ACCURACY OF POSITIONING THE CERVICAL SPINE FOR RADIATION THERAPY AND THE RELATIONSHIP TO GTV, CTV AND PTV

The purpose of the study was to evaluate the accuracy and precision of a rigid positioning device for repositioning the cervical spine accurately and precisely during conformal radiation therapy of dogs. Fifteen purpose bred research dogs in a radiation therapy study were included. The dogs were positioned using a head holder and a deflatable pillow attached to the treatment table. Port films were reviewed retrospectively, and repositioning precision was recorded by measurements in three orthogonal planes of the head, 2nd cervical vertebra and 1st thoracic spinous process. Mean treatment position was compared to the planning position for a measurement of systematic set-up error. Mean interfraction position variation of the 2nd cervical vertebra was 0.2, 0.1 and 0.2 cm for the ventrodorsal, caudocranial and laterolateral directions respectively, and the average systematic set up error was 0.2, 0.1 and 0.2 cm for the ventrodorsal, caudocranial and laterolateral directions respectively. Knowledge of the magnitude of reposition errors should be included when determining the margins around the tumor.     

COMPARISON OF INTERFRACTIONAL VARIATION IN CANINE HEAD POSITION USING PALPATION AND A HEAD‐REPOSITIONING DEVICE

Radiation treatment planning is performed on images that do not take variation in patient position into account. To compensate for expected variations in position of the patient, a three‐dimensional expansion of the clinical target volume, or set‐up margin, is added. Variations in patient position can be decreased through use of an immobilization device, allowing selection of a smaller set‐up margin. The objective of this prospective study was comparison of interfractional variation in patient position between set‐ups of the canine head region using palpation of bony landmarks and set‐ups using a head‐repositioning device. Fiducial markers were attached to the skull bones of three research dogs, and the dogs were positioned as for a typical radiation treatment of the head region using both set‐up methods. A kilovoltage on‐board imager was used to acquire orthogonal images and the difference between the x‐, y‐, and z‐axis coordinates of each fiducial marker relative to the initial reference isocenter was measured. The difference in patient position for each axis coordinate was significantly lower for set‐ups using the head‐repositioning device than for set‐ups using bony landmarks (P<0.05). Ninety‐five percent of the absolute values of the displacement vector differences were <4.62 mm for set‐up using bony landmarks, and <1.93 mm for set‐up using the head‐repositioning device. A minimum set‐up margin of 5–6 mm is recommended when patient set‐up is based on bony landmarks and of 2–3 mm when the head‐repositioning device is used.     

ASSESSMENT OF A RADIOTHERAPY PATIENT IMMOBILIZATION DEVICE USING SINGLE PLANE PORT RADIOGRAPHS AND A REMOTE COMPUTED TOMOGRAPHY SCANNER

Radiation treatment requires a precise procedure for interfraction repositioning of the patient. The purpose of this study was to determine the accuracy of our fixation device in treatment position and to evaluate the setup accuracy with two different methods. The positioning data of 19 canine patients with tumors in the head region (oral, nasal, cerebral) treated with photon or proton irradiation were included in this study. The patients were immobilized by means of an individualized fixation device. Focus was set upon interfraction displacement with systematic and random components. In one method, treatment position was evaluated using single plane port radiographs and megavoltage x-rays. In the other method, two orthogonal CT-topograms were acquired to evaluate the precision of positioning of the patient in the immobilization device. Systematic and random displacements were calculated and presented as mean values with corresponding 95% confidence intervals. In spite of a difference between both methods, the positioning seemed to be accurate within the expected range. It seems that a safety margin of 3.7 mm would be enough for both methods to take into account systematic and random position variability in the fixation device, thereby preventing geometric inaccuracies of treatment delivery. The reported immobilization protocol provides accurate patient immobilization for photon and conformal proton radiation therapy.     

