bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of antibodies reactive with porcine lymphocytes and lymphoma cells in formalin-fixed, paraffin-embedded, antigen-retrieved tissue sections
Authors: Tanimoto Tadashi , Ohtsuki Yuji
Published: January 2024

Objectives

To determine whether antibodies raised against human or porcine lymphocytes are reactive with porcine lymphocytes and lymphoma cells in formalin-fixed, paraffin-embedded, antigen-retrieved tissue sections, and to determine the conditions suitable for antigen retrieval.

Design

We evaluated reactivities of 27 anti-lymphocyte antibodies with porcine tissue sections, including those of lymphomas, treated with 11 antigen-retrieval methods.

Animals

Swine.

Procedure

We used 19 anti-human and 8 anti-porcine lymphocyte antibodies. For antigen retrieval, we tested 11 methods: heating with 6 soaking solutions in a microwave oven, heating with 2 commercially available soaking solutions in a water bath, and enzyme digestion with 3 proteases. After these treatments, sections were immunostained along with untreated sections.

Results

Of 27 anti-lymphocyte antibodies tested, CDw75 (clone LN-1), CD79α (mb-1, clone HM57), CD79β(B29, clone B29/123), HLA-DR (clone TAL. 1B5), and polyclonal CD3 were strongly reactive with porcine lymphoid tissues, including lymphomas, when sections were deparaffinized and heated in 0.1M Tns-HCI buffer (pH 8.0) in a microwave oven or with antigen-retrieval solution in a water bath. All patterns of positive staining were essentially the same as those previously described in human beings.

Conclusion

These 5 antibodies, with the antigen retrieval sequences, are helpful in identifying, distinguishing, and characterizing lymphocyte subsets in archival sections of normal and pathologic porcine lymphoid tissues including lymphomas. In addition, our findings indicated that antigen retrieval by microwave heating of the sections can appreciably expand the range of antibodies useful in paraffin immunohistochemistry, including those that otherwise work only on sections from frozen tissues or from an antigen-derived species. (Am J Vet Res 1996;57:853–859)
Allergenic cross-reactivities in flea-reactive canine serum samples
Authors: Pucheu-Haston Cherie M. , Grier Thomas J. , Esch Robert E. , Bevier Diane E.
Published: January 2024

Objectives

To determine whether flea extract could be determined (via ELISA) to share allergenic epitopes with other insects, and to determine whether sera with different reactivities to insect extracts have different cross-reactivity patterns.

Sample Population

69 canine serum samples that were selected from samples submitted for routine ELISA allergy testing and had previously been found to have high reactivities to flea.

Procedure

Each serum sample was assessed by means of a direct ELISA for IgE binding to 11 common insects. Samples that were reactive primarily to flea extract alone were designated pool 1, samples that were reactive to small numbers of insects were designated pool 2, and samples that were reactive to all or almost all insects were designated pool 3. Samples that did not have any apparent patterns of cross-reactivity were not included in the rest of the study. Inhibition ELISA techniques were used with the 3 serum pools to determine whether multiple insect extracts inhibited reactivity on flea-coated ELISA plates. Those extracts were used to coat ELISA plates, and reciprocal inhibition studies were then performed.

Results

Black fly, black ant, and cockroach extracts were capable of > 50% inhibition of flea solid-phase IgE binding with all 3 serum pools. In the ELISA inhibition studies, flea extract was able to inhibit IgE binding to each extract with all pools, confirming reciprocal inhibition.

Conclusions

Inhibition of IgE binding to solid-phase flea antigen by black ant, black fly, and cockroach extracts suggested sharing of allergenic epitopes among these species. Reciprocal inhibition studies further confirmed these findings. These results indicated in vitro cross-reactivity between flea, black ant, black fly, and cockroach extracts. These results need to be further investigated in vivo.

Clinical Relevance

It is possible that dogs may become sensitized to fleas via exposure to other insects, and that flea allergenic dogs may have signs of pruritus, in the absence of fleas, if exposed to cross-reactive insects. (Am J Vet Res 1996;57:1000–1005)
Silicate and metal dust in lungs of Greyhounds
Authors: Schoning Polly , Abraham Jerrold L. , Burnett Bryan R.
Published: January 2024

Objectives

To determine whether the amount of opaque dust in lung specimens increases with age, the composition of that dust, and whether the composition is similar for Greyhounds and pet dogs.

Design

Quantification of lung particulate dust burdens.

Animals

192 Greyhounds and 5 pet dogs.

Procedure

Lung specimens of 192 Greyhounds and 5 pet dogs were examined for dust accumulation, using light microscopy. Lung specimens from 20 Greyhounds and the 5 pet dogs were analyzed, using scanning electron microscopy and energy dispersive x-ray spectroscopy to quantify the lung burden of inorganic particles.

Results

Lung dust burden increased linearly with age. Pulmonary dust was composed of aluminum silicates, silica, and 18 metals. Silicate and metal particulate burdens were higher in Greyhounds than in the pet dogs. The 3 most common metals were iron, titanium, and chromium.

Conclusions and Clinical Relevance

Amount of opaque dust increased with age. The higher pulmonary dust burden in Greyhounds than in pet dogs suggests that environmental exposure is important. (Am J Vet Res 1996;57:1006–1009)
Immunohistochemical analysis of an equine model of synovitis-induced arthritis
Authors: Todhunter P. G. , Kincaid S. A. , Todhunter R. J. , Kammermann J. R. , Johnstone B. , Baird A. N. , Hanson R. R. , Wright J. M. , Lin H.-C. , Purohit R. C.
Published: January 2024

Objectives

To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone.

Animals

2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2).

