Use of a prestorage leukoreduction filter effectively removes leukocytes from canine whole blood while preserving red blood cell viability

Leukoreduction of blood products is a technique used to prevent leukocyte-induced transfusion reactions. Filters currently used for human blood products achieve at least a 99.9% reduction in leukocyte numbers per unit (450 mL) of blood. Goals of this study were to determine if a prestorage leukoreduction filter could effectively achieve leukoreduction of canine blood and to determine if viability of the leukoreduced red blood cell (RBC) product could be maintained after 35 days of storage. Blood collected from each dog was filtered through a leukoreduction filter at either room temperature or after cooling (4 degrees C) for 4 hours. Filtration efficacy was determined by measurement of pre- and postfiltration leukocyte counts. In vitro viability of RBCs was determined by comparing RBC adenosine triphosphate concentration and percent hemolysis before and after the storage period. In vivo viability of stored cells was determined using a biotin-streptavidin-phycoerythrin labeling technique and flow cytometry. Blood filtered within 30 minutes of collection versus blood filtered after cooling had mean reductions in leukocyte numbers of 88.90 and 99.99%, respectively. The mean ATP and hemoglobin concentrations from the in vitro analysis were comparable to those obtained in previously for canine RBC adequately stored for 35 days. The mean in vivo 24-hour survival of the stored RBC was 84.7%. The leukoreduction filter used did not adversely affect in vitro or in vivo viability of canine RBCs. The filter effectively removed leukocytes from blood, with maximal efficiency of filtration achieved with use of cooled blood.     

The use of ultrapurified bovine hemoglobin solution in the treatment of two cases of presumed red maple toxicosis in a miniature horse and a pony

Objective: To report the use of ultrapurified bovine hemoglobin (PBHg) in the treatment of red maple toxicosis in horses. Summary: The use of PBHg in a miniature horse and a pony with clinical signs of red maple toxicosis is described. Initial treatment of the horses included blood transfusion in Case 1, and intravenous crystalloid fluids, nasogastrically administered mineral oil, and activated charcoal in both cases. Both cases received PBHg (16 and 11 ml/kg, respectively) when the hematocrit dropped below 10%. Clinical stabilization of the horses (reflected by improved vital signs and mucous membrane color, increase in P_vO₂, and increase in plasma hemoglobin) was temporary, lasting 12 and 42 hours, respectively. A whole blood transfusion was given 16 hours after the infusion of PBHg in Case 1 and 42 hours after the infusion of PBHg in Case 2. The horses were discharged on days 15 and 17, respectively. Both were stable on discharge and were reported to be doing well 1 month after discharge. New information provided: Ultrapurified bovine hemoglobin is shown to be safe in the treatment of red maple toxicosis. Ultrapurified bovine hemoglobin infusion resulted in temporary improvement in oxygen (O₂) delivery as assessed by clinical and laboratory parameters. The use of PBHg may provide an ‘oxygen bridge’ in an acute hemolytic crisis in the horse. It may allow additional time in the treatment of red maple toxicity for the acute hemolysis to subside and for metabolism of the toxin to occur, thus providing a more optimal environment for an effective whole blood transfusion.     

Sucrose concentration in blood: a new method for assessment of gastric permeability in horses with gastric ulceration

