A gengivoestomatite linfoplasmocítica felina (GELF) é uma síndrome clínica frequentemente observada na medicina de felinos. Trata-se de uma síndrome provavelmente multifatorial que envolve o sistema imune dos animais, agentes infecciosos e não infecciosos, de modo que a etiologia ainda não foi totalmente determinada. Diversos agentes infecciosos como o calicivírus felino (FCV), herpesvírus felino (FHV), vírus da leucemia felina (FeLV) e vírus da imunodeficiência felina (FIV) têm sido investigados. Este trabalho apresenta uma caracterização clínico, patológica, imuno-histoquímica e molecular de 27 felinos com GELF. Quanto à raça, 85,2% eram sem raça definida, 7,4% Siamês e 7,4% Maine Coon. Os machos corresponderam a 67% dos casos e fêmeas 33%. Quanto à idade, 10% apresentavam entre 1 e 3 anos, 20% entre 4 e 6 anos, 30% entre 7 e 9 anos, 15% entre 10 e 12 anos e 25% tinham mais do que 13 anos. A idade média dos felinos foi de 8,8 anos. Os principais sinais clínicos observados foram: disfagia (83,3%), halitose (72,2%), sialorreia (44,4%), emagrecimento (38,8%), intenso desconforto oral (33,3%), sangramento oral (22,2%), pelos opacos e quebradiços (11,1%) e intensa dispneia (5,5%). Macroscopicamente, eram lesões bilaterais no arco palatino e que se estendiam até a face lateral da base da língua. Em 77,8% elas eram difusas, de aspecto proliferativo, coloração vermelho intenso, friável e que sangravam facilmente durante a manipulação e em 22,2% as lesões eram multifocais a coalescentes, por vezes formando múltiplas vesículas, no arco palatino de aspecto avermelhado e edematoso. Microscopicamente, 14,8% apresentaram uma inflamação moderada (grau 2) e 85,2% dos animais apresentaram uma inflamação grave (grau 3). Os antígenos virais do FeLV foram identificados através da técnica de IHQ no epitélio e células do infiltrado inflamatório de 29,6% dos animais que apresentavam a GELF. Os antígenos virais do FIV foram identificados nas células do infiltrado inflamatório em 3,7% dos casos. Os antígenos virais do FCV não foram identificados nas lesões de GELF. Dezoito por cento dos animais foram positivos para FIV e 37% foram positivos para FeLV no exame de PCR. / Feline chronic gingivostomatitis (FCGS) is a clinical syndrome frequently observed in feline medicine. This is probably a multifactorial syndrome which involves the immune system of animals, infectious and noninfectious agent, though the causes have not been fully determined. Several infectious agents such as feline calicivirus (FCV), feline herpesvirus (FHV), feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) have been investigated. To determine clinical, pathological, immunohistochemical (IHC) and molecular characteristics of 27 cats with FCGS was developed. Regarding race, 85.2% were mixed breed, 7.4% Siamese and 7.4% Maine Coon. Males were 67% of cases and females 33%. Regarding age, 10% were between 1 and 3 years, 20% between 4 and 6 years, 30% between 7 and 9 years old, 15% between 10 and 12 years and 25% were older than 13 years. The average age of the cats was of 8.8 years. The main clinical signs were: dysphagia (83.3%), halitosis (72.2%), sialorrhoea (44.4%), weight loss (38.8%), intense oral discomfort (33.3%), oral bleeding (22.2%), hair opaque and brittle (11.1%) and severe dyspnea (5.5%). Grossly, the lesions were bilateral in the palatal arch and extending to the lateral side of the base of the tongue. In 77.7% they were diffuse, proliferative, intense red, crispy and bled easily during handling and in 22.3% lesions were multifocal to coalescing, sometimes forming multiple vesicles, the palatal arch reddish and edematous appearance. Microscopically, 14.8% showed moderate inflammation (grade 2) and 85.2% of the animals showed severe inflammation (grade 3). FeLV viral antigens were identified by IHC reaction in the epithelium and inflammatory cells of 29.6% of the animals showing FCGS. FIV viral antigens were identified in inflammatory cells in 3.7% of cases. FCV viral antigens were not identified in lesions of FCGS. Eighteen per cent of animals were positive for FIV and 37% were positive for FeLV in the PCR.

