Tesis sobre el tema "Veterinary diagnostics"

Selected ion flow tube mass spectroscopy (SIFT-MS) is an analytical method for the investigation of volatile organic compounds (VOCs). It produces mass to charge (m/z) ratio ion counts with a range of 10-200 m/z. Current data analysis involves sifting through the spectra files one at a time looking for peaks of interest. This is time consuming and requires expert knowledge. This thesis proposes, implements and demonstrates a novel approach to the analysis of SIFT-MS data using multivariate techniques similar to those employed to analyse electronic nose and gas chromatography mass spectroscopy (GCMS) data. The methodology was developed using a set of samples created in the laboratory that belonged to two groups which contained different VOCs found in biological samples. The methodology requires the removal of the m/z peaks associated with the precursors, then principal component analysis (PCA) and partial least squares discriminant analysis (PLSDA) methods were evaluated for biomarker discovery and sample classification. Both methods produced excellent results, identifying the volatiles in the mixtures and being able to classify samples with 100% accuracy. This methodology was then tested using a variety of samples. Ammonia was found as a possible marker for bovine TB (Mycobacterium bovis) infection using serum samples taken from wild badgers. Discrimination results of an accuracy of 67%±6% were acquired. The number of sample needed to build the best performing model from this dataset was empirically shown to be 120. It was shown to be effective for the discrimination of serum samples from cattle taken before and after introduction of bovine TB (Mycobacterium bovis) bacteria in a clinical trial (accuracy of 85% achieved). A similar dataset pertaining to infection by Mannheimia haemolytica failed to produce models that performed as well as the others - this is suspect to be due to a poor experimental design. Finally, discrimination accuracies of 88% for urine samples collected from cattle from herds infected with Mycobacterium paratuberculosis and 90% for urine samples collected in the same bovine TB trial as above were achieved. The novel multivariate approach to SIFT-MS data analysis has been shown to be effective with a number of datasets but it is sensitive to the experimental design. Recommendation for the consideration required for analysis using this method have been made.

This PhD thesis is the outcome of a range of activities and experimental results aimed to a better characterization of the risk that Escherichia coli and other microrganisms and parasites may pone to the health of animals and finally humans. One of the main activity of the present project was based on the test hypothesis that the virulence profile of E. coli strains toward bovine mammary gland can be modulated by the interaction with the host cells. These hypothesis were tested through a gene expression study of some virulence factors of six E. coli strains when co-cultured with a bovine mammary cell line, since the in vitro models represent both an essential tool to investigate the biological mechanistic of mastitis, and an efficient alternative to animal experiments. Preliminarly, a meta-analysis of existing literature studies on the available bovine mammary cell lines was performed, resulting in the selection of MAC-T as the most responsive cell line to bacteria causing mastitis. The E. coli strains used for the coculture experiments with MAC-T cells were isolated from different types of bovine mastitis (acute, chronic and undetermined) and from a VTEC food-borne strain associated to human clinical disease (O157). An upregulation of the virulence factor eae (intimin) in all but one the analyzed mastitis strains following co-culture with MAC-T cell line was detected through RT-reat time PCR, and also the adherence virulence factor ycd and the b12 gene were upregulated in some strains, overall suggesting the possibility that mastitic E. coli strains can acquire a more risky molecular profile when exposed to the bovine mammary cells. This finding may have clear implications on the risk assessment related to the E. coli strains in bovine mammary tissue and milk. In addition, with the aim to improve the current methodologies for foodborne risk analysis linked to E. coli, the project activity provided a preliminary research for the setup and validation of new protocols based on real time PCR-High Resolution Melting Analysis, a widely used technique to target sequence polymorphisms of the same gene in different species without the need to perform DNA sequencing or to use species-specific probes, to help the identification of putative verocytotoxic status in E. coli strains of O26 serogroup, and other serotypes, isolated from bovine milk. Since the applications of HRMA for the characterization of microorganisms can not be limited to food safety, but can be developed for a large number of issues linked to general veterinary diagnostics, among the objectives of this PhD project some new real-time PCR-HRMA coupled methods were also developed, providing a contribution to the advancement of the existing molecular tools for sensitive and effective species identification, or variant/mutation screening, applied to different foodborne and veterinary pathogens. Thus, new HRMA-based protocols were designed and tested for the identification of Pseudomonas spp responsible for chromatic alterations in mozzarella cheese, for the detection and differentiation of Dirofilaria repens and D. immitis in canine blood samples, for the detection of the mutation site associated to FQ resistance in Staphylococcus pseudintermedius isolated from canine diagnostic samples, and for discrimination of the two most common microsporidial parasites in honeybees, Nosema apis and N. ceranae. Overall, these new HRMA-based assays could represent additional tools for epidemiological studies, routine disease assessment and therapeutical decisions. The possibility to identify the presence of risk-predictive SNPs in E. coli isolates using these newly established HRMA-based protocols is a novel, and simpler, opportunity with respect to the current, and more complex, surveillance strategies that are based on the amplification of stx genes together with other virulence factors for the evaluation of VTEC status. In the future, a possible way forward of this research is represented, on one side, by the deeper assessment of the reciprocal modulation between E. coli mastitis-derived strains and immortalized MAC-T cells using high-throughput RNA sequencing, and on the other side by a large scale validation of the HRMA-based evaluation of risk-predictive SNPs in order to improve the current approaches. And overall, the established HRMA-based protocols when extensively validated would be highly suitable for routine veterinary diagnostics applied to field investigation, as quick and sensitive single step protocols allowing specific and sensitive detection of the targets with shorter analysis time and reduced cost, in parallel or in alternative to the classical approaches.

Darbo tikslas: Įvertinti šliužo dislokacijos (ŠD) prognostinės diagnostikos galimybę, karvių fiziologinę būklę po šliužo repozicijos pagal būdingą ŠD sergamumo specifiką, pieno kiekio, elektrinio pieno laidumo, aktyvumo, kūno masės, melžimo trukmės nuokrypius. Uždaviniai: 1. Atlikti sergamumo ŠD tyrimą; 2. Įvertinti, kaip kinta pieno kiekis, elektrinis pieno laidumas, gyvulio aktyvumas, kūno masė, melžimo trukmė prieš susergant ŠD; 3. Įvertinti ŠD diagnostikos galimybę pagal kraujo serumo biocheminius rodiklius, pieno sudėtį, fiziologinių rodiklių nuokrypius, ŠD klinikinės diagnozės nustatymo dieną; 4. Įvertinti, kaip kinta pieno kiekis, elektrinis pieno laidumas, gyvulio aktyvumas, kūno masė, melžimo trukmė po ŠD repozicijos; 5. Įvertinti teoriškai pieno kiekio kitimo prognozę, atlikus ankstyvąją diagnostiką ir profilaktiką. Įvertinta karvių ŠD prognostinės diagnostikos galimybė, fiziologinė būklė po šliužo repozicijos pagal būdingą ŠD sergamumo specifiką, pieno kiekio, elektrinio pieno laidumo, aktyvumo, kūno masės, melžimo trukmės nuokrypius. Pagrįsta praktinė tiriamų parametrų panaudojimo galimybė.
The objective of this work: To evaluate the possibility of the prognostic diagnostics of the displaced abomasums, the physiological status of the cow after the reposition of abomasums according as the particularity of the displaced abomasums’ morbidity, deviation of milk yield, milk conductivity, walking activity, body weight and milking duration. The problems of this work: 1. To do the research about the morbidity of the displaced abomasums; 2. To estimate the dynamics of the milk yield, milk conductivity, cow’s walking activity, body weight, milking duration before the abomasums was displaced; 3. To estimate the possible diagnostics according as the biochemical results of the blood serum, milk composition, deviations of the physiological indicators at the day of clinical diagnosis of the displaced abomasums; 4. To evaluate the dynamics of the milk yield, milk conductivity, cow’s walking activity, body weight, milking duration after the abomasums was displaced; 5. To evaluate in theory the prognosis of the milk yield changes after the early diagnostics and prophylactics. The possibility of prognostic diagnostics of the displaced abomasums, physiological status of the cow after the reposition of abomasums according as the particularity of the displaced abomasums’ morbidity, deviation of milk yield, milk conductivity, walking activity, body weight and milking duration were evaluated. The possibility of practical appliance of the examined parameters was justified.

Il est maintenant évident que la dégradation des environnements peut favoriser la transmission des maladies infectieuses notamment en rapprochant les humains, des vecteurs ou des animaux. Concernant les Maladies Tropicales Négligées (MTN), l’OMS tente de mobiliser les institutions et la communauté scientifique en identifiant pour chacune des MTN, les lacunes existantes dans les besoins de diagnostic clinique, les critères requis pour leur développement ainsi que les stratégies à adopter pour lutter contre la maladie. Parmi elles, la schistosomiase, seconde maladie parasitaire humaine, souffre d’un manque flagrant d’outils de diagnostic alliant sensibilité et déployabilité afin de détecter les cas de faible intensité parasitaire dans les zones endémiques ; mais également pour permettre la réalisation de diagnostic animal et environnemental visant à adopter une approche intégrative dans la lutte contre la maladie. Les objectifs de cette thèse s’inscrivent dans la stratégie One Health en proposant le développement et l’application d’outils de diagnostics de terrain chez l’Homme, chez l’animal et dans l’environnement. Les efforts continus en matière de recherche, de développement de prévention, de traitement et de sensibilisation sont essentiels pour parvenir à un monde où la schistosomiase et plus généralement les MTN ne seront plus une menace pour la santé humaine
It is now evident that environmental degradation can facilitate the transmission of infectious diseases, particularly by bringing humans into closer proximity with vectors or animals. In the case of Neglected Tropical Diseases (NTDs), the World Health Organization (WHO) is endeavoring to mobilize institutions and the scientific community by identifying, for each NTD, existing gaps in clinical diagnostic needs, the criteria required for their development, and the strategies to be adopted to combat the disease. Among them, schistosomiasis, the second most prevalent human parasitic disease, suffers from a distinct lack of diagnostic tools that combine sensitivity and deployability to detect cases with low parasitic intensity in endemic areas. Additionally, such tools are needed to facilitate animal and environmental diagnosis, enabling an integrated approach to disease control. The objectives of this thesis align with the One Health strategy, proposing the development and application of field-friendly diagnostic tools for humans, animals, and the environment. Ongoing efforts in research, development, prevention, treatment, and awareness are essential to achieve a world where Schistosomiasis and other neglected tropical diseases cease to threaten human health

There is an urgent need for detection of viral respiratory pathogens to identify the causal agent(s) involved and to prevent the spread of related diseases. The first part of this dissertation focuses on development, optimization and validation of Real-time reverse transcription polymerase chain reaction (rRT-PCR) assays for the detection of several common equine viral pathogens: equine arteritis virus (EAV), equine influenza virus and equine rhinitis viruses A and B. Emphasis of the second part of this dissertation is on studying the role of EAV envelope proteins in virus attachment and entry. Using an infectious cDNA clone of EAV and reverse genetics, a panel of chimeric viruses was generated by swapping the N-terminal ectodomains and full-lengths of the two major envelope proteins (GP5 and M) from porcine reproductive and respiratory syndrome virus (PRRSV). The recombinant viruses expressing the N-terminal ectodomain of PRRSV GP5 or M or both (GP5ecto, Mecto, and GP5&Mecto, respectively) in an EAV backbone were viable and genetically stable. Compared to the parental virus, these three chimeric viruses produced lower titers and smaller plaque sizes indicating that they have a crippled phenotype. Interestingly, the three chimeric viruses could only infect EAV susceptible cell lines but not the PRRSV susceptible cell line. Therefore, the exchange of GP5 and/or M protein N-terminal ectodomains from PRRSV did not alter the cellular tropism of the chimeric viruses. We also investigated the role of one of the minor envelope proteins (E) of EAV in virus attachment and entry. The results showed that EAV infection of equine endothelial cells is heparin-dependent and the Cterminus of the E protein contains a putative heparin-binding domain. We generated a panel of arginine to glycine mutations in the conserved region of both the full-length EAV infectious cDNA clone and individual E protein expression vectors. The triple mutation R52,60,65G construct grew significantly slower and produced much smaller plaques. The double mutant R52,60G completely blocked the interaction between E protein and heparin. Taken together, these data indicated that E protein interacts with heparin to facilitate virus attachment and plays a major role in EAV infection.

