Immune function of bovine leukocytes after in vitro exposure to selected heavy metals

OBJECTIVE: To study effects of in vitro exposure of bovine leukocytes to mercury, cadmium, and lead on phagocytosis, natural killer cell activity, and lymphocyte proliferation. SAMPLE POPULATION: Leukocytes from 6 nonpregnant Holstein heifers. PROCEDURE: Leukocytes were exposed in vitro to the aforementioned metals, and leukocyte functions were assessed. RESULTS: Phagocytosis was suppressed by 10(-5) to 10(-7) M CdCl2 and by 10(-5) and 10(-6) M HgCl2, but not 10(-7) M HgCl2 nor 10(-4) to 10(-6) M PbCl2. Spontaneous and concanavalin A- or phytohemagglutinin-stimulated proliferation of metal-treated bovine blood mononuclear cells was not significantly different from that of nontreated control cells, except for enhanced spontaneous proliferation in response to 10(-5) M HgCl2. When proliferation was expressed as a stimulation index, a dose-dependent increase of spontaneous proliferation was observed in response to exposure to HgCl2 and PbCl2. Compared with response to 10(-6) or 10(-7) M CdCl2, reduction of mitogen-induced and spontaneous proliferation was observed on exposure to 10(-5) M CdCl2. Natural killer cell activity against YAC-1 target cells, evaluated by flow cytometry, was decreased only in cells exposed to 10 M HgCl2. CONCLUSION AND CLINICAL RELEVANCE: Bovine leukocytes are susceptible to the immunomodulatory effects of in vitro exposure to heavy metals at concentrations equal to or higher than those at which similar effects are seen for leukocytes from most other animal species for which data are available for comparison. Exception is phagocytosis, which is severely affected by low concentrations of CdCl2 and HgCl2 in cattle. Reduction of defense mechanisms on exposure to metals could lead to increased susceptibility to potential pathogens.     

Isolation and measurement of carbonic anhydrase isoenzymes in erythrocytes of dogs

OBJECTIVE: To purify canine carbonic anhydrase (CA) isoenzymes CA-I and CA-II and to determine concentrations of CA-I and CA-II in erythrocytes of Beagles and dogs native to Japan. SAMPLE POPULATION: Blood samples from 116 Beagles, including 24 pregnant Beagles, and blood samples from 29 dogs native to Japan. PROCEDURE: Canine CA-I and CA-II were purified by use of column chromatography. Concentrations of CA-I and CA-II in erythrocytes of dogs were determined, using an ELISA. RESULTS: Mean (+/- SD) concentrations of CA-I and CA-II in erythrocytes of Beagles were 3.21+/-0.86 and 1.63+/-0.39 mg/g of Hb, respectively. Mean concentration of CA-I was greater in male Beagles than female Beagles. In contrast, mean concentration of CA-II was greater in female Beagles than male Beagles. Furthermore, concentration of CA-II was greater in pregnant female Beagles than male or nonpregnant female Beagles. Mean concentrations of CA-I and CA-II in erythrocytes of dogs native to Japan were 11.03+/-4.39 and 3.29+/-0.91 mg/g of Hb, respectively. Mean concentration of CA-I was greater in male dogs from Japan than female dogs from Japan. CONCLUSIONS AND CLINICAL RELEVANCE: The ELISA used in this study proved to be precise and sensitive for determining CA-I and CA-II concentrations in dogs. The ELISA may enable study of changes in isoenzymes associated with hereditary or metabolic disorders of blood or other body fluids, using only a small sample. Measurement of the concentrations of CA isoenzymes in dogs may be of diagnostic value.     

Concentrations of trace minerals in the spinal cord of horses with equine motor neuron disease

OBJECTIVE: To compare concentrations of trace minerals in the spinal cord of horses with equine motor neuron disease (EMND) with those of horses without neurologic disease (control horses). ANIMALS: 24 horses with EMND and 22 control horses. PROCEDURE: Spinal cord trace mineral concentrations in horses with EMND and control horses were analyzed by use of inductively coupled plasma atomic emission spectroscopy (calcium, phosphorus, sodium, potassium, magnesium, copper, iron, manganese, nickel, zinc, aluminum, cobalt, and chromium), atomic absorption spectrophotometry (lead and cadmium), flameless atomic absorption (mercury), and fluorometry (selenium). RESULTS: Copper concentration was significantly higher in the spinal cord of horses with EMND, compared with control horses; spinal cord concentrations of all other trace minerals were similar between groups. CONCLUSION AND CLINICAL RELEVANCE: Among spinal cord trace minerals investigated in the study, only copper concentrations were significantly different between horses with EMND and horses without neurologic disease, which suggests that copper may be involved in the pathogenesis of EMND. An hypothesis of oxidative injury in this disease is supported by the finding of increased copper concentrations in the spinal cord and by low vitamin E concentrations reported by other researchers.     