VALIDATION OF AN INDEXED RADIOTHERAPY HEAD POSITIONING DEVICE FOR USE IN DOGS AND CATS

Setup variability affects the appropriate delivery of radiation and informs the setup margin required to treat radiation patients. Twenty‐four veterinary patients with head and neck cancers were enrolled in this prospective, cross‐sectional study to determine the accuracy of an indexed board immobilization device for positioning. Couch position values were defined at the first treatment based on setup films. At subsequent treatments, patients were moved to the previously defined couch location, orthogonal films were acquired, table position was modified, and displacement was recorded. The mean systematic displacement, random displacement, overall displacement, and mean displacement values of the three‐dimensional (3D) vector were calculated. Three hundred thirty‐two pairs of orthogonal setup films were analyzed for displacement in cranial–caudal, lateral, and dorsal–ventral directions. The mean systematic displacements were 0.5, 0.8, and 0.5 mm, respectively. The mean random displacements were 1.0, 1.1, and 0.7 mm, respectively. The overall displacements were 1.1, 1.4, and 0.9 mm, respectively. The mean 3D vector value was 1.6 mm with a standard deviation of 1.2 mm. Ninety‐five percent of the vectors were <3.6 mm. These values were compared to data obtained with a previously used immobilization device. A t‐test was used to compare the two devices. The 3D vector, random displacement in all directions, and overall displacement in the cranial–caudal and dorsal–ventral directions were significantly smaller than displacements with the previous device. The precision and accuracy of the indexed board device was superior to the historical head and neck device.     

VALIDATION OF NEW BITE BLOCK‐TYPE HEAD‐IMMOBILIZATION DEVICES FOR RADIOTHERAPY IN DOGS

An ideal head‐immobilization method provides a high level of accuracy and reproducibility in the immobilization. Various head‐immobilization methods for radiotherapy have been published and are excellent in terms of accuracy; however, these methods are complicated to use, and labor intensive. The present study describes two new bite block‐type head‐immobilization devices designed for higher stability and lower vertical variation. The device designed in our previous study (the bite block‐type head‐immobilization device; Device A) was modified by making a groove on the top the horizontal plate (Device B) for a stable ventral‐dorsal position, or beneath the horizontal plate (Device C) for a stable dorsal‐ventral position. The three devices were objectively compared with respect to setup time, and accuracy of the computed tomography scan images by two authors independently. Five male healthy beagles were used in this study. For each device, the setup time and the variation in the coordinates were measured five times for each dog. The mean setup times for Devices A, B, and C were 3.3, 1.5, and 2.4 min, respectively, showing the groove modifications were able to reduce the setup time (in device B, by at least 50%). Moreover, three‐dimensional analysis of the computed tomography images revealed that the measurement variability of Device A (1.6 ± 1.0 mm) was significantly higher than that of Device C (0.7 ± 0.4 mm; P < 0.001). Collectively, our results show that use of a bite block‐type head‐immobilization device with a groove improves the setup time and head‐immobilization accuracy.     

EVALUATION OF A HEAD-REPOSITIONER AND Z-PLATE SYSTEM FOR IMPROVED ACCURACY OF DOSE DELIVERY

Radiation therapy requires accurate dose delivery to targets often identifiable only on computed tomography (CT) images. Translation between the isocenter localized on CT and laser setup for radiation treatment, and interfractional head repositioning are frequent sources of positioning error. The objective was to design a simple, accurate apparatus to eliminate these sources of error. System accuracy was confirmed with phantom and in vivo measurements. A head repositioner that fixates the maxilla via dental mold with fiducial marker Z-plates attached was fabricated to facilitate the connection between the isocenter on CT and laser treatment setup. A phantom study targeting steel balls randomly located within the head repositioner was performed. The center of each ball was marked on a transverse CT slice on which six points of the Z-plate were also visible. Based on the relative position of the six Z-plate points and the ball center, the laser setup position on each Z-plate and a top plate was calculated. Based on these setup marks, orthogonal port films, directed toward each target, were evaluated for accuracy without regard to visual setup. A similar procedure was followed to confirm accuracy of in vivo treatment setups in four dogs using implanted gold seeds. Sequential port films of three dogs were made to confirm interfractional accuracy. Phantom and in vivo measurements confirmed accuracy of 2 mm between isocenter on CT and the center of the treatment dose distribution. Port films confirmed similar accuracy for interfractional treatments. The system reliably connects CT target localization to accurate initial and interfractional radiation treatment setup.     