A urine sucrose test has recently been reported to be a reliable method of detecting gastric ulcers in horses; however, technical difficulties associated with urine collection have limited the practical value of the test. The objective of this pilot study was to determine whether gastric sucrose permeability, as evaluated by serum sucrose concentration, could be used to detect gastric mucosal injury in horses. Twelve adult horses with naturally acquired gastric ulceration were studied. After a 20-hour nonfeeding period, each horse was dosed with 250 g of sucrose via nasogastric intubation. Blood samples were collected at 0, 15, 30, 45, 60, and 90 minutes, and horses underwent gastroscopy 4 hours later. The severity of gastric ulceration in each horse was defined by means of a 4-point ulcer-scoring system, and the relationship with serum sucrose concentration was analyzed by means of a linear mixed-effects model. Serum sucrose concentration was measured by liquid chromatography operating in tandem with electrospray mass spectrometry. After nasogastric administration of table sugar, horses with moderate to severe gastric ulceration had significant increase in serum sucrose concentration at 30, 45, 60, and 90 minutes, relative to earlier times (P < .05). Peak sucrose concentration was observed at 45 minutes, and was correlated with ulcer severity (Spearman's rank correlation coefficient = 0.898, P < .05). These data indicate that determination of sucrose concentration in equine serum may be a useful test for identifying horses with endoscopically visible gastric ulceration and has potential use as a noninvasive method for screening and monitoring horses engaged in racing training and other performance-related disciplines.     

Risk of hemolytic anemia with intravenous administration of famotidine to hospitalized cats

BACKGROUND: Famotidine administered IV has been associated anecdotally with hemolysis in cats, and some veterinarians recommend using injectable famotidine only by the subcutaneous (SC) route for cats. However, the actual risk of such a reaction is not known. HYPOTHESIS: We hypothesized that famotidine, when given IV slowly, would not be associated with a clinically significant drop in packed cell volume (PCV) in hospitalized cats. ANIMALS: One hundred and forty-two hospitalized cats. METHODS: A retrospective medical record review was performed for hospitalized cats prescribed famotidine IV (n = 56), famotidine SC (n = 48), or no famotidine (n = 38) at a veterinary medical teaching hospital over the period from January 2004 through December 2006. RESULTS: Baseline signalment, observation times, and famotidine dosage (in treated cats) were similar among groups. Median baseline PCVs were significantly lower in the IV (31.5%) and SC (32.0%) groups compared with the control group (35.0%; P= .04). The median percent drop in PCV (3-4%), however, was no different in cats that received famotidine by either route compared with the control group (P= .90), and no cats in either famotidine group were observed to have any clinical signs of hemolysis. CONCLUSIONS AND CLINICAL IMPORTANCE: We conclude from this retrospective study that famotidine IV was given to 56 hospitalized cats without evidence of hemolysis, and that the IV route appeared safe when famotidine was administered over 5 minutes. We could not document a safety advantage of SC versus IV administration in this group of cats.     

Use of a wireless system to measure invasive arterial blood pressure in ponies – preliminary study

Objective  To evaluate the feasibility and functionality of intra-carotid wireless device implantation in ponies, and to investigate its short-term complications.
Study design  Prospective preliminary study.
Animals  Five mixed breed, adult, intact male ponies weighing 104 ± 28.8 kg (mean ± SD) underwent surgery. Arterial blood pressure data were continuously collected from four animals.
Methods  General anesthesia was induced on two consecutive days. On the first day, an intra-arterial wireless device was implanted in the right carotid artery. On the next day, a transcutaneous intra-arterial catheter was placed in the left facial artery. Data from both sources were collected. Post-mortem examination was performed.
Results  Surgical time was 27.1 ± 11.85 minutes. All catheters remained in place with some extra vascular migration. Complications included mild seroma and hematoma.
Conclusion  The wireless system allowed continuous monitoring in ponies throughout anesthesia and at rest and may allow for the recording of arterial blood pressure and heart rate when it would be difficult to achieve with a conventional system (e.g. during recovery from anesthesia).
Clinical relevance  The wireless invasive blood pressure monitor may allow continuous measurements when only intermittent measurements would be feasible with a wired system.     

Assessment of the red blood cell proteome in a dog with unexplained hemolytic anemia

A 7‐year‐old female neutered Jack Russell Terrier was presented to Langford Vets, the University of Bristol, with a history of chronic intermittent lethargy. Investigations and clinical course were compatible with hereditary hemolysis due to a red blood cell membrane defect. Proteomics was used to explore protein alterations in the presence of a hypothesized red blood cell membrane protein deficiency. Proteomic analysis revealed downregulation of the band 3, and alpha‐ and beta‐adducin proteins, and alterations in the red blood cell proteome consistent with previous reports of changes due to the presence of reticulocytosis and ongoing hemolysis. The spectrum of protein alterations identified in the affected dog may be homologous to a band 3 protein deficiency secondary to hereditary spherocytosis, as described in people.     

