after infusion, no significant changes were measured in temperature, heart rate, or respiration rate. In addition, significant changes in PCV, total protein concentrations, estimates of platelet numbers, total RBC counts, total WBC counts, and differential WBC counts were not measured in blood specimens obtained before infusion and 24 hours after the infusion of microwave-thawed plasma.     

Study of haemostatic disorders in experimentally induced leishmaniasis in Beagle dogs

Haemostatic alterations in dogs experimentally infected with Leishmania infantumwere studied before and after therapy with meglumine antimonate. Haemostatic function tests including platelet count, collagen-induced platelet aggregation, prothrombin time, activated partial thromboplastin time, thrombin time, plasma fibrinogen determination, and serum fibrinogen/fibrin degradation products concentration were performed. In the course of infection and before treatment, moderate thrombocytopenia (P<0·00001) decreased collagen induced platelet aggregation (P=0·0003), prolonged thrombin time (P=0·0117) and increased fibrinogen/fibrin degradation products were observed. Statistically significant differences of plasma fibrinogen concentration, prothrombin time, and activated partial thromboplastin time were not encountered. Haemostatic parameters returned to normal values after therapy. The results indicate that Leishmania infection may impair haemostasis suggesting induction of disseminated intravascular coagulation (DIC), and that treating dogs in an early stage of infection may potentially avoid the possibility of developing an uncompensated DIC.     

One-stage prothrombin time, activated partial thromboplastin time, thrombin time, fibrin degradation product concentration, and antithrombin activity in healthy adult alpacas

Background: Alpacas are increasingly presented to veterinarians for evaluation and care. Reports of alpaca reference intervals for one‐stage prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), concentration of fibrin degradation products (FDP), and antithrombin (AT) activities are scarce or nonexistent. Objective: The aim of this study was to determine values for blood coagulation times (PT, aPTT, and TT), FDP concentrations, and AT activities in healthy adult alpacas. Methods: Of blood samples collected from 35 clinically healthy adult alpacas via jugular venipuncture and placed into sodium citrate and FDP tubes, 29 samples were assayable for coagulation testing. PT, aPTT, and TT were determined by physical (mechanical) clot detection; AT activity was determined using a thrombin‐specific chromogenic substrate end‐point assay; and FDP concentrations were determined by the slide agglutination method. Results: Median values and ranges (minimum–maximum) were determined for PT (8.7 seconds, 6.6–11.2 seconds), aPTT (17.3 seconds, 11.9–22.5 seconds), TT (10.2 seconds, 5.4–16.0 seconds), and AT activity (123.3%, 104.8–144.2%). The mean concentration of FDP was <8 μg/mL. Conclusion: These values for coagulation times, FDP concentration, and AT activity will provide a useful starting point in the diagnostic evaluation of ill adult alpacas.     

The Evaluation of Coagulation Profiles in Calves with Suspected Septic Shock

The purpose of the study reported here was to evaluate the haemostatic function in calves with suspected septic shock and to reflect the occurrence of disseminated intravascular coagulation (DIC). Twenty-six calves suspected of having septic shock (experimental group) and 10 clinically healthy calves (control group) were used. On admission, the experimental group of calves had been ill for an average of 2 days. Therapy was applied to the experimental group of calves. The packed cell volume (PCV), haemoglobin (Hb), white blood cell (WBC), red blood cell (RBC) and platelet (PLT) counts were determined. Blood smears for toxic neutrophil and schistocyte intensity were evaluated. For the coagulation profile, plasma activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen and fibrin/fibrinogen degradation products (FDPs) values were determined. Toxic neutrophils in blood smears were observed in 12 calves of the experimental group. APTT was prolonged in the experimental group compared with the control group (p < 0.05). Fibrinogen concentration was found to be higher in the experimental group than in the control group (p < 0.001). Total leukocyte counts were higher in the experimental group compared with the control group (p < 0.01). Platelet counts in the experimental group were lower than the control group (p < 0.001). However, when the individual values of coagulation profiles of each calf were evaluated, 8 calves had at least three abnormal coagulation profiles (APTT >72 s, PT >34.5 s, TT >33.7 s, FDPs >5 microg/ml, PLT < or = 150 x 10(3)/mm(3)) and abnormal erythrocyte morphology (schistocytes > or = 1). The most common abnormal tests in the coagulation profile were APTT and PT (7 cases), FDPs (6 cases), thrombocytopenia (4 cases), and schistocytes in blood smears (8 cases) in these 8 calves. The results of this study indicate that DIC might be a significant risk factor for mortality in calves with suspected septic shock.     

