A BIBLIOGRAPHY OF VETERINARY RADIOTHERAPY

A review of the veterinary radiotherapy literature from 1939 through June of 1989 is presented. Subject material includes: radiation effects/injury, radiation oncology, hyperthermia, phototherapy, and various combination treatment protocols. Species covered are divided into small animal (canine, feline), large animal (equine, bovine, caprine, ovine) and miscellaneous species.     

Synthesis of heat stress proteins in lymphocytes from livestock

Cultured bovine, equine, ovine and chicken lymphocytes responded to heat stress by the increased synthesis of a specific set of proteins known as heat stress proteins (HSP). Proteins with molecular weights of 70 and 90 kDa were synthesized in all species. Additional proteins were found in bovine, ovine and chicken lymphocytes. A time course of induction showed an increased synthesis of some of these proteins with only 30 min of heat stress and of several proteins with 60 min of heat stress. A specific monoclonal antibody was used to identify HSP70 as one of the stress proteins in bovine lymphocytes. Cells from these animals are capable of responding to heat stress by the production of heat stress proteins. These may be of physiological importance.     

Further biological, serological and biochemical characterization of North American, European and Southeast Asian strains of bovine herpesvirus 1 compared with other alphaherpesvirinae members

Hemagglutination activity, structural protein profiles and neutralization assays were used in a comparative study of bovine herpesvirus 1 strains from the U.S.A., Canada, Great Britain, Denmark and Malaysia with equine, feline and human herpesviruses in order to further characterize the bovine herpesvirus 1 hemagglutinin. Bovine herpesvirus 1 strains of different geographical origins all showed hemagglutinating activity for mouse erythrocytes; furthermore, feline herpesvirus 1 was also shown to hemagglutinate mouse erythrocytes. Analyses of partly purified viruses showed that a distinctive and specific polypeptides profile is associated with each species of herpesviruses used in our study; strains of bovine herpesvirus 1 from North America, Europe and Southeast Asia however, presented a remarkable similarity as to their electrophoretic protein patterns. A protein similar to the 97-kDa bovine viral hemagglutinin was not identified with the hemagglutinating feline herpesvirus. An important neutralization epitope on the bovine viral hemagglutinin was also not found on feline, equine and human herpesviruses but was identified on all bovine strains tested from North America, Europe and Southeast Asia stressing the importance of the bovine hemagglutinin for eventual prophylactic purposes.     

Effect of fetal bovine serum and heat-inactivated fetal bovine serum on microbial cell wall-induced expression of procoagulant activity by equine and canine mononuclear cells in vitro

OBJECTIVE: To determine the effect of fetal bovine serum (FBS) and heat-inactivated FBS (HI-FBS) on lipopolysaccharide (LPS)- and zymosan-induced procoagulant activity of equine and canine mononuclear cells. SAMPLE POPULATION: Mononuclear cells from 18 horses and 3 dogs. PROCEDURES: Cells were incubated with various concentrations of FBS, HI-FBS, LPS, zymosan, polymyxin B, and anti-LPS-binding protein monoclonal antibody or combinations of these constituents. A 1 stage recalcification assay was used to determine procoagulant activity. RESULTS: Addition of FBS to media significantly increased procoagulant activity; equine and canine cells were stimulated by 1% and 10% FBS, respectively. Coincubation of cells with FBS and polymyxin B did not reduce this effect, suggesting that the response was not attributable to LPS contamination. Addition of HI-FBS to media did not stimulate procoagulant activity of equine or canine cells, and the sensitivity of the equine cells to LPS was significantly increased by HI-FBS. This increased LPS sensitivity was reduced 40% with monoclonal antibody directed against human recombinant LPS-binding protein. Increasing concentrations of HIFBS significantly increased LPS- and zymosan-induced procoagulant activity of canine cells. CONCLUSION AND CLINICAL RELEVANCE: Procoagulant activity production in equine and canine mononuclear cells was significantly increased by addition of FBS, whereas heat inactivation of FBS eliminated this effect. Heat inactivation did not eliminate the function of serum proteins involved in enhancement of LPS and zymosan-induced procoagulant activity. Results suggest that HI-FBS can be used as a source of serum proteins that increase the sensitivity of mononuclear cells to bacterial and yeast cell wall components.     

