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Biomedical subjects

M L Anderson

Publications and source records attributed to M L Anderson.

At least 37 records · Page 2Linked to original sources

A bovine model of vaccine enhanced respiratory syncytial virus pathophysiology.

A critical issue has been the observation that vaccination of children with a formalin-inactivated respiratory syncytial virus (RSV) vaccine is associated with disease enhancement. We have taken advantage of bovine RSV and our experience with this disease in calves to develop a natural model that parallels human disease. Using formalin-inactivated bovine RSV vaccine calves were either sham-vaccinated/infected, vaccinated/infected, or vaccinated/sham-infected and their clinical signs, pulmonary function, and histological lung lesions quantitatively scored. Interestingly there was significantly greater disease in vaccinated/infected calves and histological lesions in calves were similar to those of affected children. Finally, we note that vaccination did not induce neutralizing antibodies, but IgG antibodies were detected by ELISA. Our model of RSV enhanced disease is important because it provides quantifiable evidence of disease severity that can be applied to evaluate the mechanisms of immunopathology and the safety of candidate RSV vaccines.

Analysis of Variance↗

Female reproductive tract immunity in bovine trichomoniasis.

PROBLEM: Mechanisms of protective immunity in the female reproductive tract are poorly understood. For sexually transmitted diseases, bovine trichomoniasis is a useful model because it resembles human trichomoniasis to some extent, and antibodies play an important role in protection against these extracellular parasites. Protective efficacy was compared in animals with genital responses of predominantly immunoglobulin G (IgG) or predominantly IgA antibodies to a purified surface antigen of Tritrichomonas foetus. METHOD OF STUDY: Immunization of mice by various routes with immunoaffinity-purified T. foetus surface antigen (TF1.17) or killed cells was used to define the best routes and antigen combinations to give predominantly IgG or IgA antibodies to TF1.17 antigens in genital secretions. Cattle were then immunized either subcutaneously (SC) two times with TF1.17 antigen and once SC with killed T. foetus or twice SC with TF1.17 antigen and once intravaginally with killed T. foetus. All immunizations were in Quil A adjuvant. Controls were not immunized. Animals were challenged intravaginally with 10(6) T. foetus 3 weeks after the third immunization. Vaginal mucus was collected weekly for culture and antibody assays. Serum was collected weekly, and uterine secretions were collected at 10 weeks post-challenge. Tissues were fixed at 10 weeks also. RESULTS: Murine studies showed systemic priming with vaginal boosting gave the highest genital IgA responses. In cattle, systemic immunization (group S) induced high IgG1 antibody levels in vaginal secretions. Systemic priming with vaginal boosting (group S/V) primed for an anamnestic vaginal IgA response after challenge with T. foetus. Cattle with predominantly IgG or predominantly IgA responses in vaginal secretions either did not become infected or cleared infection faster than controls. Uterine IgA responses at 10 weeks were highest in the vaginally boosted group, but other responses were not different from the controls at this time point. Microscopic examination of genital tissues showed subepithelial infiltration of mononuclear cells in all groups. Lymphoid aggregates or nodules were detected in vaginal sections in cattle of groups S/V and C as well as in uterine sections of all animals in all three groups. CONCLUSIONS: Both IgG and IgA antibodies to T. foetus superficial antigen were associated with protection. The timing of the response was related to the time of clearance. Lymphoid organization in the vagina and uterine tissues suggested development of mucosal inductive sites.

Animals↗

Heat shock protein 70 of the agent of human granulocytic ehrlichiosis binds to Borrelia burgdorferi antibodies.

We describe a patient with human granulocytic ehrlichiosis (HGE), a diagnosis confirmed by PCR and immunoblot analysis. Unexpectedly, immunoglobulin G (IgG) directed towards an 80-kDa ehrlichial antigen (without detectable IgM) was present in the patient's serum in the first week of illness. Lyme disease immunoblots were reactive for IgG (but not IgM), a result indicative of prior exposure to the Lyme disease spirochete. Amino-terminal sequencing revealed that the 80-kDa ehrlichial antigen was an HSP-70 homolog similar to Borrelia burgdorferi HSP-70. We conclude that antibodies against B. burgdorferi HSP-70 may cross-react with the ehrlichial heat shock protein and that this possibility must be considered when serologic test results for HGE and Lyme disease are interpreted.

