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Biomedical subjects

L W George

Publications and source records attributed to L W George.

At least 19 recordsLinked to original sources

Cloning and characterization of a Moraxella bovis cytotoxin gene.

OBJECTIVE: To identify the Moraxella bovis cytotoxin gene. PROCEDURE: Hemolytic and nonhemolytic strains of M. bovis were compared by use of western blotting to identify proteins unique to hemolytic strains. Oligonucleotide primers, designed on the basis of amino acid sequences of 2 tryptic peptides derived from 1 such protein and conserved regions of the C and B genes from members of the repeats in the structural toxin (RTX) family of bacterial toxins, were used to amplify cytotoxin-specific genes from M. bovis genomic DNA. Recombinant proteins were expressed, and antisera against these proteins were produced in rabbits. RESULTS: Several proteins ranging in molecular mass from 55 to 75 kd were unique to the hemolytic strain. An open reading frame encoding a 927-amino acid protein with a predicted molecular mass of 98.8 kd was amplified from M. bovis genomic DNA. The deduced amino acid sequence encoded by this open reading frame was homologous to RTX toxins. Antisera against the recombinant carboxy terminus encoded by this open reading frame neutralized hemolytic and cytolytic activities of native M. bovis cytotoxin. CONCLUSIONS AND CLINICAL RELEVANCE: A gene was identified in M bovis that encodes a protein with sequence homology to other RTX toxins. Results of cytotoxin neutralization assays support the hypothesis that M. bovis cytotoxin is encoded by this gene and belongs in the RTX family of bacterial exoproteins. Identification of this gene and expression of recombinant cytotoxin could facilitate the development of improved vaccines against infectious bovine keratoconjunctivitis.

Amino Acid Sequence↗

Efficacy of florfenicol for treatment of naturally occurring infectious bovine keratoconjunctivitis.

OBJECTIVE: To determine the efficacy of florfenicol for treatment of calves with naturally occurring infectious bovine keratoconjunctivitis (IBK). DESIGN: Randomized controlled field trial. ANIMALS: 63 beef calves and 80 dairy calves between 4 and 12 months of age. PROCEDURE: Calves were randomly assigned to 1 of 3 treatment groups. Calves in the SC treatment group received a single dose of florfenicol (40 mg/kg [18.2 mg/lb of body weight), SC, on day 0. Calves in the IM treatment group received florfenicol (20 mg/kg [9.1 mg/lb]), IM, on days 0 and 2. Calves in the control group received injections of saline solution (0.9% NaCl), IM, on days 0 and 2. Calves were reevaluated every other day for 20 days after treatment. RESULTS: Corneal ulcers healed by day 20 in 48 of 49 (98%) calves treated with florfenicol IM, 39 of 42 (93%) calves treated with florfenicol SC, and 33 of 52 (63%) control calves. CONCLUSIONS AND CLINICAL RELEVANCE: Florfenicol administered SC (1 dose) or IM (2 doses 48 hours apart) was effective for treatment of calves with naturally occurring IBK.

Animals↗

Treatment of hypernatremia in an acidotic neonatal calf.

A 7-day-old Jersey calf was evaluated because of diarrhea and pneumonia. The calf was hypernatremic, hypoproteinemic, and acidemic, and was treated initially with i.v. administration of fluids with sodium concentration (175 mEq/L) similar to the calf's serum sodium concentration. Sodium concentration of the administered fluids was gradually decreased over the following days, but the calf's serum sodium concentration decreased too rapidly, and the calf developed neurologic signs attributed to cerebral edema. Treatment with mannitol and i.v. administration of fluids with a higher concentration of sodium resulted in abatement of clinical signs. In calves, hypernatremia may develop over several days. Prescribing traditional isotonic or hypotonic fluids in such cases will be harmful, because during chronic hypernatremia, the brain's adaptive mechanisms involve accumulation of organic osmoles that may take several days to equilibrate across cell membranes, and cerebral edema may result. Administration of fluids containing sodium concentration approximately equal to the patient's measured serum sodium concentration is required to decrease serum sodium concentration more slowly than is possible with traditional isotonic fluids.

Acidosis↗

Efficacy of florfenicol in the treatment of experimentally induced infectious bovine keratoconjunctivitis.

