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Biomedical subjects

M M Garcia

Publications and source records attributed to M M Garcia.

At least 91 records · Page 5Linked to original sources

Campylobacteriosis in Jamaican cattle.

Infertility is a widespread problem among cattle herds in Jamaica and an intergovernmental project was initiated to investigate the problem. One aspect of the project included diagnosis of campylobacteriosis (vibriosis), which was suspected in some herds but had not been confirmed in the laboratory. Tests were undertaken in late 1978 using improved techniques for handling and transporting specimens to the laboratory. Campylobacter fetus subspecies fetus was isolated from five of six herds tested. The diagnosis of C. fetus infection has led to improvements in farm management and a better appreciation of more efficient artificial insemination and veterinary services.

Agglutination Tests↗

An evaluation of prepackaged columns for the separation of carcinoembryonic antigen from perchloric acid: concise communication.

Commerically available Sephadex buffer-exchange columns for the separation of carcinoembryonic antigen (CEA) from perchloric acid were evaluated. Elution patterns of individual columns were found to be variable, but separation of acid from CEA could be accomplished. Repeatability of replicate elutions and agreement with dialysis results were acceptable. Nevertheless, the method is excessively time-consuming and inefficient for processing large numbers of patient samples.

Buffers↗

An assessment of methods for the estimation of free thyroxine.

Three commercial kit methods for the estimation of free thyroxine (FT4) (Clinical Assays, Corning Medical, Damon Diagnostics) were evaluated. The effects of changes in thyroxine-binding globulin (TBG) and FT4 concentrations in these systems were assessed. Measurements were made in serum samples from hyperthyroid, hypothyroid, and euthyroid patients with different TBG concentrations, and results were correlated with functional thyroid status. The Clinical Assays and Damon Diagnostics assays were found to be essentially independent of binding-protein concentration effects and responded appropriately to changes in FT4 concentration. The method of Corning Medical does not measure FT4 directly but yields an FT4 index calculated from a TBG-dependent T4 uptake and the total T4 concentration. This corning index yields falsely elevated results in patients with marked elevation in TBG concentration.

Evaluation Studies as Topic↗

Contagious equine metritis: a review.

Contagious equine metritis is a highly contagious genital infection of mares, spread venereally, and was first described in 1977. Although most contagious equine metritis outbreaks involved Thoroughbreds, infection in other breeds has also occurred. The disease has been reported in Europe, Australia and the United States. In Canada, contagious equine metritis has been designated a reportable disease under the Animal Disease and Protection Act. Contagious equine metritis is characterized by an endometritis and infertility and infected mares show no signs of systemic infection. Clinical signs have not been observed in stallions. An asymptomatic carrier state exists in both mares and stallions.Infected mares respond clinically to the topical and parenteral administration of antibacterial drugs. However, a proportion of mares remain carriers of the contagious equine metritis organism. Treatment of stallions is successful. Haemophilus equigenitalis has been proposed as the species name of the Gram-negative, microaerophilic coccobacillus. Sample collection and laboratory methods for the diagnosis of contagious equine metritis are described.

Animals↗

Assessment of serum myoglobin as a marker for acute myocardial infarction.

The reliability of serum myoglobin as a marker for acute myocardial infarction was evaluated in 157 consecutive coronary-care admissions. Admission myoglobin was elevated in 47 of 52 patients with acute infarction. Excluding those patients who presented later than 24 hr after symptom onset, only one patient with acute infarct had a normal admission myoglobin. In 22 of 105 patients with no infarct, myoglobin was elevated in association with angina, congestive heart failure, arrhythmias, and renal insufficiency. The detection of acute infarction by serum myoglobin measurement equals that of serial serum creatine phosphokinase isoenzymes (CPK-MB) by electrophoresis, but an elevated myoglobin is not specific for what is now considered clinically significant myocardial infarction.

Creatine Kinase↗

Effects of contaminant radioactivity on results of 125I-radioligand assay.

Diagnostic radionuclide imaging procedures are often used in patients whose sera are later assayed by use of 125I-labeled radioligands; thus, it is important to identify those assays that potentially may be affected by contaminating radioactivity. Results obtained with assays in which specific separation methods (solid-phase primary antibody, second antibody) or small (10--25 micdo L) sample volumes are used are little affected by the presence of such contaminating radioactivity. Less-specific techniques (polyethylene glycol, charcoal, ion-exchange resin) segregate some of the contaminant activity into the bound fraction. The degree to which such activity is protein bound and the concentration of endogenous ligand then contribute to the resulting error in dose estimation. Samples for these assays should be screened for radioactivity before the assay is begun. Inclusion of nonspecific binding tubes for patients' samples when contamination is present permits the contaminating radioactivity to be evaluated and the patient's dose concentration to be more accurately estimated.

Chorionic Gonadotropin↗

Spurious underestimation of results for digoxin radioimmunoassay with a commercial kit.