Shelters often make euthanasia or adoption decisions based on the results of FeLV-FIV point-of-care tests but given the low estimated prevalence of these diseases and imperfect test performance, this might not be a good practice because of diagnostic error. The objectives of this study were to determine the true prevalence of FeLV and FIV in apparently healthy and sick shelter cats in Mississippi, estimate predictive value of the Zoetis Witness FeLV-FIV Rapid ImmunoMigration test results at the estimated true prevalences through Bayesian latent class modeling, and formulate testing recommendations for shelters. One chapter will review the literature on FeLV and FIV. The bulk of this thesis will focus on determining the true prevalence of retroviral infection in Mississippi shelter cat populations. The last chapter will use Bayesian modeling to estimate test performance and predictive value of test results in healthy and sick shelter cat populations.

Feline coronavirus (FCoV) is endemic in cat populations worldwide. Persistently, subclinically infected cats play a significant role in spreading the infection. Testing fecal samples of cats may facilitate efforts to decrease the viral burden within a population. Real-time RT-PCR is highly sensitive and specific for the detection of FCoV but must be performed in a fully equipped laboratory. A simple and accurate assay is needed to identify FCoV at the point-of-need. The aim of this study was to develop a rapid FCoV detection assay based on isothermal amplification technology, i.e., reverse transcription-recombinase polymerase amplification (RT-RPA). Primers were designed to target the highly conserved 3′ untranslated region of the 7b gene. Running on a constant temperature of 42 °C, reverse transcription as well as DNA amplification and detection was achieved in a maximum of 15 min. A probit analysis revealed a detection limit of 58.5 RNA copies/reaction. For cross-detection, nucleic acids from 19 viruses were tested. Both RT-RPA and real-time RT-PCR showed cross-detection with canine coronavirus and transmissible gastroenteritis virus, but not with other pathogens. To evaluate clinical performance, RNA was extracted from 39 fecal samples from cats. All samples were tested simultaneously with real-time RT-PCR resulting in a RT-RPA sensitivity and specificity of 90.9% and 100%, respectively. RT-RPA can be considered a promising simple method for rapid detection of FCoV.

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Cette étude prospective, randomisée et à l’aveugle a évalué la validité, la réactivité et fiabilité de l’échelle de grimaces félines (Feline Grimace Scale; FGS) chez les chatons. Trente-six jeunes chattes en santé (âgées de 10 semaines à 6 mois) étaient filmées avant puis 1 et 2 h après ovariohystérectomie. La procédure a été effectuée avec un protocole d’anesthésie injectable (sans opioïde), avec ou sans analgésie multimodale. Les chatons en douleur étaient également filmés avant et 1h après avoir reçu une analgésie de secours (buprénorphine 0.02 mg/kg IM). Quatre évaluateurs, aveugles aux conditions expérimentales, ont évalué deux fois à cinq semaines d’intervalle les expressions faciales sur 111 images extraites des vidéos. Les cinq unités d’action (action unit; AU) de la FGS ont été évaluées (position des oreilles, serrage orbital, tension du museau, position des moustaches, position de la tête; avec un score possible de 0 à 2 pour chacune). La validité de construit, la réactivité et la fiabilité inter- et intra-évaluateur de la FGS ont été analysées en utilisant un modèle linéaire avec correction Benjamini–Hochberg, un test Wilcoxon signed-rank et un coefficient de corrélation intra-classes unique (ICCsingle), respectivement (P <0.05). Les ratios des scores FGS totaux (médiane [étendue interquartile, EI]) étaient augmentés 1 et 2 h après l’ovariohystérectomie (médiane [EI] : 0.30 [0.20–0.40] et 0.30 [0.20–0.40], respectivement) comparativement à la mesure de base (médiane [EI] : 0.10 [0.00–0.30]) (P <0.001), et inférieurs après l’analgésie (médiane [EI] : 0.40 [0.20–0.50]) qu’avant son administration (médiane [EI] : 0.20 [0.10–0.38]) (P <0.001). Pour la fiabilité inter-évaluateur, les ICCsingle des ratios des scores FGS totaux étaient 0.68 et compris entre 0.35 et 0.70 pour chaque AU, individuellement. Pour la fiabilité