Os vírus entéricos causam infecções que podem ocasionar uma alta morbidade e mortalidade em cães. A diarreia se destaca como o principal sinal clínico e a subsequente desidratação pode causar a morte do animal. O Carnivore protoparvovirus 1 (CPV-2), Canine mastadenovirus A tipo 1 (CAdV-1), o Canine coronavirus (CCoV), o Canine rotavirus (CRV) e o Canine distemper virus (CDV) são considerados os principais agentes que causam gastroenterite viral aguda em cães jovens. O objetivo deste trabalho foi detectar a presença destas cinco espécies virais na população de cães do Brasil. Para isto, foram coletados 325 suabes retais de cães com ou sem diarreia, jovens (> 6 meses) e adultos (< 6 meses), com ou sem histórico de vacinação e de diversas regiões do Brasil. As amostras foram analisadas através da reação em cadeia da polimerase (PCR) ou transcrição reversa seguida de PCR (RT-PCR) utilizando iniciadores específicos para cada um dos agentes virais. Resultaram 81% (264/325) das amostras positivas para um destes vírus, das quais 30,7% (100/325) foram positivas para o CPV-2, 25,5% (83/325) para o CDV, 17,2% (56/325) para o CCoV, 4,6% (15/325) para o CRV e 2,7% (9/ 325) para o CAdV-1. Algumas amostras apresentaram co-infecções, sendo que as espécies mais predominantemente encontradas em co-infecções foram o CDV e CPV-2 em 15,4% (50/325) e 15,0% (49/325), respectivamente, seguidos do CCoV em 10,1% (33/325,), CRV em 3,0% (10/325) e CAdV-1 em 1,5% (5/325) das amostras. A associação viral mais observada foi CDV e CPV-2 em 31/325 (9,5%) amostras positivas para ambos os vírus. Em conclusão, os resultados demonstram que CPV-2, CDV e CCoV são os principais vírus entéricos patogênicos que circularam no Brasil entre os anos de 2008 e 2014, infectando mais frequentemente animais jovens.
Enteric viruses cause infections that lead to high morbidity and mortality. Diarrhea is the main clinical sign, whose subsequent dehydration can cause death of the animal. Carnivore protoparvovirus 1 (CPV-2), Canine mastadenovirus A (CAdV-1), Canine coronavirus (CCoV), Canine rotavirus (CRV) and Canine distemper virus (CDV) are considered the main agents that cause acute viral gastroenteritis in young dogs. The aim of this study was the detection of these five viral species in dogs from Brazil. Rectal swabs from 325 dogs, puppies (< six months old), adult dogs (> 6 months old), with or without a history of vaccination, were collected from various regions of the country. The samples were analyzed by polymerase chain reaction (PCR) or reverse transcription following followed by PCR (RT-PCR) using oligonucleotides specific for each one of this virus. As result, 81% (264/325) of the samples were observed to be positive for at least one virus, 30,7% (100/325) were positive for CPV-2, 25,5% (83/325) for CDV, 17,2% (56/325) for CCoV, 4,6% (14/325) for CRV and 2,7% (9/325) for CAdV-1. Some samples showed co-infection, where the species most predominantly found were CDV and CPV-2 with 15,4% (50/325) and 15,0% (49/325), respectively, followed by CCoV with 10,1% (33/325,), CRV with 3,0% (10/325) and CAdV-1 1,5% (5/325). The most observed viral association was CDV and CPV-2, with 31/325 (9,5%) positive samples for both viruses. In conclusion, the results showed that CPV-2, CDV and CCoV are the main pathogenic enteric viruses that circulated in Brazil between the years 2008 and 2014, infecting more frequently puppies.

Os vírus influenza A (VIA) e circovírus suíno tipo 2 (PCV2) são os agentes etiológicos da influenza suína (IS) e da circovirose suína (CS), respectivamente. Estas doenças têm um impacto econômico significativo na suinocultura mundial. Adicionalmente, o VIA pode ser transmitido entre animais e humanos, sendo por isso, importante para a saúde pública. O presente trabalho teve o objetivo de pesquisar a ocorrência desses vírus em suínos de abate no sul de Moçambique. As amostras foram coletadas em um abatedouro na cidade da Matola, nos períodos de dezembro de 2014 a fevereiro de 2015 e dezembro de 2015 a fevereiro de 2016. Os materiais e métodos aplicados e resultados obtidos estão apresentados em dois artigos científicos. O primeiro relata a infecção pelo VIA associada à caracterização anatomopatológica e imuno-histoquímica (IHQ) das lesões pulmonares. Foram avaliados 457 pulmões de suínos, e amostras de 38 (8.3%) pulmões, que apresentaram áreas de consolidação, foram coletadas e submetidas ao exame histopatológico e IHQ para a detecção de antígenos do VIA, PCV2 e Mycoplasma hyopneumoniae. Antígenos do VIA foram detectados em 32/38 (84.3%) dos pulmões com pneumonia através da IHQ, e os suínos positivos eram provenientes dos distritos de Matutuine (5/32), Moamba (2/32), Namaacha (21/32), Boane (3/32) e Cidade da Matola (1/32). Todos os pulmões com pneumonia foram negativos no exame de IHQ para PCV2 e M. hyopneumoniae. O segundo artigo teve o objetivo de detectar lesões histológicas, antígenos e DNA de PCV2 em linfonodos mesentéricos de suínos e realizar a caracterização filogenética de isolados de PCV2 circulantes no sul de Moçambique. Foram coletados aleatoriamente 111 linfonodos mesentéricos de suínos de abate provenientes de nove distritos do sul de Moçambique. As amostras foram submetidas ao exame histopatológico, IHQ e reação em cadeia da polimerase (PCR). Uma amostra positiva para PCV2 na PCR de cada distrito (n=9) foi selecionada aleatoriamente e submetida ao sequenciamento da região aberta de leitura ORF2. DNA de PCV2 foi detectado em 53.8% (62/111) das amostras e em 73.8% de granjas dos nove distritos. No exame de IHQ, linfonodos mesentéricos de seis suínos positivos para PCV2 na PCR apresentaram antígenos desse vírus associados à depleção linfoide e infiltrado de histiócitos e células gigantes multinucleadas. Na análise filogenética, sequências dos isolados dos distritos de Namaacha, Moamba e Maputo ficaram agrupadas no genótipo PCV2d-2; as sequências de isolados dos distritos de Manhiça e Matola, no genótipo PCV2d-1; enquanto os isolados dos distritos de Boane, Matutuine, Chibuto e Xai-Xai, no genótipo PCV2b-1A/B. Os resultados do trabalho permitem concluir que o VIA e PCV2 circulam na população suína em vários distritos da região sul de Moçambique.
Influenza A virus (IAV) and porcine circovirus type 2 (PCV2) are the etiological agents of swine influenza (SI) and porcine circovirus associated diseases (PCVAD) respectively. These diseases represent a significant economic impact on pig production worldwide. In addition, IAV can be transmitted between animals and humans with consequences for public health. The objective of this study was to investigate the occurrence of these viruses in slaughter pigs in Southern Mozambique. Samples were collected in a slaughterhouse in Matola city, from December 2014 to February 2015 and December 2015 to February 2016. The materials and methods applied and the results obtained are presented in two manuscripts. The first article reports IAV infection in pigs and characterize the anatomopathological and immunohistochemical features of the associated lung lesions. Lungs from 457 slaughtered pigs were evaluated grossly, and samples from 38 (8.3%) of these that presented pulmonary consolidation were collected and examined for histopathology and immunohistochemistry (IHC) for the presence of IAV, PCV2 and Mycoplasma hyopneumoniae antigens. IAV antigens were detected in 32/38 (84.3%) of pneumonic lungs, and positive pigs were from Matutuine district (5/32), Moamba district (2/32), Namaacha district (21/32), Boane district (3/32) and Matola City (1/32). All lung samples were immunohistochemically negative for PCV2 and M. hyopneumoniae. The second article aimed to detect histological lesions, PCV2 antigens and DNA and perform phylogenetic analysis of PCV2 strains circulating in Southern Mozambique. At slaughter, mesenteric lymph nodes were collected from 111 randomly selected pigs from nine districts of Southern Mozambique. Samples were submitted to histopathological examination, IHC and polymerase chain reaction (PCR). One PCV2 PCR positive sample from each district (n=9) was randomly selected in order to obtain sequences covering the ORF2 region. PCV2 DNA was detected in 53.8% (62/111) of the samples and 73.8% of the farms from all nine districts. PCV2 antigen was detected by IHC in six lymph nodes that were positive for PCV2 by PCR and antigens were associated with lymphoid depletion and infiltrate of histiocytes and multinucleated giant cells. Phylogenetic analysis demonstrated that three sequences from Maputo, Namaacha and Moamba were grouped with PCV2d-2, two sequences from Manhiça and Matola were grouped as PCV2d-1, and four sequences from Boane, Matutuine, Chibuto, and Xai-Xai were closely related to PCV2b-1A/B genotypes. The results of this study indicate that IAV and PCV2 circulate in the swine population in several districts of the southern region of Mozambique.