ASSESSMENT OF THE ACCURACY AND PRECISION OF A PATIENT IMMOBILIZATION DEVICE FOR RADIATION THERAPY IN CANINE HEAD AND NECK TUMORS

The positioning accuracy and precision of a head and neck immobilization device for radiation therapy of tumors in the canine skull was evaluated. Nineteen dogs with a spontaneous tumor of the head were enrolled including 12 with an intracranial mass and seven with an intranasal or maxillary tumor. Three hundred thirty-three pairs of orthogonal digital portal radiographs were analyzed to assess patient displacement in the cranial–caudal, lateral, and dorso-ventral directions. The mean systematic displacement was 0.8, 1, and 0.9 mm. The mean random displacement was 1.9, 1.6, and 1.5 mm. These values resulted in an overall displacement of 2.1 mm in the cranial–caudal direction, 1.8 mm in the lateral direction, and 1.7 mm in the dorsal–ventral direction. The mean displacement value of the three dimensional (3D) vector was 2.4 mm with a standard deviation of 2.1. Ninety-five percent of all vectors were <6.4 mm. This study quantifies the precision and accuracy of this particular positioning device. Knowing the limitations and setup variability of the system being used to set patients up for daily radiotherapy is paramount to planning and delivering appropriate radiation doses, especially as more complex treatment methods are used.     

REPEATABILITY OF A PLANNING TARGET VOLUME EXPANSION PROTOCOL FOR RADIATION THERAPY OF REGIONAL LYMPH NODES IN CANINE AND FELINE PATIENTS WITH HEAD TUMORS

For canine and feline patients with head tumors, simultaneous irradiation of the primary tumor and mandibular and retropharyngeal lymph nodes (LNs) is often indicated. The purpose of this study was to assess the repeatability of a planning target volume (PTV) expansion protocol for these LNs. Two CT image sets from 44 dogs and 37 cats that underwent radiation therapy for head tumors were compared to determine LN repositioning accuracy and precision; planning‐CT (for radiation therapy planning) and cone‐beam CT (at the time of actual treatment sessions). Eleven percent of dogs and 65% of cats received treatment to their LNs. In dogs, the mandibular LNs were positioned more caudally (P = 0.0002) and the right mandibular and right retropharyngeal LNs were positioned more to the left side of the patient (P = 0.00015 and P = 0.003, respectively). In cats, left mandibular LN was positioned higher (toward roof) than the planning‐CT (P = 0.028). In conclusion, when the patient immobilization devices and bony anatomy matching are used to align the primary head target and these LNs are treated simultaneously, an asymmetrical PTV expansion that ranges 4–9 mm (dogs) and 2–4 mm (cats), depending on the directions of couch movement, should be used to include the LNs within the PTV at least 95% of the time.     

Deslorelin acetate (Ovuplant) therapy in cycling mares: effect of implant removal on FSH secretion and ovarian function

Following induction of ovulation with deslorelin acetate (Ovuplant), gonadotrophin concentrations are reduced in the subsequent cycle, leading to increased interovulatory intervals in some mares. This study determined whether implant removal after 2 days prevented the decrease in gonadotrophin concentrations and follicular growth during the ensuing cycle. Twenty-four mares were randomised equally into 3 groups. Group 1 ovulated spontaneously, Groups 2 and 3 received the deslorelin implant to induce ovulation. Two days after treatment, the implant was removed from Group 3. On Day 10 postovulation, FSH was lower (P = 0.009) in Group 2, but not different between Groups 1 and 3. Follicular diameter on Day 14 was less (P<0.05) in Group 2 (19.0 +/- 2.1 mm) than in Groups 1 and 3 (36.6 +/- 2.5 and 30.5 +/- 2.0 mm, respectively). Interovulatory interval was longer (P<0.05) for Group 2 (25.8 +/- 2.9 days) compared to Groups 1 and 3 (18.5 +/- 0.7 and 19.4 +/- 0.3 days, respectively). Removal of the deslorelin implant eliminated the decreased FSH secretion and the increased interovulatory interval associated with implant administration. Therefore, it is recommended that the implant be removed after ovulation is detected to prevent the occurrence of a prolonged interovulatory interval.     

Effect of pen size on behavioral, endocrine, and immune responses of water buffalo (Bubalus bubalis) calves.