Estimates of digoxin in serum with a commercial redioimmunoassay kit (Schwarz/Mann) occasionally result in more counts in the patient's "bound" fraction than in the "bound" fraction for the zero standard. This phenomenon has been investigated in 26 serum samples obtained from patients who were ingesting known amounts of digoxin. The problem appears to relate to environmental differences in at least the zero standard and affects results when serum digoxin concentrations are less than 0.8 microgram/liter. Apparently dose estimates are more accurate with other methods or by calculating results after omitting the zero standard.

Digoxin↗

Intraperitoneal immunization against necrobacillosis in experimental animals.

Experiments employing recently developed mouse models indicated that intraperitoneal immunization with the cytoplasm (intracellular fraction) of Fusobacterium necrophorum protected the animals from a lethal challenge of the pathogen. The critical immunization schedule needed to achieve complete protection involved six weekly intraperitoneal doses of the intracellular antigen. Livers of immunized mice were cleared of infecting fusobacterial within 24 hours whereas those of nonimmunized mice harboured increasing numbers of hte bacteria. Sera from both groups did not protect recipient mice form developing liver abscesses after challenge. Sheep immunized intraperitoneally with 20 mg of cytoplasmic protein given in three doseases were protected against the development of abscesses induced by F. necrophorum.

Administration, Intranasal↗

Hepatic lesions and bacterial changes in mice during infection of Fusobacterium necrophorum.

Liver abscesses were induced in male albino mice within 1 week after intraperitoneal inoculation of viable Fusobacterium necrophorum LA19 culture. Fusobacteremia was transitory and reached a peak 2 h after inoculation then sharply declined until its disappearance 24 h post inoculation. By contrast, the number of fusobacteria in the liver increased rapidly during the first 4 h post inoculation and continued to do so less rapidly until the last sampling time (48 h post inoculation). There were small or large areas of necrosis, usually surrounded by inflammatory cells, small focal accumulations of lymphocytes, plasma cells, and macrophages in areas of parenchyma with no degenerations, generalized proliferation of Kupffer cells, and a few accumulations of fibrin and leukocytes on the surface. Ultrathin sections of infected liver tissues reveled both intact and partially degraded F. necrophorum cells enclosed in phagocytic and digestive vacuoles of mononuclear cells, The results indicate that macrophages play a key role in the pathogenesis of liver abscesses.

Animals↗

Spontaneous listeric encephalitis and neuritis in sheep. Light microscopic studies.

Sixteen of 17 sheep with spontaneous listeric encephalitis had neuritis characterized by diffuse and focal intrafascicular and perineural accumulations of lymphocytes, plasma cells, macrophages and neutrophils in one or more cranial nerves. Nine sheep had extensive trigeminal neuritis which was usually unilateral. Brain lesions were mainly in the stem and were foci of macrophages or neutrophils or both, malacia, neutrophilic neuronophagia, vascular cuffing, and meningitis. Lesions in the brain and trigeminal ganglia were most severe on the same side as the affected trigeminal nerve. Gram-positive bacilli were in proximal parts of cranial nerves in foci of inflammatory cells and occasionally in morphologically intact nerve fibers. Organisms in the brain were in phagocytes in areas of inflammation and in scattered neurons and axons. The results were consistent with centripetal migration of the infectious agent along one or more branches of the trigeminal nerve to the brain and dissemination in the brain stem occurring, at least partly, along fiber tracts. Intraaxonal movement of bacteria probably is a mechanism involved in the pathogenesis of this disease.

Animals↗

Characterization of endotoxin from Fusobacterium necrophorun.

Endotoxic lipopolysachharide (LPS) was obtained from phenol-water extraction of cell walls prepared from mass-cultivated Fusobacterium necrophorum. The LPS was relatively free of nucleic acids and low in protein, and constituted about 4% of the cell walls. Upon acid hydrolysis, some of the components detected were hexosamines (7.0%), neutral and reducing sugars (50.5%), heptose (6.4%), 2-keto-3-deoxyoctonate (0.8%), lipid A (21.0%), and phosphorus (1.7%). Under electron microscopy the LPS appeared mainly as ribbon-like trilaminar structures, and upon chemical treatment it displayed a behavior resembling that reported in certain enterobacterial LPS. The LPS was lethal to mice, 11-day-old chicken embryos, and rabbits. Endotoxicity in mice was enhanced at least 1,380-fold by the addition of 12.5 mug of actinomycin D. Induced tolerance to lethal effect of the endotoxin and rapidly acquired resistance to infection by F. necrophrum viable cells were also demonstrated in mice. The endotoxin produced both localized and generalized Shwartzman reactions as well as biphasic pyrogenic responses in rabbits. These results firmly establish the presence of a classical endotoxin in F. necrophorum, thus providing strong support to our recent suggestion that cell wall-associated components may contribute significantly to the pathogenicity of F. necrophorum.

