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Purpose: We questioned whether the vaccine adjuvant combination of TLR-7 ligand agonist, imiquimod, with granulocyte macrophage colony-stimulating factor (GM-CSF) would result in enhanced dendritic cell recruitment and activation with increased antigen-specific immunity as compared with either adjuvant used alone., Experimental Design: The adjuvant effects of GM-CSF and imiquimod were studied in ovalbumin (OVA) and MMTVneu transgenic mice using peptide-based vaccines. Type I immunity, serum cytokines, myeloid-derived suppressive cells (MDSC), and regulatory T cells (Treg) levels were examined., Results: Both GM-CSF and imiquimod equally induced local accumulation and activation of dendritic cells. Both adjuvants effectively enhanced OVA-specific T-cell responses. We further evaluated the antitumor efficacy of adjuvant GM-CSF and imiquimod immunizing against murine insulin-like growth factor-binding protein-2 (IGFBP-2), a nonmutated oncoprotein overexpressed in the tumors of MMTVneu transgenic mice. Tumor growth was significantly inhibited in the mice receiving IGFBP-2 peptides with GM-CSF (P = 0.000), but not in imiquimod vaccine-treated groups (P = 0.141). Moreover, the addition of imiquimod to GM-CSF negated the antitumor activity of the vaccine when GM-CSF was used as the sole adjuvant. While GM-CSF stimulated significant levels of antigen-specific T-helper cell (T(H))1, imiquimod induced elevated serum interleukin (IL)-10. Both MDSC and Tregs were increased in the imiquimod-treated but not GM-CSF-treated groups (P = 0.000 and 0.006, respectively). Depleting MDSC and Treg in animals immunized with imiquimod and IGFBP-2 peptides restored antitumor activity to the levels observed with vaccination using GM-CSF as the sole adjuvant., Conclusion: Adjuvants may induce regulatory responses in the context of a self-antigen vaccine. Adjuvant triggered immunosuppression may limit vaccine efficacy and should be evaluated in preclinical models especially when contemplating combination approaches.

Multiple TLR agonists have been shown to have antitumor effects in animal models. However, the therapeutic efficacy of TLR agonist monotherapy in cancer treatment has been limited, and the mechanisms of failure remain unknown. We demonstrate that topical treatment with a TLR-7 agonist, imiquimod, can elicit significant regression of spontaneous breast cancers in neu transgenic mice, a model of human HER-2/neu(+) breast cancer. However, tumor growth progressed once imiquimod therapy was ended. Gene expression analysis using tumor-derived RNA demonstrated that imiquimod induced high levels of IL-10 in addition to TNF-alpha and IFN-gamma. Elevated levels of circulating IL-10 were also detected in sera from imiquimod-treated mice. Elevated serum IL-10 appeared to be derived from IL-10 and dual cytokine secreting (IFN-gamma(+) and IL-10(+)) CD4(+) T cells rather than CD4(+)CD25(+)Foxp3(+) T regulatory cells, which were also induced by imiquimod treatment. Blockade of IL-10, but not TGF-beta, enhanced the antitumor effect of imiquimod by significantly prolonging survival in treated mice. These data suggest that the excessive inflammation induced by TLR agonists may result in a self-regulatory immunosuppression via IL-10 induction and that blocking IL-10 could enhance the therapeutic efficacy of these agents.

The normal sonographic appearance of the adult canine gastrointestinal tract has been described. Interpretation of abdominal ultrasonographic findings in puppies is difficult due to the lack of information on normal ultrasonographic findings. The gastrointestinal tract, jejunal lymph node size and the presence and appearance of abdominal fluid were investigated in 23 normal, 7-12-week-old Beagle puppies. The duodenal wall thickness was greater than in other parts of the gastrointestinal tract (mean 3.8 +/- standard deviation [SD] 5 mm, range 3.2-4.8 mm). The mean stomach wall thickness was 2.7 +/- SD 0.4 mm (range 2.2-3.7 mm), the mean jejunal wall thickness was 2.5 +/- SD 0.5 mm (range 1.2-3.4 mm), and the mean colonic wall thickness was 1.3 +/- SD 0.3 mm (range 0.7-2.0 mm). In addition, mean duodenal and jejunal mucosal layer thicknesses measured 2.7 +/- SD 0.5 mm (range 2.0-3.8 mm) and 1.5 +/- SD 0.4 mm (range 0.6-2.5 mm), respectively. Homogenous, hypoechoic jejunal lymph nodes were easily found and the mean thickness was 7.1 +/- SD 2.2 mm (range 1.5-12.5 mm). A mild amount of anechoic free peritoneal fluid was present in all puppies.

