MicroRNAs (miRNAs) play a vital role in differentiation, proliferation and tumorigenesis by binding to messenger RNAs (mRNA) and inhibiting translation. To initiate an investigation into the identification of miRNAs in the domestic dog, an emerging model for human disease, a comparison of the human and canine genetic databases was conducted. The bioinformatics work revealed significant conservation of miRNA genes between the two species. Proof of principle experiments, including serial dilutions and sequencing, were performed to verify that primers made to amplify human mature miRNAs can be used to amplify canine miRNAs, providing that the mature sequences are conserved. TaqMan® Real-time RT-PCR, a sensitive and specific method, was used to isolate the first miRNA mature products from canine tissues. The expression levels of miR-17-3p, miR-17-5p, miR-18, miR-19a, miR-19b, miR-20, and miR-92 were evaluated in five canine tissues (heart, lung, brain, kidney, and liver). Because miRNAs have been found to act as both tumor suppressors and oncogenes in several different cancers, expression patterns of ten miRNAs (miR-15a, miR-16, miR-17-5p, miR-21, miR-29b, miR-125b, miR-145, miR-155, miR-181b, let-7f) known to be associated with human breast cancer were compared between malignant canine mammary tumors (n=6) and normal canine mammary tissue (n=10). Resulting data revealed miR-29b and miR-21 to have a statistically significant (p<0.05) up-regulation in cancerous samples. Overall expression patterns showed nine of the ten miRNAs follow the same pattern of expression in the domestic dog as the human, while the miR-145 expression does not show a difference between the normal and cancerous samples.

Driven by both applied and theoretical goals, scientific interest in canine cognition has experienced a rapid surge in popularity, especially over the last 15 years (Morell, 2009). Here we provide the most comprehensive review to date of dog cognition research, capturing all the articles (285) we could find on the subject going back to 1911. We begin by summarizing general research trends, first documenting the rapid recent growth in dog cognition research (particularly in the domain of social cognition), and then identifying a number of trends in terms of the cognition topics and dog populations studied. Next, we summarize and synthesize the substantive conclusions emerging from research on non-social (discrimination learning, object permanence, object learning, categorization, object manipulation, quantitative understanding, spatial cognition, memory) and social (responses to human cues, perspective taking, dog-human communication, social learning) cognition. In light of burgeoning research on individual differences in cognition and on the biological organization of cognitive domains, we highlight the potential impact of these topics on dog cognition. Finally, based on our syntheses, we outline some ideas for future research, including recommendations that studies focus on: (1) incorporating multiple sensory modalities (most notably olfaction); (2) using more diverse populations of subjects; (3) replicating studies where current knowledge is based on small study sets or on small samples; (4) identifying fundamental developmental patterns of cognitive development; (5) identifying individual differences in cognitive ability; and (6) identifying potential cognitive constraints (e.g., cognitive abilities that are non-independent due to pleiotropic biological organization). / text

An inflammatory reaction is induced after release of proinflammatory mediators such asinterleukin 1 and 6 and tumour necrosis factor α. These mediators stimulate the liver tosuppress the syntheses of albumin and endure the syntheses of acute phase protein forinstance C-reactive protein. The aim of this paper was to perform a method validation on animmune turbidimetric assay to quantify C-reactive protein in canine serum at the laboratory atSkara Animals Hospital, Skara, Sweden. The validation involved evaluation of the assaylinearity, precision, stability and recovery.The method was proved to be linear for both TruLab control and Medinor control. TheTruLab control is based on human C-reactive protein while the Medinor control is based oncanine C-reactive protein. The precision of the method had a CV of 2.26 % (n=40). Themethod is considered stable and has good precision, under the circumstance that the TruLabcontrol is used. It was concluded that a Canine CRP-control is required and should beincluded in routine analysis.