Coagulation and fibrinolysis in the dog.

A range of tests of coagulation and fibrinolysis was measured in "normal" dogs and compared with values obtained in "normal" humans by the same methods. The hematocrit platelet count, fibrinogen and plasminogen were similar in dogs and in humans. The prothrombin and partial thromboplastin times were considerably shorter in the dog than in man but the thrombin clotting time was comparable. Fibrinolysis was more active in dogs but the levels of fibrin degradation products were low, suggesting that there was no significant fibrin deposition and lysis occurring in vivo.     

Haemorrhagic syndrome of cattle associated with the feeding of sweet vernal (Anthoxanthum odoratum) hay containing dicoumarol

An outbreak of a haemorrhagic diathesis in cattle fed home produced hay is described. A similar syndrome was reproduced experimentally in calves by feeding them the hay. The experimental disease was characterised by increased prothrombin and partial thromboplastin times while the leucocyte and erythrocyte counts remained normal until the terminal haemorrhage. The calves ate well and grew well until the rapid onset of progressive weakness, stiff gait, mucosal pallor, tachycardia, tachypnoea and haematomata ending in sudden death. The absence of blood coagulation was seen at necropsy while petechial, ecchymotic and free haemorrhages were found in most organs. Particularly striking were massive ecchymotic haemorrhages on the peritoneal surface of the rumen, a bloody, gelatinous mass enveloping each kidney and extensive bruising, haemorrhage and haematomata in the subcutis of the limbs. In a second feeding trial the effects of various preparations of vitamin K1 and vitamin K3 were investigated. Oral administration of large quantities of vitamin K1 reduced the elevated prothrombin time; vitamin K3 acted less consistently. Analysis of the hay for trichothecene mycotoxins was negative but floral analysis revealed that sweet vernal grass (Anthoxanthum odoratum) comprised about 80 per cent of the hay. Dicoumarol was detected in the hay and in the serum and ruminal contents of the experimental calves. The diagnosis, treatment, control and importance of this syndrome in the United Kingdom are discussed.     

Effect of carprofen on hemostatic variables in dogs

OBJECTIVE: To evaluate the effect of carprofen on hemostatic variables in clinically normal dogs. ANIMALS: 12 clinically normal Labrador Retrievers. PROCEDURE: 10 dogs (6 females, 4 males) received carprofen (2.2 mg/kg of body weight, PO, q 12 h) for 5 days. Two dogs (untreated control group; 1 female, 1 male) did not receive carprofen. Hemostatic variables (platelet count, activated partial thromboplastin time, prothrombin time, fibrinogen, platelet aggregation, and bleeding time) were assessed for all dogs prior to treatment, on day 5 of treatment, and 2 and 7 days after discontinuation of the drug (days 7 and 12). Serum biochemical variables and Hct were assessed prior to treatment and on days 5 and 12. RESULTS: In dogs receiving carprofen, platelet aggregation was significantly decreased, and onset of aggregation was significantly delayed on days 5, 7, and 12, compared with pretreatment values. Activated partial thromboplastin time was significantly increased on days 5, 7, and 12 over pretreatment values in treated dogs, but values remained within reference ranges. Significant differences were not detected in buccal mucosal bleeding time, other serum biochemical and hemostatic variables, or Hct, compared with pretreatment values and the internal control group. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of carprofen for 5 days causes minor but not clinically important alterations in hemostatic and serum biochemical variables in clinically normal Labrador Retrievers. Carprofen is commonly used to treat osteoarthritis and chronic pain in dogs, but prior to this study, its effect on platelet aggregation and hemostatic variables was unknown.     