The ultrasonographic appearance of the gastrointestinal tract of puppies suffering from parvoviral enteritis was characterized. Forty puppies between 6 and 24 weeks of age with confirmed canine parvoviral enteritis were examined ultrasonographically within 24 h of admission. Sonographic findings included fluid-filled small intestines in 92.5% of subjects, and stomach and colon in 80% and 62.5% of subjects, respectively. Generalized atony was present in 30 subjects and weak peristaltic contractions indicative of functional ileus observed in the remaining 10 subjects. The duodenal and jejunal mucosal layer thicknesses were significantly reduced when compared with normal puppies with mean duodenal mucosal layer measuring 1.7 mm and jejunal mucosal layer 1.0 mm. Additionally, a mucosal layer with diffuse hyperechoic speckles was seen in the duodenum (15% of subjects) and the jejunum (50% of subjects). The luminal surface of the duodenal mucosa was irregular in 22.5% of subjects and the jejunal mucosa in 42.5% of subjects. In all of these subjects, changes were accompanied by generalized indistinct wall layering. Small intestinal corrugations were seen within the duodenum in 35% of subjects and within the jejunum in 7.5%. A mild amount of anechoic free peritoneal fluid was observed in 26 subjects and was considered within normal limits and a moderate amount of anechoic free peritoneal fluid was observed in six subjects. The jejunal lymph node size was within normal limits. None of the above changes are pathognomonic for canine parvoviral enteritis but finding them in combination is highly suggestive.

Objective: To evaluate the accuracy of specific magnetic resonance imaging (MRI) sequences in determining the site, lateralization, and extent of extruded intervertebral disk material (EIDM), compared with surgical findings, in Dachshunds with thoracolumbar intervertebral disk extrusion (TLIDE)., Design: Prospective clinical study., Sample Population: 16 Dachshunds with clinical signs of intervertebral disk disease., Procedures: Preoperative T1-weighted, T2-weighted, and short tau inversion recovery (STIR) MRI measurements and description of the location of EIDM were compared with intraoperative measurements and determination of the EIDM position., Results: The T12-13 intervertebral disk space was the most frequent site of EIDM (6/16 dogs). The EIDM lateralized with equal frequency to the left and right sides; no central extrusions were seen. There was moderate to substantial agreement (kappa, 0.59) between MRI and surgical findings for evaluation of the craniocaudal distribution of the EIDM. For measurement of the length of EIDM, the T1-weighted, T2-weighted, and STIR sequences had a mean error of -1.15, -0.38, and -1.93 mm, respectively; concordance correlation coefficients were 0.666, 0.904, and 0.458, respectively. Mean absolute errors were 2.54, 1.35, and 2.90 mm, respectively; these values did not differ significantly., Conclusions and Clinical Relevance: In the thoracolumbar vertebral column of Dachshunds with clinical signs of intervertebral disk disease, MRI is a valuable technique for determining location and craniocaudal length of EIDM. Compared with T1-weighted and STIR images, T2-weighted images appeared to be more accurate and precise and are potentially more reliable for determination of the length of EIDM in those dogs.

Studies have shown that the immune system can recognize self-antigens under conditions (eg, cell injury) in which the self-tissue might elaborate immune-activating endogenous danger signals. Uric acid (UA) is an endogenous danger signal recently identified to be released from dying cells. Prior work has shown that UA activates immune effectors of both the innate and adaptive immune system, including neutrophils and cytotoxic T-cell immunity. However, it was unclear whether UA could enhance antibody immunity, which was examined in this study. When added to dying tumor cells or with whole protein antigen, UA increased IgG1-based humoral immunity. Further, UA blocked growth of tumor in subsequent tumor challenge experiments, which depended on CD4, but not CD8, T cells. Sera derived from UA-treated animals enhanced tumor growth, suggesting it had little role in the antitumor response. UA did not signal for T-cell expansion or altered tumor-infiltrating leukocyte populations. Consistent with the lack of T-cell expansion, when applied to dendritic cells, UA suppressed T-cell growth factors but up-regulated B cell-activating cytokines. Understanding the nature of endogenous danger signals released from dying cells may aid in a better understanding of mechanisms of immune recognition of self.

