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- Author or Editor: M. R. Lappin x
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Summary
Sixteen pregnant queens were inoculated orally with tissue cysts of Toxoplasma gondii, and fetal membranes and offspring were examined for Τ gondii infection by bioassay in mice. Queens appeared clinically normal, although all shed Τ gondii oocysts. Toxoplasma gondii was isolated from tissues of 7 of 33 fetuses or kittens from 5 litters (at 13, 23, 26, 27, and 29 postinaculation days) from 8 queens euthanatized between 10 and 31 postinoculation days. Infection with Τ gondii was found In kittens from all 8 litters from the 8 queens that were allowed to undergo parturition and nurse their kittens. A total of 43 kittens were born to these 8 quean.
Toxoplasma gondii was isolated from tissues of 26 of 40 kittens bioassayed; in 3 kittens, tissues were not available for bioassay. Toxoplasmosis was severe in full-term kittens born to 5 queens; all 25 kittens from these litters died or were ill by 24 days of age. Anorexia, lethargy, hypothermia, and sudden death were the most common manifestations. Cytologic examination of peritoneal fluid aspirate samples and determination of hepatic-associated enzyme concentrations in affected kittens, as well as measurement of anti-T gondii antibodies in serum of kittens and queens, were helpful in the diagnosis of neonatal toxoplasmosis. Transplacental transfer of anti-T gondii antibodies was not observed in cats. Toxoplasma gondii oocysts were found in fecal samples of 3 kittens from different litters at 16, 24, and 63 days of age.
Abstract
Objective
To evaluate antigen recognition patterns of serum IgM, IgG, and IgA from queens and their kittens as a method of diagnosing neonatal toxoplasmosis.
Animals
5 pregnant queens were inoculated orally with Toxoplasma gondii tissue cysts during gestation (18 to 44 days). On various days after parturition (0 to 97), serum was obtained from queens and kittens (n = 19).
Procedure
Tissues from most kittens were assessed for T gondii infection by bioassay in mice. Serum samples were evaluated by IgM, IgG, and IgA western blot immunoassays for the presence of T gondii antibodies. Antigens recognized by kitten serum samples, but not by the corresponding queen serum sample, were considered to indicate neonatal infection with T gondii.
Results
Using the results of western blot immunoassay, 8 of 19 kittens (age, 2 to 97 days) were determined to be infected with T gondii. Western blot immunoassay results correlated well with bioassay results, identifying 7 of 8 bioassay-positive kittens. Western blot immunoassay additionally identified 1 kitten as infected, but tissues from the kitten had not been bioassayed. In each of the 5 kittens that developed clinical signs of toxoplasmosis, the diagnosis of neonatal toxoplasmosis was supported by results of the western blot immunoassays.
Conclusion and Clinical Relevance
Comparison of queen and kitten T gondii antigen recognition patterns of IgM, IgG, and IgA can be used for antemortem diagnosis of neonatal toxoplasmosis. (Am J Vet Res 1996;57: 1327-1330)
Summary
We decided to determine whether Toxoplasma gondii-specific IgM or IgG is produced locally or is deposited in the aqueous humor of T gondii-naive cats after primary or secondary inoculation with T gondii. Cats were orally inoculated with T gondii tissue cysts during weeks 0 and 36. Aqueous humor and serum Tgondii-specific IgM and IgG were measured, using ELISA, during weeks 0, 2, 4, 6, 8, 12, 16, 20, 26, 34, 38, 40, 42, 44, 48, 56, 62, 66, and 72 after primary oral inoculation. Total immunoglobulin-based Goldmann-Witmer coefficients were calculated to verify intraocular antibody production.
Toxoplasmagondii-specific IgM was not detected in the aqueous humor of any cat. Data indicate that cats have transient local production of T gondii-specific IgG in the aqueous humor after primary and secondary oral inoculations with T gondii tissue cysts.
The intraocular immune response to systemic T gondii infection may signal ocular recruitment of antigen-specific lymphocytes that function independently from the general humoral response to T gondii infection. Cautious interpretation of results that suggest intraocular production of Tgondii-specific IgG in cats with uveitis is warranted.