Procedure

Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 μg/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articular cartilage sections and attached synovial membrane from the third carpal bones were examined by immunohistochemical distribution of interleukin 1β, tumor necrosis factor (TNF)-α, TNF receptors (P55, P75) and 3-B-3(–) epitopes, and by localization of proteoglycans (metachromatic staining). Proteoglycan extracts were assessed by metachromatic staining or western blotting and immunohistochemical staining, using anti-3-B-3 antibodies.

Results

Enhanced immunoreactivity for the cytokines and receptors was found in inflamed synovial membrane and noncalcified cartilage (group 2a more than 2b). Metachromasia of the noncalcified cartilage was greater in group-1 than in group-2a and group-2b specimens. In group 2a, chondrocyte hypertrophy and enhanced immunoreactivity for 3-B-3(–) epitope in areas of increased cytokine immunoreactivity suggested possible phenotypic change of the chondrocytes in response to synovitis. Immunohistochemical analysis by western blotting of proteoglycan extracts indicated strong 3-B-3(–) epitope immunolocalization in group-2a, weaker staining in group-2b, and barely detectable stain in group-1 specimens, which correlated with in situ immunolocalization.

Conclusions

Intra-articular administration of LPS may be used to induce a synovial environment conducive to increased immunoreactivity of interleukin 1β, TNF-α, and its receptors in equine synovial membrane and articular cartilage. These cytokines may be involved in the early phenotypic change of chondrocytes that is believed to occur in osteoarthritis and is characterized in this study by enhanced 3-B-3(–) epitope immunoreactivity and chondrocyte hypertrophy. (Am J Vet Res 1996;57:1080–1093)
Kinematic gait analysis in dogs with hip dysplasia
Authors: Bennett Russell L. , DeCamp Charles E. , Flo Gretchen L. , Hauptman Joe G. , Stajich Michael
Published: January 2024

Objective

To define alterations of movement in dogs with hip dysplasia by use of noninvasive, 3-dimensional, computer-assisted kinematic gait analysis.

Design

Kinematic and force plate data were collected at the trot from clinically normal dogs and from dogs with hip dysplasia.

Animals

12 large adult dogs of various breeds with clinical and radiographic evidence of hip dysplasia, and 12 clinically normal adult large dogs of various breeds with body weight similar to that of the dogs with hip dysplasia.

Procedure

Dynamic flexion and extension angles and angular velocities were calculated for the coxofemoral, femorotibial, and tarsal joints. Temporal and distance variables were also computed. Essential Fourier coefficients were determined and used to reconstruct mean dynamic flexion and extension curves for all joints, and to compare differences in movement between dogs with hip dysplasia and clinically normal dogs.

Results

Dogs with hip dysplasia had subtle characteristic changes in dynamic flexion and extension angles and angular velocities of each joint, verified by significant differences in essential Fourier coefficients between the 2 study groups. Stride length was increased and peak vertical force was decreased in dogs with hip dysplasia. Subject velocity, maximal foot velocity, stance duration, stride frequency, and impulse area did not differ between the 2 groups.

Conclusions

Kinematic gait analysis indicated that hip dysplasia is associated with alterations in movement of the coxofemoral, femorotibial, and tarsal joints. Computer-assisted kinematic gait analysis provided a noninvasive, objective tool with which to evaluate these complex motion alterations.

Clinical Relevance

The information obtained may be useful in future evaluations of various modes of treatment for hip dysplasia. (Am J Vet Res 1996;57:966–971)
Sequence analysis and polymerase chain reaction amplification of small subunit ribosomal DNA from Sarcocystis neurona
Authors: Marsh Antoinette E. , Barr Bradd C. , Madigan John , Lakritz Jeffrey , Conrad Patricia A.
Published: January 2024

Objectives

To identify Sarcocystis neurona-specific DNA sequences in the nuclear small subunit ribosomal RNA (nss-rRNA) gene that could be used to distinguish S neurona from other closely related protozoal parasites, and to evaluate a polymerase chain reaction (PCR) test, using broad based primers and a unique species-specific probe on CSF for detection of S neurona in equids.

Procedures

Sequencing of the nuclear small subunit ribosomal RNA gene from a new S neurona isolate (UCD 1) was performed. The sequence was compared with that of other closely related Sarcocystidae parasites. From this sequence, conserved DNA sequence primers were selected and an oligonucleotide probe was designed to hybridize with a unique region of the S neurona gene. For clinical evaluation, horses were considered test positive for S neurona infection on the basis of immunohistochemical detection of the parasites in the CNS.

Results

Sensitivity of this PCR and probe-based detection system was approximately 1 to 5 merozoites. Cerebrospinal fluid from 2 of 5 horses with histologic lesions consistent with equine protozoal myeloencephalitis were PCR- and probe-positive in a blind test of this procedure, and all uninfected horses were test negative.

Conclusion and Clinical Relevance

This PCR- based system is a useful method of confirming S neurona in CSF and has the advantage of facilitating detection of other apicomplexan protozoans that may be infective for horses. The usefulness of this test is limited by the presence of parasites free in the CSF of clinically affected horses. (Am J Vet Res 1996;57:975–981)
Evaluation of chromogenic substrate assays for fibrinolytic analytes in dogs
Authors: Lanevschi Anne , Kramer John W. , Greene Stephen A. , Meyers Kenneth M.
Published: January 2024

Objective

To evaluate the ability of commercial, chromogenic kits designed to measure human fibrinolytic pathway components to measure the canine plasma fibrinolytic pathway enzymes, tissue plasminogen activator (tPA) and plasminogen (PLG), and their respective inhibitors, plasminogen activator inhibitor 1 (PAI) and α2-anti-plasmim (AP).