OBJECTIVE: To evaluate efficacy of florfenicol in an induced model of infectious bovine keratoconjunctivitis, using a blinded randomized, controlled trial. ANIMALS: 48 male Holstein calves, 2 to 4 months old. PROCEDURE: Moraxella bovis infection was induced in all calves. When corneal ulcers developed, each calf was assigned randomly to 1 of 3 treatment groups, using a block design determined by corneal ulcer size (day 0). Calves were treated with florfenicol (20 mg/kg of body weight, IM) on days 0 and 2 (IM group; n = 16). Calves of a second group received a single dose of florfenicol (40 mg/kg, SC) on day 0 (SC group; n = 16). The third group of calves was not treated (control group; n = 16). Corneal ulcers were photographed, and each calf was assessed for 30 days after treatment for 10 clinical signs of infection. Corneal ulcer surface areas were measured, and clinical scores were calculated. Ocular secretions for microbiologic culture were obtained weekly from each eye. RESULTS: A Cox regression model indicated that, after adjustment for initial ulcer size, healing rates were 6.2 and 4.8 times greater in calves of the IM and SC groups, respectively, compared with the control group. Clinical scores and surface area measurements for treatment groups were significantly smaller than those for controls during posttreatment weeks 1 through 4. From day 8 through day 29, M bovis was isolated from ocular secretions of 14 of 16 control calves and 1 of 32 treated calves. CONCLUSIONS AND CLINICAL RELEVANCE: Parenterally administered florfenicol reduces corneal ulcer healing time, lessens clinical severity, and reduces the amount of bacterial shedding from calves infected with M bovis.

Animals↗

Combined parenteral and oral administration of oxytetracycline for control of infectious bovine keratoconjunctivitis.

OBJECTIVE: To determine whether combined parenteral and oral administration of oxytetracycline would ameliorate a herd outbreak of infectious bovine keratoconjunctivitis (IBK) and to compare efficacy of this regimen with that of subconjunctival administration of procaine penicillin G. DESIGN: Randomized field trial. ANIMALS: 119 Hereford calves in a herd undergoing a naturally occurring outbreak of IBK. PROCEDURE: Calves were randomly assigned to 1 of the following 3 groups: oxytetracycline treatment, procaine penicillin G treatment, and control. After initial treatment, calves were examined 3 times/wk for 7 weeks. The surface area of all corneal ulcers was measured during each examination. Ocular secretions were collected from all calves at least weekly and were tested for Moraxella bovis. RESULTS: Calves treated with oxytetracycline had a lower prevalence of IBK than did calves treated with procaine penicillin G or control calves and required fewer additional treatments than did calves treated with procaine penicillin G. Mean time for healing of corneal ulcers was significantly less for calves that received oxytetracycline or procaine penicillin G than for control calves. Calves treated with oxytetracycline developed fewer corneal ulcers and fewer recurrent ulcers than did calves in the other groups. Moraxella bovis was isolated less often from ocular secretions collected from calves in the oxytetracycline group than from calves in the other groups. CLINICAL IMPLICATIONS: Combined parenteral and oral administration of oxytetracycline appears to be an effective method of reducing severity of a herd outbreak of IBK and may be superior to treatment of affected animals with procaine penicillin G.

Administration, Oral↗

Primary photosensitization related to ingestion of alfalfa silage by cattle.

A herd of 650 Holstein cows was examined for skin disease. Approximately 400 of the lactating adults were affected, but heifers, calves, and nonlactating cows were clinically normal. The condition was characteristic of primary photosensitization. Milk production of the affected cows was normal. Affected cows did not appear to be ill, and none of the cows was icteric. Three of 7 cows had high serum gamma-glutamyltransferase activities, but in the other 4 cows, activity was within the reference range. Serum activities of other hepatic enzymes were within reference ranges in the 7 cows that were examined. Hepatic biopsy specimens from 3 cows were normal. Specimens from 4 other cows had changes that ranged from minimal to mild, chronic, lymphoplasmacytic periportal hepatitis to acute, random, necrotizing hepatitis. Development of photosensitivity was related to ingestion of alfalfa silage. Acetone extracts of the alfalfa silage, but not of other feedstuffs, were found to inhibit growth of Candida albicans under ultraviolet light. Cows experimentally fed a diet composed exclusively of the alfalfa silage developed skin lesions after 6 days, but did not have detectable serum concentrations of phylloerythrin.

Animals↗

Alterations in blood lymphocyte subpopulations and hematologic values in neonatal calves after administration of a combination of multiple-antigen vaccines.