Aged↗

A modified agglutination test for Neospora caninum: development, optimization, and comparison to the indirect fluorescent-antibody test and enzyme-linked immunosorbent assay.

Current serologic tests used to detect antibodies to Neospora caninum require species-specific secondary antibodies, limiting the number of species that can be tested. In order to examine a wide variety of animal species that may be infected with N. caninum, a modified direct agglutination test (N-MAT) similar to the Toxoplasma gondii modified direct agglutination test (T-MAT) was developed. This test measures the direct agglutination of parasites by N. caninum-specific antibodies in serum, thus eliminating the need for secondary host-specific anti-isotype sera. The N-MAT was compared to the indirect fluorescent-antibody test (IFAT) and the enzyme-linked immunosorbent assay (ELISA) with a "gold standard" serum panel from species for which secondary antibodies were available (n = 547). All positive samples tested were from animals with histologically confirmed infections. Up to 16 different species were tested. The N-MAT gave a higher sensitivity (100%) and specificity (97%) than the ELISA (74 and 94%, respectively) and had a higher sensitivity but a lower specificity than the IFAT (98 and 99%, respectively). The reduced specificity of the N-MAT was due to false-positive reactions in testing fetal fluids with particulate matter or severely hemolyzed serum. Overall, the N-MAT proved to be highly sensitive and specific for both naturally and experimentally infected animals, highly reproducible between and within readers, easy to use on large sample sizes without requiring special equipment, and useful in testing serum from any species without modification.

Abortion, Veterinary↗

Hoof size, shape, and balance as possible risk factors for catastrophic musculoskeletal injury of Thoroughbred racehorses.

OBJECTIVE: To evaluate hoof size, shape, and balance as risk factors for catastrophic musculoskeletal injuries (CMI), including suspensory apparatus failure (SAF) and cannon bone condylar fracture (CDY) in Thoroughbred racehorses. ANIMALS: 95 Thoroughbred racehorses that died between 1994 and 1996. PROCEDURE: 38 quantitative measures of hoof size, shape, and balance were obtained from orthogonal digital images of the hoof and were compared between case horses with forelimb CMI (70), SAF (43), and CDY (10) injuries and control horses whose death was unrelated to the musculoskeletal system (non-CMI, 25). Comparison of group means between cases and controls was done using ANOVA, and multivariable logistic regression was used to estimate odds ratios. RESULTS: Odds of CMI were 0.62 times lower for a 5mm increase in ground surface width difference and 0.49 times lower for a 100-mm2 increase in sole area difference. Odds of SAF were 6.75 times greater with a 10 degrees increase in toe-heel angle difference and 0.58 times lower with a 100-mm2 increase in sole area difference. Odds of CDY were 0.26 times lower with a 3 degrees increase in toe angle, 0.15 times lower with a 5-mm increase in lateral ground surface width, and 0.35 times lower with a 100-mm2 increase in sole area difference. CLINICAL RELEVANCE: Decreasing the difference between toe and heel angles should decrease risk of SAF for Thoroughbred racehorses and should be considered in addition to increasing toe angle alone to help prevent catastrophic injury. Trimming the hoof to perfect mediolateral symmetry may not be a sound approach to avoiding injury.

Animals↗

Microcephaly with agenesis of corticospinal tracts and arthrogryposis, hypospadias, single umbilical artery, hypertelorism, and renal and adrenal hypoplasia--previously undescribed syndrome.

We describe a small, term, male infant with corticospinal tract aplasia secondary to motor cortex dysplasia from a neuronal proliferation and/or migrational defect. The infant also had microdolichocephaly, sloping forehead, hypertelorism, flat nose, apparently low-set ears, micrognathia, arthrogryposis without muscle wasting, cortical thumbs, rocker-bottom feet, scoliosis, single umbilical artery, and hypospadias with chordee. Oligohydramnios was present prenatally. Neurologic examination showed a comatose state, seizures, minimal spontaneous movement, minimal response to pain, and absent primitive reflexes. At autopsy, hypoplasia of kidneys and adrenal glands was found. There was no aqueductal stenosis or pulmonary hypoplasia. Chromosomes were apparently normal. These manifestations do not correspond to those of any recognized syndrome; therefore, this patient may represent a previously undefined syndrome.