Animals

20 healthy dogs of various ages and breeds.

Procedure

The commercial procedure was adapted to a microtitration plate. Standard curves were generated by use of a canine plasma pool.

Results

Modifications of the commercial kit consisted of change in incubation periods and the substitution of urokinase for the streptokinase. Plasminogen and AP procedures yielded intra- and interassay coefficients of variation (CV) ranging from 2 to 6.4%. The tPA activity gave an acceptable intra-assay CV of 4.2%, but an equivocal interassay CV of 18%. The PAI assay gave unacceptable intra-assay and interassay CV of 59 and 66%, respectively.

Conclusions

Modifications of the commercial PLG and AP procedures were appropriate for use with fresh and frozen canine plasma. However, equivocal results were obtained for canine plasma tPA. Although the PAI assay was able to detect the inhibitor, it gave unacceptable quantifiable results. Human and canine plasma contained similar amounts of PLG and AP, but 25% more tPA was found in canine plasma than human plasma.

Clinical Relevance

With modifications, the commercial human PLG and AP chromogenic kits may serve to elucidate such canine fibrinolytic disorders as disseminated coagulopathy. The high cost of the chromogenic substrate limits its application. (Am J Vet Res 1996;57: 1124-1130)
Effects of age at vaccination on efficacy of Brucella abortus strain RB51 to protect cattle against brucellosis
Authors: Cheville Norman F. , Olsen Steven C. , Jensen Allen E. , Stevens Mark G. , Palmer Mitchell V. , Florance Allison M.
Published: January 2024

Objective

To establish that female calves vaccinated with Brucella abortus strain RB51 at 3, 5, and 7 months of age are protected against infection and abortion when challenged exposed during their first pregnancy.

Animals

Polled Hereford heifer calves obtained from a brucellosis-free herd.

Procedure

Calves were inoculated SC at 3, 5, or 7 months of age with strain RB51 (n = 26), strain 19 (n = 16), or sterile saline solution (n = 15). Calves were bred at 16 to 17 months of age and challenged exposed during the first pregnancy with virulent B abortus strain 2308.

Results

After vaccination, none of the heifers given strain RB51 developed serum antibodies that reacted in the standard tube agglutination test, but reacted in a dotblot assay, using RB51 antigen. B abortus was cultured from biopsy specimens of superficial cervical lymph nodes in the RB51 and S19 vaccinates at 10 weeks, but not at 12 weeks after vaccination. All 4 heifers that had been vaccinated with RB51 at 3 months of age were protected against infection and abortion when challenged exposed. Vaccination at 5 and 7 months of age gave equivalent protection. Heifers given strain 19 were 95% protected and controls (given saline solution) had a high incidence of infection and abortion.

Conclusions

Strain RB51 is protective at doses comparable to those of strain 19 in calves 3 to 10 months of age.

Clinical Relevance

Immunogenicity and failure to induce antibodies that interfere with the serologic diagnosis of field infections of B abortus make strain RB51 an effective vaccine. (Am J Vet Res 1996;57:1153—1156)
Development of monoclonal antibodies for measurement of immunoglobulin G antibody responses in Blue and Gold Macaws (Ara ararauna)
Authors: Lung Nancy P. , Thompson James P. , Kollias George V. , Klein Paul A.
Published: January 2024

Objectives

To produce monoclonal antibodies (MAB) with specificity for the heavy chain of macaw IgG; to incorporate these MAB into an ELISA to measure IgG responses of macaws inoculated with bovine serum albumin (BSA); and to evaluate the antigenicity of BSA in Blue and Gold Macaws.

Animals

Four 1-year-old Blue and Gold Macaws, 2 males and 2 females.

Procedure

1 male and 1 female 1 were randomly assigned to each of 2 study groups. Group-1 birds were inoculated with 200 μg of BSA on days 0, 14, 28, and 42. Group-2 birds were inoculated with 200 μg of BSA on days 0 and 28. Blood was collected weekly for measurement of anti-BSA titer. Hybridomas were prepared from mice immunized with Scarlet Macaw (Ara macao) IgG purified by salt precipitation and gel chromatography. Specificity for IgG of Scarlet Macaw and other macaw species was confirmed by ELISA and western blot analysis. Hybridoma HL544 was cloned and the antibody purified. Following biotinylation, MAB HL544 was incorporated into an ELISA that measured IgG responses of macaws inoculated with BSA.

Results

Adult Blue and Gold Macaws developed strong primary and secondary anti-BSA antibody titers 14 days after inoculation with 200 μg of BSA. An inoculation interval of 28 days resulted in stronger secondary responses than an interval of only 14 days.

Conclusions

MAB specific for macaw immunoglobulins can be used in ELISA to evaluate the humoral immune responses of macaws. 1-year-old Blue and Gold Macaws developed strong anti-BSA titer when inoculated with 200 μg of BSA. An inoculation interval of 28 days resulted in stronger secondary responses than did an interval of only 14 days.

Clinical Relevance

These MAB, the first reported to have specificity for a psittacine antibody class, will be useful in the evaluation of psittacine antibody responses and in the development of psittacine diagnostic assays. (Am J Vet Res 1996; 57:1157-1161)
Noninvasive ultrasonic subtotal ablation of the prostate in dogs
Authors: Kincaide L. F. , Sanghvi N. T. , Cummings O. , Bihrle R. , Foster R. S. , Zaitsev A. , Phillips M. , Syrus J. , Hennige C.
Published: January 2024

Objective

To determine whether high-intensity focused ultrasound (HIFU) can be used for subtotal ablation of the prostate gland in dogs without causing damage to surrounding tissues.