This thesis is an investigation of Pseudomonas aeruginosa bacterial biofilms. The objective of the first study was to evaluate the biofilm-forming capacity of canine otitis isolates of P. aeruginosa and to compare the minimum inhibitory concentrations (MICs) of antimicrobials for planktonic versus biofilm-embedded bacteria. Biofilm forming ability was assessed using a microtitre plate assay. Broth microdilution was used to assess the MICs of neomycin, polymyxin B, enrofloxacin and gentamicin for the planktonic and biofilm-embedded bacteria of eighty-three isolates. Thirty-three (40%) isolates were biofilm producers and MICs for biofilm-embedded bacteria were significantly higher than their planktonic counterparts for all antimicrobials (all P<0.05). The objective of the second study was to evaluate the impact of Tromethamine edetate disodium dihydrate (Triz-EDTA®) in combination with antimicrobials on antimicrobial susceptibility of P. aeruginosa biofilm-embedded bacteria. MICs of the four antimicrobials for the biofilm embedded bacteria and biofilm-embedded bacteria with added Triz-EDTA® were assessed with broth microdilution for thirty-one biofilm-producing isolates. Addition of Triz-EDTA® significantly reduced MICs for neomycin (P < 0.008) and gentamicin (P < 0.04) but not enrofloxacin (P = 0.7), or polymyxin B (P = 0.5). The objective of the third study was to determine the presence of biofilm-associated genes in biofilm forming and non-biofilm forming isolates. Four genes involved with carbohydrate matrix production (pelA), irreversible attachment (sadB) and quorum sensing (lasB, rhlA) were selected. DNA was extracted and polymerase chain reaction (PCR) was performed for all isolates. All isolates possessed lasB and sadB, 74 (90%) possessed pelA and 74 (90%) possessed rhlA. All thirty-two (100%) isolates that were classified as biofilm producers contained all genes. There was an association between the presence of pelA and rhlA and biofilm production (P < 0.017) and between the presence of rhlA and pelA and the quantity of biofilm produced (both P < 0.001). These results highlight that biofilm formation of Pseudomonas aeruginosa otic isolates does occur and can impact antimicrobial therapy. Certain compounds can also influence antimicrobial susceptibility of biofilm-embedded bacteria. Genetics may also play a role in biofilm formation.

No description available.

The four objectives of this study were to determine: 1) the incidence of Dirofilaria immitis and Dipetalonema reconditum in dogs of East Central Indiana, 2) the variables significantly affecting the incidence of each parasite, 3) the correlation of heartworm infections with clinical symptoms, and 4) to determine whether Dirofilaria immitis and Dipetalonema reconditum are endemic in Indiana.From December, 1973 to April, 1974, blood samples were taken from 335 dogs in East Central Indiana and were examined for microfilariae using the direct smear and the modified Knott concentration techniques. At the same time, complete information on each dog was collected on a "SURVEY INFORMATION” form. The data was statistically analyzed by crosstabulation of each variable with the results of the blood tests for each parasite. Levels of significance were computed using chi square values with appropriate degrees of freedom. Values of 0.05 were considered to be significant.In this study, the overall incidence of Dirofilaria immitis was found to be 3°% which included one dog infected with both parasites. Of the 131 hunting dogs, 6.1% were infected, while only 1% of the 204 non-hunting dogs were infected. Furthermore, all of the dogs infected with heart-worms were those which were principally outdoor dogs. These differences were significant and they reflect thehigher exposure risk of hunting dogs and dogs living out-doors to the mosquito vectors.The incidence of Dipetalonema reconditum was 6% overall with 11.5 % of the hunting dogs and 2.5% of the non-hunting dogs infected. It was most prevalent in dogs three to six years of age. After age seven the prevalence declined sharply. This decrease with age may be the result of an immune response to the infective larvae which prevents reinfection.Clinical symptoms could not be correlated with infections with Dirofilaria immitis since 90% of the dogs infected with heartworms had no symptoms of the disease. This indicates that most of the heartworm infections in dogs of this area are light, probably involving few adult worms. The results of this study indicate that Dirofilaria imnitis and Dipetalonema reconditum are endemic in Indiana, as 60% of the dogs with heart-worms and 35% of the dogs with Dipetalonema reconditum had never been out of the state.

