The objectives of this study were to characterize the ocular and systemic adverse effects, and systemic pharmacokinetics of topical 0.1% diclofenac. This was investigated in 8 healthy cats using a blinded, randomized, placebo-controlled, cross-over design. Drops were administered bilaterally 4 times daily for 7 days. Ocular, hepatic and renal variables were measured at various timepoints. Pharmacokinetic sampling occurred on Days 1 and 7. Treated animals were 8 times more likely to develop conjunctival hyperemia than control animals (p=0.0161). Pharmacokinetic analysis showed that accumulation occurs with repeated dosing. Topical 0.1% diclofenac treatment did not have any significant effect on hepatic or renal function, other than reduction GFR in the second phase of the study (p=0.0013). In conclusion, topical 0.1% diclofenac appears to be safe in healthy cats causing only mild ocular irritation. Careful patient selection may be indicated as systemically-absorbed diclofenac may be associated with reduction in GFR in volume-contracted states. / Ontario Veterinary College Pet Trust Fund

The exact role of mycoplasma in feline ocular and respiratory disease is not yet understood. The results of previous studies are contradictory in this regard. There is some evidence to suggest that M. felis has a pathogenic role in such diseases, but it is inconclusive. / The aim of this study was to investigate the prevalence and anatomical distribution of mycoplasmas in a population of shelter cats, to determine which species were present, and establish the association of their presence with ocular or respiratory disease. / The prevalence of mycoplasma in the 110 cats examined was 71.8%, as determined by in vitro culture. Mycoplasma was most commonly isolated from the pharynx, followed by the bronchus and conjunctiva. In infected cats, mycoplasmas were likely to be isolated from multiple anatomical sites. / The polymerase chain reaction (PCR) was used to amplify part of the 16S rRNA gene, and the mutation scanning technique non-isotopic single-strand conformation polymorphism (SSCP) was utilised to delineate mycoplasma isolates based on nucleotide sequence variation. PCR-SSCP proved to be a useful method to screen large numbers of samples for variation and to group them according to species. / The species of mycoplasma identified by nucleotide sequencing were M. felis and M. gateae. It was not determined whether it was possible to differentiate between M. gateae and M. arginini based on SSCP profile results with the target DNA region used due to their almost identical nucleotide sequence. This group of M. gateae/M. arginini served as a useful non-pathogenic comparison group to M. felis. / There was no statistically significant difference between M. felis and the M. gateae/M. arginini group with respect to prevalence or anatomic distribution. There was no evidence of any association of mycoplasma with disease linked to any of the anatomic locations studied. / Mycoplasmas were isolated from the lower respiratory tract in 42.7% of cats. The isolation of mycoplasmas from the lower respiratory tract of healthy cats has been reported once, but this is the first report of M. felis being isolated from this location in healthy cats. This finding indicates that the isolation of mycoplasmas from the lower respiratory tract is not sufficient evidence to implicate a role in respiratory disease. / Mycoplasmas were not significantly involved in ocular or respiratory disease in the population of cats studied. More likely, they are commensal organisms in the conjunctiva, pharynx and bronchus. Whether they are capable of playing an opportunistic role in disease, or what conditions may facilitate such a role remains to be determined.

Zugl.: Giessen, Univ., Diss., 2009

Thesis (Ph.D.) - University of Queensland, 2004. / Includes bibliography.