SUMMARY
An elisa for detection of Toxoplasma gondii-specific IgA in feline serum was developed. A group of cats (n = 7) was inoculated orally with T gondii bradyzoites. Toxoplasma gondii-specific serum IgM, IgG, and IgA responses were followed sequentially by use of the ELISA for 34 weeks. Serum IgA was detected later than IgM or IgG, and was detected in most cats on week 34 after inoculation. None of the cats was seropositive for IgA during the oocyst-shedding period. A group of client-owned cats with suspected clinical toxoplasmosis and a group of healthy cats were tested for Toxoplasma gondii-specific IgA in serum. A trend toward association of T gondii-specific IgA in serum of cats with ocular disease was observed.
Abstract
OBJECTIVE
To assess whether hyperinoculation of cats with a feline herpesvirus-1, calicivirus, and panleukopenia virus (FVRCP) vaccine could be used as a model to study interstitial nephritis and to assess humoral and cell-mediated immune responses toward vaccinal α-enolase.
ANIMALS
6 healthy young adult purpose-bred research cats.
PROCEDURES
Baseline renal cortical biopsies, whole blood, serum, and urine were collected prior to administration of a commercial FVRCP parenteral vaccine. Vaccine hyperinoculation was defined as a total of 8 vaccinations given at 2-week intervals over a 14-week period. Blood samples were collected immediately prior to each vaccination, and a second renal biopsy was performed 2 weeks after hyperinoculation (week 16). Renal histopathology, renal α-enolase immunohistochemistry, and assays to detect humoral and cell-mediated immune reactions against Crandell-Rees feline kidney (CRFK) cell lysates and α-enolase were performed. An α-enolase immunoreactivity score for renal tubules and glomeruli based on signal intensity was determined by a blinded pathologist.
RESULTS
Hyperinoculation with the vaccine was not associated with clinicopathologic evidence of renal dysfunction, and interstitial nephritis was not recognized by light microscopy in the time studied. The mean serum absorbance values for antibodies against CRFK antigen and α-enolase were significantly (P < 0.001) higher at weeks 4, 8, and 16 versus week 0. Renal tubular and glomerular α-enolase immunoreactivity scores were higher at week 16 compared to baseline.
CLINICAL RELEVANCE
Findings suggested that systemic immunological reactions occurred and renal tissues were affected by vaccine hyperinoculation; however, short-term FVRCP vaccine hyperinoculation cannot be used to study interstitial nephritis in cats.
Abstract
Case Description—A 16-week-old female Boxer that had been treated for 5 weeks with trimethoprim-sulfamethoxazole and chloramphenicol because of aspiration pneumonia was evaluated for bilaterally symmetric masses in the subcutaneous tissues of the ventral neck, in the region of the larynx.
Clinical Findings—Fine-needle aspirates were obtained from the neck masses; cytologic examination revealed well-differentiated thyroid epithelial tissue. A blood sample was collected for serum biochemical and thyroid function analyses. Mild hyperphosphatemia, severe hypercholesterolemia, mild hyperkalemia, and a mild increase in creatine kinase activity were identified. Serum concentration of total thyroxine was less than the lower reference limit, and that of thyroid-stimulating hormone was greater than the upper reference limit. Findings were consistent with a diagnosis of clinical hypothyroidism in a skeletally immature dog.
Treatment and Outcome—Treatment with trimethoprim-sulfamethoxazole was discontinued. The dog was reevaluated 3 weeks later, at which time the neck masses were markedly decreased in size. Serum concentrations of cholesterol and potassium were lower; serum concentrations of total thyroxine and thyroid-stimulating hormone were near or within respective reference ranges. Age-appropriate increases in serum phosphorus concentration and serum alkaline phosphatase activity were also detected.
Clinical Relevance—To the authors' knowledge, this is the first report of antimicrobial-induced goiter in a dog. Cytologic examination of fine-needle aspirates and interpretation of data from serum biochemical and thyroid function analyses were needed to obtain a definitive diagnosis. Practitioners should include goiter among the differential diagnoses for ventral neck swellings in young dogs receiving potentiated sulfonamide antimicrobials.
Abstract
Objective—To determine whether Ctenocephalides felis can transmit Mycoplasma haemofelis (Mhf) and Candidatus Mycoplasma haemominutum (Mhm) through hematophagous activity between cats.
Animals—11 cats.