Design

Experimental trial.

Animals

Adult hounds ≥5 years old and weighing between 20 and 30 kg.

Procedure

Prostatic ablation was performed in all dogs, using a transrectal HIFU probe. Acute effects of HIFU treatment were evaluated in 4 dogs. These dogs were euthanatized and necropsied 4 hours after the procedure. Chronic effects were evaluated in the other 3 dogs. Serial CBC, serum biochemical analyses, urinalyses, and transrectal ultrasonography were performed. Dogs were euthanatized and necropsied 3 months (1 dog) or 1 year (2 dogs) after HIFU treatment.

Results

Histologic examination of the prostate glands from the 4 dogs euthanatized 4 hours after treatment revealed that 80 to 90% of the gland had undergone hemorrhagic, liquefactive necrosis. Only slight discoloration of the prostatic capsule was detected, and there were not any gross or histologic lesions of the rectal mucosa or urinary bladder. All 3 dogs followed up after HIFU treatment developed cystic cavities within the prostate. Clinicopathologic testing did not indicate any long-term adverse effects.

Conclusions and Clinical Relevance

This method was effective in causing subtotal ablation of prostatic tissue in dogs. Further study of morbidity is required before the technique can be used clinically. (Am J Vet Res 1996; 57:1225-1227)
Sonographic observations of the gastroduodenal junction of dogs
Authors: Agut A. , Wood A. K. W. , Martin I. C. A.
Published: January 2024

Objectives

To determine the sonographic dimensions and an imaging protocol for the pyloric region of the normal canine stomach, and whether sonography could be used to demonstrate antroduodenal motility.

Animals

20 clinically normal, mixed-breed dogs (mean ± SD, 15.1 ± 7.1 kg).

Procedure

In vitro sonographic observations of the gastroduodenal junction were made after each had been mounted on an acrylic rack placed in an organ bath. The in vivo studies were performed in anesthetized dogs.

Results

The wall of the pyloric antrum was 4 to 5 mm thick, that of the pylorus was 3 to 4 mm thick, and that of the cranial part of the duodenum was 2 to 3 mm thick. The pyloric canal had a length of 16 to 17 mm and overall in vivo diameter of 22 mm, and the muscular part of its wall was 2 to 3 mm thick. In vivo sonographic images were best made with the dog in dorsal recumbency, tilted 30° to the right-the feasibility study clearly demonstrated the form and function of antroduodenal motility, including gastric and duodenal peristalsis, opening and closing of the pyloric canal, and duodenogastric reflux of fluid and gas.

Conclusions and Clinical Relevance

The use of a routine imaging protocol, accompanied by mensuration of the gastroduodenal junction, will permit an opinion to be given as to whether the structure and function of the region are normal or whether pathologic change exists. (Am J Vet Res 1996;57:1266-1273)
Temporal variation and factors affecting measurement of canine von Willebrand factor
Authors: Moser Janet , Meyers Kenneth M. , Meinkoth James H. , Brassard Jaqueline A.
Published: January 2024

Objective

To determine whether canine plasma von Willebrand factor (vWf) varies between and within individuals over time and with different blood sample collection and processing procedures.

Animals

26 adult dogs and 6 pups.

Procedure

Blood was obtained from the jugular or cephalic vein daily for 8 to 19 days and weekly for 9 to 23 weeks in adult dogs and periodically up to 180 days of age in pups. Temporal variation in vWf concentration and the effect of vascular occlusion, venipuncture site, lipemia, hemolysis, anticoagulant, storage time, freeze-thawing, and centrifugation speed on plasma vWf concentration, measured by ELISA, were determined.

Results

Plasma vWf concentration varied over time. In dogs with mean vWf concentration ≥ 79 U/dl, the largest intraindividual range in vWf spanned 64 U/dl with daily and 53 U/dl with weekly sample collection. In dogs with mean vWf concentration ≤ 24 U/dl, the largest individual variation was 12 U/dl with daily and weekly sample collection. In dogs with mean vWf concentration ≥ 53 and ≤ 74 U/dl, the largest intraindividual range spanned 35 U/dl. Mean vWf concentration of pups from 3 to 180 days of age did not change. Sample hemolysis decreased mean vWf by 37%. Mean vWf concentration was 9% higher in cephalic than jugular vein samples (P = 0.056). Other sample collection/preparation procedures did not affect vWf concentration.

Conclusion

There was substantial temporal variation in vWf concentration within individual dogs.

Clinical Relevance

Multiple tests may be necessary to obtain a reliable estimate of vWf concentration in dogs. (Am J Vet Res 1996;57:1288-1293)
Toxicity and kinetics of amitraz in dogs
Authors: Hugnet Christophe , Buronfosse Florence , Pineau Xavier , Cadore J. -L. , Lorgue Guy , Berny Philippe J.
Published: January 2024

Objective

To evaluate the toxic effects of amitraz in dogs and their reversal by various doses of atipamezole.

Animals

6 male 1-year-old Beagles.

Procedure

Dogs were given 100 mg of amitraz/kg of body weight, PO. Atipamezole was administered at 3 dose rates. Clinical examination and blood sample collection were performed regularly for 48 hours to examine biological parameters and determine the toxicokinetics of amitraz as well as the efficacy of the antidote. A specific high-performance thin layer chromatographic method was developed to determine plasma amitraz concentrations.

Results

Clinical signs of toxicosis included sedation, bradycardia, polyuria, hypothermia, and hyperglycemia, all of which could be related to the α2-agonist activity of amitraz, and were reversed by low doses of atipamezole (50 μg/kg, IM), a potent α2-antagonist, within 10 minutes after injection. Peak plasma concentrations were observed after 5 hours, and the elimination half-life was long (about 24 hours).