Use of a Grading System to Facilitate Treatment and Prognosis in Horses with Negative Palmar Angle Syndrome (Heel Collapse): 107 Cases

Negative palmar angle syndrome refers to the condition of progressive heel collapse and its consequences on gait and performance. Treatment and prognosis are facilitated by grading the severity of biomechanical disorder according to physical and radiographic features. Although negative palmar angle syndrome in grades I (mild) and II (moderate) can be corrected with trimming and routine shoeing, grades III (severe) and IV (complicated by flexor contracture) require more intensive mechanical intervention and patience—however, comfort and function can be improved immediately.     

Quantification of surface EMG signals to monitor the effect of a Botox treatment in six healthy ponies and two horses with stringhalt: Preliminary study

Reasons for performing the study: Therapeutic options for stringhalt in horses are limited, whereas medical experiences with botulinum toxin type A (Botox) have been positive. To evaluate its effectiveness in horses, surface electromyography (sEMG) signals before and after injection need to be quantified. Hypothesis: Treatment of healthy ponies and cases with Botox should reduce muscle activity in injected muscles and reduce spastic movements without adverse side effects. Methods: Unilaterally, the extensor digitorum longus, extensor digitorum lateralis and lateral vastus muscles of 6 healthy mature Shetland ponies and 2 talented Dutch Warmblood dressage horses with stringhalt were injected (maximum of 400 iu per pony and 700 iu per case; 100 iu in 5 ml NaCl divided into 5 injections) with Botox under needle EMG guidance. Surface EMG data were evaluated using customised software, and in the individuals gait was analysed using Proreflex. Statistical analysis was performed using mixed models and independent sample t test (P<0.05). Results: Surface EMG signals were quantified using customised software. The area under the curve (integrated EMG) in time was used as variable. It became significantly reduced in injected muscles after injection of Botox in normal ponies (P<0.05). This effect was present from Day 1 until Day 84 after injection. In the 2 cases, after injection of 3 muscles, the integrated EMG in time became significantly reduced in all 3 muscles. Kinematic measurements confirmed reduction of frequency and amplitude of hyperflexing or hyperabducting strides of the affected hindlimbs. The duration of effect was also seen in the cases until around 12 weeks after injection. Conclusions and potential relevance: After EMG guided injections of Botox, sEMG signals recorded from injected muscle were reduced, which proves this to be a useful tool in statistically evaluating a treatment effect. The positive results of this pilot study encourage further research with a larger group of clinical cases.     

Effects of isovolemic resuscitation with hemoglobin-based oxygen carrier Hemoglobin glutamer-200 (bovine) on systemic and mesenteric perfusion and oxygenation in a canine model of hemorrhagic shock: a comparison with 6% hetastarch solution and shed blood