Diseases of the abdomen of the cheetah (Acinonyx jubatus) include those affecting the liver, spleen, and urinary tract. The most common diseases of captive-bred cheetah are gastritis, gastric ulceration, glomerulosclerosis, and hepatic veno-occlusive disease, and are the most frequent causes of mortality in these animals. The purpose of this study was to describe the ultrasonographic anatomy of the normal liver, spleen, kidney, and urinary bladder of the anesthetized captive-bred cheetah. Twenty-one cheetahs were examined. Eight of the 21 animals had subclinical evidence of either gastritis or chronic renal disease. The ultrasonographic appearances of the liver, gall bladder, common bile duct, and spleen were evaluated and various measurements made. Statistical analyses of the measurements were performed on all the healthy and subclinically ill animals taking sex, age, mass, and anesthetic protocol into account. There were no significant differences in any parameters between the healthy and subclinically ill animals (P > 0.25) and data were combined for statistical analyses. The mean mass was 41.1kg ( +/- 8.8) and the mean age was 5.0 years (+/- 2.2). The mean thickness of the liver medial to the gall bladder was 67.0 mm (+/- 14.8) and the liver was within the left costal arch in 75% of animals, extended caudal to the right costal arch in 50% of animals for an average of 30 mm, and extended caudal to the sternum in 63% of animals for an average of 32.5 mm. The maximum mean hepatic vein diameter at the entrance to the caudal vena cava was 8.6 +/- 2.8 mm; the mean diameters of the portal vein at the hilus and that of the caudal vena cava as it entered the liver were 7.5 +/- 1.6 and 9.9 +/- 4.1 mm, respectively. The mean diameter of the caudal vena cava was significantly affected by the type of anesthetic used (P < 0.10). The mass of the animals was significant in explaining the variance in maximum portal vein diameters (P < 0.10). The mean maximum velocity of the hepatic vein flow at the entrance to the caudal vena cava was 25.3 +/- 2.8 cm/s (n=4), the hilar portal vein was 11.7 +/- 3.3 cm/s (n=7), and the caudal vena cava was 33.8 +/- 19.8 cm/s (n=5). The mean maximum gall bladder length and width, and the mean common bile duct diameters were 44.6 mm (+/- 10.4), 23.3 mm (+/- 5.0), and 8.1 mm (+/- 2.4), respectively. Age was significant in explaining the variance in gall bladder lengths (P<0.10). Urinary tract ultrasonography was performed only in animals that had normal urea and creatinine levels (n=13). Renal cortico-medullary distinction was present in all kidneys and a cortico-medullary rim sign was seen in 21 of 26 kidneys. Mean kidney length, height, and width was 63.9 +/- 5.7, 38.1 +/- 5.2, and 42.1 +/- 5 mm, respectively. The average resistivity index was 0.58 (n=5). Mean urinary bladder length, height, and width were 57.0, 19.2, and 34.9 mm, respectively.

Thoracic and pelvic limbs from 15 euthanized free-ranging lions (Panthera leo), ranging in age from 16 to 144 months, underwent standard radiographic evaluation. All lions had tested positive for Mycobacterium bovis by means of a modified intradermal tuberculn test. The radiographs of six lions were normal and nine had incidental findings of which six had more than one lesion. Seven lions had lesions suspected to be associated with tuberculosis, which was confirmed in specific joints in two lions. Incidental pathology was classified as traumatic injuries and degenerative or trauma-associated joint disease. The traumatic lesions were fractures of which the most remarkable was a femur malunion. Four lions had fibula and another three lions had metacarpal/tarsal and phalangeal fractures. Joint lesions included glenoid, humeral head, and accessory carpal bone osteophytes. There was evidence of a cranial cruciate ligament rupture in an 8-year-old male. Trauma induced joint lesions were seen in four stifles (fragmented or displaced sesamoid bones, fragmented meniscal ossicle, or mineralized fragments). Radiological abnormalities believed to be caused by M. bovis were present in one stifle, one radiocarpal three tibiotarsal, and one tarsometatarsal joints. These had evidence of septic arthritis with extensive bone formation and capsular mineralization. In one 20-month-old lion, changes typical of a bone abscess were found in a proximal tibia. Radiologic evidence of elbow hygromas were seen in three elbows, all believed to be caused by M. bovis. Lions appeared to cope fairly well with a variety of traumatic injuries and were also susceptible to some of the aging/incidental radiologic findings seen in dogs and cats. The suspected M. bovis osseous lesions were more likely to involve the joints, particularly the tarsal joint and were mainly proliferative.