Conclusions

All clinical and biological effects observed during the course of amitraz poisoning could be attributed to the parent compound itself and were reversed by low doses of atipamezole. The half-life of amitraz was substantially longer than that in other studies because of the high dose administered.

Clinical Relevance

Atipamezole can be administered IM to dogs with severe amitraz poisoning to reverse all the effects observed. (Am J Vet Res 1996;57:1506-1510)
Polymerase chain reaction for the detection of Toxoplasma gondii in aqueous humor of cats
Authors: Lappin Michael R. , Burney Derek P. , Dow Steven W. , Potter Terry A.
Published: January 2024

Objectives

To develop Toxoplasma gondii B1 gene polymerase chain reaction (PCR) for use with aqueous humor of cats, and to report PCR and antibody detection results in naturally exposed cats with and without uveitis.

Sample Population

Serum and aqueous humor samples from client-owned, healthy cats (n = 23) and client-owned cats with uveitis (n = 43).

Procedure

T gondii-specific IgM and IgG were measured in serum and aqueous humor from all cats. The Goldman-Witmer coefficient for ocular antibody production was calculated for cats positive for T gondii-specific IgM or IgG in aqueous humor. Aqueous humor from all cats was assessed by the B1 gene PCR.

Results

T gondii was detected in aqueous humor by PCR from 2 of 23 (8.7%) healthy cats and 8 of 43 (18.6%) cats with uveitis. T gondii-specific IgM in either serum or aqueous humor was detected in 5 of 8 (62.5%) cats with uveitis and T gondii in aqueous humor. All cats with uveitis and T gondii in aqueous humor had anterior segment disease. In 5 of 8 (62.5%) cats with uveitis and T gondii in aqueous humor, ocular production of T gondii antibodies was not detected. T gondii was not detected in aqueous humor from 14 of 17 (82.4%) cats with ocular production of T gondii-specific antibody.

Conclusions

The presence of T gondii in aqueous humor may correlate to clinical disease in some, but not all, cats.

Clinical Relevance

T gondii-specific aqueous humor antibody tests and PCR should be used together to aid in the diagnosis of ocular toxoplasmosis in cats. (Am J Vet Res 1996;57:1589–1593)
Linkage of a microsatellite marker to the canine copper toxicosis locus in Bedlington Terriers
Authors: Yuzbasiyan-Gurkan Vilma , Blanton Susan Halloran , Cao Yueying , Ferguson Paul , Li Jianping , Venta Patrick J. , Brewer George J.
Published: January 2024

Objective

To identify a DNA marker for the copper toxicosis (CT) locus in Bedlington Terriers (BT).

Animals

77 BT, of which 25 were affected. Diagnosis of affected or unaffected with CT was made in all cases by quantitative copper determinations on liver biopsy samples by use of established criteria.

Procedure

BT pedigrees segregating for CT were identified. Linkage studies were carried out using polymorphic microsatellite markers developed for the canine genome in these pedigrees. DNA was isolated from blood samples of pedigree members. Polymerase chain reaction was used to amplify and type alleles at 213 microsatellite loci in each dog, and findings were subjected to linkage analysis.

Results

One microsatellite marker was identified to be closely linked to CT with logarithm of odds score of 5.96 at a recombination fraction of zero.

Conclusions

Using the linked marker, it has become possible to distinguish affected, homozygous normal, and carrier dogs in some BT pedigrees.

Clinical Relevance

In informative pedigrees where the marker is variable in the parents, it is possible to identify which dogs will require anticopper therapy and provide breeders with sound scientific advice about breeding strategies. (Am J Vet Res 1997;58:23–27)
Altered biological activity of equine chondrocytes cultured in a three-dimensional fibrin matrix and supplemented with transforming growth factor β-1
Authors: Fortier Lisa A. , Nixon Alan J. , Mohammed Hussni O. , Lust George
Published: January 2024

Objective

To determine the effects of transforming growth factor-β1 (TGF-β1) on the synthesis of DNA, collagen, and proteoglycans (PG) by equine chondrocytes.

Sample Population

Articular cartilage obtained from multiple joints of a 4-month-old foal.

Procedure

Chondrocytes were isolated by collagenase digestion, cultured in monolayer, trypsinized, and implanted at a cellular density of 10 × 106 chondrocytes/ml in a three-dimensional fibrin matrix. Chondrocytes in culture were supplemented with TGF-β1 at concentrations of 0, 1, 5, or 10 ng/ml in serum-free medium or medium containing fetal bovine serum (FBS). Total PG accumulation, [35S]-labeled PG synthesis, PG monomer hydrodynamic size, type II collagen production, total DNA content, and [3H]thymidine incorporation into DNA were determined at 7 and 14 days of culture.

Results

Chondrocytes maintained a rounded phenotype, dedifferentiating slightly to a more fibroblastic appearance only in medium containing FBS and 10 ng of TGF-β1/ml. Type II collagen immunoreaction on day 14 was decreased in the pericellular matrix in cultures containing FBS and 1, 5, and 10 ng of TGF-β1/ml, and in all serum-free culture conditions compared to FBS and 0 ng of TGF-β1/ml. Total proteoglycan accumulation and [35S]-labeled proteoglycan synthesis in cultures on days 7 and 14 were increased by the addition of exogenous TGF-β1 in serum-free conditions and decreased by TGF-β1 in FBS-supplemented conditions. Calculation of the partition coefficients for PG indicated that there was synthesis of low molecular weight PG in serum-free conditions and larger sized proteoglycans in FBS-supplemented conditions. Proteoglycan molecular size was unchanged by the addition of TGF-β1. Total DNA content of chondrocytes increased with the addition of TGF-β1 in FBS-supplemented conditions and decreased in serum-free conditions.