La prima parte del nostro studio riguarda la tecnica LAMP (Loop-mediated isothermal amplification), una tecnica di amplificazione isotermica recentemente inventata (Notomi et al., 2000). Essa presenta notevoli vantaggi rispetto alle tradizionali PCR: non necessita di strumentazioni sofisticate come i termociclatori, può essere eseguita da personale non specializzato, è una tecnica altamente sensibile e specifica ed è molto tollerante agli inibitori. Tutte queste caratteristiche fanno sì che essa possa essere utilizzata al di fuori dei laboratori diagnostici, come POCT (Point of care testing), con il vantaggio di non dover gestire la spedizione del campione e di avere in tempi molto brevi risultati paragonabili a quelli ottenuti con la tradizionale PCR. Sono state prese in considerazione malattie infettive sostenute da batteri che richiedono tempi molto lunghi per la coltivazione o che non sono addirittura coltivabili. Sono stati disegnati dei saggi per la diagnosi di patologie virali che necessitano di diagnosi tempestiva. Altri test messi a punto riguardano malattie genetiche del cane e due batteri d’interesse agro-alimentare. Tutte le prove sono state condotte con tecnica real-time per diminuire il rischio di cross-contaminazione pur riuscendo a comprendere in maniera approfondita l’andamento delle reazioni. Infine è stato messo a punto un metodo di visualizzazione colorimetrico utilizzabile con tutti i saggi messi a punto, che svincola completamente la reazione LAMP dall’esecuzione in un laboratorio specializzato. Il secondo capitolo riguarda lo studio dal punto di vista molecolare di un soggetto che presenza totale assenza di attività mieloperossidasica all’analisi di citochimica automatica (ADVIA® 2120 Hematology System). Lo studio è stato condotto attraverso amplificazione e confronto dei prodotti di PCR ottenuti sul soggetto patologico e su due soggetti con fenotipo wild-type. Si è poi provveduto al sequenziamento dei prodotti di PCR su sequenziatore automatico al fine di ricercare la mutazione responsabile della carenza di MPO nel soggetto indicato.
The first part of the present study concerns the LAMP technique (Loop-Mediated Isothermal Amplification), an isothermal amplification technique recently developed (Notomi et al., 2000). LAMP has many advantages over traditional PCR: it doesn’t require sophisticated instruments like thermal cyclers, it can be performed by unskilled staff, it is a highly sensitive and specific technique and it is very tolerant to inhibitors. All these characteristics make it suitable to be used outside diagnostic laboratories, as POCT (Point-of-care testing), with the advantage of not having to send the sample and obtaining results as accurate as PCR tests and in very short times. We designed and optimized assays to detect bacteria that require a very long time for cultivation or that are not even cultivable. We drew assays for the diagnosis of viral diseases that require to be diagnosed as soon as possible. We developed a test to assess two genetic diseases of the dog and two food contaminating bacteria. All tests were carried out using real-time technique to decrease the risk of cross-contamination. Finally, we developed a colorimetric method for showing results which can be applied to all of the assays we optimized. The second section presents the molecular study of a subject who had myeloperoxidase deficiency at the automated cytochemistry analysis (ADVIA ® 2120 Hematology System). The study was conducted through amplification and comparison of the PCR products obtained from the pathological subject and on two subjects with wild-type phenotype. The products were sequenced using an automated sequencer in order to find the responsible mutation for the MPO deficiency in the indicated subject.

La prima parte del nostro studio riguarda la tecnica LAMP (Loop-mediated isothermal amplification), una tecnica di amplificazione isotermica recentemente inventata (Notomi et al., 2000). Essa presenta notevoli vantaggi rispetto alle tradizionali PCR: non necessita di strumentazioni sofisticate come i termociclatori, può essere eseguita da personale non specializzato, è una tecnica altamente sensibile e specifica ed è molto tollerante agli inibitori. Tutte queste caratteristiche fanno sì che essa possa essere utilizzata al di fuori dei laboratori diagnostici, come POCT (Point of care testing), con il vantaggio di non dover gestire la spedizione del campione e di avere in tempi molto brevi risultati paragonabili a quelli ottenuti con la tradizionale PCR. Sono state prese in considerazione malattie infettive sostenute da batteri che richiedono tempi molto lunghi per la coltivazione o che non sono addirittura coltivabili. Sono stati disegnati dei saggi per la diagnosi di patologie virali che necessitano di diagnosi tempestiva. Altri test messi a punto riguardano malattie genetiche del cane e due batteri d’interesse agro-alimentare. Tutte le prove sono state condotte con tecnica real-time per diminuire il rischio di cross-contaminazione pur riuscendo a comprendere in maniera approfondita l’andamento delle reazioni. Infine è stato messo a punto un metodo di visualizzazione colorimetrico utilizzabile con tutti i saggi messi a punto, che svincola completamente la reazione LAMP dall’esecuzione in un laboratorio specializzato. Il secondo capitolo riguarda lo studio dal punto di vista molecolare di un soggetto che presenza totale assenza di attività mieloperossidasica all’analisi di citochimica automatica (ADVIA® 2120 Hematology System). Lo studio è stato condotto attraverso amplificazione e confronto dei prodotti di PCR ottenuti sul soggetto patologico e su due soggetti con fenotipo wild-type. Si è poi provveduto al sequenziamento dei prodotti di PCR su sequenziatore automatico al fine di ricercare la mutazione responsabile della carenza di MPO nel soggetto indicato.
The first part of the present study concerns the LAMP technique (Loop-Mediated Isothermal Amplification), an isothermal amplification technique recently developed (Notomi et al., 2000). LAMP has many advantages over traditional PCR: it doesn’t require sophisticated instruments like thermal cyclers, it can be performed by unskilled staff, it is a highly sensitive and specific technique and it is very tolerant to inhibitors. All these characteristics make it suitable to be used outside diagnostic laboratories, as POCT (Point-of-care testing), with the advantage of not having to send the sample and obtaining results as accurate as PCR tests and in very short times. We designed and optimized assays to detect bacteria that require a very long time for cultivation or that are not even cultivable. We drew assays for the diagnosis of viral diseases that require to be diagnosed as soon as possible. We developed a test to assess two genetic diseases of the dog and two food contaminating bacteria. All tests were carried out using real-time technique to decrease the risk of cross-contamination. Finally, we developed a colorimetric method for showing results which can be applied to all of the assays we optimized. The second section presents the molecular study of a subject who had myeloperoxidase deficiency at the automated cytochemistry analysis (ADVIA ® 2120 Hematology System). The study was conducted through amplification and comparison of the PCR products obtained from the pathological subject and on two subjects with wild-type phenotype. The products were sequenced using an automated sequencer in order to find the responsible mutation for the MPO deficiency in the indicated subject.

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Tendo em vista a alta incidência das neoplasias mamárias e metástases em cadelas e objetivando contribuir com a oncologia veterinária na padronização de um teste simples, eficaz e acessível no diagnóstico precoce destas metástases, esse estudo teve como meta avaliar o marcador tumoral CA 15.3 no soro de cadelas saudáveis e investigar a viabilidade da quimioluminescência e do kit de reagentes desenvolvido para a espécie humana como ferramenta neste processo. Numa primeira etapa avaliou-se o soro de 100 cadelas saudáveis, de raças e idades variadas. Numa segunda etapa o soro de 10 cadelas saudáveis foi submetido ao processo de concentração de proteínas e posteriormente avaliado. Nas amostras não concentradas os níveis do marcador ficaram abaixo do limiar de detecção do kit. Nas amostras concentradas, o valor médio e respectivo desvio padrão observados foram 0,4140 ± 0,25 U/mL nas amostras com o dobro da concentração inicial e 0,5170 ± 0,25 U/mL nas amostras quatro vezes mais concentradas. Com base nos resultados obtidos neste estudo concluiu-se que o método empregado, ensaio imunoluminométrico por quimioluminescência e o kit de reagentes desenvolvido para a detecção do CA 15.3 na espécie humana mostram-se viáveis para utilização na espécie canina, necessitando entretanto de adequação da técnica
Given the high incidence of mammary tumors and metastases in female dogs and trying to contribute with the veterinary oncology in standardization of a simple, effective and affordable technique in the early diagnosis of these metastases, this study was aimed to evaluate the tumor marker CA 15.3 in serum of healthy dogs and to investigate the feasibility and the chemiluminescence reagent kit designed for the human species as a tool in this process. In a first step we evaluated the serum of 100 healthy female dogs of different breeds and ages. In a second step, the serum of 10 healthy female dogs was subjected to the process of proteins concentration and subsequently evaluated. In samples not concentrated, the levels were below the detection of the kit. In concentrated samples, the average value and its standard deviation observed were 0.4140 ± 0.25 U/mL in the samples with at double the initial concentration, and 0.5170 ± 0.25 U/mL in samples four times more concentrated. Based on the results of this study it is concluded that the method employed, chemiluminescent immunoluminometric assay, and the reagents kit designed for detection of the CA 15.3 in humans appear to be viable for use in dogs, however requiring adjustment of the technique

Orientador: Aureo Evangelista Santana
Banca: Maria Angélica Dias
Banca: Marcia Ferreira da Rosa Sobreira
Resumo: Tendo em vista a alta incidência das neoplasias mamárias e metástases em cadelas e objetivando contribuir com a oncologia veterinária na padronização de um teste simples, eficaz e acessível no diagnóstico precoce destas metástases, esse estudo teve como meta avaliar o marcador tumoral CA 15.3 no soro de cadelas saudáveis e investigar a viabilidade da quimioluminescência e do kit de reagentes desenvolvido para a espécie humana como ferramenta neste processo. Numa primeira etapa avaliou-se o soro de 100 cadelas saudáveis, de raças e idades variadas. Numa segunda etapa o soro de 10 cadelas saudáveis foi submetido ao processo de concentração de proteínas e posteriormente avaliado. Nas amostras não concentradas os níveis do marcador ficaram abaixo do limiar de detecção do kit. Nas amostras concentradas, o valor médio e respectivo desvio padrão observados foram 0,4140 ± 0,25 U/mL nas amostras com o dobro da concentração inicial e 0,5170 ± 0,25 U/mL nas amostras quatro vezes mais concentradas. Com base nos resultados obtidos neste estudo concluiu-se que o método empregado, ensaio imunoluminométrico por quimioluminescência e o kit de reagentes desenvolvido para a detecção do CA 15.3 na espécie humana mostram-se viáveis para utilização na espécie canina, necessitando entretanto de adequação da técnica
Abstract: Given the high incidence of mammary tumors and metastases in female dogs and trying to contribute with the veterinary oncology in standardization of a simple, effective and affordable technique in the early diagnosis of these metastases, this study was aimed to evaluate the tumor marker CA 15.3 in serum of healthy dogs and to investigate the feasibility and the chemiluminescence reagent kit designed for the human species as a tool in this process. In a first step we evaluated the serum of 100 healthy female dogs of different breeds and ages. In a second step, the serum of 10 healthy female dogs was subjected to the process of proteins concentration and subsequently evaluated. In samples not concentrated, the levels were below the detection of the kit. In concentrated samples, the average value and its standard deviation observed were 0.4140 ± 0.25 U/mL in the samples with at double the initial concentration, and 0.5170 ± 0.25 U/mL in samples four times more concentrated. Based on the results of this study it is concluded that the method employed, chemiluminescent immunoluminometric assay, and the reagents kit designed for detection of the CA 15.3 in humans appear to be viable for use in dogs, however requiring adjustment of the technique
Mestre