intra-évaluateur, les ICCsingle des ratios des scores FGS totaux étaient compris entre 0.77–0.91 et 0.55–1.00 pour chaque AU. La FGS est un outil d’évaluation de la douleur aiguë valide et réactif chez les chatons avec une fiabilité inter-évaluateur modérée et intra-évaluateur bonne à excellente. / This prospective, randomized, blinded study investigated the construct validity, responsiveness and reliability of the Feline Grimace Scale (FGS) in kittens. Thirty-six healthy female kittens (aged 10 weeks to 6 months) were video recorded before, 1 and 2 h after ovariohysterectomy using an opioid-free injectable anesthetic protocol with or without multimodal analgesia. Painful kittens were additionally filmed before and 1 h after administration of rescue analgesia (buprenorphine 0.02 mg/kg IM). One hundred eleven facial images collected from video recordings were randomly scored by 4 observers, blinded to treatment groups and time points, twice with a 5 weeks interval using the FGS. The five action units (AU) of the FGS were scored (ear position, orbital tightening, muzzle tension, whiskers position and head position; 0–2 each). Construct validity, responsiveness, inter- and intra-rater reliability of the FGS were evaluated using linear models with Benjamini–Hochberg correction, Wilcoxon signed-rank test and single intra-class correlation coefficients (ICCsingle), respectively (P <0.05). The FGS total ratio scores were higher 1 and 2 h after ovariohysterectomy (median [interquartile range, IQR]: 0.3 [0.20–0.40], and 0.30 [0.20–0.40], respectively) than at baseline (median [QR]: 0.10 [0.00–0.30]) (P <0.001), and lower after the administration of rescue analgesia (median [QR]: 0.40 [0.20–0.50]) than before (median [QR]: 0.20 [0.10–0.38]) (P <0.001). The inter-rater ICCsingle was 0.68 for the FGS total ratio scores and 0.35–0.70 for AUs considered individually. The intra-rater ICCsingle was 0.77–0.91 for the FGS total ratio scores and 0.55–1.00 for AUs considered individually. The FGS is a valid and responsive acute pain scoring instrument with moderate inter-rater reliability and good to excellent intra-rater reliability in kittens.

Master of Science / Department of Clinical Sciences / Amy Rankin / Objective - To establish the aqueous humor flow rate in normal cats via fluorophotometry, utilizing a noninvasive method previously established in other species and to evaluate the effect of topical 2% dorzolamide on aqueous humor flow rate and intraocular pressure (IOP) in normal cats. Animals - 20 clinically normal domestic shorthair cats. Procedures – Topical administration of 10% sodium fluorescein was performed using a 3-drop protocol to establish its use in this species. Aqueous flow was measured using fluorophotometry in the right and left eyes. The subjects were then divided into 2 groups: the first received topical 2% dorzolamide (Trusopt®) and the second received topical artificial tear solution (control). The study was divided into two phases: a 3 day acclimation phase (no treatments given) and a 5 day treatment phase (treatments given three times daily). IOP measurements were taken at 7am, 10am, 1pm, 5pm, and 9pm throughout all phases of the study. Fluorophotometry was performed to measure the aqueous flow rate just prior to and at the end of the treatment phase (days 3 and 9 respectively). Results - The calculated aqueous humor flow rate for normal cats in the right, left, and both eyes was 5.94 ± 2.30 μl/min, 5.05 ± 2.06 μl/min, and 5.51 ± 2.21 μl/min, respectively. No significant differences were noted between the right and left eyes. In the dorzolamide group, the average flow rate during treatment (3.47 + 1.50 μl/min ) was significantly lower than prior to treatment (5.9 + 2.20 μl/min)(P < 0.001). The mean IOP during treatment (11 + 3 mmHg) was significantly lower than the mean IOP prior to treatment (15 + 3 mmHg)(P < 0.001). In the control group, there were no significant differences in aqueous humor flow or IOP values before or during treatment. Conclusions - The technique utilized for this study met the standard for accurate fluorophotometric calculation of aqueous humor flow. The average aqueous humor flow rate for normal cats calculated in this study was 5.51 ± 2.21 μl/min. Topical 2% dorzolamide significantly lowers aqueous humor flow (a 41% reduction) and IOP (a 26% reduction) in normal cats.