Conclusions

In a solid three-dimensional fibrin matrix, the effects of TGF-β1 on chondrocyte biological activity depend on the culture duration and on the presence of FBS in the medium. Stimulatory effects of TGF-β1 were most pronounced in serum-free culture conditions with high concentration of TGF-β1 (5 and 10 ng/ml) on day 7 and with low concentration of TGF-β1 (1 ng/ml) on day 14.

Clinical Relevance

TGF-β1 may not be a suitable growth factor for enhancement of equine articular grafting in sites exposed to serum. (Am J Vet Res 1997;58:66–70)
Effects of choreito and takushya consumption on in vitro and in vivo struvite solubility in cat urine
Authors: Buffington C. A. Tony , Blaisdell Jean L. , Komatsu Y. , Kawase K.
Published: January 2024

Objective

To determine the effects of the takushya portion of choreito, a traditional Chinese treatment for urolithiasis, on urine and struvite crystal variables in cats fed diets containing takushya.

Sample Population

6 male and 6 female adult cats, all considered to be clinically normal on the basis of physical examination findings, results of CBC, serum biochemical analyses, urinalyses, and urine cultures; and freedom from urolithiasis on the basis of urethrocystoscopic (females) or urethrocystographic (males) findings.

Procedure

Cats were fed a commercial canned diet supplemented with 0.1-mg of takushya/kg of body weight, or with 0.5 mg of choreito/kg. Diets were fed, using a Latin-square design, to 3 groups of 4 cats (2 male, 2 female) each for 2 weeks, followed by blood and 24-hour urine sample collections.

Results

Consumption of takushya, which comprises 20% by weight of choreito, was not associated with adverse effects in cats at the amounts provided during the period of study. Moreover, takushya was responsible for most of the effect of choreito consumption on reduction of urine pH, and approximately half its ability to reduce struvite crystal formation in cat urine.

Clinical Relevance

Alternative treatments for struvite urolithiasis in cats may be feasible. (Am J Vet Res 1997;58:150–152)
Effect of carprofen on sulfated glycosaminoglycan metabolism, protein synthesis, and prostaglandin release by cultured osteoarthritic canine chondrocytes
Authors: Benton Hilary P. , Vasseur Philip B. , Broderick-Villa Gregory A. , Koolpe Mitchell
Published: January 2024

Objective

To determine whether the nonsteroidal anti-inflammatory drug carprofen directly influences canine chondrocyte metabolism.

Animals

Cartilage from the femoral heads of 13 dogs undergoing total hip replacement.

Procedure

Rates of glycosaminoglycan (GAG) synthesis and degradation, protein synthesis, cell viability, and prostaglandin release were determined in canine expiant cartilage or monolayer canine chondrocyte cultures in the presence of 0 to 100 µg of carprofen/ml. Rate of GAG synthesis was assessed as incorporation of [35S]sulfate into cartilage matrix during a 3-hour pulse label. Degradation of cartilage GAG was assessed as rate of release of [35S]sulfate from prelabeled expiant cultures. Rates of total protein synthesis were assessed as incorporation of [35S]methionine into trichloracetic acid precipitable material during a 3-hour pulse label. Radiolabeled chondrocyte proteins were separated by Polyacrylamide gel electrophoresis and visualized by fluorography. Rates of prostaglandin E2 release were assessed by radioimmunoassay.

Results

Carprofen stimulated a significant increase in the rate of GAG synthesis at concentrations of 1 and 10 µg/ml, with no change in total protein synthesis, pattern of new protein synthesis, or cell viability. At concentration ≥ 20 µg/ml, inhibition of GAG synthesis and total protein synthesis was observed. There was no significant change in rate of release of GAG from cartilage explants, but potent inhibition of prostaglandin release was observed.

Conclusions

Carprofen has a direct influence on chondrocyte activity, resulting in changes in rate of production of cartilage matrix.

Clinical Relevance

In determining the optimal therapeutic dose of carprofen for arthritic conditions in dogs, it is important to consider potential influences on cartilage, as well as anti-inflammatory actions. (Am J Vet Res 1997;58:286–292)
Body weight, fluid, electrolyte, and hormonal changes in horses competing in 50- and 100-mile endurance rides
Authors: Schott H. C. , McGlade K. S. , Molander H. A. , Leroux A. J. , Hines M. T.
Published: January 2024

Objective

To investigate effects of prolonged exercise on fluid and electrolyte losses in horses competing in 50- and 100-mile endurance competitions, with emphasis on recovery.

Animals

Changes in body weight (BW); PCV; serum osmolality; plasma total protein, lactate, aldosterone, and serum electrolyte concentrations; and exchangeable cation content were measured in 12 and 7 horses before and after and before, during, and after successful completion of 50- and 100-mile endurance rides, respectively.

Procedure

BW was measured by use of a portable load bar scale, and blood samples were collected during the hour before ride start, at ride finish, and after approximately 2 and 18 hours (overnight) of recovery for horses competing in the 50-mile ride. For horses competing in the 100-mile ride, BW was measured and blood samples were collected at the start; after 50, 67, and 84 miles of the ride; at the finish; and after approximately 12 hours (overnight) of recovery.

Results

BW decreased by (mean ± SEM) 3.6 ± 0.6% and 4.9 ± 0.8% in horses that successfully completed rides of 50 and 100 miles, respectively. After the overnight recovery period, BW was 4.3 ± 0.5% and 3.9 ± 0.8% lower than preride values for horses performing the 50- and 100-mile rides, respectively. A decrease in plasma volume during the ride was reflected by an increase in plasma total protein concentration, but both measures returned to preride values after overnight recovery. Serum osmolality and serum electrolyte concentrations decreased and aldosterone concentration increased during prolonged exercise. Aldosterone concentration peaked after overnight recovery.