Female water buffalo (Bubalus bubalis) calves (n = 28) aged 7 to 10 d were divided into four groups of seven animals each to examine the effects of space allowance (Group A: 2.6 indoor m2 + 2.0 outdoor m2/calf; Group B: 2.6 indoor m2/calf; Group C: 1.5 indoor m2/calf; Group D: 1.0 indoor m2/calf) on behavioral, endocrine, and immune variables for a period of 60 d. Animals were offered 7 L/d of a commercial acidified milk substitute. The calves averaged 45.9 kg initially and 92.4 kg finally. The behavior observations were conducted 7 d after grouping and fortnightly thereafter. At wk 4 and 8, the phytohemagglutinin (PHA) skin test was performed to induce aspecific delayed hypersensitivity. At wk. 1 and 3, calves were injected i.m. with keyhole limpet hemocyanin. Antibody titers were determined at weekly intervals for 7 wk. Calves in pens with greater space allowance (Groups A and B) were less active than Groups C and D (P<.001). The latter groups were also observed feeding more often at wk 7 (P<.01). Calves provided with an outdoor paddock spent less time standing than Groups C and D (P<.01), and lay with a greater number of outstretched legs (P<.001). Groups C and D showed a lower reaction to PHA in both skin tests than did Groups A and B (P<.001 and P<.05, respectively). Group A showed an antibody response consistently higher than groups B, C, and D (P<.01, P<.05, and P<.05, respectively). At the end of the experimental period, the calves were subjected to an isolation test lasting 10 min. Group D showed a longer duration of movement with respect to Groups A and B (P<.01); animals from Group C walked more than did Group A (P<.05). Cortisol concentration evaluated 0, 10, 45, 90, 150, and 225 min after separation from the group was higher in Groups C and D than in Groups A and B (P<.01). For all animals, the highest cortisol level was observed immediately after the isolation test (P<.001). Space restriction resulted in evidence of stress in the animals as shown by alterations in a number of physiological responses. However, the use of small groups of only seven animals per pen may have affected their reactions to space restriction. It is possible that using larger groups could change these conclusions.     

A VACUUM-FORMABLE MATTRESS FOR VETERINARY RADIOTHERAPY POSITIONING: COMPARISON WITH CONVENTIONAL METHODS

This prospective study was undertaken to compare the positioning repeatability and setup time of a rigid immobilization device (Vac-Lok mattress) to conventional positioning methods (sandbags, tape, foam wedges) in the clinical veterinary radiotherapy setting. Positioning repeatability was determined by using port films to verify appropriate patient positioning. Setup time was determined by recording the time required to set up each patient using each positioning method. Sixty-seven patients receiving radiotherapy were positioned using both the Vac-Lok mattresses and conventional positioning methods during their treatments. Seventy-eight total sites were treated. Forty-eight were treated daily (Monday through Friday, 2 to 4 weeks) and 30 were treated once weekly (4 weeks). Patients were grouped according to the site treated: head (29), neck/body (24), and limb (25). Vac-Lok mattresses were similar to conventional means in positioning repeatability and setup time. Vac-Lok mattresses are potentially advantageous in specific situations, including use during pre-radiotherapy tumor imaging. These mattresses are not recommended for distal limb positioning.     

Immune response to Neospora caninum native antigens formulated with immune stimulating complexes in calves

The aim of this study was to compare the immune responses to live Neospora caninum tachyzoites and N. caninum native antigens formulated with immune stimulating complexes matrix (ISCOM-matrix) in calves. Fifteen calves were used in this study: 3 were intravenously inoculated with 1 × 10(8) live tachyzoites (Group A), 3 were inoculated twice with N. caninum native antigens formulated with ISCOMs (Group B); 3 with N. caninum native antigens in phosphate-buffered saline (PBS) (Group C); 3 received ISCOM-matrix (ISCOMs without antigen) (Group D) and 3 were negative controls receiving PBS (Group E). The last four groups were inoculated subcutaneously. The specific total IgG and its subtypes were analyzed by an indirect enzyme-linked immunosorbent assays (ELISAs) and by Western blot. IFN-γ levels in plasma was quantified using a commercial kit. All calves were challenged intravenously with 1 × 10(8) live tachyzoites at week 11 after receiving the first dose. Parasitemia was assessed in plasma samples by semi-nested PCR. Neospora-specific antibodies were detected in animals from Groups A and B in the week 2 after inoculation. The ELISA OD values were higher in Group B compared with Group A from weeks 6 to 11 (P<0.05). Analysis of the subisotype specific antibodies in experimentally infected calves revealed a predominant IgG(2) response; however, a predominant IgG(1) response was observed in animals inoculated with N. caninum native antigens formulated with ISCOM-matrix. Control calves remained seronegative until challenge infection. The pattern of bands by Western blot was similar when testing sera from animals in Groups A and B. The levels of IFN-γ production after respective immunization schedules were similar between Groups A and B. Neospora-DNA was detected in plasma samples shortly after intravenous challenge in calves from all groups including those receiving the experimental vaccine formulation. The duration of the parasitemia was similar in all groups.     