Procedure—2 cats were carriers of either Mhf or Mhm. Nine cats had negative results via polymerase chain reaction (PCR) assay for Mhf and Mhm DNA; 3 of those cats were infected from the chronic carriers via IV inoculation of blood. At the time of maximum organism count for each of the Mycoplasma spp, 1 chamber containing 100 C felis was bandaged to the amplifier cats. Five days later, fleas, feces, larvae, or eggs from each chamber were analyzed for Mycoplasma spp DNA. Viable fleas from the chambers were allocated into new chambers (3 Mhm and 6 Mhf) and attached to naïve cats for 5 days. Cats were monitored daily for clinical signs and weekly via CBC and PCR assay for infection with Mhf or Mhm for a minimum of 8 weeks.
Results—Uptake of Mhf and Mhm DNA into fleas, feces, and, potentially, eggs and larvae was detected. Of the naïve cats fed on by Mhf-infected fleas, 1 cat transiently yielded positive PCR assay results for Mhf on 1 sampling date without clinical or hematologic changes consistent with Mhf infection.
Conclusions and Clinical Relevance—Results suggest that hematophagous transfer of Mhm and Mhf into fleas occurred and that C felis is a possible vector for Mhf via hematophagous activity. (Am J Vet Res 2005;66:1008–1012)
Abstract
Objective—To develop a broad-range 28S ribosomal DNA quantitative PCR (qPCR) assay for detection of fungal DNA in equine endometrial samples.
Sample—12 fungal samples from a clinical diagnostic laboratory and 29 samples obtained from 17 mares.
Procedures—The qPCR assay was optimized with commercially acquired fungal organisms and validated with samples obtained from the clinical diagnostic laboratory. Subsequently, 29 samples from 17 mares suspected of having fungal endometritis were evaluated via the qPCR assay and via traditional fungal culture and endometrial cytology. Amplicons from the qPCR assay were subjected to genetic sequencing to identify the organisms.
Results—The qPCR assay theoretically had a detection threshold of 2 organisms of Candida albicans. Fungal DNA was amplified from all 12 fungal samples from the commercial diagnostic laboratory. Fungal identification by use of genetic sequencing was successful for 34 of 36 amplicons from the 12 samples assayed. A fungal agent was identified via qPCR assay and genetic sequencing in all 12 samples; in contrast, a fungal agent was identified in only 8 of 12 samples via standard fungal culture and biochemical analysis. The qPCR assay detected fungal DNA in samples from 12 of 17 mares suspected of having fungal endometritis.
Conclusions and Clinical Relevance—A rapid, sensitive, and repeatable qPCR assay was developed for detection of fungal DNA from equine endometrial samples. The qPCR may prove to be clinically useful as an adjunct to microbial culture and cytologic examination to provide identification of fungal organisms in a timely manner.
Abstract
Objective—To evaluate orally administered famciclovir for treatment of cats with experimentally induced disease attributable to feline herpesvirus type-1 (FHV-1).
Animals—16 nonvaccinated specific-pathogen-free cats.
Procedures—Cats were treated orally with famciclovir (90 mg/kg; n = 10) or a similar volume of lactose (400 mg; 6) 3 times/d for 21 days. Cats were inoculated with FHV-1 and administered the first treatment dose on day 0. Disease score; weight; results of urinalysis, serum biochemical analysis, and CBC; histologic conjunctivitis score; herpetic DNA shedding; goblet cell density; anti-FHV-1 antibody concentration; and plasma penciclovir concentration were measured.
Results—On days 4 to 18 following inoculation, disease scores were lower in famciclovir-treated cats than in lactose-treated cats. Lactose-treated cats decreased in weight during the first 7 days after inoculation, but famciclovir-treated cats increased in weight throughout the study. Percentage change in weight was greater in famciclovir-treated cats on days 7 and 14 than in lactose-treated cats. Serum globulin concentration was lower on days 3 through 9, conjunctivitis histologic score was lower on day 14, herpetic DNA was shed less frequently throughout the study, goblet cell density was greater on day 21, and circulating anti-FHV-1 antibody concentration at study end was lower in famciclovir-treated cats, compared with these measurements in lactose-treated cats. Approximate peak plasma penciclovir concentration was 2.0 μg/mL.
Conclusions and Clinical Relevance—Famciclovir administration improved outcomes for systemic, ophthalmic, clinicopathologic, virologic, and histologic variables in cats experimentally infected with FHV-1. Adjunctive topical mucinomimetic and antimicrobial treatments may also be necessary.