Orientador: Luiz Carlos Marques
Banca: Eduardo Harry Birgel Junior
Banca: Paola Castro Moraes
Resumo: Devido à escassez de estudos mielográficos em bovinos e relatos de complicações no procedimento, o presente estudo teve por objetivo estudar a eficácia e segurança dos meios de contrastes utilizados a fim de nortear a escolha mais adequada para o diagnóstico de afecções vertebrais e medulares em bovinos. Foram utilizados dez bezerros holandeses, hígidos de sete dias a dois meses de idade. Os bezerros formaram dois grupos, um deles recebeu Ioexol e o outro Iopamidol, na cisterna cerebelomedular. Foi realizado estudo mielográfico da coluna vertebral na posição látero-lateral, que foram reproduzidas em tempos determinados em minutos totalizando 20 tempos, para posterior análise da opacidade, detalhes da imagem, distensão do canal medular e progressão da linha de contraste. Após a mielografia os animais foram observados durante a recuperação até deambulação. Após intervalo de sete dias foi realizado segundo período experimental, que compreendeu na inversão do meio de contraste dentro de cada grupo. Diferenças significativas em relação à qualidade da imagem e velocidade do preenchimento da coluna medular entre os dois meios de contrastes estudados não foram observadas. Não foram observadas anormalidades clínica após o exame mielográfico. Conclui-se que para a obtenção de imagens mielográficas com excelente radiopacidade, rica em detalhes, com distensão do espaço subaracnoide adequada e completa progressão da linha do contraste é necessário que as tomadas radiográficas sejam no seg... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Due to the shortage of myelographic studies in cattle and reports of complications in the procedure, this study aimed to study the efficacy and safety of contrast media used to guide the most appropriate choice for the diagnosis of vertebral and spinal cord diseases in cattle. Ten Holstein calves, healthy seven days to two months of age were used. Calves formed two groups, one received iohexol and iopamidol other in cerebelomedular tank. Myelographic study was conducted on the spine side-to-side position, they are reproduced in time determined 20 minutes total time for analysis of the opacity image, details, distension medullary canal and contrast line progression. After myelography animals were observed during recovery to ambulation. After seven days interval was performed according to experimental period understood that the inversion of the contrast medium within each group. Significant differences in image quality and speed of fulfillment of the spinal column between the two means of studied contrasts were not observed. There were no clinical abnormalities after myelographic examination. We conclude that to obtain myelographic images with excellent radiopacity, rich in detail, with distension of subarachnoid space adequate and complete progression of contrast is line necessary that the radiographs are in the cervical segment 6 to 8 minutes after contrast administration in the thoracic segment eighty minutes after inoculation contrast and lumbar segments, sacral and cauda e... (Complete abstract click electronic access below)
Mestre

L'imaging diagnostico è una parte fondamentale nella valutazione del paziente, anche per un soggetto animale questa tecnica è di usuale utilizzo. Non sostituisce però un esame fisico approfondito: da solo fornisce raramente un’indicazione specifica della causa di una particolare malattia tuttavia può dare un’accurata base di partenza per arrivare alle sue cause finali. L’imaging diagnostico è utilizzato per confermare o confutare una lesione clinicamente sospetta, per suggerire o documentare il sito di una presunta lesione, caratterizzare la natura e l'estensione di una lesione nota, per seguire la progressione della malattia o la guarigione, stabilire la prognosi, pianificare o valutare terapie chirurgiche, suggerire o guidare procedure diagnostiche aggiuntive e controllare le malattie con segni clinici oscuri. La selezione dello studio di imaging diagnostico appropriato è determinata dalla struttura anatomica da valutare e dal tipo di informazioni richieste. Con l'avvento di nuove modalità di imaging, le informazioni anatomiche e funzionali sul sistema muscolo-scheletrico possono essere meglio determinate con l'aumentare della precisione diagnostica e della risoluzione anatomica. L’imaging diagnostico, termine moderno per indicare tutte le modalità di formazione dell’immagine biomediche utilizzate a scopo diagnostico e terapeutico, comprende radiologia, tomografia, ecografia e risonanza magnetica, ed è parte fondamentale nella valutazione del paziente sia umano che animale.

Resumo: Avaliou-se o trato respiratório de 129 potros PSI de corrida, 59 machos e 70 fêmeas, entre 18 e 24 meses de idade, filhos de 44 garanhões, antes do início dos treinamentos, em 7 haras das regiões criatórias de Piraquara, São José dos Pinhais, Tijucas do Sul e Araucária, no Estado do Paraná. Empregou-se um questionário para avaliar a vida dos potros até o momento do exame, com respeito ao manejo de cocheira, alimentação, sanidade e filiação. Os potros foram examinados clinicamente. O exame endoscópico foi realizado com um video-endoscópio Welch- Allyn de 200 cm de comprimento e 9 mm de diâmetro, avaliando-se o trato respiratório superior e o trato respiratório inferior até a bifurcação da traquéia. O aspirado traqueal foi procedido quando necessário, corando-se o esfregaço pela técnica de Pappenhein para o exame citológico da secreção. Observou-se um aumento do número de potros mantidos em regime de liberdade após os 6 meses de idade. As auscultações traqueal e pulmonar não mostraram correlação com a presença de secreção traqueo-bronquial. HFL grau II ou mais foram observadas em 97,5% dos potros. Observou-se DDPM intermitente e DDPM persistente em 14,0% e 2,5% dos potros, respectivamente. A epiglote foi considerada comprida em 8,3% dos potros e, 4,2% e 0,8% apresentaram epiglote curta e frágil e curta, frágil e com extremidade desviada para a esquerda, respectivamente. Observou-se NLP II esquerda em 23,1% dos potros e NLP II direita em 2,5%. Alguns garanhões apresentaram um número maior de potros com alterações na forma da epiglote e no funcionamento das cartilagens aritenóides. As espirais de Curshmann apareceram no aspirado de 16 potros e, as células gigantes de corpo estranho, os eosinófilos e os macrófagos espumosos estiveram presentes em 4, 61 e 70 potros, respectivamente. Os potros mantidos em cocheiras por um tempo maior e aqueles que receberam alfafa fenada na alimentação mostraram maiores alterações do aspirado traqueal. Evidenciou-se que potros PSI de corrida podem apresentar anormalidades das vias aéreas superiores e comprometimento das vias aéreas inferiores antes de iniciarem os treinamentos, o que provavelmente pode resultar num rendimento atlético abaixo do seu potencial.

Šunų ausų ligos gana dažnai pasitaiko veterinarijos gydytojų praktikoje. Netiksliai diagnozavus ausies uždegimą gali būti pasirinktas netinkamas gydymo būdas, liga gali komplikuotis, sparčiai progresuoti, pasidaro lėtinė. Dažnai gyvūnai pradedami gydyti pasireiškus klinikiniams požymiams – vietiniam skausmui, niežėjimui, eritemai, epitelio pleiskanojimui ir susikaupus nemalonaus kvapo išskyroms. Ausų uždegimą gali sukelti labai daug priežasčių, kurios skirstomos į predisponuojančias, pirmines ir antrines. Predisponuojantiems veiksniams priskiriama šuns veislė, anatominė ausų stuktūra, jatrogeninės traumos, netinkamas gydymas, auglių obstrukcijos ar polypai. Infekciją gali sukelti ir į ausį patekę svetimkūniai, išoriniai dirgikliai: sieros sankaupos, kai ausys ilgai nevalomos, vanduo, patekęs į šuns ausis. Pirminiai veiksniai tiesiogiai sukelia išorinės ausies uždegimą. Tai ausų erkutės (Otodectes cynotis) ir odos parazitai (Demodex canis, Otodectes, Sarcoptes ir kt.), alergija, autoimuninės ligos, pirminė seborėja, blusos, hormonų pusiausvyros sutrikimas, endokrininiai sutrikimai. Antrinėms priežastims priskiriamos sąlyginai patogeninių bakterijų asociacijos ir grybeliai, kurie ausyje gyvena simbiozėje ir dauginasi. Bakterinį ausų uždegimą dažniausiai sukelia Staphylococcus spp., Pseudomonas aeruginosa, Proteus spp., rečiau – Bacillus ir Corynebacterium genčių mikroorganizmai bei Malassezia spp. mielės. Šio darbo tikslas buvo atlikti veiksnių, įtakojančių ausų uždegimo... [toliau žr. visą tekstą]
Ear diseases in dogs are common in veterinary medicine. Inaccurate diagnoses and inappropriate treatment of otitis, can rapidly cause serious progressive complications and the disease will be chronic. Frequently, animal treatments start when symptoms have been displayed: pain, head shaking, scratching at the pinnae, and malodorous ears, unpleasant smells and erythema are common complaints. Otitis can be caused by many reasons. There are distributive to predisposing, primary and secondary reasons. Predisposing factors are abnormal or breed-related conformation of the ear canal, including stenosis, yatrogenic injuries, inappropriate treatment hirsutism, and pendulous pinnae that restrict proper air flow into the ear canal. Excessive moisture, due to swimming or to frequent cleanings with improper solutions, can also lead to infections. Primary causes directly make inflammation of otitis externa. Parasites (Otodectes cynotis) and skin parasites (Demodex canis, Otodectes, Sarcoptes etc.), atopy, food allergy, auto-immune diseases primary seborrhea, fleas, and endocrine disorders. Secondary causes may be assigned to conditioned pathogenic bacteria as well as fungus which lives in ear with symbiosis and it is reproductive. Bacterial infections are often infected by Staphylococcus spp., Pseudomonas aeruginosa, Proteus spp., more rarely – Bacillus and Corynebacterium spp.and Malassezia spp. barm. The objective of the research work was to accomplish inflammation factors of otitis and... [to full text]

A mortalidade fetal é uma causa importante de perdas reprodutivas em animais domésticos e tem impacto significativo na rentabilidade de um sistema de produção animal. No Brasil, poucas informações são disponíveis sobre as principais causa de aborto em bovinos. O presente estudo resultou em cinco artigos. O primeiro artigo descreve as principais causas de aborto bovino diagnosticadas no Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul no período 2003-2011. Neste período um total de 490 fetos bovinos foi analisado. Causas específicas de aborto foram encontradas em 46,7 % dos casos. Infecções por protozoários, em especial Neospora caninum acometeram 33 % dos casos (162/490). Bactérias em 6,3 % (31/490), seguidas por fungos 0,8% (4/490) foram causas adicionais de abortos. Em dois fetos (0,4 %), coinfecções por dois agentes foram identificadas. Causas não infecciosas foram observadas em 3 % dos abortos e malformações congênitas a 2,6%. No segundo artigo estima-se a ocorrência de abortos por Brucella abortus em bovinos na região sul, caracterizando os achados macroscópicos, histopatológicos, bacteriológicos, imuno-histoquímicos e de PCR desta enfermidade. No terceiro artigo relata-se a infecção por Geotrichum candidum em feto bovino abortado associado a lesões de pele e pulmonares. A ocorrência de malformações congênitas múltiplas em um feto bovino abortado é relatada no quarto artigo e casos de condrodisplasia tipo Dexter em fetos bovinos abortados são descritos no quinto.
The fetal mortality is an important cause of reproductive failure in domestic animals and has a significant impact on the profitability of a livestock production system. In Brazil, little information is available on the main cause of abortion in cattle. This study resulted in five articles. The first article describes the main causes of abortion in cattle diagnosed in the period January 2003 to december 2011. A total of 490 fetuses, from several Brazilian states, were evaluated. Specific causes of abortion were found in 46,7 % of cases. Protozoan abortions especially Neospora caninum were detected in 33 % (162/490) of the cases. Bacterial abortions corresponded to 6,3 % (31/490), followed by fungal 0,8 % (4/490). In two aborted fetuses (0,4 %), a co-infection with two agents could be identified. Noninfectious diseases could be associated with 3 % of the abortions and congenital malformations with 2,6%. In the second article estimated the incidence of abortion by Brucella abortus in cattle in the southern region, characterizing the macroscopic findings, histological, bacteriological, immunohistochemical and molecular aspects of this disease. The third article relates a Geotrichum candidum infection in aborted bovine fetus associated with skin and lung lesions. The occurrence of multiple congenital malformations in an aborted bovine fetus is reported in the fourth article and cases of Dexter chondrodysplasia type of aborted fetuses are described in the fifth.