OBJECTIVE: To study Hemoglobin glutamer-200 bovine (Hb-200), 6% hetastarch (HES) and shed whole blood (WB) resuscitation in canine hemorrhagic shock. STUDY DESIGN: Prospective laboratory investigation. Animals Twelve adult dogs [29 +/- 1 kg (mean +/- SD)]. METHODS: Anesthetized dogs were instrumented for recording systemic and mesenteric hemodynamic parameters and withdrawal of arterial, mixed and mesenteric venous blood, in which hematological, oxygenation, blood gas and acid-bases variables were determined. Recordings were made before [baseline (BL)], after 1 hour of hypovolemia and immediately and 3 hours post-resuscitation with 30 mL kg(-1) of either Hb-200, HES, or WB. RESULTS: Blood withdrawal (average 34 +/- 2 mL kg(-1)) caused significant hemodynamic changes, metabolic acidosis and hyperlactatemia characteristic for hemorrhagic shock. Only WB transfusion restored all variables. Hemoglobin glutamer-200 bovine infusion returned most hemodynamic parameters including cardiac output and mesenteric arterial blood flow to BL but increased mean arterial pressure above BL (p < 0.05). However, Hb-200 failed to restore total Hb and arterial oxygen content (CaO2), leaving systemic (DO2I) and mesenteric O2 delivery (DO2Im) below BL (p < 0.05). Nevertheless, acid-base variables recovered completely after Hb-200 resuscitation, and met-hemoglobin (Met-Hb) levels increased (p < 0.05). Hetastarch resuscitation returned hemodynamic variables to or above BL but further decreased total Hb and CaO2, preventing recovery of sDO2I and mDO2I (p < 0.05). Thus, systemic and mesenteric O2 extraction stayed above BL (p < 0.05) while acid-base variables recovered to BL, although slower than in Hb-200 and WB groups (p < 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: Resuscitation with Hb-200 seemed to resolve metabolic acidosis and lactatemia more rapidly than HES, but not WB; yet it is not superior to HES in improving DO2I and DO2Im. The hyperoncotic property of solutions like Hb-200 that results in rapid volume expansion with more homogenous microvascular perfusion and the ability to facilitate diffusive O2 transfer accelerating metabolic recovery may be the key mechanisms underlying their beneficial effects as resuscitants.     

Weaning management of newly received beef calves with or without continuous exposure to a persistently infected bovine viral diarrhea virus pen mate: effects on health, performance, bovine viral diarrhea virus titers, and peripheral blood leukocytes

Exposure to animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) results in immunomodulation of cohorts that may have health and growth consequences; however, effects may differ in low-risk, preconditioned (PC) vs. high-risk, auction market (AM) beef cattle. Our objective was to compare health and performance of PC or AM management systems with (PI) or without (CON) presence of a PI-BVDV pen mate using a 2 × 2 factorial arrangement. Four shipment blocks of crossbred PC steers (n = 236) from 3 ranch-origins were weaned, dewormed, vaccinated, tested for PI-BVDV, and kept on the ranch for ≥42 d. Subsequently, PC steers were transported to a stocker receiving unit (RU), weighed (251 ± 2 kg), blood sampled, stratified by d -1 BW, and assigned randomly to treatment (PCPI or PCCON) with no additional processing. Simultaneously, 4 blocks of crossbred AM calves (n = 292) were assembled from regional auction markets and transported to the RU ± 36 h from PC arrival. The AM calves were weighed (245 ± 1.3 kg), stratified by gender and d -1 BW, processed under the same regimen used for PC steers at their origin ranch except bull calves were castrated, and then assigned randomly to treatment (AMPI or AMCON). Treatment pens (0.45 ha) were arranged spatially such that PI did not have fence-line or water source contact with CON. Calves were fed identically and followed the same antibiotic treatment protocol. Daily BW gain for the entire 42-d receiving trial was greater (P < 0.001) for PC (1.2 kg) compared with AM (0.85 kg). There was an exposure effect (P = 0.002) on ADG from d 28 to 42; CON gained 1.12 kg vs. 0.90 kg BW for PI cohort. Morbidity was markedly greater (P < 0.001) in AM (70%) vs. PC (7%), resulting in (P < 0.001) an antibiotic treatment cost of $20.52 and $2.48/animal, respectively. Treatment with a third antibiotic occurred more often (P = 0.04) for PI cohort, and the percentage of chronically ill cattle was greatest (P = 0.06) for AMPI. Upon arrival, BVDV type 1a, 1b, and 2a titers were greater for PC (treatment × day, P < 0.001), and the percentage seropositive to BVDV type 1a on d 0 was 100% for PC vs. 23% in AM. Platelets increased transiently (P < 0.001) with greater platelets observed in AM (P < 0.001). Results indicate that PC calves gain faster and require fewer antibiotic treatments during the receiving period. Exposure to PI reduced BW gain from d 28 to 42, increased the number of calves treated thrice, and increased chronically ill cattle for AM.     