Feline oral squamous cell carcinomas (FOSCCs) are locally aggressive tumours and a common cause of mortality and morbidity. Current treatment options are rarely successful and animals are frequently euthanised upon diagnosis due to their grave prognosis. Epidermal Growth Factor Receptor (EGFR) is a tyrosine kinase receptor which is frequently dysregulated in SCC of the head and neck (HNSCC) in man. Recent advances in human medicine have identified EGFR as a therapeutic target in HNSCC. In this study the role of EGFR in FOSCC was investigated. Sixty seven biopsy samples were immunohistochemically labelled for EGFR and Ki67, a proliferation marker. The tyrosine kinase region of feline EGFR was cloned and sequenced, and six small interfering RNAs (siRNAs) targeting the tyrosine kinase region were developed. The most effective siRNA as well as an EGFR specific tyrosine kinase inhibitor, gefitinib, was then used on a feline SCC cell line (SCCF1), and the effect of EGFR targeting alone, or in combination with irradiation, on the cell line was determined. The majority of the biopsy samples were labelled positively for EGFR and Ki67, and high proliferation corresponded with poor prognosis. The siRNA caused reduction in EGFR mRNA by Real-Time Polymerase Chain Reaction and protein levels as assessed by western blot analysis. Reduced cell proliferation and migration were also observed by proliferation assays and scratch assays respectively. Combining EGFR knockdown with irradiation caused an additive effect on the ability of the cell line to form colonies. These results support the role of EGFR as a potential therapeutic target in FOSCCs.

This research focused on three areas of interest regarding feline amputees; owner perception of how their cats adapt to limb amputation, the possibility of phantom sensation, and changes in gait. In general owners felt their cats had an acceptable quality of life; however a proportion believed their cat experienced pain. Anecdotally, owners reported that their cats continued to attempt to use the missing limb following amputation. This prompted the investigation of noninnate forelimb behaviours potentially indicative of phantom sensation; such behaviours apparently persist for months or years after amputation. The impact of phantom sensation on feline welfare is not known, although in humans phantom sensation is a risk factor for phantom pain. Alterations in gait and posture in humans are associated with pain and osteoarthritis. The kinetic changes in feline gait were assessed using a pressure sensitive walkway; this provided its own challenges as the software was designed for large bipeds. As such, a proportion of this work was devoted to developing methods to reliably extract data for small quadrupeds. The results detail how feline amputees alter their weight distribution and paw placement when moving. Observation of amputee cats suggests that they move their limbs differently to those of normal cats, and this was confirmed in a kinematic study using markers to track the motion of each limb. Prior to this research very little was known about how cats coped with limb amputation. The results will better inform the veterinary profession and owners of feline amputees about expected changes, and potentially inform future work on the impact of limb amputation on the welfare of cats. On balance, although there may be some areas of concern, the welfare of cats is acceptable following amputation.

Una de las principales complicaciones durante la anestesia de un proceso quirúrgico es el monitoreo de los problemas de oxigenación del paciente; la falta de oxigenación desencadena un estado de hipoventilación que consecuentemente produce un estado de hipoxemia, lo cual conllevará a un severo deterioro en la salud del paciente y, en el peor de los casos, a la muerte del mismo. El objetivo de este estudio fue determinar el porcentaje de accidentes hipóxicos en pacientes del curso de Cirugía de Animales Menores de la Universidad Ricardo Palma en la ciudad de Lima. Se evaluaron 45 fichas clínicas de registro anestésico entre Agosto del 2012 a Julio del 2014; con los protocolos de Xilacina - Ketamina (Protocolo 1); Xilacina - Ketoprofeno - Ketamina (Protocolo 2) y Xilacina - Tramadol - Ketamina (Protocolo 3). Las fichas de record anestésico se analizaron mediante los programas SPSS y Microsoft Excel. Los resultados promedio indicaron que del total del tiempo de cirugía en cada paciente, el 55% de dicho tiempo el paciente se encontraba en hipoxemia, con un valor de saturación de oxígeno de 92 ± 4.4. Los resultados por protocolo anestésico mostraron un mayor porcentaje de hipoxemia con el Protocolo 3. Los resultados por American Society of Anesthesiologists (ASA) presento un mayor porcentaje de hipoxemia en los ASA 1. Por otro lado se mostraron un mayor porcentaje de hipoxemia en la especie felina. Asimismo se demostró un mayor porcentaje de