No description available.

Canine tick-borne diseases are an emerging problem within Australia and throughout the world. This thesis investigates Babesia gibsoni and Anaplasma platys infections in dogs in Australia and also explores the evolutionary relationships and taxonomy of the canine piroplasm species and the members of the order Piroplasmida. A nested PCR-RFLP assay was developed for the detection and differentiation of the canine piroplasm species and was found to have a high detection limit, capable of detecting a 2.7 x 10-7 % parasitaemia or the equivalent of 1.2 molecules of target DNA. Detection of piroplasm DNA applied to Whatman FTA“ classic cards using nested-PCR was found to have a lower detection limit than when using DNA extracted from whole blood but higher than IsoCode‘ Stix or QIAamp extraction from filter paper based techniques. The nested PCR-RFLP assay was further evaluated for the detection of B. gibsoni infection in dogs being exported from Australia to New Zealand and compared to the current screening methods, the Immunofluorescent Antibody Test (IFAT) and microscopy. Of 235 dogs screened, 11 were IFAT positive, 1 was microscopy positive and 3 were PCR positive for B. gibsoni, highlighting the discordance that exists between various detection techniques. Replacing microscopic examination of blood smears with PCR-RFLP is suggested for screening dogs entering New Zealand, in addition to revising the current IFAT cut-off titre to minimize false positive results. The first case of B. gibsoni in New South Wales is also reported. A study was also conducted to further investigate the recent discovery of B. gibsoni in Australia and the association of this infection with American Pit Bull Terriers in an epidemiological study. Both American Pit Bull Terriers (n = 100) and other dog breeds (n = 51) were screened for B. gibsoni using IFAT and PCR-RFLP. A questionnaire was also completed by each dog owner regarding thethe husbandry and habits these dogs. Fourteen dogs were positive for B. gibsoni using IFAT and/or PCR-RFLP and all were American Pit Bull Terriers. Dogs that were male and/or were bitten by or were biters of other American Pit Bull Terriers were statistically more likely to be B. gibsoni positive, thus suggesting that blood-to-blood transmission may contribute to the spread of this disease. Experimental B. gibsoni infections were established in vivo to investigate the efficacy of combined atovaquone and azithromycin therapy and to determine the detection limits of PCR, IFAT and microscopy during various stages of infection. While atovaquone and azithromycin produced a reduction in circulating parasite levels, it did not cause total eradication, and possible drug resistance also developed in one dog. PCR was found to be most useful in detecting early and acute stage infections, while IFAT was most useful during chronic and acute infections. Microscopy is suggested to be only useful for detecting acute stage infections. This study also describes the detection of B. gibsoni in tissue samples during chronic infection for the first time, suggesting possible sequestration of this parasite. Anaplasma platys has also only recently been reported in Australia and the distribution, molecular-charcterisation, pathogenesis, co-infection with Babesia canis vogeli and treatment of infection with doxycycline were investigated. For the first time, A. platys is reported in Western Australia, Queensland and Victoria, with each isolate found to be genetically identical on the basis of the 16S rRNA gene. No correlation could be established between A. platys infection and the development of clinical signs or pathogenesis and definitive treatment using doxycycline could not be determined. Isolates of canine piroplasms from various geographical locations worldwide (n = 46), including Australia were characterised on the basis of multiple gene loci to explore the distribution, genetic variation and possible phylogeographical relationships of these species. Separate genotypes of B. canis vogeli, B. canis canis and B. gibsoni are suggested and may be correlated to different geographical origins. Characterization of B. canis vogeli, B. canis canis and B. canis rossi on the basis of the HSP 70 gene and B. gibsoni on the basis of the ITS 1, 5.8S rRNA gene and ITS 2 is described for the first time. Elevation of each of the B. canis subspecies, with the exclusion of B. canis presentii, to separate species is also proposed. The current paraphyly and taxonomic confusion associated with the members of the order Piroplasmida led to a review of the phylogenetic and taxonomic status of this group of organisms. Phylogenetic relationships are determined using 18S rRNA gene, 5.8S rRNA gene, HSP 70 gene and combined loci analyses. Rearrangement of the Piroplasmida into three families, including the new family Piroplasmiidae is proposed, in addition to the establishment of two new genera, the Piroplasma (Patton, 1895) and the Achromaticus (Dionisi, 1899). Other proposed schemes of classification and the limitations of phenotypic characteristics in taxonomic classification within the Piroplasmida are also discussed.