Tritrichomonas foetus: a scanning electron microscopy study of erythrocyte adhesion associated with hemolytic activity

The in vitro hemolytic activity of Tritrichomonas foetus was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of nine adult animals of different species (the rabbit, rat, chicken, cat, dog, swine, horse, bovine, and sheep). The results showed that T. foetus strains (ATCC KV1, K, PAL, 5022, RJ, 90) did not present any hemolytic activity against any human erythrocyte group nor against rabbit, rat, chicken, cat, dog and swine erythrocytes. T. foetus strains, however, lysed horse, bovine, and sheep erythrocytes. No hemolysin released by the parasites could be identified. Hemolysis did not occur with trichomonad culture supernatants, with sonicated extracts of T. foetus, nor with killed organisms. Scanning electron microscopy (SEM) showed that human erythrocytes did not adhere to the trophozoites, in contrast horse erythrocytes adhered to the surface of the parasites and were phagocytosed for up to 90 min. The parasites are able to exert their cytopathic effects through: (a) physical contact established between the two cell surfaces, (b) toxins released from parasites into the interaction media, or (c) the association of both mechanisms. Further studies are necessary to clarify the importance of the hemolytic activity in the biology of T. foetus.     

Propagation and quantitation of animal herpesviruses in eight cell culture systems

A comparative study was carried out to determine the relative sensitivities of eight different cell culture systems to six different herpesviruses of animals. The cells used were: OFL (ovine fetal lung), ML (mink lung), FK (ferret kidney), PTK-2 (potoroo kidney), TEK (turkey embryo kidney), ED (equine dermal), BT (bovine turbinate), and PK15 (porcine kidney). The viruses tested were: PRV (pseudorabies) of swine, CPHV (caprine herpesvirus), IBRV (infectious bovine rhinotracheitis virus), DN-599 strain of bovine herpesvirus type 4, EHV-1 (equine herpesvirus), and CHV (canine herpesvirus). On the basis of virus titers obtained and the time of appearance of CPE (cytopathic effects), ML cells were found to be the most useful because of their sensitivity to all six viruses tested. BT and OFL cells were also found to be highly sensitive to all viruses with the exception of CHV.     

High-performance liquid chromatography determination of erythrocyte membrane phospholipid composition in several animal species

High-performance liquid chromatography (HPLC) was used to determine the phospholipid (PL) composition of ovine, equine, bovine, porcine, and canine RBC membranes. Procedural modifications of established techniques provided for separation of 7 PL within a 15- to 20-minute sample run. Significant (P less than 0.05) differences were detected in RBC membrane PL composition among the various species. The concern for physiologic properties associated with hemolysis and/or sedimentation rate must include evaluation of differences in the PL bilayer structure.     

Cross-reactivity between a monoclonal antibody that recognizes a tumor-associated antigen on bovine lymphosarcoma cells and blood lymphocytes from various mammalian species.

Tumor-associated antigens that are expressed in lymphosarcoma B cells of cattle with enzootic bovine leukosis had been analyzed in terms of their reactivity with 13 monoclonal antibodies (MAB). By use of flow cytometry and radioimmunoprecipitation, 1 of the MAB (c143) that recognized a tumor-associated antigen cross-reacted with blood lymphocytes (BL) from various mammalian species. By use of flow cytometry, the c143 MAB reacted with 10 to 49% of BL derived from human beings, mice, dogs, horses, pigs, llamas, sheep, goats, and cattle. Titer of the c143 MAB with BL from horses, pigs, human beings, and llamas ranged between 1:6.0 x 10(4) and 1:5.3 x 10(5); titer associated with BL of goats and sheep was 1:1.6 x 10(6); and that associated with BL of cattle was 1:4.3 x 10(7). The c143 MAB specifically immunoprecipitated 3 homologous proteins from cell extracts of caprine, ovine, and bovine BL (32-, 34-, and 36- to 37-kDa bovine proteins; 31-, 32-, and 36- to 37-kDa caprine proteins; and 31.5-, 33-, and 36- to 37-kDa ovine proteins), but none was immunoprecipitated from human, murine, canine, porcine, and llama BL. These results indicate that the avidity of the c143 MAB in binding to BL from ruminants (eg, goats, sheep, and cattle) is higher than that to BL from human beings, mice, dogs, horses, pigs, and llamas. In sheep, the c143 MAB could immunoprecipitate the aforementioned proteins from BL of the Suffolk breed, but not BL from the Corriedale breed, whereas the c143 MAB immunoprecipitated apparently identical proteins from BL of 4 breeds of cattle.     