A 22-year-old male European brown bear (Ursus arctos arctos) was presented to the Onderstepoort Veterinary Academic Hospital after an acute onset of hind limb paralysis 4 days earlier. Previous radiographs revealed marked degenerative joint disease of the stifles, tarsi and digits. The clinical findings were consistent with acute disc prolapse. Lateral radiographs of the entire vertebral column were made as well as ventrodorsal pelvic radiographs. The latter were within normal limits. The vertebral column revealed multiple lesions consistent with chronic and acute disc herniations. Lateral compression of the caudal lumbar nerve roots could not be ruled out. Owing to multiple significant findings of the vertebral column and the poor prognosis for full recovery after surgery, the bear was euthanased. The diagnosis of an acute disc prolapse and multiple chronic disc herniations was confirmed on necropsy.

Ultrasonographic examinations were performed on 17 clinically healthy adult common marmosets to gain information about the normal abdominal echoanatomy. The marmosets were 1.5-9 years of age and weighed between 328 and 506g. Marked species-specific differences compared with the cat or dog were noted. Good images of the kidneys, bladder, spleen, adrenal glands, liver, and the gastrointestinal tract could be obtained. The pancreas, caecum, and abdominal lymph nodes were not seen. The spleen was the least echogenic organ, followed by the medium echogenic liver and the sometimes isoechoic, but mostly hyperechoic renal cortex. The kidneys had a poor corticomedullary distinction. The prominent right lobes of the liver extended caudally far beyond the costal arch. The gallbladder had a bi- to multi-lobed appearance with a wide, tortuous cystic duct. The pylorus was centrally positioned. The adrenal glands were readily seen, but should not be confused with the adjacent spleen. A statistically significant (P<0.05) difference between female and male kidney, and right adrenal gland length was present.

A standard radiographic procedure was developed for the thorax and abdomen in the common marmoset. A description and reference values for the corresponding radiographic anatomy are given. Radiographs were obtained from 17 anaesthetized healthy mature marmosets ranging from 1.5 to 9 years and 328 to 506 g. Left-to-right lateral recumbent and ventrodorsal whole-body radiographs made at end inspiration are recommended. Images of the heart, lungs, liver, gastric axis, and at least one kidney could be evaluated consistently. A generalized interstitial/peribronchial pattern was normally present. The mean of the vertebral heart size +/- SD on dorsoventral or ventrodorsal views was 9.42 (+/- 0.44), ranging from 8.8 to 10.6. Abdominal detail was generally poor. The gastrointestinal structures could often only be identified because of their luminal gas. The right liver lobes were prominent and extended caudally beyond the costal arch. The pylorus was in a central position and the spleen could not be distinguished. Additionally, pancreas, lymph nodes, urinary bladder, and ureters were not identified. A statistically significant difference (P < or = 0.05) between female and male kidney length existed. This study emphasizes that significant species differences exist, and simply applying canine or feline radiographic interpretation will result in misinterpretation.

A 14-month-old female Eclectus parrot (Eclectus roratus) was presented with a 4-week history of bloody diarrhea and depression. No additional information could be gained from physical examination. Only selected diagnostic tests (faecal examination, haematocrit, aspartate aminotransferase, and uric acid) could be performed due to financial constraints, but all where within reference range. Unspecific antibiotic treatment was started and the bird responded well initially, but had to be readmitted 2.5 weeks after initial presentation. Four weeks after initial presentation the owner finally consented to taking whole body radiographs and a diagnosis of an intestinal foreign body could be made. The foreign body was surgically removed 2 days later. The bird recovered uneventfully after surgery and was still in good health 1 year after surgery. This article emphasises the importance of diagnostic imaging in the avian patient. A brief review of avian gastrointestinal foreign bodies is given (concentrating on the psittacine patient) and the importance of distinguishing metallic from non-metallic gastrointestinal foreign bodies are discussed.

Large clonal expansions of peripheral CD8(+) T cells carrying receptors for single epitopes of CMV are common in the elderly and may be associated with an immune risk phenotype predicting mortality. To study the effect of ageing on the ability of CMV-specific CD8(+) T cells to produce type 1- and type 2-cytokines, interferon-gamma-and IL-10-producing, CD8(+) T cell responses in the presence of CMV peptide antigen were measured in CMV-seropositive old and young donors. We found that large expansions of A2/NLV-specific CD8(+) T lymphocytes in the elderly are accompanied by a partial loss of antigen responsiveness as reflected in a greatly decreased frequency of antigen-specific IFN-gamma-and IL-10-producing cells. Thus, despite carrying specific antigen receptors, the majority of the clonally expanded CMV-specific CD8(+) cells in the elderly was dysfunctional according to these criteria. Our data indicated a bias towards a more anti-inflammatory response in the elderly. The accumulation of dysfunctional CMV-specific cells might fill the 'immunological space' and decrease the available repertoire of T cells for novel antigens. This might account for the increased incidence of many infectious diseases in the elderly.