Resumo: A neoplasia intra-epitelial prostática (HGPIN) e a atrofia inflamatória proliferativa (PIA) são lesões potencialmente pré-malignas encontradas na próstata humana (De Marzo et aI., 1999; Bostwick e Qian, 2004). HGPIN tem sido relatada na próstata canina com características semelhantes às observadas em humanos (Waters et aI., 1997), porém a PIA ainda não foi caracterizada nesta espécie. O presente trabalho objetivou a avaliação imunoistoquímica do índice proliferativo (PCNA e Ki67), da expressão de proteínas pró-apoptose (caspase-3), genes supressores de tumores (p-53), genes inibidores de apoptose (bcl-2) e de moléculas de adesão (E-caderina), além da avaliação histoquímica do estado proliferativo (AgNOR) e da determinação de mensurações nucleares por análise quantitativa computadorizada de imagens (AQCI) em focos HGPIN, PIA e em ácinos normais da próstata canina. Considerando os resultados de ácinos normais, constatou-se que as lesões estudadas apresentam alto índice proliferativo; capacidade proliferativa no compartimento epitelial secretor; ausência de expressão de p-53; perda de expressão de E-caderina; padrão de AgNOR semelhante ao de ácinos normais; e núcleos celulares maiores e com fator de circunferência maior. Focos de PIA exibem ainda índice apoptótico semelhante ao de ácinos normais e predomínio do linfócitos T, quando considerado o infiltrado Iinfocítico. Tais achados aproximam estas duas lesões entre si e sugerem que possam estar envolvidas no processo de transformação neoplásica da próstata canina. / Abstract: Prostatic intraepithelial neoplasia (HGPIN) and proliferative inflammatory atrophy (PIA) are potentially premalignant lesions, found in human prostate (De Marzo et aI., 1999; Bostwick e Qian, 2004). HGPIN have been reported in canine prostate (Waters et aI., 1997), and share the same aspects with the humans, and PIA hadn't been described in the dogs. The aim of this work was to evaluate, by immunohistochemistry proliferative index (PCNA and KI-67), proapoptotic proteins expression (caspase 3), tumour suppressor gene (p-53), adhesion molecules (Ecadherin), histochemical proliferative status (AgNOR) and nuclear measurement by image computer quantitative analysis (AQCI) in HGPIN focus, PIA e normal acinus in canine prostate. Comparing the normal acinus results, the lesions showed higher proliferative index, secretory epithelial cells capable of proliferation, absence of p-53 expression, loss of E-cadherin expression, AgNOR patterns similar to normal acinus, cellular nucleus bigger and with higher nuclear round factor. PIA had the same apoptotic index as the normal acinus and mainly T Iymphocytes in the inflammatory infiltrate. Our findings allow us to consider these two lesions close to each other, and they may be involved in the process of neoplastic transformation of canine prostate. / Orientador: Enio Pedone Bandarra / Coorientador: Luiz Henrique de Araújo Machado / Banca: Renée Laufer Amorim / Banca: Sérgio Luis Felisbino / Banca: Ana Paula F. R. L. Bracarense / Banca: Maria Lúcia Zaidan Dagli / Doutor