Este trabalho tem como objetivo descrever as principais doenças diagnosticadas em caprinos no Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul no período de 2000 a 2016. No primeiro artigo foi realizado um estudo retrospectivo das causas de morte em caprinos diagnosticadas de 2000 a 2016. Do total de 322 caprinos necropsiados neste período, 290 (90%) obtiveram um diagnostico conclusivo. Dos casos conclusos, 167 (57,6%) corresponderam a enfermidades de origem infecciosa e toxi-infecciosas e 123 (42,4%) enquadrados em causas não infecciosas. Entre as doenças infecciosas foram contabilizados 54 casos de origem bacteriana, 60 casos com envolvimento parasitário, 14 casos de origem viral, além de 39 casos toxi-infecciosos. As doenças de caráter não infeccioso foram ainda agrupadas em doenças metabólicas (44 casos), intoxicações por plantas ou substâncias tóxicas (36), deficiências minerais e nutricionais (20), neoplasias e distúrbios no desenvolvimento (5). A hemoncose, eimeriose, pleuropneumonias e a enterotoxemia foram as doenças mais frequentemente diagnosticadas neste período. O segundo artigo descreve um surto de tuberculose em caprinos jovens. Onze de um total de 15 caprinos, de cinco a 15 dias de idade, foram positivos ao teste de tuberculina. Na necropsia, o parênquima pulmonar de todos os caprinos positivos apresentavam nódulos de 0,3 a 10 cm de diâmetro, coloração brancacenta a amarelada, ocasionalmente, também observados no fígado e baço Os linfonodos retrofaríngeos, mediastínicos e traqueobrônquicos apresentavam-se acentuadamente aumentados de tamanho e aspecto caseoso. Na avaliação histológica, a lesão era caracterizada por intensa necrose caseosa, com áreas de mineralização distrófica, associados a acentuado infiltrado inflamatório granulomatoso. A coloração de Ziehl-Neelsen e a marcação por imuno-histoquímica anti-complexo Micobacterium tuberculosis evidenciou discreta a moderada quantidade de bacilos álcool-ácido resistentes. O cultivo microbiológico e a análise molecular confirmaram o agente etiológico M. bovis. O terceiro artigo descreve dermatite e hepatopatia tóxica crônica natural e experimental em caprinos associadas ao consumo de farelo de arroz desengordurado. Caprinos jovens, de um a quatro meses de idade, apresentavam alopecia e formações crostosas na pele, apatia, emagrecimento, prurido discreto e, vinham a óbito em um período de 30-40 dias. À necropsia, o fígado apresentava irregularidades na superfície capsular, coloração alaranjada a avermelhada, além de rins com múltiplas áreas circulares brancacentas na superfície capsular. À análise microscópica, acentuada atrofia de hepatócitos em região periportal hepática e moderada degeneração hepatocelular microvacuolar. No estudo experimental comprovou-se a etiologia dos casos, através da manifestação de lesões de pele, hepática e renais similares ao dos casos naturais, entretanto em menor intensidade.
This work aims to describe the main diseases diagnosed in goats in the Sector of Veterinary Pathology of the Federal University of Rio Grande do Sul from 2000 to 2016. The first article describes the main causes of death in goats diagnosed between 2000 and 2016. A conclusive diagnosis was obtained in 290 (90%) cases from a total of 322 goats necropsied. Of these cases, 167 (57.6%) corresponded to infectious and toxi-infectious diseases, and 123 (42.4%) included non-infectious causes. Among the infectious diseases 54 cases were of bacterial origin, 60 cases were caused by parasite agents, 14 cases of viral origin, and 39 toxi-infectious cases. Non-infectious diseases were also grouped into metabolic diseases (44 cases), poisoning by plants or toxic substances (36), mineral and nutritional deficiencies (20), neoplasms and developmental disorders (5). Haemonchosis, eimeriosis, pleuropneumonia and enterotoxemia remain as one of the major control obstacles in goat farms. The second article describes an outbreak of tuberculosis in goat kids. Eleven of a total of 15 kids, from 5 to 15 days old, were positive to tuberculin. At necropsy, the pulmonary parenchyma of all positive goats had white to yellowish nodules of 0.3 to 10 cm in diameter, that were occasionally also observed in the liver and spleen The retropharyngeal, mediastinal and tracheobronchial lymph nodes were markedly enlarged and with a caseous aspect. Histologically, the lesion was characterized by an intense caseous necrosis, with areas of dystrophic mineralization, associated to a marked granulomatous inflammatory infiltrate. Ziehl-Neelsen histochemistry exam and immunohistochemical anti-Micobacterium tuberculosis complex evidenced mild to moderate amount of bacilli. Microbiological culture and molecular analysis confirmed M. bovis as the etiological agent. The third article describes a natural and an experimental toxic liver disease associated with the consumption of defatted rice bran in goats. These presented with alopecia and crusted formations on the skin, apathy, weight loss, mild pruritus, and death within a period of 30-40 days. At necropsy, the liver presented multifocal to coalescing orange to reddish irregular areas on the capsular surface, and the kidneys presented multiple white circular areas on the capsular surface. Microscopic analysis revealed a marked hepatocyte atrophy at the hepatic periportal region, and a moderate microvacuolar hepatocellular degeneration. In the experimental study, the etiology of the cases was demonstrated through the manifestation of lower intensity skin, liver and kidney lesions similar to those of the natural cases.

Publically-administered veterinary diagnostic laboratories are accredited to an industry-developed quality standard, AAVLD, which is similar to the ISO 17025 international standard. Laboratories wanting to improve their client base of private industry partners must increasingly consider ISO 17025 accreditation to be successful, as it goes beyond the AAVLD standard by verifying the laboratory's technical competence to perform testing within its accreditation scope.

The research considers the differences between the AAVLD and ISO 17025 standards and strategies used by other laboratories to successfully implement a quality program, and provides a gap analysis between the two standards. The plan considers project management and change management strategies, details roles and responsibilities for the project team, and provides a range of aids including commonly used quality assurance tools to successfully develop the ISO 17025 quality system. The project plan can be successfully implemented using the laboratory's existing AAVLD quality system as a starting point.

The expanding aquaculture industry continues to encounter major challenges in the form of highly contagious aquatic viruses. Control and eradication measures targeting the most lethal and economically damaging virus-induced diseases, some of which are notifiable, currently involve ‘stamping out’ policies and surveillance strategies. These approaches to disease control are performed through mass-culling followed by restriction in the movement of fish and fish products, resulting in considerable impacts on trade. Although effective, these expensive, ethically complex measures threaten the sustainability and reputation of the aquatic food sector, and could possibly be reduced by emulating innovative vaccination strategies that have proved pivotal in maintaining the success of the terrestrial livestock industry. DIVA ‘differentiating infected from vaccinated animal’ strategies provide a basis to vaccinate and contain disease outbreaks without compromising ‘disease-free’ status, as antibodies induced specifically to infection can be distinguished from those induced in vaccinated animals. Various approaches were carried out in this study to assess the feasibility of marker/DIVA vaccination for two of the most important disease threats to the global Atlantic salmon and common carp/koi industries, i.e. infectious salmon anaemia (ISA) and koi herpesvirus disease (KHVD), respectively. Antibody responses of Atlantic salmon (Salmo salar L.), following immunisation with an ISA vaccine, administered with foreign immunogenic marker antigens (tetanus toxoid (TT), fluorescein isothiocyanate (FITC) and keyhole limpet hemocyanin (KLH)) were assessed by antigen-specific enzyme linked immunosorbent assay (ELISA). Although antibodies were induced to some markers, these were unreliable and may have been affected by temperature and smoltification. Detectable antibodies to ISAV antigen were also largely inconsistent despite low serum dilutions of 1/20 being employed for serological analysis. The poor antibody responses of salmon to the inactivated ISA vaccine suggested that DIVA vaccination is not feasible for ISA. A similar approach for KHV, utilising green fluorescent protein (GFP) as the marker, similarly failed to induce sufficiently detectable antibody responses in vaccinated carp (Cyprinus carpio L.). However, as high anti-KHV antibody titres were obtained with an inactivated KHV vaccine (≥1/3200), alternative approaches were carried out to assess the feasibility of DIVA vaccination for carp. Investigations of early KHV pathogenesis in vivo and antigen expression kinetics in vitro (0-10 days post infection (dpi)) provided valuable data for the diagnostics necessary for DIVA surveillance strategies. Following viral infection, molecular methods were shown to be the most effective approach for early detection of KHV infected fish prior to sero-conversion, during which time antibodies are not detectable. An experimental immersion challenge with KHV, however, revealed complications in molecular detection during early infection. The KHV DNA was detected in external biopsies of skin and gills, but also internally in gut and peripheral blood leukocytes ≤ 6 hours post infection (hpi), suggesting rapid virus uptake by the host. The gills and gut appeared to be possible portals of entry, supported by detection of DNA in cells by in situ hybridisation (ISH). However, many false negative results using organ biopsies occurred during the first 4 dpi. The gills were the most reliable lethal biopsy for KHV detection by various polymerase chain reaction (PCR) assays, with a PCR targeting a glycoprotein-gene (ORF56) and a real-time PCR assay being the most sensitive of the 7 methods investigated. Importantly, non-lethal mucus samples reduced the number of false negative results obtained by all KHV PCR assays during the earliest infection stages with large levels of viral DNA being detected in mucus (up to 80,000 KHV DNA genomic equivalents 200 μL-1). KHV DNA was consistently detected in the mucus as a consequence of virus being shed from the skin. Determining the expression kinetics of different viral structural proteins can be useful for DIVA serological tests. Analysis of KHV antigen expression in tissues by immunohistochemistry and indirect fluorescent antibody test was inconclusive, therefore 2 novel semi-quantitative immunofluorescence techniques were developed for determining KHV antigen expression kinetics in susceptible cell lines. During the course of KHV infection in vitro, a greater abundance of capsid antigen was produced in infected cells compared to a glycoprotein antigen (ORF56), as determined by detection with antigen-specific monoclonal antibodies (MAbs). The capsid antigen was characterised as a ~100 kDa protein by SDS-PAGE and identified as a product of KHV ORF84 by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF/TOF MS). This antigen was subsequently detected in the serum of >25% of KHV infected/exposed carp (6/17), as well as in carp vaccinated with a live attenuated vaccine (3/4), but not with an inactivated vaccine (0/7), by Western blot making it a potential DIVA target for an inactivated vaccine. Attempts were made to improve the sensitivity of KHV serological testing by taking advantage of recombinant proteins specific for KHV (CyHV-3), rORF62 and rORF68 and eliminating any interference by cross-reacting antibodies to carp pox (CyHV-1). These proteins successfully reacted with anti-KHV antibodies. The feasibility of DIVA strategies for KHVD was determined using these recombinant antigens to coat ELISA plates. Differential antibody responses were detected from carp sera to an internal virus tegument protein (rORF62) and external region of a transmembrane protein (rORF68). Fish vaccinated with an inactivated vaccine produced significantly lower antibody responses to rORF62 than to rORF68, whereas infected, exposed and live attenuated vaccinated fish recognised both proteins allowing differentiation between vaccinated and infected carp. However, the sensitivity of the assay was limited, possibly by high levels of natural antibodies detected at the relatively low serum dilutions (1/200) used. As the capsid antigen (ORF84) and tegument protein (ORF62) are derived from internal KHV structural proteins, they induce non-neutralising antibodies, which may be useful for DIVA strategies. Such antibodies are longer lasting than neutralising antibodies and often comprise the majority of fish anti-viral antibodies. This was noted in a fish surviving experimental challenge, which had an antibody titre of 1/10,000, but neutralising titre of 1/45. Such antigens may therefore hold potential for developing effective serological diagnostic tests for KHV and provide the potential for DIVA strategies against KHVD. Natural antibodies will, however, continue to present a challenge to the development of sensitive and reliable KHV serological tests, and hence the application of DIVA strategies.