Abnormalities, Multiple↗

Evidence of vertical transmission of Neospora sp infection in dairy cattle.

OBJECTIVE: To determine whether heifers with naturally acquired congenital exposure to Neospora sp would transmit the infection to their offspring during gestation. DESIGN: Prospective cohort study. ANIMALS: Neonatal heifers on a dairy with a history of Neospora sp infections were selected for the study on the basis of their serum titers to Neospora sp, as determined by the use of indirect fluorescent antibody testing. Seropositive heifers (n = 25) had titers > or = 1:5,120 and seronegative heifers (25) had titers < or = 1:80. All heifers were raised and bred on the dairy, and samples were obtained from heifers and their calves at the time of calving. PROCEDURE: Blood samples were tested for Neospora sp antibodies. Histologic evaluations, Neospora sp immunohistochemical examinations, and protozoal culturing were performed on samples obtained from selected offspring (second-generation calves). RESULTS: Seropositive heifers gave birth to calves with titers > or = 1:1,280 to Neospora sp. All offspring from seropositive heifers that were necropsied had evidence of Neospora sp infection. All seronegative heifers and their offspring had titers < 1:80 to Neospora sp. CLINICAL IMPLICATIONS: Congenitally acquired Neospora sp infection can persist in clinically normal heifers and be transmitted transplacentally to their offspring. Vertical transmission can be a way by which neosporosis is maintained in herds.

Animals↗

Lack of effect of fluoxetine on the hypoprothrombinemic response of warfarin.

The purpose of this study was to assess the effect of fluoxetine on the hypoprothrombinemic response of warfarin in patients chronically anticoagulated. Patients receiving low-intensity anticoagulation with warfarin were recruited. All patients were taking stable dosages of warfarin and had two baseline prothrombin times (PTs) within 10% of each other. Each patient received fluoxetine (20 mg daily) for 21 days. PTs were measured on days 2, 5, 8, 12, 15, 19, and 22 of fluoxetine administration. Six patients completed the study. There was no significant difference in mean PTs before and during fluoxetine administration. Fluoxetine at the dosage studied does not predictably effect the hypoprothrombinemic response of warfarin.

Aged↗

Cloning and characterization of two recombinant Neospora protein fragments and their use in serodiagnosis of bovine neosporosis.

Bovine neosporosis causes fetal abortion and/or congenital neurologic disease in cattle. For the serodiagnosis of this parasitic disease, two immunodominant clones from a bovine Neospora lambda gt11 library were identified, characterized, and expressed as recombinant proteins for the development of an enzyme-linked immunosorbent assay (ELISA). These two clones, designated N54 and N57, were 29 and 20 kDa, respectively, when expressed as histidine fusion proteins from the pRSET expression vector. Antibodies to recombinant protein N54 recognized five major bands from a Neospora tachyzoite lysate with molecular masses of 97, 87, 77, 67, and 64 kDa. Antibodies to recombinant protein N57 recognized four primary bands with molecular masses of 34, 31, 30, and 28 kDa. When a defined "gold standard" panel of bovine sera from confirmed Neospora-positive and Neospora-negative cattle were characterized by immunoblotting, 57 of the 60 Neospora-positive serum samples recognized proteins with the molecular masses of the N54 heptuplet. Binding to the N57 quadruplet was more variable. The same gold standard panel was used to evaluate and compare an N54-based ELISA, an N57-based ELISA, and a whole-tachyzoite lysate-based ELISA. The sensitivities and specificities were 95 and 96% (N54 ELISA), 82 and 93% (N57 ELISA), and 74 and 93% (lysate ELISA). Thus, compared to the whole-tachyzoite lysate-based ELISA, both recombinant-protein-based ELISAs had higher sensitivities and higher or the same specificities and can be used to replace the whole-tachyzoite lysate ELISA for the serodiagnosis of bovine neosporosis.

Animals↗

Detection of Neospora sp. from infected bovine tissues by PCR and probe hybridization.