Conclusion

Despite apparent rapid return of plasma volume and ionic composition to near normal values, substantial depletion of body fluid and electrolyte stores persists after an overnight recovery period in horses that successfully complete 50 or 100 miles of endurance competition. (Am J Vet Res 1997;58:303–309)
Characterization of eosinophil progenitor cells in feline bone marrow
Authors: Young Karen M. , Monello Karen A. , Peickert Heide
Published: January 2024

Objective

To identify eosinophil progenitor cells in feline bone marrow, establishing an assay method to use in studies of eosinophilopoiesis and eosinophilopoietic factors in cats.

Animals

Healthy, laboratory animal source cats.

Procedure

Sources of colony-stimulating activity were prepared by conditioning media with bone marrow, spleen, and blood mononuclear cells from cats infected with Toxocara canis. Bone marrow cells were aspirated and cultured to develop the eosinophil progenitor cell assay and to test cells from 9 healthy cats in the assay.

Results

Optimal conditions for identifying colonyforming units-eosinophil and cluster-forming units-eosinophil were as follows. Bone marrow mononuclear cells (105) were plated in 1 ml of supplemented medium, fetal bovine serum, and agar. The source of eosinophil growth factor(s) was bone marrow-conditioned medium made in the presence of 2.5 µg of concanavalin A/ml; other conditioned media also supported eosinophil colony growth. Dishes were incubated for 7 days at 37 C and 7% CO2. The colonyforming units-eosinophil formed aggregates of > 50 Luxol fast blue-positive cells and had dispersed morphology; the cluster-forming units-eosinophil formed aggregates of < 50 cells.

Conclusion and Clinical Relevance

Similar to other species, cats have separate and distinct eosinophil progenitor cells. The eosinophil progenitor assay may be used to characterize altered kinetics of eosinophilopoiesis, to assess eosinophil growth factors, and to evaluate therapeutic regimens that might be useful in the management of excess eosinophil production. (Am J Vet Res 1997; 58:348-353)
Cerebrospinal fluid γ-aminobutyric acid and glutamate values in dogs with epilepsy
Authors: Podell Michael , Hadjiconstantinou Maria
Published: January 2024

Objectives

To investigate changes in CSF concentrations of inhibitory and excitatory neurotransmitters in dogs with confirmed idiopathic epilepsy, and to evaluate them with regard to the clinical characteristics of the sample population and of the seizures.

Animals

13 (8 male and 5 female) drug-naive dogs with an initial generalized seizure, 6 (4 male and 2 female) drug-naive dogs with an initial partial seizure, and 10 clinically normal (5 male and 5 female) control dogs.

Procedure

At least 24 hours after the last observed seizure, CSF was collected aseptically from the cisterna cerebellomedullaris, and a portion was immediately aliquoted into vials, placed on dry ice, then stored at −80 C. The CSF glutamate (GLU) and γ-aminobutyric acid (GABA) concentrations were estimated by use of HPLC with electrochemical detection. Cerebellomedullary cisternal CSF glutamate and GABA values were analyzed in dogs with noninduced idiopathic epilepsy. Comparisons were determined for differences attributable to weight, sex, age at first seizure, seizure type, and total time of past seizure history.

Results

Mean (range) age at onset of the first seizure was 3.33 (0.1 to 11), 3.5 (0.1 to 11 ), and 3.25 (0.5 to 9) years for all dogs, dogs with initial partial seizure, and dogs with initial generalized seizure, respectively. Low GABA and high GLU values were found in the CSF of epileptic dogs, and were independent of time elapsed between the first observed seizure and CSF sample collection. The GABA value was inversely related to body weight in epileptic dogs, independent of age. Changes in GABA and GLU concentrations were not related to seizure type.

Conclusions

Altered GABA and GLU values in CSF might be indicative of a state of chronic overexcitation in the brain of dogs with primary or idiopathic epilepsy.

Clinical Relevance

CSF GABA and GLU may serve as important markers in epileptic dogs for potential response to antiepileptic drugs. (Am J Vet Res 1997;58:451–456)
In vitro effects of an acidic by-product feed on bovine teeth
Authors: Rogers G. M. , Poore M. H. , Ferko B. L. , Kusy R. P. , Deaton T. G. , Bawden J. W.
Published: January 2024

Objective

To evaluate in vitro erosive effects of sweet potato cannery waste (SPCW) on bovine incisor enamel.

Sample Population

20 bovine mandibles.

Procedure

Mandibles were collected and incisors were classified into 3 categories: lacking observable wear advanced normal wear, or abnormal wear associated with feeding SPCW. Intact mandibles were radiographed. Contralateral normal teeth from the same jaw were used to compare Ca2+ loss (etching) with SPCW, lactic acid (pH 3.2), or SPCW neutralized with NaOH to pH 5.0 or 5.5. Scanning electron microscopy was performed to compare etched and unetched specimens. Two abnormally worn teeth were evaluated histologically. Knoop hardness testing was conducted on unexposed areas of surface enamel and enamel exposed to SPCW.

Results

Radiography revealed large periapical abscesses in the mandibles exposed to SPCW. Nearly identical amounts of Ca2+ were removed by SPCW and lactic acid solution at the same pH. Scanning electron microscopy did not indicate consistent differences between etch patterns resulting from exposure to SPCW or lactic acid. Mean rate of calcium removal was 56% higher in deciduous than permanent teeth. Knoop hardness data suggested that softening occurred in enamel exposed to SPCW. Neutralizing SPCW to pH 5.5 eliminated calcium removal. Histologic examination of sections indicated that SPCW degraded and removed some dentin matrix proteins.