Baigiamojo darbo tikslas: išanalizuoti kačių astmos pasireiškimo dažnumą, ligos atsiradimo priežastis, dažniausiai taikomas diagnostines priemones ir gydymą. Apibūdinimas: kačių astma – tai galimai alerginės kilmės susirgimas, kurio metu atsiranda kvėpavimo takų uždegimas, padidėjusi gleivių sekrecija, spontaniška bronchokonstrikcija ir kvėpavimo takų remodeliacijos. Šie pokyčiai sukelia kosulį, dusulį, švokštimą ir kvėpavimo nepakankamumą. Dažniausiai praktikoje susiduriama su diagnostikos problematika. Baigiamojo darbo metodikos: tyrimas buvo atliekamas 2013 metais, jame dalyvavo 9 astma sergančios katės ir 5 sveikos katės. Buvo atlikta kačių, įtariamų sergant astma, šeimininkų apklausa, kurios metu vertinami galimai astmos pasireiškimui turėję įtakos veiksniai. Tuomet atliekamos kitos diagnostinės procedūros, kurių metu gaunama diagnozė, patvirtinanti kačių astmą ir skiriamas tolimesnis gydymas. Pagrindiniai tiriamojo darbo rezultatai ir išvados: kačių astma nėra dažnai pasitaikanti kvėpavimo takų liga. Tarp astma sergančių kačių nėra ženklių lyties skirtumų, 56 % sirgusiųjų sudarė patelės, o patinai – 44 %. Astma dažniau pasireiškė katėms, kurių amžius svyravo nuo 1 iki 5 metų (45 %) ir toms, kurios buvo negrynaveislės (45 %). Apklausos metu nustatyti pagrindiniai potencialūs astmą sukeliantys veiksniai, t.y. šeimininkų rūkymas (12 %), parfumuotų valiklių (12 %), oro gaiviklių bei dezodorantų (12 %) naudojimas katės gyvenamojoje aplinkoje. Astmos diagnostikoje takyti... [toliau žr. visą tekstą]
The aim: to analyze the manifestation frequency of feline asthma, diagnostic methods, treatment and identify the factors, which have influence in manifestation of this disease. Description: feline asthma has an allergic etiology. The hallmark features include airway inflammation, mucus hypersecretion, spontaneous bronchoconstriction and airway remodeling. These changes lead to clinical signs of cough, dyspnea, wheeze and respiratory distress. The diagnosis of asthma can be complicated, because there are no single test for diagnose feline asthma. Methodology: the study was carried out in a small animal clinic X in 2013 year. 9 asthmatic cats and 5 healthy cats were included in this study. There was composed questionnaire form for the owners, whose cats were possibly asthmatic. All the received material was used for statistic analysis about the factors, which had more impact in causing asthma. The other diagnostic manipulations were performed to get this certain diagnosis and find the best treatment options. There were reviewed epidemiology, clinical manifestations, patogenesis and factors, which influence asthma, blood results, diagnosis and treatment of feline asthma. Results and conclusions: feline asthma is not common respiratory disease in cats. The study revealed that there is no clear sex predilection (females – 56 % and males – 44 %). Middle-aged cats (1 to 5 years) were seen more affected (45 %). 45 % of the cats were mixed breed. The main asthma predisposing factors... [to full text]

Diagnostic test validation is challenge for the clinical pathologist when a new instrument or a new analyzer is introduced in the lab. The aim of this work is to assess the analytical and/or clinical performances of novel biomarker to evaluate a possible application as a useful marker in veterinary medicine. Four different objectives are investigate: 1. Haematological parameters: two new gates are draw, in particular a “high fluorescent region” to identify canine blood samples with cell of probable neoplastic origin that need further investigation and a gate to identify “high fluorescent platelet fraction” in macrothrombocytopenic Norfolk terrier. Both gates are useful to perform a correct diagnosis with a better outcome for the patients. 2. Inflammatory markers: the measurement of dROMs (derivative of reactive oxygen metabolites) in serum of dogs with leishmaniasis showe a strong involvement of these compounds in the symptomatic or the asymptomatic Leishmania infections, and could be use to identify dogs with sub-clinical forms. Another studies involve the validation of a new test for IFN-γ measurement after incubation of feline leukocytes with immunogenic peptides. This study is from one side a draw up of a new test with consequently assessment of analytical performances, and from another side a pathogenic study about FIP infection in cats. The paraoxonase 1 activity in dogs is the last inflammatory maker investigated, and results show a probable biological behaviour as a negative acute phase protein. 3. Hypertension markers: homocysteine and entothelin-1 (ET-1). Hypertension usually is secondary to diseases that indirectly could affect the systemic pressure, mainly renal and heart diseases. Homocysteine appears increased in dogs with both heart and kidney disease, but the clinical usefulness and the sensibility is not enough for positive impact on patient management. ET-1 appeared with higher sensibility and specificity in dogs with chronic kidney diseases and with hypertension, and its related with the severity of disease and could be a useful marker for the monitoring the follow up. 4. Development of new methods to investigate sialic acid characteristics and to assess if the different molecules bind to sialic acid molecules could interact with the development of the pathology. Results show an increased metastatic potential when sialic acid has α2,6 conformation in dogs with mammary tumour, and in cats the total sialic acid serum concentration is related to the immune response of cats during Coronavirus infections. 5. At last, evaluation of analytical performances of urine protein-to-creatinine ratio: it’s a commonly used test, but this study revealed a high imprecision that could affect test results.

Pagrindinis darbo tikslas buvo išsiaiškinti, kokie yra dažniausiai pasitaikantys šunų širdies ritmo sutrikimai, kokie pagrindiniai diagnostiniai metodai naudojami aritmijoms nustatyti ir diferencijuoti, bei aprašyti medikamentinius preparatus, naudojamus gydant šunis nuo aritmijų. Darbo metu buvo analizuojami LSMU VA Dr. L. Kiaučeliūno smulkiųjų gyvūnų klinikoje, nuo 2008 iki 2013 metų, surinkti gydytų šunų duomenys, kuriems pasireiškė širdies ritmo sutrikimai. Buvo analizuojami pacientų anamnezės duomenys, kuriuos pateikė gyvūnų savininkai, ieškoma aritmijų pasireiškimo priklausomybių nuo šunų lyties, veislės, amžiaus, kilmės, svorio. Širdies ritmo sutrikimai buvo nustatyti 145 šunims, iš kurių 103 buvo patinai ir 42 patelės. Širdies ritmo sutrikimai buvo diagnozuojami auskultuojant širdies darbą, atliekant echokardiografiją, rentgeninį tyrimą bei užrašant elektrokardiogramą – tai yra pagrindinis aritmijų tyrimo metodas. Buvo nustatyta, jog dažniausiai pasitaikantis širdies ritmo sutrikimas šunims yra sinusinės aritmijos, kurios pasireiškė 61 pacientui, tai sudarė 42 proc. šunų, kuriems buvo nustatytos aritmijos. Be sinusinių aritmijų, tiriamojoje šunų grupėje, buvo nustatyti dar 6 rūšių širdies ritmo sutrikimai: supraventrikulinės ekstrasistolijos buvo diagnozuotos 21 (14,5 proc.) šuniui, ventrikulinės ekstrasistolijos – 34 (23,4 proc.), Hiso pluošto kojyčių blokados – 9 (6,2 proc.), Io AV blokados – 10 (6,9 proc.), IIo AV blokados – 6 (4,1 proc.), o IIIo AV blokados - 3 (2... [toliau žr. visą tekstą]
The main aim of work – to determine what are the most common canine cardiac rhythm disorders, analyse diagnostic methods and describe basic medical drugs used in the treatment of arrhythmias. Data of dogs morbidity of cardiac rhythm disorders in 2008 – 2013 has been collected at Dr. L. Kriaučeliūnas Small Animal Clinic of Veterinary Academy of Lithuanian University of Health Sciences. Cardiac rhythm disorders have been diagnosed to 145 dogs: 103 of them were males, and 42 – were females. Clinical diagnosis has been approved according to results of diagnostic methods like: auscultation of heart rate and sounds, X-ray, cardioechoscopy and the main diagnostic method of cardiac arrhythmias – EKG. Sinus arrhythmia has been determined as the most common cardiac rhythm disorder of dogs. This type of arrhythmia has been diagnosed to 61 (42 %) dogs. There were more types of arrhytmias diagnosed like: premature ventricular contractions – 34 (23,4 %) cases, premature atrial contractions - 21 (14,5 %) cases, Hiss bundle branch blocks – 9 (6,2 %) cases and AV blocks – 17 (13,1 %) cases (p<0.001). Male dogs were ill with the cardiac rhythm disorders more often compared to females (71,0% vs. 29,0 %). Arrhythmias were diagnosed to 109 (75,2 %) purebred dogs and 36 (24,8 %) mixed breed dogs (p<0.01). Cardiac rhythm disorders have been diagnosed to 13 Rottweilers (11,9%) and 11 German Shepherds (10,0%), which were most common breeds (p<0.01). In addition, dogs from eight years old and older... [to full text]