Neospora sp. can cause fetal abortion or neurological disease in congenitally infected calves. Latent tissue stages in infected cows may contribute to vertical transmission of Neospora sp. from dam to offspring in multiple pregnancies. In this investigation, the polymerase chain reaction (PCR) and Neospora-specific assay were employed to detect Neospora sp. by amplification of nuclear small-subunit rRNA gene sequences in infected cattle tissues. Tissues from 11 cattle, including 6 experimentally and 2 naturally infected cows, 1 naturally infected newborn calf, and 2 uninfected control cows, were evaluated in this study. Neospora-specific PCR products were amplified from DNAs of different bovine tissues, including brain, spinal cord, heart, lung, kidney, diaphragm, skeletal muscle, and placenta, as well as amniotic fluid samples of infected cattle. The PCR-based amplification and probe hybridization system proved useful in assessing the location of tissue-stage parasites in naturally and experimentally infected cattle, even when Neospora sp. antibody titers fall below normal cut-off values by an indirect immunofluorescent antibody test.

Abortion, Veterinary↗

Fatal musculoskeletal injuries incurred during racing and training in thoroughbreds.

OBJECTIVE: To characterize and contrast data from Thoroughbreds that incurred a fatal musculoskeletal injury (FMI; injury resulting in death or euthanasia) during racing or training and data from all California race entrants during a 9-month period in 1991. DESIGN: Case-control study. ANIMALS: Thoroughbreds that incurred a FMI during racing or training at a California race-meet and all California race entrants from January through June and October through December 1991. PROCEDURE: Age and sex were compared with chi 2 and Fisher's exact tests among horses fatally injured while racing and training. A log-linear model was fit to assess the relationship between race-meet and age and sex of California race entrants. Incidence risk of racing FMI was estimated per 1,000 race entrants, and the relationship between the occurrence of FMI during racing with race-meet, age, and sex was evaluated by logistic regression. RESULTS: Injury type and sex-specific age distributions differed among the horses fatally injured during racing and training. Age and sex distributions of the race entrants were not independent and varied among race-meets. Overall incidence risk of racing FMI was estimated at 1.7/1,000 race entrants. Risk of racing FMI in male horses was about twofold that in female horses, and in 4-year-olds was twofold that in 3-year-olds. CLINICAL IMPLICATIONS: Age and sex-related differences in risk of incurring a FMI during racing should be considered when comparing fatal injury rates among race-meets.

Age Distribution↗

Identification of bovine Neospora parasites by PCR amplification and specific small-subunit rRNA sequence probe hybridization.

Neospora is a newly recognized genus of pathogenic coccidia, closely related to Toxoplasma gondii, that can cause abortion or congenital disease in a variety of domestic animal hosts. On the basis of the small-subunit rRNA gene sequences of Neospora spp. and other apicomplexa coccidia, oligonucleotide primers COC-1 and COC-2 were used for PCR amplification of conserved sequences of approximately 300 bp in size. A Neospora-specific chemiluminescent probe hybridized to Southern blots of amplification products from Neospora DNA but not to Southern blots with amplified DNA from the other coccidian parasites tested. A Toxoplasma-specific probe whose sequence differed from that of the probe for Neospora spp. by a single base pair was used to distinguish these parasites by specific Southern blot hybridization. The PCR system detected as few as one Neospora tachyzoite in the culture medium or five tachyzoites in samples of whole blood or amniotic fluid spiked with Neospora parasites. In addition, Neospora PCR products were successfully amplified from whole blood and amniotic fluid samples of experimentally infected bovine and rhesus macaque fetuses. These results indicate that this PCR and probe hybridization system could be a valuable adjunct to serology and immunohistochemistry for the diagnosis of Neospora infections in bovine or primate fetuses.

Abortion, Veterinary↗

Epitheliocystis infection in cultured white sturgeon (Acipenser transmontanus): antigenic and ultrastructural similarities of the causative agent to the chlamydiae.