OBJECTIVE: To evaluate alterations in lymphocyte subpopulations, CBC results, and clinical signs in neonatal calves inoculated with 3 commercially available proprietary multiple-antigen vaccines containing known quantities of endotoxin. DESIGN: Prospective, randomized controlled field trial. ANIMALS: 36 healthy Holstein heifer calves between 3 and 31 days old. PROCEDURE: Vaccines were administered to 18 calves according to label instructions, except for the recommended age of administration. The 18 other calves served as unvaccinated controls. Two weeks after entry into the study, calves were given secondary doses of the same vaccines. Calves in both groups were examined and blood samples were collected for determination of lymphocyte subpopulations and hematological parameters once daily for 5 days beginning on the day that both the primary and the secondary vaccinations were given. Lymphocyte subpopulations, including BoCD2+, BoCD4+, BoCD8+, B cells, and gamma/delta T cells, were determined by use of flow cytometry, using monoclonal antibodies as markers. RESULTS: Vaccinated calves did not develop clinical signs of illness. There were no significant differences in absolute numbers of lymphocyte subpopulations between vaccinated and unvaccinated calves. Vaccinated calves had significantly higher rectal temperatures, total WBC counts, and absolute neutrophil counts than did control calves. These differences persisted for 3 to 4 days after vaccination. CLINICAL IMPLICATIONS: Findings confirm empirical observations that vaccination with multiple products at the same time may induce evidence of an inflammatory response in most calves. Additional research is indicated to further evaluate the safety of using multiple vaccines simultaneously.

Animals↗

Effect of penicillin or penicillin and dexamethasone in cattle with infectious bovine keratoconjunctivitis.

To evaluate the efficacy of penicillin or penicillin and dexamethasone for treatment of infectious bovine keratoconjunctivitis, 6- to 8-month-old beef heifers with clinical signs of infectious bovine keratoconjunctivitis were randomly assigned to 1 of 3 treatment groups: penicillin only, penicillin and dexamethasone, or control. Cattle assigned to the penicillin group (n = 18) were treated with 3 daily subconjunctival injections of procaine penicillin G. Cattle assigned to the penicillin/dexamethasone group (n = 13) were treated with 3 daily subconjunctival injections of procaine penicillin G and dexamethasone sodium phosphate. Control cattle (n = 14) were not treated. Healing times and frequency of recurrence for corneal ulcers; severity, diameter, and surface area measurements of corneal ulcers; and clinical scores did not differ among the 3 groups. Frequency of Moraxella bovis isolation from specimens of ocular secretions from ulcerated and non-ulcerated eyes was similar in all groups. Minimum inhibitory concentration of penicillin G for 95 of the 102 tested M bovis isolates was 0.3 U/ml, and for 7 others was 0.03 U/ml. When first and last specimens from 42 of 45 calves with isolation of M bovis on serial microbial cultures were compared, the susceptibility of each last isolate was similar to that of the corresponding first isolate.

Animals↗

Seizures and acute death attributable to hypovitaminosis A and suspected hypovitaminosis D in feeder pigs.

Clinical signs that included lethargy, inappetence, diarrhea, and vomiting and that progressed to seizures were observed in 40 feeder pigs that were approximately 70 days old. The pigs were fed ground red wheat and whole milk and were housed in a barn that did not allow exposure to direct sunlight. Analysis of samples of feed obtained from the farm indicated inadequate quantities of calcium and phosphorus as well as a low ratio of these 2 nutrients. Serum and tissue concentrations of vitamin A were less than normal. Low serum calcium concentrations, high serum phosphorus concentrations, and high alkaline phosphatase and creatine kinase activities were compatible with low vitamin D concentrations.

Animal Feed↗

Topically applied benzathine cloxacillin for treatment of experimentally induced infectious bovine keratoconjunctivitis.