Hematological changes in calves exposed to a mixture of lipopolysaccharide and crude leukotoxin of Pasteurella haemolytica.

The purpose of this investigation was to determine if culture supernatants of Pasteurella haemolytica containing crude leukotoxin and lipopolysaccharide (CLCL) causes disseminated intravascular coagulopathy (DIC) when injected into calves. The effect of intraduodenal (ID) exposure followed by a subsequent subcutaneous (SC) inoculation of either heat-treated or untreated CLCL was evaluated. The relative contribution of the crude leukotoxin and lipopolysaccharide (LPS) to the virulence of P. haemolytica was evaluated. One group of calves received an ID inoculation of CLCL followed two weeks later by a SC inoculation of CLCL; one group received an ID inoculation of tissue culture medium followed two weeks later by a SC inoculation of CLCL; and a third group received an ID inoculation of CLCL followed two weeks later by a SC inoculation of heat-treated CLCL. Hematological parameters used to evaluate DIC included white cell count, platelet count, neutrophil number, fibrinogen, fibrin degradation products, one stage prothrombin time (OSPT), activated partial thromboplastin time, body temperature and clinical signs. Each parameter was measured in calves at 0, 2, 4, 6, 12 and 24 h following the SC inoculation of CLCL. Each group had significant changes over time in all parameters except body temperature. Calves that received a SC inoculation of heat-treated CLCL had smaller changes in all parameters except OSPT compared to the other groups. Results suggest that the LPS and leukotoxin of P. haemolytica exert additive effects on the coagulation cascade and number of peripheral leukocytes, and that the ID inoculation of CLCL does not affect the response of calves to a SC inoculation of toxin.     

An epidemiological study of natural in utero infection with bovine leukemia virus.

The purpose of this study was to examine rates of natural in utero infection with bovine leukemia virus for association with breed, sex, dam age, dam parity and time of maternal seroconversion. Analyses conducted for breed and sex, dam age and parity and time of maternal seroconversion were the FUNCAT procedure for categorical data, Wilcoxon Rank Sums test and Fisher's exact test, respectively. A total of 223 calves born between July 1979, and September 1980, to cows infected with bovine leukemia virus in the University of Florida Dairy Research Unit herd were tested for detectable bovine leukemia virus antibodies prior to the consumption of colostrum. Sera were tested for antibodies by agar-gel immunodiffusion and radioimmunoprecipitation using the glycoprotein-51 antigen. In a group of 125 calves in which in utero infection could be confirmed through serological follow-up (group A), eight calves (6.4%) had precolostral bovine leukemia virus antibodies. For all 223 calves (group B), 18 (8.1%) had detectable bovine leukemia virus antibodies. For calves in group A, no associations were detected between precolostral bovine leukemia virus antibodies and breed (p = 0.66), dam age (p = 0.86), dam parity (p = 0.83), or time of maternal seroconversion to bovine leukemia virus (p = 0.50). However, precolostral bovine leukemia virus antibodies were found in 17.4% of the males and 3.6% of the females in group A (p = 0.11) and in 12.4% of the males and 3.6% of the females in group B (p = 0.04).     

Evaluation of antithrombin-III activity as a coindicator of disseminated intravascular coagulation in cats with induced feline infectious peritonitis virus infection

Six adult specific-pathogen-free cats were inoculated intraperitoneally with a cell culture-adapted strain of feline infectious peritonitis virus. Plasma samples were evaluated for antithrombin-III (AT-III) activities at post-inoculation days (PID) 0, 4, and 11 and at termination on PID 16 (1 cat) or 21 (5 cats). Other hemostatic values evaluated were activated partial thromboplastin times, prothrombin times, thrombin times, fibrinogen, platelet counts, and fibrin/fibrinogen degradation products. Antithrombin-III activity remained within normal or above normal range (89 to 246%) in all cats, with the exception of one cat on PID 4 (AT-III, 70%). Mean baseline AT-III activity for 6 cats at PID 0 was 123%. Mean AT-III activity on PID 4, 11, and 16 or 21 was 98, 162, and 130%, respectively. On PID 4 and 16 or 21, results of coagulation screening tests indicated that all cats had disseminated intravascular coagulation. Histologically, cats also had severe fibrinonecrotizing thrombovasculitis.     