Conclusions

Exposure to SPCW results in enamel erosion in vitro; low pH is the most likely cause of erosion. Neutralizing SPCW to pH 5.5 eliminated erosive effects.

Clinical Relevance

Confirmation of SPCW's erosive effects on enamel in vitro supported the field diagnosis. (Am J Vet Res 1997;58:498–503)
Resazurin reduction as a function of respiratory burst in bovine neutrophils
Authors: Fang Weihuan , Myllys Vesa , Sandholm Markus
Published: January 2024

Objective

To determine whether the respiratory burst of neutrophils from bovine blood and milk can be analyzed by use of a fluorometric resazurin reduction assay.

Sample Population

Neutrophils were obtained from EDTA-anticoagulated blood of 7 dairy cows. Neutrophils also were isolated from milk samples of a cow intramammarily challenge exposed with Escherichia coli lipopolysaccharide.

Procedure

The respiratory burst of neutrophils was analyzed in parallel, using the conventional luminol-enhanced luminometric procedure and a novel fluorometric procedure with resazurin as the fluorogenic substrate. Opsonized zymosan and phorbol myristate acetate were used as stimulants. The mechanism of the fluorescent response was analyzed, using metabolic inhibitors to various cell functions. Luminometry and fluorometry were carried out in parallel, using microtitration tray-reading instruments.

Results

Stimulation of neutrophils induced resazurin reduction to resorufin and a fluorescent response. The luminescent response was transient, but the fluorescent response (build-up of fluorescent resorufin) was cumulative. Therefore, a single end-point measurement can be used for the fluorometric assay.

Conclusions

The proposed fluorometric microtitration tray technology is simple and has a high throughput capacity. The fluorometric and luminometric assays seem to have similar potential in the analysis of phagocyte functions. (Am J Vet Res 1997;58:601–607)
Effect of furosemide and subsequent intravenous fluid administration on right atrial pressure of splenectomized horses
Authors: Rivas L. J. , Hinchcliff K. W.
Published: January 2024

Objective

To investigate the effect of IV administration of fluids on the furosemide-induced reduction in right atrial pressure (RAP) and relative change in blood volume (BV) of splenectomized mares.

Animals

5 splenectomized mares.

Procedure

RAP was measured by use of a micromanometer placed in the right atrium. Jugular venous blood was collected for measurement of hematocrit, plasma total protein concentration, and hemoglobin concentration. Right atrial pressure was recorded and blood samples were collected immediately before furosemide (1 mg/kg of body weight, IV) administration, then every 15 minutes for 240 minutes. Beginning 120 minutes after furosemide administration, polyionic fluids (lactated Ringer's solution) were administered (2 L q 15 min) for 120 minutes.

Results

Furosemide induced a significant (P < 0.05) decrease in mean RAP (7.6 ± 1.5 and 3.2 ± 1.2 mm of Hg before and 15 minutes after furosemide administration, respectively), and BV (8.4 ± 1.1 % by 15 minutes). Polyionic fluid administration restored RAP and BV The volume of polyionic fluids administered (32 ± 2 ml/kg) was not significantly different from the volume of urine produced (38 ± 7.8 ml/kg). Difference was not apparent in the relation between change in BV and RAP before or after fluid administration.

Conclusions

The effect of furosemide on RAP of horses is mediated in large part by furosemide-induced reduction in BV. However, an effect of furosemide on venous compliance cannot be excluded as contributing to the reduction in RAP. (Am J Vet Res 1997;58:632–635)
Effects of training on the development of exercise-induced arterial hypoxemia in horses
Authors: Christley Robert M. , Hodgson David R. , Evans David L. , Rose Reuben J.
Published: January 2024

Objectives

To compare the development of exercise-induced arterial hypoxemia in horses before and after training, and to determine whether increases in maximum oxygen uptake (V˙O2max) following training results in a greater degree of exercise-induced arterial hypoxemia.

Animals

13 three- to five-year-old Standardbred geldings without clinical signs of respiratory or cardiovascular disorders.

Procedure

Horses were rested for 4 months prior to commencing a 16-week training program. Arterial blood was collected from the transverse facial artery during standardized exercise tests performed before and after 8 and 16 weeks of training. Variables measured during exercise tests included arterial blood gas tensions and V˙O2max. Training and testing was performed on a treadmill set at a 10% slope.

Results

Minimum arterial partial pressures of oxygen (PaO2) during exercise decreased from 83.3 ± 1.5 mm of Hg before training to 77.8 ± 1.0 mm of Hg after 16 weeks of training. Maximum arterial oxygen content increased from 239.2 ± 3.1 to 257.9 ± 3.8 ml/L, which resulted from an increase in hemoglobin concentration. The V˙O2max increased by 19% following training. Minimum values of PaO2 and arterial oxygen saturation were significantly correlated with V˙O2max when data from each stage of training were pooled. Calculated values for minimum alveolar oxygen tension decreased after 8 weeks of training, and alveolar ventilation increased at the end of training. Alveolar-arterial oxygen tension difference increased by 4 mm of Hg following training.

Conclusions

In trained horses, an increase V˙O2max is associated with a decrease in minimum PaO2 during intense exercise and is mostly attributable to an increase in the alveolar-arterial oxygen tension difference. Therefore, interpretation of blood gas data from exercising horses should take into consideration V˙O2max. (Am J Vet Res 1997;58:653–657)