Submitted by Beatrice Ingrid Macente null (beatrice.vetuel@yahoo.com.br) on 2017-12-13T17:24:54Z No. of bitstreams: 1 Tese de doutorado - FINAL.pdf: 1497835 bytes, checksum: ec09acb806dd08292a1d4ec818d596e7 (MD5) / Approved for entry into archive by Karina Gimenes Fernandes null (karinagi@fcav.unesp.br) on 2017-12-13T18:29:08Z (GMT) No. of bitstreams: 1 macente_bi_dr_jabo.pdf: 1454157 bytes, checksum: 970c2725b03ed803688cfef9261d801c (MD5) / Made available in DSpace on 2017-12-13T18:29:08Z (GMT). No. of bitstreams: 1 macente_bi_dr_jabo.pdf: 1454157 bytes, checksum: 970c2725b03ed803688cfef9261d801c (MD5) Previous issue date: 2017-11-06 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Esta pesquisa teve por justificativa ampliar os estudos acerca da criopreservação de tecido gonadal e células espermáticas de gatos domésticos, podendo servir de modelo experimental para felinos selvagens. Objetivou-se comparar os efeitos da criopreservação sobre diferentes tamanhos de fragmentos testiculares, empregando dois crioprotetores. Utilizaram-se os testículos de 31 gatos domésticos, submetidos à orquiectomia eletiva. Os testículos foram pesados e apenas os que apresentarem pesos entre 1 e 2 g foram empregados. O tecido testicular foi dissecado e cortado em fragmentos de tamanhos pré-determinados (0,3cm3 e 0,5cm3). Uma amostra foi direcionada para as avaliações a fresco: integridade de membrana e do DNA; histopatologia; incidência de apoptoses por imuno-histoquímica; e índice de peroxidação lipídica - TBARS. A criopreservação foi feita em meio Tris-gema Equex STM suplementado com 3% de crioprotetores (glicerol e propanediol) através da técnica congelação rápida. Após a descongelação, foram realizados os mesmos testes iniciais. Os primeiros resultados obtidos com os 12 gatos iniciais apontaram na avaliação histomorfológica e pelo teste de lipoperoxidação lipídica que o glicerol foi mais eficiente que o propanediol na criopreservação de fragmentos testiculares de gatos domésticos de 0,5cm3 (Capítulo 2). No experimento seguinte, avaliaram-se os fragmentos testiculares de outros 31 gatos domésticos, seccionados nos mesmos tamanhos e criopreservados nas mesmas condições do experimento anterior, empregando-se 3% glicerol ou 3% propanediol no meio diluente Tris-gema Equex STM, utilizando a técnica de congelamento rápido, seguindo das avaliações pelos testes para dano ao DNA dos espermatozoides por meio da acridina laranja; e análise dos fragmentos teciduais semi-quantitativamente por histomorfologia e imuno-histoquímica. A avaliação imuno-histoquímica foi mais eficiente em verificar os danos às células tubulares seminíferas, sendo possível observar a marcação diferenciada da caspase presente no citoplasma e núcleo das células espermáticas, bem como na cabeça dos espermatozoides. Por meio da caspase foram verificado que os fragmentos frescos já apresentavam danos consideráveis as células teciduais e que não diferiram dos achados para os fragmentos criopreservados, demonstrando a eficiência do protocolo de congelação adotado, sem a observação de superioridade entre os dois tamanhos de fragmentos ou entre os crioprotetores. Contudo, a avaliação apenas do espermatozoides tanto pela técnica de acridina laranja, como pela imuno-histoquímica, pode-se verificar a inferioridade do propanediol em relação ao glicerol para fragmentos de 0,5 cm³ (Capítulo 3). Novas pesquisas que possam complementar os testes realizados neste estudo, como o cultivo dos fragmentos, a fertilização in vitro por meio de ICSI, ou ainda, o uso dos fragmentos para xenotrasnplantes poderiam avaliar com maior acurácia os achados das pesquisas reportadas nesta tese. / This research had the justification to broaden the studies about the cryopreservation of gonadal tissue and sperm cells of domestic cats, and could serve as an experimental model for wild cats. The objective of this study was to compare the effects of cryopreservation on different sizes of testicular fragments using two cryoprotectants. The testicles of 31 domestic cats submitted to elective orchiectomy were used. The testes were weighed and only those weighing between 1 and 2 g were used. The testicular tissue was dissected and cut into fragments of predetermined sizes (0.3cm3 and 0.5cm3). A sample was directed to fresh evaluations: membrane integrity and DNA; histopathology; incidence of apoptosis by immunohistochemistry; and lipid peroxidation index - TBARS. Cryopreservation was done on Tris-egg yolk Equex STM medium supplemented with 3% cryoprotectants (glycerol and propanediol) by the rapid freezing technique. After thawing, the same initial tests were performed. The first results obtained with the 12 initial cats showed that glycerol was more efficient than propanediol in the cryopreservation of testicular fragments of domestic cats of 0.5 cm3 (Chapter 2). In the following experiment, the testicular fragments from 31 other domestic cats, sectioned in the same sizes and cryopreserved under the same conditions as in the previous experiment, were evaluated using 3% glycerol or 3% propanediol in the Tris- egg yolk Equex STM medium, using rapid freezing technique, followed by evaluations for DNA damage of the spermatozoa by acridine orange; and analysis of the tissue fragments semi-quantitatively by histomorphology and immunohistochemistry. The immunohistochemical evaluation was more efficient in verifying the damage to the seminiferous tubular cells, being possible to observe the differentiated marking of the caspase present in the cytoplasm and nucleus of the sperm cells, as well as in the spermatozoa head. By caspase, it was verified that the fresh fragments already presented considerable damage to the tissue cells and did not differ from the findings for the cryopreserved fragments, demonstrating the efficiency of the adopted freezing protocol, without the observation of superiority between the two sizes of fragments or between the cryoprotectants. However, the evaluation of spermatozoa by both acridine orange and immunohistochemistry, the inferiority of propanediol to glycerol can be verified for fragments of 0.5 cm³ (Chapter 3). New research that could complement the tests performed in this study, such as the culture of fragments, in vitro fertilization using ICSI, or the use of fragments for xenotransplantation could more accurately assess the findings of the research reported in this thesis.