Foetal protection against bovine virus diarrhoea virus after two-step vaccination

In order to assess the efficacy of a two-step vaccination protocol with respect to foetal protection against transplacental infections with bovine virus diarrhoea virus (BVDV) with special attention to BVDV-2 seronegative heifers were vaccinated with an inactivated BVDV-1 vaccine and boostered with a modified live BVDV-1 vaccine after 4 weeks. A second group was left unvaccinated as control. Between days 30 and 120 of pregnancy the heifers of both groups were intranasally challenged with a mixture of BVDV-1 and -2. All heifers of the vaccinated group gave birth to nine clinically healthy, seronegative (precolostral) and BVDV-free calves. In contrast in the control group four BVDV viraemic underdeveloped calves were born. Additionally, one calf was stillborn and another viraemic calf was not viable and died 2 days after birth. All six calves of the control group were viraemic with BVDV-2. This study demonstrated for the first time that two-step vaccination of breeding cattle with a modified live BVDV vaccine 4 weeks after application of an inactivated BVDV vaccine was capable of providing a foetal protection against transplacental infection with BVDV-2.     

Bovine anaplasmosis: transplacental transmission as it relates to stage of gestation

Six mature, pregnant, anaplasmosis-susceptible and 3 anaplasmosis-carrier beef cows were used in Anaplasma in utero transmission studies. Susceptible cows were randomly allotted into 3 groups of 2 cows each and were inoculated with a Virginia A marginale stabilate. Each group was Anaplasma-exposed once during 1 of the 3 trimesters of pregnancy. Blood samples were obtained from each fetus at various stages of development for evaluation and for subinoculation into splenectomized calves once during gestation. Precolostral blood from neonates was also subinoculated into individual susceptible calves. Two of the 9 splenectomized calves given fetal blood inoculations developed acute anaplasmosis. The dam of 1 fetus with infective blood was Anaplasma-exposed during the 2nd trimester of pregnancy, and the dam of the second fetus was exposed during her 3rd trimester. Infective fetal blood was obtained during the same trimester of gestation in which the dams were inoculated. Calves given neonatal precolostral blood did not develop anaplasmosis.     

The combined effect of fenbendazole treatment and a move to aftermath 7 or 9 weeks after turnout on Dictyocaulus viviparus infections in calves

A grazing study was performed with the main objective of examining the effect of fenbendazole (FBZ) in a ‘dose and move’ system on nematode infections in calves with special emphasis on Dictyocaulus viviparus.

Three groups of six calves were grazed from May to October 1993. One group (DM7) was treated with FBZ and moved to aftermath (pasture which had only been mown) 7 weeks after turnout. The second group (DM9) was similarly treated and moved 9 weeks after turnout and the third group served as untreated pasture control group (PC) and was moved to aftermath 9 weeks after turnout.

FBZ treatment removed adult lungworms from DM7 and DM9. Tracer calves grazed during the first 7 or the first 9 weeks after turnout acquired mean burdens of 18 and 125 lungworms, respectively. In PC faecal larval counts increased until the end of August. Most of the animals in this group were then suffering from lungworm disease and emergency treatment with ivermectin was given. In both FBZ-treated groups, larvae reappeared in the faeces of some of the calves 4–5 weeks after treatment. Subsequent reinfection resulted in higher mean faecal larval counts in both groups 2 months after treatment, although variation in faecal larval counts was high. In DM7 values tended to be higher than in DM9. These higher larval counts were associated with mild signs of parasitic bronchitis in some calves of DM7, whereas no signs were seen in DM9.

At the end of the experiment, all calves, and also a group of six permanently housed non-infected control calves (HC), were infected experimentally with 5000 D. viviparus larvae to evaluate development of immunity. The worm counts at necropsy showed that all calves on pasture had developed immunity.