Development of a polymerase chain reaction-based method to identify species-specific components in dog food

OBJECTIVES: To determine whether there is a relationship between species-specific mitochondrial DNA (mtDNA), especially canine and feline mtDNA, and detectable amounts of pentobarbital in previously analyzed dog food samples. SAMPLE POPULATION: 31 dog food samples previously analyzed for pentobarbital (limit of detection, 1 microg/kg). PROCEDURE: Polymerase chain reaction (PCR) analysis was performed on dog food samples by use of PCR primers specific for either canine, feline, equine, bovine, porcine, ovine, or poultry mtDNA. RESULTS: PCR amplicons specific for feline or canine mtDNA at a 0.007% (70 microg/g [wt/wt basis]) or 0.0007% (7 microg/g) level, respectively, were not found in the 31 dog food samples. Most of the 31 dog food samples had a PCR amplicon on PCR analysis when a PCR primer set capable of simultaneously detecting mtDNA of cows, pigs, sheep, goats, deer, elk, and horses was used. Results of PCR analysis by use of primers specific for bovine, swine, sheep and goat, or horse mtDNA revealed amplicons specific for bovine or swine mtDNA only in 27 of the 31 samples. Analysis of the remaining 4 samples failed to yield amplicons for any mammalian mtDNA. Pentobarbital was detected in 2 of these 4 samples. Results of PCR analysis correlated with the stated ingredient list for most, but not all samples. CONCLUSIONS AND CLINICAL RELEVANCE: Because canine and feline mtDNA were not found in a set of retail dog food samples, these results indicate that the source of pentobarbital in dog food is something other than proteins from rendered pet remains.     

Effect of storage on measurement of ionized calcium and acid-base variables in equine, bovine, ovine, and canine venous blood.

The stability of blood ionized calcium (Ca2+) and acid-base variables in equine, bovine, ovine, and canine venous blood samples (n = 15, in each group) stored at 4 C for 3, 6, 9, 24, or 48 hours was studied. Variables included blood Ca2+ and standard ionized calcium (Ca2+ corrected to pH 7.4) concentrations, pH, blood carbon dioxide and oxygen tensions, base excess, bicarbonate concentration, and total carbon dioxide content. Results indicate that storage of blood samples at 4 C for up to 48 hours, despite appreciable acid-base changes, is associated with less than 1.5% change in equine, bovine, and ovine blood Ca2+ concentrations. Similar changes were observed in canine blood during the first 9 hours' storage. After 24 and 48 hours' storage, clinically relevant decrease (10.5 and 15.5%) in canine blood Ca2+ concentration was measured. Therefore, Ca2+ concentration in equine, bovine, and ovine venous blood samples stored up to 48 hours, and in canine blood samples stored up to 9 hours at 4 C is of diagnostic use.     

Preliminary Observations on the Isolation of Escherichia coli Bacteriophages from the Intestinal Contents of Chickens

Eschericia coli bacteriophages were isolated from the intestines of chickens. These phages had different lytic patterns, and were propagated in nutrient broth containing 0.4 gm calcium chloride/litre. The agar layer technique was used to determine the routine test dilution (RTD) and plaque morphology. The phages differed in their 1) morphology, 2) RTD values, and their ability to lyse E. coli strains from various animals. All phages isolated lysed human K12 E. coli strains, whereas only two phages lysed the chicken E. coli strain. Phages isolated lysed E. coli from chicken, bovine, ovine, equine, and human, but not from porcine, canine and other avian species.     

Rat, caprine, equine and bovine erythrocyte ghosts exposed to t-butyl hydroperoxide as a model to study lipid peroxidation using a chemiluminescence assay

The aim of the present study was to analyze the time-course of t-butyl hydroperoxide-induced changes in lipid peroxidation, fatty acid composition and chemiluminescence intensity in rat, caprine, equine and bovine erythrocyte ghosts. A relatively high content of arachidonic acid (C20:4 n6) and docosahexaenoic acid (C22:6 n3) was characteristic of the rat erythrocyte ghosts. The fatty acid composition of native erythrocyte ghosts obtained from caprine, equine and bovine was characterized by a high content of oleic acid (C18:1 n9) and a low content of the peroxidable polyunsaturated fatty acids (C20:4 n6 and C22:6 n3). The proportion of linoleic acid (C18:2 n6) was higher in equine and bovine compared to rat and caprine. Increase in lipid peroxidation in rat erythrocyte ghosts was maximal within 12 min of incubation, t-butyl hydroperoxide concentration dependent and was paralleled by a decrease in C18:2 n6, C20:4 n6 and C22:6 n3 and an increase in chemiluminescence formation. Polyunsaturated fatty acids (PUFAs) present in rat erythrocyte ghosts exhibit the highest sensitivity to oxidative damaged and their sensitivity increases as a power function of the number of double bonds per fatty acid molecule. Light emission in caprine, equine and bovine erythrocyte ghosts was very low, t-butyl hydroperoxide concentration-dependent but changes in fatty acid composition were not observed. The main conclusion of this work is that a low unsaturation degree of fatty acids in erythrocyte ghosts of caprine, equine and bovine prevent the lipid peroxidation on those membranes when they are incubated with t-butyl hydroperoxide.     