Effects of long- or short-term exposure to a calf identified as persistently infected with bovine viral diarrhea virus on feedlot performance of freshly weaned, transport-stressed beef heifers

A single experiment with a completely randomized design was conducted to evaluate the effects of long- or short-term exposure to a calf identified as persistently infected with bovine viral diarrhea virus (PI-BVD) on feedlot performance and carcass characteristics of freshly weaned, transport-stressed beef heifers. Two hundred eighty-eight heifers that had been vaccinated for BVD before weaning and transport were processed and given a metaphylactic antibiotic treatment at arrival and were fed common receiving, growing, and finishing diets for a 215-d period. Treatments were designed to directly or adjacently expose the cattle to a PI-BVD heifer. Directly exposed treatments were 1) negative control with no PI-BVD calf exposure (control), 2) PI-BVD calf commingled in the pen for 60 h and then removed (short-term exposure), and 3) PI-BVD calf commingled in the pen for the duration of the study (long-term exposure); and spatially exposed treatments were 1) negative control with no PI-BVD calf exposure (adjacent pen control), 2) PI-BVD calf commingled in the adjacent pen for 60 h and then removed (adjacent pen short-term exposure), and 3) PI-BVD calf commingled in the adjacent pen for the duration of the study (adjacent pen long-term exposure). Exposure to a PI calf transiently (60 h) or for the duration of the feeding period (215 d) did not affect (P > or = 0.25) final BW compared with heifers that were not exposed. Neither period nor overall DMI was affected (P > or = 0.37) by PI-BVD calf exposure, and no differences (P > or = 0.44) were observed between short- and long-term exposed heifers in the direct or spatially exposed groups. Likewise, total trial ADG was not affected (P > or = 0.36) and overall efficiency of gain (P > or = 0.19) was unaffected by PI-BVD calf exposure in the direct or spatially exposed groups. The results from this study suggest that exposing previously vaccinated, freshly weaned, transport- stressed beef calves to a calf that is persistently infected with bovine viral diarrhea virus has little, if any, marked effects on health, performance, or carcass characteristics.     

Effects of excessive selenium supplementation to diet contaminated with deoxynivalenol on blood phagocytic activity and antioxidative status of broilers

This study examined the effects of excessive dietary supplementation with organic selenium on phagocytic activity and antioxidative status of chickens for fattening fed diet contaminated with deoxynivalenol (DON). Sixty chickens of Ross 308 hybrids were at day of hatching divided into four groups with 15 birds in each. The background DON dietary levels in both negative and positive control groups were 0.2 mg/kg. The complete feed for positive control group was supplemented with Se dose 1 mg/kg in the form of Se-yeast. Group 3 was fed diet with DON level 3 mg/kg while diet for group 4 combined DON level 3 mg/kg with a excessive supplement of Se-yeast (Se dose 1 mg/kg). After 6 weeks of dietary intake, six randomly-chosen chickens from each group were sampled. Feeding of contaminated diet resulted in significantly reduced blood phagocytic activity (19.5 ± 1.1% in the negative control vs. 12.8 ± 0.8% in the DON-treated group, p < 0.05). Se-yeast supplemented to the DON contaminated diet prevented suppression of phagocytic activity. Dietary intake of DON at levels 3 mg/kg did not influence the plasma α-tocopherol level while excessive dietary Se dose reduced it in both Se supplemented groups. Neither the birds of DON-treated group nor the birds from group 4 with DON and Se-yeast showed any response in plasma γ-glutamyltransferase (GGT) activity. Subtoxic dietary level of DON significantly increased the activity of glutathione peroxidase (GPx) in the duodenal mucosa, while additional Se supplementation prevented such a response to mycotoxin. On the other hand, both Se supplemented groups showed significantly elevated GPx activities in blood, liver and kidney, (p < 0.05). The results suggest a potential ability of excessive supplementation of organic selenium to prevent the blood phagocytic activity suppression and changes in GPx activity in duodenal tissue induced in broilers by subtoxic dietary levels of DON.     