Animals↗

Biological characterization of Fusobacterium necrophorum. Cell fractions in preparation for toxin and immunization studies.

Fusobacterium necrophorum isolated from bovine liver abscesses was grown in bulk at 37 C for 24 h under a strict anaerobic atmosphere. Harvested washed cells were disrupted ultrasonically and fractionated by differential centrifugation into the intracellular (cytoplasm) and cell wall fractions. Both intact cells and cell fractions induced generalized cytopathic effect on primary pig kidney cultures and caused a variety of signs of illness and/or death of intraperitoneally injected mice. The intact cells, disrupted cells, and cell walls produced necrotic lesions and erythema on intradermally injected guinea pigs and rabbits, whereas the cytoplasm mainly erythema. By contrast, the used culture medium (culture filtrate) of F. necrophorum did not show any detectable toxicity. The toxic component of the cytoplasm appears to be associated with nondialyzable, hemolytic, high-molecular-weight proteins and its toxicity is reduced by trypsin and pronase. Heating at 60 C for 10 min decreased markedly its erythemal and cytotoxic ability, wheras the toxicity of the cell walls appeared to be only slightly affected even when heated at 100 C for 1 h. These results suggest that at leasttwo distinct cell-bound toxic factors are present in F. necrophorum cells.

Amino Acids↗

Results of a preliminary trial with Sphaerophorus necrophorus toxoids to control liver abscesses in feedlot cattle.

A preliminary field experiment was undertaken to evaluate the efficacy of alum precipitated toxoids of Sphaerophorus necrophorus prepared from sonicated whole cells and cell fractions to reduce the incidence of bovine abscesses. A total of 108 calves were divided into five groups and treated as follows: I. uninoculated control, II. adjuvant inoculated control, III. 15.5 mg protein of sonicated (fragmented cells) toxoid, IV. 10.5 mg protein of cytoplasmic toxoid. V. 15.5 mg protein of cytoplasmic toxoid. All animals were maintained under similar conditions to those prevailing in feedlots in Alberta. Livers were examined at slaughter. The most promising result was achieved with the injection of 15.5 mg protein of cytoplasmic toxoid. In this treatment group, no scars (healed lesions) were found in the liver and the incidence of liver abscesses was reduced to 10% from the average 35% liver abscesses and scars found in the uninoculated and adjuvant inoculated groups. The toxoid from sonicated whole cells did not reduce liver abscess incidence. These data suggest that the incidence of liver abscesses in cattle fattened in feedlots may be reduced by immunization.

Adjuvants, Immunologic↗

Evaluation of various media and the use of soil infusion in the production of Brucella abortus antigen.

Brucella abortus (ADRI 413) was grown on various synthetic, commercial, laboratory-formulated and soil infusion-supplemented agar media. Of 22 media tested, three soil infusion-supplemented media and a serum dextrose agar yielded more brucella antigen than the meat infusion agar, the standard reference medium. Results from agglutination and stability tests indicated that antigens produced from most of the laboratory-formulated media and the soil infusion-supplemented agars compared favourably with that derived from the meat infusion agar. The presence of meat infusion or commercial beef extract appeared to be a basic requirement for producing satisfactory and stable antigen. The growth enhancing effect of certain soil infusions seemed to be associated with the soil organic matter fraction. A significant correlation (p>.05) was obtained between the yield of brucella antigen and the glutamic acid content of the soil infusions. With proper screening, media containing, among other components, beef extract and soil infusions may be of value particularly in areas where the limited supply of fresh meat would make meat infusion agar uneconomical in the production of B. abortus (ADRI 413) antigen.

Agar↗

Application of immunofluorescence to studies on the ecology of Sphaerophorus necrophorus.

Sphaerophorus necrophorus (bovine liver abscess isolates) antiserum was fractionated and labeled with fluorescein isothiocyanate. The fluorescent-antibody (FA) conjugate proved to be species-specific and facilitated the detection of S. necrophorus cells in liver abscesses, viscera, and ruminal contents of cattle. Brightly fluorescing S. necrophorus cells were observed in experimentally inoculated soil incubated anaerobically at 37 and 4 C for 8 and 10 months, respectively. When incubated under moist conditions (80% water holding capacity) at 37 C, the test organism survived in both sterile and unsterile soil for as long as 8 weeks. Results strongly support the feasibility of using FA techniques concurrent with cultural methods for rapid detection of S. necrophorus infections.

Animals↗

Pathogenic microorganisms in soil: an old problem in a new perspective.

Early investigations on the survival of pathogenic bacteria in soil have been directed mainly to changes in the numbers of viable populations. More reliable techniques to assess the physiological, biochemical and pathogenic capabilities of such organisms are necessary to gain a better understanding of the nature of the diseases they cause and ultimately of the means to control them.

Actinomyces↗