Granulocyte isolation from whole blood of goat, sheep, cattle, horse, dog, pig, and man

A simple two step procedure for the isolation of caprine, ovine, bovine, equine, canine, porcine and human peripheral blood granulocytes is described. After enrichment of granulocytes by centrifugation, contaminating erythrocytes are lysed hypotonically. Recovery, purity, and viability of the granulocyte suspensions are determined. FACScan analysis of the cell suspensions measuring cellular size by forward and sideward light scatter is compared with the corresponding analysis of whole blood leukocytes. Constituencies of the isolated cell suspensions and loss of granulocyte subpopulations through isolation procedure is discussed with regard to granulocyte function assays.     

Hemagglutinating factor in an extract of Sarcoptes scabiei var. suis (De Geer)

A naturally occurring hemagglutinating factor to tanned human O positive, ovine and porcine erythrocytes was found in extracts from Sarcoptes scabiei var. suis. This hemagglutinating factor did not react with bovine, equine or avian erythrocytes. This factor was demonstrated by microscopic examination of the tanned erythrocytes and by the passive hemagglutination assay.     

Cloning and sequence analysis of the complementary DNA for feline preproparathyroid hormone

OBJECTIVES: To clone and sequence the cDNA for feline preproparathyroid hormone (preproPTH) and to compare that sequence with other known parathyroid hormone (PTH) sequences. SAMPLE POPULATION: Parathyroid glands from 1 healthy cat. PROCEDURES: A cDNA library was constructed in lambda phage from feline parathyroid gland mRNA and screened with a radiolabeled canine PTH probe. Positive clones were sequenced, and nucleic acid and deduced amino acid sequences were analyzed and compared with known preproPTH and PTH sequences. RESULTS: Screening of approximately 2 X 10(5) recombinant plaques revealed 3 that hybridized with the canine PTH probe; 2 clones comprised the complete sequence for feline preproPTH. Feline preproPTH cDNA consisted of a 63-base pair (bp) 5'-untranslated region (UTR), a 348-bp coding region, and a 326-bp 3'-UTR. The coding region encoded a 115-amino acid peptide. Mature feline PTH consisted of 84 amino acids. Amino acid sequence analysis revealed that feline PTH was > 83% identical to canine, bovine, swine, equine, human, and macaque PTH and 69, 71, and 44% identical to mouse, rat, and chicken PTH, respectively. Within the region responsible for hormonal activity (amino acids 1 to 34), feline PTH was > 79% identical to other mammalian PTH sequences and 64% identical to the chicken sequence. CONCLUSIONS AND CLINICAL RELEVANCE: The amino acid sequence of PTH is conserved among mammalian species. Knowledge of the cDNA sequence for feline PTH may be useful to investigate disturbances of calcium metabolism and alterations in PTH expression in cats.     

Cloning and characterisation of the Pasteurella multocida ahpA gene responsible for a haemolytic phenotype in Escherichia coli

Haemolysins are membrane-damaging agents which have been described as bacterial virulence factors due to their ability to lyse erythrocytes and other host cells, and therefore inducing a greater inflammatory response (Elliott et al., 1998). Pasteurella multocida was found to be haemolytic under anaerobic conditions. In this study, we cloned and characterised a P. multocida gene, designated ahpA, which conferred a haemolytic phenotype on Escherichia coli when incubated under anaerobic conditions. A deletion was introduced into the ahpA open reading frame which abolished the haemolytic phenotype. The clone containing ahpA showed erythrocyte specificity, causing haemolysis of bovine and equine erythrocytes, and demonstrated weak haemolysis on ovine erythrocytes. Upon further investigation, AhpA was found to affect the expression of the E. coli K-12 latent haemolysin, SheA, under anaerobic conditions.     