Components of electroencephalographic responses to slaughter in halothane-anaesthetised calves: Effects of cutting neck tissues compared with major blood vessels

AIM: To identify whether cutting neck tissues or cutting major blood vessels initiates the mechanisms responsible for electroencephalographic (EEG) responses to slaughter by ventral-neck incision without prior stunning in halothane-anaesthetised calves.

METHODS: Calves were assigned to two groups, viz transection of neck tissues with intact blood circulation through the brain (n=10), or transection of the major blood vessels of the neck but not most other neck tissues (n=7). They were minimally anaesthetised with halothane, using an established anaesthesia protocol. The animals in the neck-tissue transection group had their carotid arteries and jugular veins exposed and cannulated proximal and distal to the proposed site of subsequent ventral-neck incision; this diverted blood fl ow through these vessels so that cerebral perfusion and drainage were preserved. In animals in the blood-vessel transection group, the carotid arteries and jugular veins were exposed bilaterally by surgical dissection. They were then transected without further damage to the remaining structures of the neck. Changes in the median frequency (F50), 95% spectral edge frequency (F95), total power of the EEG (Ptot), and arterial blood pressure were compared within each group before and after neck-tissue or blood-vessel transection, and between groups following treatments.

RESULTS: Neck-tissue transection resulted in little overall change in the F50, an increase in the F95, and an initial increase in Ptot followed by a transient decrease and eventual return to pre-treatment values. There was between-animal variation in these EEG parameters. Transection of the major blood vessels of the neck resulted in a decrease in F50 in most animals; changes in F95 were highly variable, and there was a decrease in Ptot.

CONCLUSIONS: The EEG responses seen following necktissue and blood-vessel transection were qualitatively distinct, and suggested that cutting neck tissues caused greater noxious sensory input than transection of only the major blood vessels of the neck. These observations support the conclusion that the EEG responses seen after ventral-neck incision in intact animals are primarily due to noxious stimulation as a result of incision of ventral-neck tissues and not mainly as a result of loss of blood flow through the brain.     

Comparison of gel column agglutination with monoclonal antibodies and card agglutination methods for assessing the feline AB group system and a frequency study of feline blood types in northern Italy

Background: A new commercial gel column agglutination system is reported to have high sensitivity in detecting cats with blood type AB. Objectives: The aims of this study were to compare gel column agglutination and card agglutination methods for feline blood‐typing and to determine the frequency distribution of feline blood types in northern Italy. Methods: Blood‐typing was performed on 120 cats using both a commercial gel column containing monoclonal antibodies (ID Gel‐Test Micro Typing System) and a card agglutination method (RapidVet‐H Feline). Results were confirmed with back‐typing. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated for the 2 methods. A second group of 140 Domestic Shorthair (DSH) cats was blood‐typed using the gel column technique to determine the frequency distribution of feline blood types in northern Italy. Results: The card agglutination method demonstrated poor sensitivity in identification of type‐AB cats (61%) and was only 95% specific when identifying type‐B cats. The gel column agglutination technique demonstrated 100% sensitivity and specificity for typing all 3 blood types (A, B, and AB). The frequency distribution study of 140 cats demonstrated that 127 (90.7%) cats were type A, 10 (7.1%) were type B, and 3 (2.1%) were type AB. Conclusion: When blood‐typing cats of breeds with a relatively high frequency of blood types B and AB, methods that use monoclonal antibodies for detection of blood types B and AB are recommended. Alternatively, blood type can be confirmed by more sensitive supplemental testing, such as back‐typing. The high frequency of blood type A in DSH cats in northern Italy was comparable to previously reported frequencies in Italy and world‐wide.