Orientador: Antonio Fernandes Pestana de Castro
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
Made available in DSpace on 2018-07-13T23:19:32Z (GMT). No. of bitstreams: 1 Arita_GoncalaMariaMartins_M.pdf: 9310353 bytes, checksum: f2ade0de335e487237939e6bbb772414 (MD5) Previous issue date: 1990
Resumo: No presente estudo o virus da lingua azul-sorotipo 4 (VLA-S4) adaptado às linhagens celulares BHK-21, clone 13 e VERO apresentou efeito citopático característico entre 72 e 96h pos-inoculação, com títulos que variaram entre 10 'POT. 3.6¿ a 10 'POT. 5.6¿ DICC 50%/ml. A partir do sedimento das células infectadas as partículas virais foram purificadas em ultracentrifugações em gradiente de CsCI, e apresentavam densidade de cerca de 1,38 g/cm 'POT. 3¿. Ao microscópio eletrônico foram visualizadas partículas com forma aproximadamente esférica e diâmetro médio de 54,99 '+ ou ¿' 0,12 nm, correspondentes ao virion e de 49,68 '+ ou ¿' 0,08 nm, ao nucleocapsideo. O perfil eletroforético do ARN do VLA-S4 mostrou a presença de dez segmentos com valores de PM 2,92 x 10 'POT. 6¿ d (segmento 1) e 0,49 X 10 'POT. 6¿ d (segmento 10) e o das proteínas, as sete proteínas estruturais -VP1 a VP7- com PM variando de 103,69 x 10 'POT. 3¿ d (VP1) a 30,24 x 10 'POT. 3¿ d ( VP7 ) . O antígeno solúvel (AS), produzido a partir do sobrenadante de culturas de células VERO infectadas pelo VLA e concentrado por ultrafiltração sequencial em membranas com CNL de 10.000 e 25.000, apresentou em teste de identidade com os antígenos do NADC e comercial (COM), uma única linha de precipitação, nítida e confluente. Na análise do perfil eletroforético dos três antígenos observou-se um padrão muito semelhante, o que sugere que as três preparações antigênicas contém proteínas semelhantes. A avaliação semi-quantitativa deste antígeno em imunodifusão radial simples apresentou uma potência relativa 127 ligeiramente superior à do NADC e igual à do COM, e na titulação em bloco por imunodifusão dupla. titulo de uso 1/2, evidenciando-se que o AS poderia ser utilizado em provas rotineiras de 1DGA. para o diagnóstico soro lógico da LA. O componente protéico do AS, principal responsável pela linha de precipitação em 1DGA, apresentou PM 60,00 x 10 'POT. 3' d, sugerindo tratar-se, provavelmente, da proteína NS1 (P5a) ou da VP5. A imunofluorescência indireta (IF1), executada em células VERO adicionadas de soros e conjugados anti-1gG de bovinos ou de ovinos, apresentou fluorescência intracitoplasmática, com aspecto granular, e em algumas células foi observada apenas na membrana celular. Foram testadas 190 amostras de soros sanguíneos de bovinos e 72 de ovinos, pelas técnicas de IDGA e 1FI. Em IDGA obteve-se um total de 134 amostras positivas e 128 amostras negativas, enquanto que em 1FI observou-se 137 amostras positivas e 125 negativas. A análise estatística destes dados registrou uma alta concordância para os soros de bovinos e para os soros de ovinos, entre as duas técnicas. Em 1FI encontrou-se alta sensibilidade, especificidade e valores preditivos dos resultados positivos e dos resultados negativos, em comparação com a IDGA. o que nos permite afirmar que as duas técnicas podem ser utilizadas na rotina laboratorial como instrumento para uma avaliação real da situação epidemiológica da LA no nosso meio. A 1FI oferece a vantagem adicional da possibilidade de ser utilizada para detecção de antígenos virais em cultivos celulares inoculados pelo VLA
Abstract: In the present study a strain of Bluetongue virus, serotype 4 (BTV-S4), when adapted to BHK 21, clone 13 and to VERO cell lines has shown a distinctive cytophatic effect between 72 h and 96 h after inoculation, with titres ranging from 10 'POT. 3.6¿ to 10 'POT. 3.6¿ DICT 50%/ml. After a high speed centrifugation of the packed infected cells the viral particles were purified by ultracentrifugation using CsCIgradient, showing density around 1.38 g/cm 'POT. 3¿. In the electron microscope, two types of spherical particles were observed one with a mean diameter of 54.99 ' + ou ¿' 0,12 run corresponding to the virion and the other of 49.68 ' + ou ¿' 0,08 nm, corresponding to the nucleocapsid. The electrophoretic profiles of the RNAextracted from BTV-S4 showed ten segments with MW between 2.96 x 10 'POT. 6¿ d (segment 1) to 0.49 X 10 'POT. 6¿ d (aegment 10). With regard to the atructural proteina, the MW a180 varied from 103.69 X 10 'POT. 3¿ d (VP1) to 30.24 x 10 'POT. 3¿ d (VP7). The aoluble antigen (SA) produced from culture supernatanta of BTV-infected VERO cells and concentrated by aequential ultrafiltration with membranes with cut-off values 10.000 and 25.000, when compared with the antigens produced by by the agar gel the NADC and commercially, showed total identity immunodiffusion (AGID) test forming a nitid and confluent precipitation line without any spur. The electrophoretic protein profile of the 3 antigens was quite ainÜlar suggesting an identical antigen preparation. A semiquantitative evaluation of thia antigen by aingle radial immune diffusion test showed a relative potency slightly higher than the NADC and equal to the commercial one. Furthermore checking block titration revealed that routine dilution of this antigen to be used in AG1D for the serological diagnosis of BT should be 1/2. The proteie eomponent of the AS. main responsible for the IDGA reaction, showed a MW around 60.00 x 10 'POT. 3¿ d suggesting that this might be, probably, the NS1 (P5a) protein or the VP5. The indireet immunofluorescent technique (IIFT) carried out with BTVinfected VERO cells using bovine and ovine sera and anti-1gG conjugates showed a granular intracytoplasmatie fluorescenee and in some cells this fluorescence was observed only on cellular membranes. Sera from 190 bovine and 72 ovine were examined by AG1D and 11FT. In the AG1D 134 samples were positive for BTV antibodies and 128 were negative, whereas in the 11FT 137 sera were positive for BTV antibodies and 125 were negative. Statistical analysis of these data showed close agreement between the two techniques, regardless of the kind of sera examined. 1n the 11FT, high sensitivity, specificity and predictable values for positive and negative results were found compared with the 1DGA, which means, that both techniques can routinely be used in epidemiologic evaluation studies of BT in our country. The 11FT offers an additional advantage as it may be used to detect viral antigens in BT-infected cell lines
Mestrado
Microbiologia
Mestre em Ciências Biológicas