Expressed gene sequence of the IFNγ-response chemokine CXCL9 of cattle, horses, and swine

This report describes the cloning and characterization of expressed gene sequences of bovine, equine, and swine CXCL9 from RNA obtained from peripheral blood mononuclear cells (PBMC) and other tissues. The bovine coding region was 378 nucleotides in length, while the equine and swine coding regions were 381 nucleotides. Mapping showed that all three sequences were coded for in four exons in the genome, as are the human and mouse genes. The bovine, equine, and swine coding regions shared 83%, 86%, and 84% homology with human CXCL9, respectively, and all three were 74% homologous with mouse CXCL9. Cladogram comparison of the nucleotide sequences of CXCL9 showed that the bovine, equine and swine sequences were more closely related to one another than to either the human or the mouse sequences. However, the human sequence was more closely related to them than it was to the mouse sequence. These relationships were preserved when the deduced amino acid sequences were evaluated and all sequences showed conservation of the characteristic four cysteines. This work sets the stage for further work with these molecules; an integral goal of the U.S. Veterinary Immune Reagent Network is to develop reagents for investigating diseases in livestock species, poultry, and fish.     

Comparison of two automated multi-channel blood cell counting systems for analysis of blood of common domestic animals

An automated, multi-channel blood cell counting system (S-Plus) was compared to a reference counting system using blood samples from 187 animals of four species. The standard red cell bath aperture current of 150 volts (V) was used during analysis of 75% of the samples. At this setting, all samples with a Mean Corpuscular Volume (MCV) greater than 50 fl had accurate erythrocyte counts. As the MCV decreased below 50 fl, the severity of false low erythrocyte counts and false high MCV values increased. The remaining 25% of samples were analyzed with the red cell bath aperture current increased to 200 V. At this setting, only 5% or less of erythrocytes from animals with normal MCV values(>36 fl)were below the erythrocyte threshold. The red cell distribution width values provided by the S-Plus indicated that equine and bovine erythrocytes have greater anisocytosis than canine and feline erythrocytes. Leukocyte counts were significantly lower on the S-Plus (p<0.01). Canine and equine samples most frequently had platelet size distribution within the S-Plus platelet counting threshold window. Electronic whole blood platelet counting appeared unsatisfactory in cats due to large platelet size and erythrocyte-platelet size overlap. Small platelet size in cattle indicated that further modifications of the red cell bath aperture current would be required to count and size platelets in this species. Following electronic modifications, this state-of-the-art system appears adaptable to hematologic profiling in most species.     

The inhibitory effects of colchicine, vinblastine and cytochalasin D on lymphocyte blastogenic responses

The inhibitory effects of colchicine, vinblastine and cytochalasin D on blastogenic responses of equine, bovine and canine peripheral blood lymphocytes (PBL) were investigated. These drugs inhibited blastogenic responses of each PBL. The concentrations for 50% recovery in PBL blastogenic response of colchicine and vinblastine were lower in equine and canine PBL than in bovine PBL. This suggested that microtubules may be more concerned in blastogenic response of equine and canine PBL than of bovine PBL. On the other hand, the concentration for 50% recovery in PBL blastogenic response of cytochalasin D was almost the same in the PBL of each animal. This meant that microfilaments made only a small contribution to the lymphocyte blastogenic response.     

Relation between reticulocyte count and characteristics of erythrocyte 5'-nucleotidase in dogs,cats, cattle and humans

To examine substrate specificity and susceptibility to lead, erythrocyte 5'-nucleotidase was measured in dogs, cats, cattle and humans, and its relationship to the reticulocyte count in these species was determined. The reticulocyte count in dogs was similar to that in humans, but the count in cats was higher than that in humans. Reticulocytes were not observed in cattle. The activities of canine erythrocyte 5'-nucleotidase measured using cytidine and uridine 5'-monophosphates, which are preferentially catalyzed by one of the human pyrimidine 5'-nucleotidase isozymes (P5N-I), were similar to those of the human enzyme. The canine enzyme preferentially catalyzed thymidine 3'-monophosphate, which is catalyzed only by human P5N-II, more strongly than the human enzyme. This suggests that canine erythrocytes have two isozymes similar to human P5N-I and P5N-II, and a higher P5N-II-like activity than human erythrocytes. Feline erythrocytes had the highest level of P5N-I-like activity among the species examined, and the bovine enzymic activities including those of P5N-I and II were the lowest among these species. According to these observations, the reticulocyte count was approximately proportional to the P5N-I-like activity in these species. Therefore, the P5N-I-like activity may be involved in the morphological maturation of mammalian erythrocytes. The canine and feline erythrocytes had markedly high activity and preferentially catalyzed purine 5'-monophosphate suggesting the presence of a purine-specific 5'-nucleotidase as in human erythrocytes. In addition, the canine and feline P5N-I-like activity showed less susceptibility to lead than the human P5N-I. This may be a reason why there are few case reports of lead-induced anemia in dogs and cats.