Made available in DSpace on 2014-06-11T19:30:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-26Bitstream added on 2014-06-13T19:19:19Z : No. of bitstreams: 1 rodrigues_mmp_dr_botfmvz.pdf: 907066 bytes, checksum: 46839f5f715b3cba912be1b604d7f3fa (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / No homem e no cão, as lesões displásicas são a neoplasia intra-epitelial prostática (PIN) e a atrofia inflamatória proliferativa (PIA), as quais vêm sendo estudadas devido seu potencial pré-maligno. Diversos fatores, bem como o processo inflamatório, estão relacionados à carcinogênese prostática. A cicloxigenase-2 (COX-2) apresenta papel fundamental na resposta inflamatória e apresenta relação com o desenvolvimento do câncer de próstata. Este trabalho teve por objetivos caracterizar a proliferação e morte celular (apoptose) das células basais e secretoras do tecido prostático normal e atípico (PIA, PIN e carcinoma) dos cães, avaliar a integridade da camada de células basais por meio do anticorpo p63 e, analisar a expressão gênica de COX-2 nos diferentes componentes teciduais da PIA canina por qRT-PCR. Avaliaram-se, pela técnica de dupla marcação imunoistoquímica, os índices de proliferação e apoptose com os marcadores Ki-67 e caspase-3, respectivamente, das células secretoras (PSA positivas) e das células basais (34âE12 positivas). Para isso, foram estudadas 130 próstatas de cães adultos, inteiros e sem histórico de lesão prostática. Observou-se aumento da proliferação e diminuição da apoptose das células secretoras da PIA e do carcinoma, entretanto, as células basais apresentaram maior índice proliferativo e menor taxa de apoptose apenas nos focos de PIA. Na PIN, a proliferação celular mostrou-se mais evidente em relação à morte celular. Maior quantidade de células basais que expressaram p63 foi constatada na PIA e no carcinoma. Os focos de PIA apresentaram maior expressão gênica de COX-2 no estroma em relação às células epiteliais. Conclui-se que os compartimentos secretor e basal exibem maior proliferação celular e menor taxa apoptótica na PIA. O aumento da marcação de p63 na PIA e no carcinoma sugere participação das... / Dysplastic lesions in human and dogs are prostatic intraepithelial neoplasia (PIN) and proliferative inflammatory atrophy (PIA), which have been investigated because of their premalignant potential. Inflammatory process is related to prostate carcinogenesis. Cyclooxygenase-2 (COX-2) plays a role in the inflammatory response and is correlated with the development of prostate cancer. This study aimed to characterize the proliferation and apoptosis of basal and secretory cells of normal and atypical prostate tissue (PIA, PIN and carcinoma) of dogs, to assess the integrity of the basal cell layer through the p63 immunohistochemistry. In addition, we analyzed the gene expression of COX-2 in different tissue components of canine PIA by qRT-PCR. Immunohistochemical double staining technique was performed using Ki-67 and caspase-3 antibodies to asses cellular proliferation and apoptosis, respectively. PSA was used to characterize the secretory cells and 34âE12 to characterize the basal cells. Were analyzed 130 prostates of adult dogs with or without history of prostatic lesion. Increased proliferation and decreased apoptosis of the secretory cells of the PIA and the carcinoma was observed, however, the basal cells showed higher proliferative index and lower rate of apoptosis only in foci of PIA. PIA and carcinomas showed high expression of p63. It was observed higher gene expression of COX-2 in PIN stroma compared to epithelial cells. We conclude that basal and secretory compartments exhibit higher proliferative and lower apoptotic rate in PIA. The higher expression of p63 in PIA and carcinomas suggests involvement of the basal cells in atypical lesions of the prostate. Furthermore, the expression of COX-2 in PIA is higher in the stromal compartment, revealing important role of this enzyme in the canine prostate carcinogenesis