FC‐14 Clonality studies of feline cutaneous lymphocytosis

A previous study described cutaneous lymphocytosis (CL) in 23 cats. The process resembles cutaneous pseudolymphoma in humans, a heterogeneous group of benign reactive proliferations of well‐differentiated lymphocytes in the skin of humans. Morphological and immunophenotypic characteristics do not offer reliable criteria to accurately predict the clinical outcome of feline CL or pseudolymphoma in humans. Presence of clonal cell populations is more consistent with a neoplastic process. In a previous study, feline CL lesions (20 cats) were evaluated for clonality using PCR, and only two cats had monoclonal T‐cell populations. Because false‐negative results may occur, the purpose of this study was to repeat the PCR using a revised primer set based on analysis of additional feline T‐cell receptor γ (TCRγ) sequences. DNA was isolated from 29 skin lesions and six internal organs of 20 cats. DNA integrity was assessed by glyceraldehyde‐3‐phosphate dehydrogenase PCR. Polymerase chain reaction clonality was performed using the revised primer set specific for feline TCRγ, and duplicate samples were evaluated. The PCR products were assessed by heteroduplex analysis. Clonal rearrangement of TCRγ was detected in 14 cats (24 of 35 tissues: 21 of 29 skin lesions and three of six internal organs); eight of these cats are still alive and six were euthanized. Monoclonal populations were seen in three of five cats that had involvement of internal organs. These findings indicate that feline CL is best considered as a slowly progressive process which may be reactive, but often evolves into a low‐grade indolent lymphoma. Funding: George H. Muller Fund for Research in Dermatology.     

Prevalence of selected zoonotic and vector-borne agents in dogs and cats in Costa Rica

To estimate the prevalence of enteric parasites and selected vector-borne agents of dogs and cats in San Isidro de El General, Costa Rica, fecal and serum samples were collected from animals voluntarily undergoing sterilization. Each fecal sample was examined for parasites by microscopic examination after fecal flotation and for Giardia and Cryptosporidium using an immunofluorescence assay (IFA). Giardia and Cryptosporidium IFA positive samples were genotyped after PCR amplification of specific DNA if possible. The seroprevalence rates for the vector-borne agents (Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis, and Anaplasma phagocytophilum) were estimated based on results from a commercially available ELISA. Enteric parasites were detected in samples from 75% of the dogs; Ancylostoma caninum, Trichuris vulpis, Giardia, and Toxocara canis were detected. Of the cats, 67.5% harbored Giardia spp., Cryptosporidium spp., Ancylostoma tubaeforme, or Toxocara cati. Both Cryptosporidium spp. isolates that could be sequenced were Cryptosporidium parvum (one dog isolate and one cat isolate). Of the Giardia spp. isolates that were successfully sequenced, the 2 cat isolates were assemblage A and the 2 dog isolates were assemblage D. D. immitis antigen and E. canis antibodies were identified in 2.3% and 3.5% of the serum samples, respectively. The prevalence of enteric zoonotic parasites in San Isidro de El General in Costa Rica is high in companion animals and this information should be used to mitigate public health risks.     

Acquired Thermo‐Tolerance and Trans‐Generational Heat Stress Response at Flowering in Rice

In rice, pre‐exposure to sublethal treatment followed by harsh lethal treatment is known to improve tolerance of different abiotic stresses at the vegetative stage within and across generations. Our major aim was to test the phenomenon of thermo‐tolerance at flowering across (trans)‐generations and within generation using rice cultivars contrasting for heat stress tolerance at flowering. To test trans‐generational response, plants were exposed to higher temperature at flowering stage and seeds obtained from previous generations were exposed to heat stress during flowering, which recorded significantly lower fertility when exposed to the same degree of stress in their subsequent generations. A pre‐acclimation to moderately high acclimating temperatures imposed over three different durations during the vegetative and initial reproductive stage showed positive response in the tolerant N22, particularly under severe heat stress (40 °C). This finding indicates the possibility of acquiring ameliorative thermo‐tolerant mechanisms at anthesis, restricted to tolerant genetic backgrounds to combat subsequent harsh conditions within the same generation. However, trans‐generational memory was ineffective in mitigating spikelet sterility losses in both tolerant and susceptible backgrounds. Rice is extremely sensitive to heat stress during flowering; hence, similar exercise across other crops of interest needs to be carried out before generalizing conclusions.     

Humoral immune responses of black-footed penguins (Spheniscus demersus) after DNA-mediated immunization with a beta-galactosidase reporter gene.

Humoral immune responses of black-footed penguins (Spheniscus demersus) to DNA-mediated immunization with a beta-galactosidase reporter gene expression plasmid were evaluated. Six male and 6 female adult penguins received either test plasmid, pCMV-beta, containing the beta-galactosidase gene or control plasmid, pCI, lacking a gene for expression. Three birds from each group were used previously in a diluent control group and given one injection of sterile saline. All samples were screened for anti-beta-galactosidase antibodies by indirect enzyme-linked immunosorbent assay with anti-chicken immunoglobulin G as secondary antibody. Antibodies to beta-galactosidase were detected in the sera of pCMV-beta-inoculated penguins, with a peak response on day 21. Antibody titers of the test plasmid group versus both control groups on days 21, 28, and 42 differed significantly. These results demonstrate that black-footed penguins can be safely transfected with the gene encoding beta-galactosidase and will mount a humoral response against the in vivo-expressed protein. Knowledge from this initial study can be applied to the development of DNA-mediated vaccines against specific infectious diseases of penguins.     

Pharmacokinetic evaluation of a slow-release cefotaxime suspension in the dog and in sheep

Three Merino ewes were given cefotaxime IM, and 3 were given cefotaxime subcutaneously (50 mg/kg of body weight each); each dose was suspended in 6 ml of oil. Five dogs were also given an oily suspension of cefotaxime subcutaneously (SC) (50 mg/kg of body weight). The plasma concentrations (Cp) and pharmacokinetic data obtained after cefotaxime in the oily suspension was injected IM and SC were compared with data from the same animals after they were given an aqueous solution of cefotaxime by the same routes. Key pharmacokinetic values obtained after cefotaxime was administered IV to sheep and to dogs are discussed. Mean peak Cp (Cpeak) in sheep when given the oily suspension IM was approximately 53 micrograms/ml at 0.18 to 0.40 hour, and that value in sheep given the aqueous preparation was 62 micrograms/ml 0.08 to 0.18 hour. Mean Cpeak values after the oily suspension and the aqueous preparation were injected SC were 11.0 micrograms/ml (between 0.8 and 1 hour) and 51 micrograms/ml (between 0.25 and 1 hour), respectively. Bioavailabilities were approximately 70% after IM injection was done and 90% after SC injection was done. The beta-plasma half-lives were 0.7 hour after IM injection was done and 2.9 hours after SC injection was done. Mean Cpeak in dogs when given the oily suspension SC was 30 micrograms/ml at 1.0 hour, and when dogs were given the aqueous preparation SC, Cpeak was 27 micrograms/ml at 0.6 hour. Absorption was virtually complete after the oily suspension and aqueous preparations were given.(ABSTRACT TRUNCATED AT 250 WORDS)