Ten gastrointestinal contrast studies were performed with barium on seven clinically healthy ostriches. Concentrations of 25-100% w/vol liquid barium sulfate at 7 and 10 ml/kg were administered by stomach tube after withholding food for 16 h. A 6-frame technique for left-to-right lateral views in standing and a 3-frame technique for the dorsoventral views in sternally recumbent adult ostriches were used for survey and contrast radiographs. Objectives were to describe the appearance of the normal gastrointestinal tract with contrast radiography and to provide a guideline for optimal dosage and concentration of barium sulfate as well as a reliable protocol for frequency of radiographs. Structures that were consistently identified included the esophagus, proventriculus, ventriculus, duodenum, jejunum, proximal, and distal rectum. Due to the superimposition of the remainder of the small intestine, individual components were difficult to differentiate. The caeca were inconsistently highlighted and only for a short time. The ventral pouch of the coprodeum never filled with contrast medium.

Large clonal expansions of peripheral CD8+ T cells carrying receptors for single epitopes of CMV and EBV are common in the elderly and may be associated with an immune risk phenotype predicting mortality. Here we show that the frequency of CD8+ T cells expressing the inhibitory killer cell lectin-like receptor G1 (KLRG1), a marker of cells unable to undergo further clonal expansion, was markedly elevated in CD8+ T cells from old donors. Moreover, tetramer staining revealed that the elevated frequency of CMV-specific CD8+ T cells in the elderly was due to an accumulation of cells bearing this dominant negative receptor. The fraction of CMV-specific T cells able to secrete interferon-gamma after specific antigenic stimulation was significantly lower in the elderly than in the young, although the total number of functional cells was comparable. Therefore, the majority of the clonally expanded virus-specific CD8+ cells in the elderly was dysfunctional. Thus, T cell responses are altered in the aged by an accumulation of replicatively senescent dysfunctional T cells carrying receptors for persistent herpes viruses. The presence of clonal expansions of such virus-specific cells may shrink the available repertoire for other antigens and contribute to the increased incidence of infectious disease in the elderly.

Longitudinal studies suggest that a set of immune parameters including high percentages of peripheral CD8+, CD28-, CD57+ T lymphocytes, low CD4 and B cell counts, and poor T cell proliferative responses to mitogens is associated with decreased remaining longevity in the free-living very elderly (> 85 years). This combination of immune parameters was also significantly associated with an inverted CD4/CD8 ratio and cytomegalovirus seropositivity. Here, using tetramer technology, we show markedly increased numbers of CD8+ T cells bearing receptors for one single CMV epitope in the very elderly. Moreover, the fraction of these tetramer-reactive cells secreting interferon-gamma after specific antigenic stimulation was significantly lower in the old than in the young, as was the percentage of CD28-positive cells in this population. Therefore, we conclude that marked expansions of CMV-specific CD8+ T cells have occurred and that the obsession of a large fraction of the entire CD8+ T cell subset with one single viral epitope may contribute to the increased incidence of infectious disease in the elderly by shrinking the T cell repertoire available for responses to other antigens.

The aim of this study was to provide a basis for investigating the effects of one very common environmental factor, Epstein-Barr virus (EBV), on age-related changes in the immune system. To this end, the frequency of CD8(+) T cells carrying receptors for an immunodominant EBV lytic epitope was assessed by direct staining with HLA-peptide tetrameric complexes in 19 very old (>87 years) and 12 young (20-40 years) EBV carriers. The frequency of EBV-tetramer-positive cells within the CD8(+) subset was significantly greater in the old compared to the young group (P=0.001). However, the frequency of EBV antigen-specific IFN-gamma producing T cells, as determined by ELISPOT, was significantly lower in the old (P=0.001). Therefore, the absolute number of functional EBV-specific T cells in the elderly and the young was probably similar. These data suggest CD8 clonal expansions in the elderly, resulting in an accumulation of dysfunctional EBV-specific cells which possibly fill the 'immunological space' and could lead to a shrinking of the T cell repertoire for other novel antigens. This may help to explain the increased incidence and case-fatality caused by viruses and intracellular pathogens in the elderly.