A mild to moderate branchial epitheliocystis infection was diagnosed in subyearling (11 months old, 250-300 g) white sturgeon (Acipenser transmontanus) from a private culture facility with a 4-8% mortality in the population. Infected branchial epithelial cells contained the coccoid to coccobacillary epitheliocystis organisms, which appeared as cytoplasmic inclusions composed of a fine, homogeneous, dense, basophilic, granular material. The infected cells were variably enlarged with spherical to oval profiles and were randomly distributed throughout the branchial epithelium. The cytoplasmic inclusions stained positive with Macchiavello stain but negative with Brown and Brenn, periodic acid-Schiff, and Gimenez stains. Expression of chlamydial antigen was demonstrated within the cytoplasmic inclusions using a standard peroxidase-antiperoxidase immunohistochemical technique. Three stages of coordinated intracellular development were recognized by electron microscopy. The reticulate bodies were oval to spherical and 0.4-0.8 x 0.5-1.4 microns but often exhibited a pleomorphic and convoluted appearance because of variable membrane invaginations and evaginations suggestive of uneven fission and budding. Separate host cells contained intermediate bodies that were spherical to oval and 0.2-0.4 x 0.3-0.6 microns although often observed in the process of apparent uneven division. The presence of a cap or plaque composed of hexagonally arrayed fibrillar surface projections was initially recognized in this stage. A homogeneous population of 0.3-0.4 microns oval elementary bodies were observed separately in individual host cells. This developmental stage had a single, dense, compact, eccentrically located cytoplasmic condensation that occurred opposite to the location of the cap of hexagonally arrayed fibrillar surface projections. Morphologic characteristics of the epitheliocystis organism in these white sturgeon were similar to those previously described in other teleosts and expands the species catalogue of epitheliocystis infection. Furthermore, the ultrastructural similarities to the chalmydiae and the immunohistochemical detection of chlamydial antigen provides further evidence that the epitheliocystis agent is related to members of the Chlamydiales. Although the infection was considered mild to moderate and could not be definitively attributed to the mortality in this population, the potential adverse impact of epitheliocystis infection on sturgeon culture should be considered especially in intensive fish culture operations.

Animals↗

Dairy cattle abortion in California: evaluation of diagnostic laboratory data.

A descriptive study was undertaken on 595 dairy cattle abortion submissions to the California Veterinary Diagnostic Laboratory System from July 1, 1987, to December 31, 1989, to determine the etiologic nature and distribution (seasonal and geographical) of dairy cattle abortion in California as reflected by laboratory submissions. Univariate analysis was performed to characterize abortion-related submissions by farm and laboratory variables, and logistic regression analysis was performed to determine factors that may influence success of abortion diagnosis in the laboratory. The proportions of dairies that submitted abortion-related specimens from northern, central, and southern milksheds during the 2.5-year period were 20.3%, 15.7%, and 13.1%, respectively, and 60% of submissions were from medium-sized (200-999 cows) dairies. Submissions consisted of fetus (58%), placenta (2%), fetus and placenta (12%), and fetus, placenta, and maternal blood (0.84%); fetal tissues and uterine fluid constituted the rest. An apparent pattern in abortion submissions was indicated by a peak in submissions during the winter and summer of 1988 and 1989. Infectious agents were associated with 37.1% of submissions; noninfectious causes, 5.5%, and undetermined etiology, 57.3%. Bacterial abortion accounted for 18% of etiologic diagnoses; protozoal, 14.6%; viral, 3.2%; and fungal, 1.3%. Submissions comprising fetus, placenta, maternal blood, or their combinations were associated with a higher likelihood of definitive diagnosis for abortion than tissues, as were fresher specimens and submissions associated with the second trimester of fetal gestation.

Abortion, Veterinary↗

Evidence suggesting a point source exposure in an outbreak of bovine abortion due to neosporosis.

A Holstein dairy farm suffered an abortion outbreak due to neosporosis. Abortion losses were > 18%. Cows with the highest Neospora antibody titers were at the greatest risk of aborting. Mummified fetuses were found after the 43rd day of the outbreak. The epidemic curve was suggestive of a point source exposure, which is consistent with the hypothesis that Neospora can be spread by a definitive host.

Abortion, Veterinary↗

Diagnosis of oleander poisoning in livestock.

Since mid-1989, 37 cases of oleander poisoning in livestock have been diagnosed at the California Veterinary Diagnostic Laboratory System. The most frequent source for oleander exposure was plant clippings. Sudden death was the most common presenting complaint. Other signs reported included diarrhea, pulmonary edema, tachycardia, cardiac arrhythmias, colic, and lethargy. In the past, a presumptive diagnosis of oleander poisoning could be based only on matching clinical signs with evidence of consumption of oleander. A new 2 dimensional Thin-layer chromatography analysis of ingesta for oleandrin and an awareness of lesions in heart muscle have greatly improved the ability to diagnose oleander toxicosis.

Animal Feed↗