Behavioral and physiological responses of horses (Equus caballus) to head lowering

Horse trainers often report that lowering the height of a horse’s head so the poll is below the height of the withers can induce a calming effect during training. Four groups of horses were used in a 2-part study to investigate the behavioral and physiological effects of head lowering in horses. In Part 1, Group A had no experimental stimuli applied and horses in Group B were trained to lower their heads when presented with a specific stimulus by the handler. The stimulus for head lowering was the application of downward pressure on the headcollar via the lead rope until the horse lowered its head such that its lips were approximately at mid-cannon (third metacarpal) height, whereupon the pressure was released. The stimulus was applied again if the horse raised its head during the 300-second test period. In Part 2, Groups C and D were aroused until their heart rates exceeded 100 beats per minute (bpm). Group C had no further experimental stimuli applied whereas Group D lowered their heads as a response to the above stimulus for a period of 300 seconds. Repeated measures analysis showed that there was no difference between the heart rate of Groups A and B or Groups C and D but that the heart rate of Groups A and B were lower than Groups C and D during the 300-second post-arousal (P < 0.001). The horses in Groups A and B were more likely to contact the handler (P < 0.001), exhibit licking and chewing (P < 0.001), rest a hindleg (P < 0.001), and sniff the ground (P < 0.001) than those in Groups C and D. The number of stimuli required to maintain the head in a lowered position was greatest during the first 30 seconds (P = 0.012 and P < 0.001, Parts 1 and 2, respectively). The current study has shown that head lowering in horses does not influence cardiac responses, even after the horses had been aroused to have their heart rates above 100 bpm. Therefore, it is not a method that will aid in calming an aroused horse in training. Contrary to popular belief, there was no association with licking-and-chewing and head lowering, nor with these behaviors and response acquisition.     

The early phase of the immune response to live and killed Staphylococcus aureus vaccines in sheep — Cellular and immunoglobulin parameters

Various cellular and protein parameters in efferent lymph were monitored during early stages of immune responses in the draining popliteal lymph nodes of sheep following injection with either live or killed $\text{Staphylococcus aureus}$ vaccines. Significantly higher outputs of leucocytes and lymphoblasts were measured in lymph from animals injected with the live vaccine (Group 1) compared with animals given the killed vaccine (Group 2) and non-immunised controls (Group 3). At 48 h post-injection the mean output of leucocytes in lymph for Group 1 was 9.5 × 10⁷ cells/h, and for Groups 2 and 3 comparable mean outputs were 5.2 × 10⁷ and 2.2 × 10⁷ respectively.There was a marked increase in IgM-containing cells in Group 1 animals, significant differences between Group 1 and 2 occurring 96 h post-injection. A difference was observed between the two immunised groups in the kinetics of output of cells containing antibody to a staphylococcal antigen extract. There was a slower increase in antibody-containing cells for Group 1 than for Group 2 animals. The mean proportion of antibody-containing cells reached 0.09% (of total leucocytes) at 72 h post-injection for Group 2 and 0.10% at 120 h post-injection for Group 1.Rapid increases in flow rate of lymph were recorded in animals in Group 1. These results were in contrast to lower corresponding values observed in sheep in Groups 2 and 3. There was a twofold increase in the lymph: serum (L:S) concentration ratio for IgG2 in animals in Group 1 and the evidence suggested that this was due to local synthesis of IgG2 by lymphoid cells in the abscess at the site of injection and/or in the draining popliteal lymph node. This change in L:S ratio for IgG2 was not recorded for sheep in Groups 2 and 3.     