Use of canine red blood cell with high concentrations of potassium, reduced glutathione, and free amino acid as host cells for in vitro cultivation of Babesia gibsoni

OBJECTIVE: To determine the usefulness of canine RBC with high concentrations of potassium, reduced glutathione (GSH), and amino acid(i.e., HK cells) for in vitro cultivation of Babesia gibsoni. ANIMALS: RBC were obtained from 3 dogs that had inherited HK cells and from 3 genetically unaffected dogs that, therefore, had RBC with lower potassium (LK) concentrations (i.e., LK cells). PROCEDURES: First, B. gibsoni were cultivated using HK or LK cells in alpha-modification of Eagle medium, consisting of Earle salts with glutamine and without ribosides, deoxyribosides, and sodium bicarbonate under a humidified atmosphere containing 5% CO2 at 37 C. Second, parasites were cultivated with LK- or HK-cell lysates. Finally, HK cells were separated into 3 fractions (bottom, middle, top layers) by density gradient centrifugation, and B. gibsoni were cultivated with each of the HK-cell fractions. In addition, the concentrations of free amino acids and reduced glutathione (GSH) in each HK-cell fraction were measured. RESULTS: B. gibsoni preferentially multiplied in HK-cell cultures rather than in LK-cell cultures. Furthermore, the addition of HK-cell lysate to the culture medium resulted in enhanced multiplication of the parasites. Higher multiplication of the parasites was observed in HK cells from the top layer, compared with HK cells from the middle and bottom layers. The HK cells from the top layer had higher concentrations of glutamate, aspartate, and GSH, compared with HK cells from the middle and bottom layer. CONCLUSIONS: Canine HK cells are useful host cells for in vitro cultivation of B. gibsoni, and the high concentrations of glutamate, aspartate, and GSH may result in enhancement of multiplication of the parasites in HK cells.     

Incidence of and breed-related risk factors for gastric dilatation-volvulus in dogs

OBJECTIVE: To compare incidence of and breed-related risk factors for gastric dilatation-volvulus (GDV) among 11 dog breeds (Akita, Bloodhound, Collie, Great Dane, Irish Setter, Irish Wolfhound, Newfoundland, Rottweiler, Saint Bernard, Standard Poodle, and Weimaraner). DESIGN: Prospective cohort study. ANIMALS: 1,914 dogs. PROCEDURE: Owners of dogs that did not have a history of GDV were recruited at dog shows, and the dog's length and height and depth and width of the thorax and abdomen were measured. Information concerning the dogs' medical history, genetic background, personality, and diet was obtained from owners, and owners were contacted by mail and telephone at approximately 1-year intervals to determine whether dogs had developed GDV or died. Incidence of GDV based on the number of dog-years at risk was calculated for each breed, and breed-related risk factors were identified. RESULTS AND CLINICAL RELEVANCE: Incidence of GDV for the 7 large (23 to 45 kg [50 to 99 lb]) and 4 giant (> 45 kg [> 99 lb]) breeds was 23 and 26 cases/1,000 dog-years at risk, respectively. Of the 105 dogs that developed GDV, 30 (28.6%) died. Incidence of GDV increased with increasing age. Cumulative incidence of GDV was 5.7% for all breeds. The only breed-specific characteristic significantly associated with a decreased incidence of GDV was an owner-perceived personality trait of happiness.     

Coronavirus-associated epizootic catarrhal enteritis in ferrets

OBJECTIVE: To characterize clinical signs and lesions and identify the etiologic agent associated with epizootic catarrhal enteritis in domestic ferrets. DESIGN: Cross-sectional study. ANIMALS: 119 ferrets with epizootic diarrhea of presumed viral cause and 5 control ferrets. PROCEDURE: Clinical records and biopsy or necropsy specimens of ferrets with presumed epizootic catarrhal enteritis were reviewed. Immunohistochemical staining for coronavirus antigen was performed on paraffin-embedded tissues from approximately 10% of affected ferrets to identify viral antigen and determine its distribution. Transmission electron microscopy was performed on fecal samples and sections of jejunum. Virus isolation studies as well as immunofluorescent tests for other similar viruses were performed. RESULTS: Characteristic microscopic lesions consistent with intestinal coronavirus infection (vacuolar degeneration and necrosis of villus enterocytes; villus atrophy, fusion, and blunting; and lymphocytic enteritis) were consistently detected in affected ferrets. Coronavirus particles were identified in feces and jejunal enterocytes by use of transmission electron microscopy. Immunohistochemical staining of jejunal sections revealed coronavirus antigens. Antigen staining was not detected in healthy ferrets or ferrets with other gastrointestinal tract diseases. Virus isolation was unsuccessful, and other similar viruses were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results strongly implicate a coronavirus as the causative agent of epizootic catarrhal enteritis in ferrets. Diagnosis may be made on the basis of a combination of historical, clinical, and microscopic findings.     

Mycobacterium infection in a captive-reared capercaillie (Tetrao urogallus)

One captive male capercaillie (Tetrao urogallus) was found dead on December 1993 at the breeding center of capercaillie in Catalonia, Spain. The bird was emaciated and, at necropsy, had numerous nodules of various sizes subcutaneously in the cervical region, pleura, lungs, liver, spleen, and mesentery. Microscopic examination revealed granulomatous lesions with central caseous necrosis, epithelioid cells, giant cells, and few lymphocytes in all affected organs. Numerous acid-fast bacilli were demonstrated in the tubercles with Ziehl-Nielsen stain.