hipoxemia en ovariohisterectomías. Se comprobó la existencia de estados hipoxémicos en los procedimientos quirúrgicos de la Sala de Cirugía de Animales Menores recomendándose realizar un monitoreo más exhaustivo de constantes vitales en los diferentes procedimientos quirúrgicos y clasificaciones ASA, empleando las diferentes fichas de monitoreo para cada especie. One of the main complications of anesthesia during a surgical procedure, it´s the monitoring of patient oxygenation problems, the lack of oxygen trigger a state of hypoventilation which consequently produces a state of hypoxia, which it would give rise to a severe deterioration in the health of the patient and in the worst cases, death of it self. The aim of this study was to determine the percentage of hypoxic injury in patients during surgery of small animals in the Ricardo Palma´s University in Lima. 45 anesthesic records registered from August 2012 to July 2014; with the protocols of Xylazine – Ketamine (Protocol 1); Xylazine – Ketoprofen – Ketamine (Protocol 2) and Xylazine – Tramadol – Ketamine (Protocol 3) were analyzed using SPSS and Microsoft excel. The average results indicated that the total operation time for each patient, 55% of the time the patient was in hypoxemia with oxygen saturation value 92 +- 4.4. the results for anesthesic protocol demonstrated higher percentage of hypoxemia with Protocol 3. The results for ASA demonstrated a higher percentage of hypoxemia in the ASA 1. On the other hand a higher percentage of hypoxemia were shown in the feline species. A higher percentage of hypoxemia in ovariohysterectomiy is also demonstrated. The existence of hypoxemic states in surgical procedures for the surgery of small animals recommending more comprehensive monitoring of vital signs in different surgical procedures and ASA classifications using different monitoring records for each species registered

A gengivite crônica e intratável observada em gatos infectados pelo vírus da imunodeficiência felina (FIV) é um problema bastante freqüente na clínica de pequenos animais. O papel do FIV na etiologia da estomatite persistente ainda está por ser determinado. As manifestações orais são freqüentemente os primeiros sintomas observados em pacientes humanos infectados pelo HIV e podem ser usadas como indicadores da progressão da doença. O objetivo do presente estudo foi quantificar os linfócitos T CD4+, T CD8+ e a razão CD4+/CD8+ em uma colônia de gatos com gengivite crônica e naturalmente infectados pelo FIV. Para tanto, foram utilizados 20 gatos, todos apresentando gengivite com graus variando de 1 a 4. Desse total, 10 gatos não eram infectados pelo FIV e os outros 10 felinos eram infectados pelo FIV. Utilizou-se como controle 20 gatos sem gengivite, sendo 10 infectados pelo FIV e outros 10 não infectados pelo Retrovírus. As contagens dos linfócitos T CD4+ e CD8+ foram realizadas utilizando-se a técnica de citometria de fluxo. Os resultados obtidos demonstraram que os gatos com gengivite e infectados pelo FIV apresentaram uma contagem significativamente menor de linfócitos T CD4+ quando comparado aos gatos com gengivite e não infectados pelo FIV. Não houve diferença significativa na contagem de linfócitos T CD8+ entre os gatos com gengivite, infectados ou não pelo FIV. A razão CD4+/CD8+ também se mostrou em declínio nos gatos com gengivite e infectados pelo FIV. Concluiu-se que nas condições do presente estudo, a infecção pelo FIV compromete a resposta imunológica de felino diante da inflamação gengival. / Chronic and intractable gingivitis in FIV-infected cats is a relatively common clinical problem in veterinary practice. The role of FIV in the etiology of persistent stomatitis is still undetermined. Oral manifestations often found in HIV-infected people are frequently the first clinical sign of the infection and can be considered as an indicator of the progression of the HIV infection. The purpose of this study was to evaluate the CD4+ and CD8+ T-lymphocytes count and CD4+:CD8+ ratio in a colony of cats with chronic gingivitis. To achieve these goals, a colony of twenty domestic shorthair cats was used. All cats had some degree of gingival inflammation with scores ranging from 1 through 4. Ten cats were FIV-positive and ten were FIV-negative. As a control, twenty cats without gingivitis were used (ten cats were FIV-positive and ten were FIV-negative). CD4+ and CD8+ T-lymphocytes counts were performed by means of flow cytometry in all forty cats and results compared. The results showed that cats with gingivitis and FIV-infected had a lower CD4+ T cells count than cats with gingivitis but not FIV-infected. There was no difference in CD8+ T lymphocytes count among the cats with gingivitis infected or not with the FIV. The CD4+:CD8+ ratio was lower in cats with gingivitis and FIV-infected. One can conclude that FIV infection induces immunological disorders in cats with gingival inflammation.