Influence of rapeseed meal on productivity and health of broiler chicks

Research was focussed on investigating the influence of different quantities (0, 10, 20 and 30%) of rapeseed meal (RM), 00-cultivar Silvia on production results, as well as on the morphological and pathohistological changes in the internal organs of chicks during the course of the experiment, measured on the 21st and 42nd days of the 42-day experiment. The experiment involved 120 chicks divided into four groups: a control group (C) and three experimental groups (E1-3). It was found that chicks in Groups C, E1 and E2 realised significantly (P < 0.05) higher gains than those in Group E3, both after the test period and at the end of the experiment. No significant differences with regard to feed conversion were found between groups of chicks. Throughout the experiment chicks in Groups E1-3 were found to have a significantly (P < 0.05) heavier liver. Compared to Groups E1-3, Group C chicks had a significantly (P < 0.05) heavier gizzard after 21 days, but following the finisher diet Group E3 had a significantly (P < 0.05) lighter gizzard. Compared with birds in Group C, those in Groups E2-3 had significantly heavier (P < 0.05) unevacuated intestines when fed the starter diet, and those in Groups E1-2 showed a similar result when fed the finisher diet. Chicks in Group E3 had a significantly lower grill weight than those in Groups C, E1 and E2, both in the first half of the experiment and at the end. The starter diet did not result in any differences in the quantity of abdominal fat among groups, but following the finisher diet Groups E2-3 showed significantly less abdominal fat in comparison to Groups C and E1 (P < 0.05). Group C showed a significantly higher carcass yield than Groups E1-3 (P < 0.05) in the first half of the experiment and at the end. Groups E1-3 manifested a slight to medium hypertrophy of thyroid epithelial tissue as well as slight thymus hypertrophy and slight atrophy of the cloacal bursa follicles.     

Equal calcitonin response in paretic and non-paretic cows after intravenous calcium infusion

Seventeen cows divided into 3 groups were studied: Group 1 consisting of newly delivered cows (1–3 days post partum) suffering from parturient paresis; Group 2 comprising newly delivered (1–4 days post parturn) healthy cows and Group 3 consisting of non-pregnant, non-lactating cattle. All animals were given an intravenous standard Ca infusion, and blood samples were obtained before treatment and at intervals during a 3-hour period thereafter. Plasma levels of calcitonin (CT), calcium (Ca) and inorg. phosphate (P) were determined. No significant difference for the mean CT response existed between animals in Groups 1 and 2. Blood levels of inorg. P increased significantly with time in animals of Groups 1 and 2 but not in animals of Group 3.The finding here of a similar CT response after Ca stimulation in both paretic and non-paretic cows demonstrates that the C-cells in both groups of animals have the same capacity to secrete CT.     

Prophylactic efficacy of buparvaquone in experimentally induced Theileria annulata infection in calves

The antitheilerial activity of buparvaquone (BW 720C) was evaluated in experimentally induced Theileria annulata infections in cross-bred male calves. T. annulata infections were induced by injecting a suspension of infected ground tick tissue suspension (GUTTS) equivalent to two ticks subcutaneously into each calf. Buparvaquone at a dose of 2.5 mg kg-1 body weight was given as a single injection (intramuscularly) on Day 0 (Group 1), Day 8 (Group 2) and Day 12 (Group 3) post-infection. The animals in Groups 4 and 5 were untreated and challenged controls, respectively. All of the recovered animals from Groups 1-4 were challenged with a lethal dose of T. annulata at 6 weeks post-infection. The immunized animals were resistant to the homologous challenge, which killed three of four control animals (Group 5); the controls showed typical antemortem and post-mortem lesions of theileriosis.     

Effect of head position on intraocular pressure in horses

OBJECTIVE: To evaluate the effect of head position on intraocular pressure (IOP) in horses. ANIMALS: 30 horses. PROCEDURES: Horses were sedated with detomidine HCl (0.01 mg/kg, IV). Auriculopalpebral nerve blocks were applied bilaterally with 2% lidocaine HCl. The corneas of both eyes were anesthetized with ophthalmic 0.5% proparacaine solution. Intraocular pressures were measured with an applanation tonometer with the head positioned below and above heart level. The mean of 3 readings was taken for each eye at each position for data analysis. The effect of head position on IOP was assessed and generalized estimating equations were used to adjust for the correlation from repeated measures of the same eye and intereye correlation from the same horse. RESULTS: Of the 60 eyes, 52 (87%) had increased IOP when measured below the heart level. A significant difference (mean +/- SE, 8.20 +/- 1.01 mm Hg) was seen in the mean IOP when the head was above (17.5 +/- 0.8 mm Hg) or below (25.7 +/- 1.2 mm Hg) heart level. No significant effect of sex, age, or neck length on IOP change was found. CONCLUSIONS AND CLINICAL RELEVANCE: Head position has a significant effect on the IOP of horses. Failure to maintain a consistent head position between IOP measurements could potentially prevent the meaningful interpretation of perceived aberrations or changes in IOP.     