Master of Science in Biomedical Sciences / Department of Clinical Sciences / Jessica Meekins / Objectives- To determine the effect of oral robenacoxib on experimentally-induced anterior uveitis, and to evaluate the ability of robenacoxib to cross an intact blood-aqueous barrier. Animals- Twelve healthy adult domestic shorthair cats. Procedures- Cats in the treatment group (n=6) received oral robenacoxib (1.51 ± 0.36 mg/kg ) once daily beginning 1 day before experimental induction of uveitis by anterior chamber paracentesis (ACP) and continuing 1 day after paracentesis. Anterior chamber paracentesis was performed using a 30 g needle attached to a 1 mL syringe, and 100 µL of aqueous humor were aspirated over 3-5 seconds. Anterior chamber fluorophotometry was performed in both eyes of each cat immediately before ACP (time 0), and at 6, 24, and 48 hours after ACP. An independent t-test was used to compare percent fluorescein increase in treatment versus control cats at each time point. Values of p<0.05 were considered significant. Concentrations of robenacoxib in aqueous humor were measured using liquid chromatography and mass spectrometry. Results- There was no statistically significant difference between the ACP and control eye at time 0 (p=0.322). When comparing the percent fluorescein increase between treatment and control groups, there was no statistically significant difference at any time point (p>0.05). Robenacoxib was present in small but detectable levels in 5/6 cats in the treatment group. Conclusions and clinical relevance- Administration of oral robenacoxib did not significantly lessen experimentally-induced anterior uveitis in normal cats, as assessed by fluorophotometry. Low concentrations of aqueous humor robenacoxib were detectable in the majority of cats receiving the drug.

O coronavírus felino (FCoV) ocorre sob uma grande diversidade gênica de amostras e é classificado em dois patotipos: o coronavírus felino entérico (FECoV) e o vírus da peritonite infecciosa felina (FIPV). O patotipo FIPV é altamente virulento e responsável pelo desenvolvimento de uma doença altamente fatal, denominada de peritonite infecciosa felina (PIF). Já o FECoV apresenta-se amplamente disseminado na população felina e é responsável na maioria das vezes por infecção assintomática. Atualmente, nenhum marcador gênico conhecido é capaz de diferenciar os patotipos FECoV de FIPV. O presente estudo foi dividido em dois capítulos. No primeiro capítulo, objetivou-se avaliar a diversidade molecular do gene da membrana (M) em 190 amostras provenientes de 5 gatos sem manifestações de PIF (PIF-) e de 10 gatos com manifestações clínicas e histopatológicas de PIF (PIF+). Com esse estudo, conclui-se que tanto a hipótese de mutação in vivo do FECoV para FIPV, quanto a hipótese de transmissão entre gatos do patotipo FIPV são plausíveis. No segundo capítulo, com o objetivo de avaliar a diversidade dos genes 3a-c, E e M foram sequenciados clones de amplicons para estes genes obtidos, de 6 gatos PIF+ e 2 gatos PIF-. Os genes 3a-c, E e M apresentaram diversidade gênica que confere a constituição das quasiespécies de coronavírus felino com probabilidade de emergência do patotipo de alta virulência, mas de um modo hospedeiro-específico. Com o segundo estudo, conclui-se que as linhagens FIPV de coronavírus felino apresentaram a proteína 3c truncada, sendo o gene 3c o único marcador de patotipo dos FCoVs observado dentre os genes estudados. / Feline coronavirus (FCoV) occurs as a large genic diversity of strains and is classified as two pathotypes: feline enteric coronavirus (FECoV) and feline infectious peritonitis virus (FIPV). The FIPV pathotype is highly virulent and responsible for the onset of a highly fatal disease named feline infectious peritonitis (FIP), while the FECoV pathotype is widely disseminated in feline populations leading mostly to asymptomatic infections. No genic marker is currently known to differentiate the FECoV and FIPV pathotypes. This study has been divided in two chapters. In the first chapter, the aim was to evaluate the molecular diversity of the membrane (M) gene in 190 samples from 5 cats without FIP (FIP-) and 10 cats with clinical and histopathological evidence of FIP (FIP+). The conclusion of this study is that both the in vivo mutation hypothesis in the FECoV-to-FIP direction and the hypothesis of FIPV transmission amongst cats are plausible. In the second chapter, aimed to evaluate the diversity of genes 3a-c, E and M, clones of amplicons for these genes were obtained and sequenced from samples from six FIP+ and 2 FIP- cats. Genes 3a-c, E and M show a genic diversity that results in a quasispecies constitution of FCoV that leads to the probability of the emergence of the highly virulent pathotype in a host-specific way. The conclusion of this second study is that FIPV lineages show a truncated form of the 3C protein, making the 3c gene the only pathotype marker for FCoV observed amongst the genes studied herein.

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