The efficacy of an ophthalmic ointment containing benzathine cloxacillin for treatment of infectious bovine keratoconjunctivitis was determined in 2 experiments. In the first experiment, Holstein calves (n = 6/group) were inoculated with Moraxella bovis and treated on postinoculation days 3 and 6 with either topically applied benzathine cloxacillin (250 mg/eye) or long-acting oxytetracycline formulation (20 mg/kg of body weight, IM). A third group of inoculated calves remained untreated as controls. For the second experiment, 4 groups of calves (n = 6/group) were inoculated and treated on postinoculation days 3 and 6 with 50, 125, 250, or 375 mg of benzathine cloxacillin; a fifth untreated group served as controls. Ocular specimens were obtained for microbiologic culture, and eyes were observed and assigned a clinical score daily. Eyes were photographed on alternate days. Ulcer surface area was measured, using a planimeter. In experiment 1, the week-2 ulcer surface area measurements for both groups of treated calves were smaller than those for controls. There was a greater frequency of M bovis isolation from the ocular secretions of controls than from those of benzathine cloxacillin-treated calves during postinoculation weeks 2 and 3. The number of M bovis isolations from the benzathine cloxacillin- and oxytetracycline-treated calves was not significantly different at any sample collection interval. On week 3, the scores of the benzathine cloxacillin-treated calves were smaller than those of controls. In experiment 2, calves of the 250- and 375-mg groups had smaller ulcer surface area measurements than did controls on week 2. By week 3, calves of the 375-mg group had smaller scores than did controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Topical↗

Ocular and serum disposition kinetics of cloxacillin after topical administration of benzathine cloxacillin and intravenous administration of sodium cloxacillin to calves.

Disposition kinetics of cloxacillin were examined in calves after topical administration of benzathine cloxacillin and single IV administration of sodium cloxacillin, and the susceptibility of 17 field isolates of Moraxella bovis was measured. For the IV pharmacokinetic phase, sodium cloxacillin was administered at dosage of 10 mg/kg of body weight to male Holstein calves (n = 6, weighing 146 to 170 kg), and serum concentration of cloxacillin was measured thereafter for 10 hours. For the ocular pharmacokinetic phase, 6 calves were given either of 4 benzathine cloxacillin topical formulations consisting of 50-, 125-, 250-, or 375-mg doses. Treatment was repeated every 10 days until all 4 benzathine cloxacillin dosages were tested in the same 6 calves. Blood and tears were collected for 72 hours after each benzathine cloxacillin formulation was administered, and the concentration of cloxacillin in each specimen was measured, using a bioassay. The minimal inhibitory concentration of cloxacillin for 17 field isolates of M bovis was determined by use of an agar pour-plate dilution assay. After single IV administration of sodium cloxacillin, its half-life, body clearance, and volume of distribution were 19.5 +/- 12.8 minutes, 18.3 +/- 2.2 ml/min.kg, and 496 +/- 290 ml/kg, respectively. After topical administration of benzathine cloxacillin, cloxacillin concentration in lacrimal fluid peaked between 30 and 45 minutes and ranged between 963 micrograms/ml and 3,256 micrograms/ml for the 125- and 375-mg doses, respectively. There was no detectable cloxacillin activity in the lacrimal fluid of any calf by 36 hours after topical administration of benzathine cloxacillin, and cloxacillin was not detected in the serum at any time.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Topical↗

Phagocytic and nitroblue tetrazolium reductive properties of mature and immature neutrophils and eosinophils from blood and bone marrow from cows.

Functional capabilities of morphologically mature (segmented) and immature granulocytes (neutrophils and eosinophils) from bone marrow from cows were studied and compared with similar activities of segmented granulocytes from blood. Phagocytosis of Escherichia coli and postphagocytic oxidative metabolic stimulation, measured by nitroblue tetrazolium (NBT) reduction, were evaluated simultaneously. Phagocytosis was observed readily in segmented neutrophils, neutrophilic bands, and metamyelocytes and rarely in myelocytes. Phagocytosis was not seen in promyelocytes and myeloblasts. Neutrophilic bands and metamyelocytes were phagocytically less active than were segmented neutrophils. Washed segmented bone marrow neutrophils possessed phagocytic activity similar to that of blood neutrophils, whereas the activity of unwashed segmented bone marrow neutrophils was markedly less than that of blood neutrophils. Reduction of NBT was observed only in blood segmented neutrophils and bone marrow segmented neutrophils; the magnitude of NBT reduction was significantly (P = less than 0.005) less in bone marrow neutrophils than in blood neutrophils. Eosinophils were phagocytically less competent than were neutrophils. The NBT reduction was observed only in eosinophils from blood, but not in eosinophils from bone marrow.

Animals↗

Survival analysis for evaluation of corneal ulcer healing times in calves with naturally acquired infectious bovine keratoconjunctivitis.