Immune responses of pony foals during repeated infections of Strongylus vulgaris and regular ivermectin treatments.

Ten helminth-free pony foals divided into three groups were used in this study. Eight foals were each experimentally infected per os with 50 Strongylus vulgaris infective larvae weekly for 4 weeks, at which time one foal died of acute verminous arteritis. The remaining seven foals subsequently received 50 S. vulgaris infective larvae every 2 weeks for an additional 20 weeks. Four of the infected foals remained untreated (Group 1) and three of the infected foals were given ivermectin at 8, 16 and 24 weeks post initial infection (Group 2). Two foals served as controls (Group 3). Foals in Group 1 developed eosinophilia, which was sustained throughout the course of infection. A mild eosinophilia also developed in Group 2 foals; however, the eosinophil numbers were markedly reduced for 3 weeks after each ivermectin treatment. Only foals in Group 1 developed significant (P less than 0.05) hyperproteinemia, hyperbetaglobulinemia and a reversal of the albumin/globulin (A/G) ratio 4 weeks after initial infection. Significant (P less than 0.05) IgG anti-S. vulgaris ELISA titers developed in foals in Groups 1 and 2 3 weeks after infection and were sustained for the duration of the experiment. Western blot analysis of soluble somatic antigens of S. vulgaris adult female and male worms probed with sera from foals in Groups 1 and 2 revealed only subtle differences between these animals. The blastogenic reactivity of peripheral blood mononuclear cells (PBMC) to phytohemagglutinin and concanavalin A was not significantly different between groups. Peripheral blood mononuclear cells from foals in Groups 1 and 2 developed significant (P less than 0.05) blastogenic reactivity to S. vulgaris soluble adult somatic antigen when examined at 25 weeks after infection. Mesenteric lymph node cells from foals in Group 2, although not statistically significant, were more reactive to antigen than were the mesenteric lymph node cells from foals in Group 1 when examined at 27 weeks after infection. These results suggest that significant alterations in the immune response of ponies to S. vulgaris does not occur after intravascular killing of larvae by ivermectin treatments.     

The recombinant rabbit hemorrhagic disease virus VP60 protein obtained from Pichia pastoris induces a strong humoral and cell-mediated immune response following intranasal immunization in mice

Rabbit hemorrhagic disease (RHD) is a contagious and highly lethal viral disease of rabbits that spreads rapidly and infects animals by nasal, conjunctival and oral routes. Therefore, this experiment was undertaken to study the immune response generated after intranasal (i.n.) vaccination with the recombinant VP60 capsid protein from rabbit hemorrhagic disease virus (RHDV) expressed at high levels in Pichia pastoris. Groups of BALB/c mice were immunized with three doses of purified VP60 protein (Group 1), VP60 formulated within the cell debris fraction of the transformed yeast (Group 2) and placebo (Group 3) by intranasal route. Mice were also intramuscularly injected with purified VP60 protein (Group 4). A rapid antibody response specific against rabbit hemorrhagic disease virus was observed in all the experimental groups, except in Group 3, as detected by ELISA. The highest titers were found 60 days after the first immunization. Mice from Group 1 showed the highest IgG response (p<0.05) and the most balanced profile of IgG1, IgG2a and IgG2b subclasses. IgA titers specific to the virus were found only in animals from this group, which also developed the highest specific lymphocyte proliferative response. Interferon-gamma (IFN-gamma) and interleukin-12 (IL-12) gene expression was also detected after an ex vivo-specific stimulation of mice from Groups 1 and 4. These data demonstrated the capacity of VP60 protein expressed in P. pastoris to elicit a potent humoral and cell-mediated immune response following an intranasal immunization scheme.