Chronic myelogenous leukemia (CML) is characterized by a reciprocal translocation leading to the Philadelphia chromosome. Two fusion genes are created by this translocation: bcr/abl and abl/bcr. The fusion regions of both translocation products are unique and strictly limited to leukemia cells, giving rise to potential tumor-specific antigens. Although several studies on the immunogenicity of peptides spanning the bcr/abl fusion region have been reported, little is known about the corresponding reciprocal translocation product abl/bcr. Here we report that synthetic peptides representing the fusion region of the abl/bcr forms a1bb3 and a1bb4 can be specifically recognized by HLA-A2-restricted cytotoxic T lymphocytes from healthy donors. Furthermore, HLA-matched a1bb3-expressing CML cells can be recognized by a1bb3-specific HLA-A2-restricted T cells, indicating natural processing and presentation of abl/bcr protein by leukemia cells. Moreover, a 19-mer peptide encompassing this class I-binding sequence also elicited a1bb3-specific class II-restricted T-cell responses. Thus, both class I- and class II-restricted T-cell responses can be stimulated in healthy donors by abl/bcr peptides in vitro. Because abl/bcr is expressed in the majority of CML patients, it may represent a highly leukemia-specific antigen with potential use in immunotherapy.

The elderly suffer from an increased incidence of infectious disease, accompanied by increased morbidity and mortality. Interferon-gamma plays an important role in defense against intracellular pathogens such as mycobacteria and viruses. A reduced capacity to produce this cytokine in the elderly, as demonstrated by our findings of significant decreases in IFN-gamma production in vitro on stimulation with bacterial products (LPS) or viral antigens (influenza vaccine), might therefore contribute to disease susceptibility. Moreover, accumulating data suggest that persistent infection with EBV and particularly CMV impacts upon the immune system in aging and may contribute to the immune risk phenotype (IRP), which predicts remaining longevity in the very elderly. Using tetramer technology and IFN-gamma ELISPOT assays, we found that the commonly-observed clonal expansions of CD8+ T-cells in the elderly were for the most part poorly-functional CMV- and EBV-specific cells, expressing little CD28. The resulting accumulation of dysfunctional cells may lead to a reduction of the repertoire of functional T-cells available for responses to novel antigens. Further longitudinal studies are needed to demonstrate whether cytokines such as IFN-g are also part of the IRP. Improving the definition of the IRP will help to understand and thus prevent or reverse age-associated immune dysfunction.

This study provides a reference for the radiographic anatomy of the thoraco-abdominal cavity of female ostriches as a representative of ratites. One ostrich cadaver, 2 adult and 2 growing ostriches were used. Right lateral radiographs produced by a 6-frame technique and 2 dorsoventral radiographs produced by an adapted 3-frame technique were selected and schematic illustrations of these were labelled to illustrate normal radiographic anatomy. Differences from other avian species and unique features of the ostrich are briefly discussed.

Ultrasonographic examinations were performed on clinically healthy growing and adult nonbreeding female ostriches. Multiple acoustic windows and the normal ultrasonographic appearance of the coelomic viscera were described. Good images of the heart and its major vessels, proventriculus, ventriculus, intestines, liver, and kidneys could be obtained. Additionally, an anechoic structure, believed to function as an urinary bladder, could be imaged in the cloacal region. The pancreas, spleen, thyroid glands, ovary, and adrenals could not be seen in this study. General limitations were the size of the ostriches, massive leg and dorsal muscles, large sternum, the extensive air sac system, compact convoluted intestines, and varying amounts of gastrointestinal gas. Ostriches do not posses a gall bladder, and thus it could not be used as a landmark or acoustic window. The extensive air sac system and feathers did not limit the use of ultrasonography as much as anticipated. Imaging of air sacs should, however, be considered to detect pathology such as air sacculitis, which may result in consolidation. Veterinary

A standard radiographic procedure was developed for the thoraco-abdominal cavity of female nonbreeding ostriches. Positioning, collimation, centering and a technique chart were defined to give reproducible and consistently good quality radiographs. Radiographs were obtained from one adult ostrich cadaver, two adult female ostriches as well as two growing ostriches at various stages. A 6-frame technique was established for lateral views taking the topographic tissue distribution into consideration and using easily palpable landmarks as centering points. Standing true right lateral radiographs are recommended for standard procedures. For dorsoventral exposures a 3-frame technique in the recumbent ostrich was found to be optimal. Birds should be fasted if possible. A technique chart for lateral exposures is provided.