A clinical trial examining the efficacy of 2 drugs for treatment of a natural epizootic of infectious bovine keratoconjunctivitis was performed. The study was conducted in 103 grazing Hereford calves during the summer of 1985. The calves were prospectively and randomly assigned to 1 of 3 groups at the beginning of the study on June 17, and were examined 3 times weekly thereafter until the final observation on August 6. Calves in group 1 (n = 34) were not treated and were used as controls. Calves of group 2 (n = 34) with corneal ulcers were treated with a long-acting oxytetracycline formulation (OTC group). The parenteral treatment was repeated in 72 hours. Affected calves of group 3 (n = 35) were treated topically with furazolidone spray when they developed new corneal ulcers, or when existing lesions worsened during subsequent examination periods (NFZ group). Healing times of the corneal ulcers were reported in 3 ways: the combined times for ulcers present in both eyes of a calf simultaneously (method A), independent times of each ulcer on a calf (method B), and time of the first ulcer for each calf (method C). Censored healing times were examined as left censored (ulcer present at the beginning of the study), right censored (ulcer not healed at the end of the study), or uncensored (true) healing times. The effect that the treatments had on healing times were investigated by use of notched box and whisker plots, life tables, and Cox regression models. The analysis indicated that treatment of calves with either antimicrobial reduced the healing time of corneal ulcers, compared with untreated controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effectiveness of two benzathine cloxacillin formulations for treatment of naturally occurring infectious bovine keratoconjunctivitis.

A field study was performed to determine the effectiveness of benzathine cloxacillin for the treatment of infectious bovine keratoconjunctivitis in cattle from 2 farms located in northern California. The study was performed between June and September of 1987. Affected calves ranging from 2 to 9 months of age were selected from the main herd when signs of corneal ulceration were observed. The study was conducted in 2 phases. For phase I, the affected calves of herd 1 (n = 21; Holsteins) and herd 2 (n = 43 Angus crossbred), were randomly assigned to 1 of 3 groups, and were either treated with 250 (n = 23) or 375 mg (n = 21) of benzathine cloxacillin, or mineral oil (n = 20) on days 1 and 4. For phase II, affected calves (n = 16; Angus crossbred, 3 to 9 months of age) from herd 2 were treated with benzathine cloxacillin (250 mg). Eight of these calves were retreated on day 4. After treatment, all calves were examined every 72 hours for 16 days. For examinations, a clinical score was assigned to each eye, and the surface areas of photographed corneal ulcers were measured. The ocular secretions were collected and examined culturally for Moraxella bovis. On days 7, 10, and 13, the calves treated with benzathine cloxacillin had significantly (P less than 0.05) lower lesion scores, compared with the controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cytopathic effects of Moraxella bovis on cultured bovine neutrophils and corneal epithelial cells.

The effects of Moraxella bovis on the morphologic features of purified bovine neutrophils and bovine corneal epithelial cells were examined, using transmission and scanning electron microscopy and light microscopy. Within 2 minutes after incubation of bovine neutrophils with living M bovis, electron microscopic cellular changes included vacuolation, swelling, and loss of microplicae. Most of the neutrophils were lysed by 10 minutes of incubation. Human neutrophils phagocytosed the M bovis and remained intact, even after 30 minutes of incubation with the bacteria. Living M bovis killed bovine corneal epithelial cells in vitro. Sterile filtrates prepared from 6-hour shaker cultures of M bovis also killed bovine corneal epithelial cells, but the cytotoxic activity was less than that produced by the living bacteria. Cellular changes were first observed in specimens collected 1 hour after corneal cell monolayers were inoculated with sterile culture filtrates. The changes in these cells included pit-like lesions on the cellular surface, cellular separation, and vacuolation.

Animals↗

Abomasal dilatation and emptying defect in a flock of Suffolk ewes.

Abomasal dilatation and emptying defect was identified in 14 of 130 Suffolk sheep from a single flock in northern California. Abomasal dilatation and emptying defect was diagnosed in 13 of the sheep between November 1986 and April 1987. All affected sheep were ewes and in all, the condition developed during the lambing season. Clinical signs consisted of chronic anorexia and weight loss. Laboratory data from affected ewes reflected concurrent disease and was not helpful in diagnosis of the disease. Necropsy of 13 affected ewes revealed a distended, ingesta-filled abomasum in all of them. The brain stem of 6 ewes was examined, and lesions suggestive of scrapie were not observed. Pedigree analysis determined that in this flock, abomasal dilatation and emptying defect development was independent of sire. A relationship between the age of affected ewes and the disease was not found. Treatment was unsuccessful in 13 ewes, but 1 recovered after treatment with metoclopramide.

Abomasum↗