Thermography is the infrared imaging (or thermal imaging) obtained with an infrared camera. Based on these thermal images it is possibile to perform accurate temperature measurements, to detect even small temperature differences. Scientific applications of infrared thermography are numerous and located in many different areas. In medicine, the temperature of the body is a complex phenomenon. The body produces heat that must be dispersed into the environment. The skin functions as the interface between heat production and the environment. This dynamic body has adapted to balance the internal and external conditions, in response to physiological needs of the body. Infrared are widely accepted as accurate and reliable tool to perform medical examinations and diagnostic tests. Many veterinarians already use infrared thermography as a noninvasive method, reliable and fast, to identify hot spots. This technique provides in fact, in real time, a visual map of gradients in temperature of the skin. Because heat is one of the main signals of approaching of inflammation or injury, it is possibile to use infrared camera to detect and diagnose these problems in advance. This technology also allows to easily locate damaged nerves and muscles. Among the most common applications: axis deviation, diseases of joints, fractures, tendinopathy and many other inflammatory conditions. In addition to the wide use in the horse sports, is growing strongly the use of infrared materials and in other fields of veterinary medicine, as its use in farms of small and large ruminants, wild animals and into zoo, as a tool for diagnosis of diseases or evaluation of alterations of animal welfare. Trial 1: Infrared thermography, allowing you to perform evaluations of temperature away from animals and without contact, can be very important in determining their acute stress. The purpose of this study is, therefore, assess the potential of infrared thermography as a technique to detect stress in the horses in order to obtain a noninvasive method of diagnostic and complementary to other diagnostic techniques. The experimental test is to subject various horses to acute stress and to examine certain physiological parameters (blood corticotropin and cortisol, changes in rectal temperature and heart rate) related to changes in temperature, detected by infrared thermography to eye level and the trigone lip. The first report of stress was induced by banding of the eyes of animals (proof of banding), then another test was conducted in which the stressful event was the administration of food so that the horses were able to view it without reaching and smell (proof of frustration). Our study also provided as a comparison of the response to acute stress in two groups of animals, one of which was placed in enriched environment, the other served as control. In this experiment participated in 9 horses (aged between 5 and 20 years, belonging to different races) For statistical analysis, given the relatively small number of subjects who have been part of the experimental evidence, has chosen to use non-parametric statistical tests. The Wilcoxon test was used for comparison between baseline values and those before the proof and subsequently statistical analysis within the same groups of "enriched" and "control", while the Spearman's rank test was used for the correlation between them. Baseline analysis of results, in general, we can claim to have observed that the surveys carried out by using the camera, do not significantly influence the parameters considered in animals. There are not major differences between the baseline and the pre-test, this may indicate that our manual and our presence does not have varied far the physiological conditions of the horses. In the course of each stress test, however, we have noticed that the infrared camera detects temperature changes, increasing or decreasing, and therefore we have always noticed a difference in temperature between the initial and final values of the tests, both level of eye and lip trigone. Finally, considering our data between enriched and control group, we can see that even in this case, there is a difference that depends on the type of stress test implementation. Descriptive analysis of results obtained in enriched and control groups, as has been previously analyzed each group individually, we noted that the proof bandage influence the temperature of the eye, increasing it, while the trigone is variable. The test of frustration, however, decreases the temperature at eye and attrigone lip. Trial 2: A proper operation of the milking is essential both for the welfare of the buffalo and for an optimize productivity, maintaining a healthy breast. Rising the vaccum level, while allowing more rapid extraction of milk, the other affects the integrity of the breast tissue. The increase of the vaccum increases the rate of flow of milk by lowering the time of issuance, but may also be a cause of occurrence of diseases borne apparatus in different species from breast milk. The mechanical action exerted by the compound during the milking vacuum, and the weight of the sheath, induces changes in the tissues of the nipple that may alter the resistance to the entry of pathogenic bacteria causing infections. The objective of this study was to evaluate the influence of different levels of high and low vacuum on temperature changes of the nipple in Buffalo, before, during and after the milking process, by infrared thermography. Within the herd were considered two groups of 14 buffaloes, which were milked at 42kPa and after three weeks of adaptation to progressively lower levels of vacuum were milked at 36kPa, using the same parameters of pulsation. Thermographic images of the back of the udder and teats were captured before milking, during milking, immediately after milking and up to 5 minutes after milking. The camera was a practical method for detecting changes at surface temperature of the nipple and analyzing the results obtained using the two tests of milking, we highlighted the important data. At first observation of the data reveals that the low vacuum influence greater than the high elevation of the nipple thermal vacuum. Another very interesting observation is related to the time of milking: in fact are very similar and therefore the gap found between the temperatures taken after milking, is not due to the time of milking, but probably on the level of vacuum applied. All these tests have shown a higher sensitivity of the middle and tip teat from the base, applying equally high or low vacuum. This implies a greater influence of the nipple at both anatomical and thermal. Trial 3: Subacute rumen acidosis (SARA) represent one of the most important metabolic disorders in intensive dairy farms that affects rumen fermentations, animal welfare, productivity and profitability. The determination of ruminal pH is a key factor for the diagnosis of SARA. Different methods are available for the collection of rumen fluid for pH analysis. According several researcher rumenocentesis may be useful for the collection of rumen fluid for pH determination. A group of at least 12 cows (early or middle lactation) is defined as having SARA when more than 30% of them show a ruminal pH lower than 5.5. The purpose of this study was to verify the applicability of thermography as a tool as an aid to diagnosis of this disorder fermentative. Also, apply the thermographic evaluation of point of needle insertion from rumenocentesis in order to further confirm the absence of negative implications of this technique on the health and animal welfare. The results of this study may show how infrared thermography can be of great importance as a tool as an aid to diagnosing subacute rumen acidosis because the alterations of ruminal fermentations leads to a change in temperature of the rumen and/or surface circulation in relation to the mechanisms of heat loss. Moreover, since some authors still consider rumenocentesis as an invasive technique and various professionals deem too difficult to use in routine clinical investigations, this study has again demostrated that no animal has shown particular problems during this diagnostic procedure, and not have been identified local or systemic reactions following the ruminocentesis
La termografia è la produzione di immagini a infrarossi (o immagini termiche) ottenute con una telecamera a infrarossi, detta termocamera. Sulla base di tali immagini termiche si possono eseguire accurate misurazioni della temperatura, per individuare differenze di temperatura anche minime. Le applicazioni scientifiche della termografia a infrarossi sono numerose e localizzate nei campi più disparati. In campo medico la temperatura del corpo è un fenomeno complesso. L'organismo produce calore che si deve disperdere nell'ambiente. La cute funziona da interfaccia tra la produzione di calore e l'ambiente. Questo organo dinamico si adegua costantemente per bilanciare le condizioni interne ed esterne, rispondendo alle esigenze fisiologiche del corpo. Gli infrarossi sono ampiamente accettati come strumento accurato e affidabile per effettuare esami medici e diagnostici. Numerosi medici veterinari utlizzano già  la termografia a infrarossi come metodo non invasivo, affidabile e rapido, per individuare punti caldi. Questa tecnica fornisce infatti, in tempo reale, una mappa visiva dei gradienti della temperatura della cute. Poichè il calore è uno dei segnali principali dell'approssimarsi di infiammazioni o infortuni, si possono usare gli infrarossi per individuare e diagnosticare con largo anticipo tali problemi. Questa tecnologia consente anche di individuare facilmente nervi e muscoli lesionati. Tra le applicazioni più comuni: deviazioni assiali, patologie articolari, fratture, tendinopatie e molti altri stati infiammatori. Oltre al largo impiego nel cavallo sportivo, sta crescendo fortemente l'utilizzo degli infrarossi anche in altri campi e materie della medicina veterinaria, come il suo utilizzo negli allevamenti di piccoli e grandi ruminanti, selvatici e zoo, come strumento di diagnosi di patologie e valutazione di alterazioni di benessere animale. Lavoro 1: La termografia ad infrarossi, permettendo di eseguire valutazioni di temperatura lontano dagli animali e senza contatto, può essere molto importante nella determinazione di un loro stress acuto. Lo scopo di questo studio è, perciò, valutare le potenzialità  della termografia ad infrarossi come tecnica per individuare lo stress nel cavallo sportivo, al fine di ottenere un metodo non invasivo e complementare ad altre tecniche diagnostiche. La prova sperimentale consiste nel sottoporre diversi soggetti ad uno stress acuto ed esaminare se alcuni parametri fisiologici (corticotropina ematica, cortisolo ematico, variazioni della temperatura rettale e frequenza cardiaca) ed altri di tipo comportamentale potessero essere in relazione alle variazioni della temperatura, rilevate con l'ausilio della termografia ad infrarossi, a livello dell'occhio e del trigono labiale. La prima situazione di stress è stata indotta mediante il bendaggio degli occhi degli animali (prova di bendaggio); successivamente è stata svolta un'altra prova nella quale l'evento stressante è stato la somministrazione di cibo in modo tale che i cavalli riuscissero a vederlo ed annusarlo senza raggiungerlo (prova di frustrazione). Il nostro studio prevedeva, inoltre, il confronto della reazione ad uno stress acuto di due gruppi di animali; uno di questi è stato posto in ambiente arricchito, l'altro fungeva da controllo. L'arricchimento ambientale da noi messo in pratica è stato di tipo alimentare. A questo esperimento hanno partecipato 9 cavalli (di età compresa tra 5 e 20 anni, appartenenti a varie razze). Per l'analisi statistica, dato il numero relativamente ridotto dei soggetti che hanno fatto parte della prova sperimentale, si è scelto di usare test statistici di tipo non parametrico. Il test di Wilcoxon è stato utilizzato per il confronto tra i valori basali e quelli antecedenti ogni prova stressogena e successivamente per l'analisi statistica all'interno degli stessi gruppi di "arricchito" e di "controllo"; mentre il test per ranghi di Spearman è stato utilizzato per le correlazione tra gli stessi. Nell'analisi dei risultati basali, in generale, possiamo affermare di aver osservato che le rilevazioni effettuate mediante l'uso della termocamera, non influenzano in maniera significativa i parametri considerati negli animali. Abbiamo potuto osservare che, infatti, nonostante le correlazioni e le differenze significative riportate per alcuni parametri in alcuni gruppi, non vi sono differenze rilevanti tra i valori basali e quelli pre prova; questo può indicare che le nostre manualità  e la nostra presenza non hanno variato di molto le condizioni fisiologiche dei cavalli. Nel corso di ogni singola prova di stress, invece, abbiamo potuto notare che la termocamera ad infrarossi rileva variazioni della temperatura, in aumento o in diminuzione, ed abbiamo perciò sempre rilevato una differenza della temperatura tra i valori iniziali e finali delle prova, sia a livello di occhio sia di trigono labiale. Infine, Esaminando i nostri dati tra gruppo arricchiti e controllo, possiamo notare che, anche in questo caso, vi è una differenza che dipende dal tipo di prova di stress messa in atto. Dall'analisi descrittiva dei risultati ottenuti nei gruppi arricchiti e di controllo, così come constatato analizzando in precedenza ogni gruppo singolarmente, abbiamo notato che la prova di bendaggio influenza la temperatura dell'occhio, aumentandola, mentre quella del trigono risulta variabile; la prova di frustrazione, invece, diminuisce la temperatura sia a livello oculare sia di trigono labiale. Lavoro 2: Un corretto funzionamento dell'impianto di mungitura è essenziale sia per il benessere della bufala che per ottimizzare la produttività  e mantenere una mammella sana. L'innalzamento del vuoto di lavoro se da un lato consente un'estrazione più rapida del latte, dall'altro condiziona l'integrità dei tessuti mammari. L'aumento del vuoto operativo accresce la velocità  di deflusso del latte diminuendo i tempi di emissione, ma può anche essere causa dell'insorgenza di patologie a carico dell'apparato mammario nelle diverse specie da latte. L'azione meccanica esercitata durante la mungitura dall'effetto combinato del vuoto, delle guaine e del peso del gruppo, induce dei cambiamenti nei tessuti del capezzolo che ne possono alterare la resistenza all'ingresso dei batteri patogeni causa di infezioni mastitiche. L'obiettivo di questa ricerca è stato quello di valutare l'influenza dei diversi livelli di alto e basso vuoto sui cambiamenti di temperatura del capezzolo nella bufala, prima, durante e dopo il processo di mungitura, attraverso la termografia ad infrarossi. All'interno della mandria sono stati considerati due gruppi di 14 bufale, le quali venivano munte a 42kPa e dopo tre settimane di adattamento a livelli di vuoto progressivamente più bassi, erano munte a 36kPa, usando gli stessi parametri di pulsazione. Le immagini termografiche dell'area posteriore della mammella e dei capezzoli erano catturate prima della mungitura, durante la mungitura, immediatamente dopo la mungitura e fino a 5 minuti dopo la mungitura. La termocamera è risultata un metodo pratico per la rilevazione di alterazioni della temperatura superficiale del capezzolo ed analizzando i risultati ottenuti eseguendo le due prove di mungitura, abbiamo evidenziato dei dati importanti. Ad una prima osservazione dei dati si nota che il basso vuoto influenza maggiormente l'innalzamento termico del capezzolo rispetto all'alto vuoto. Un'altra osservazione piuttosto interessante è relativa ai tempi di mungitura: infatti sono molto simili e dunque il gap riscontrato tra le temperature prese dopo la mungitura, non è dovuto ai tempi di mungitura, ma probabilmente dal livello di vuoto applicato. Tutte queste analisi hanno dimostrato una superiore sensibilità  del middle teat e del tip teat rispetto alla base teat, applicando indistintamente alto o basso vuoto. Ciò implica una maggiore influenza del capezzolo sia a livello anatomico, che termico. Lavoro 3: L'acidosi ruminale subacuta (SARA dall'Anglosassone: subacute ruminal acidosis) rappresenta una delle principali problematiche nell'allevamento della bovina da latte, in grado di provocare notevoli ripercussioni economiche, sia dirette che indirette. Dalle ricerche effettuate in questi ultimi anni, si può affermare che la tecnica più idonea e sicura per il prelievo del liquido ruminale al fine di una diagnosi presuntiva di SARA è la ruminocentesi: in un'azienda si effettua la ruminocentesi in un gruppo di 12 bovine nelle prime fasi di lattazione e si emette diagnosi di SARA quando almeno il 30% delle bovine in esame presenta un pH ruminale inferiore a 5.5. Scopo del presente lavoro è stato quello di verificare l'applicabilità  della termografia ad infrarosso quale mezzo d'ausilio per la diagnosi di tale disturbo fermentativo; inoltre è stata valutata l'applicabilità  della termografia nel punto di inserzione dell'ago da ruminocentesi al fine di valutare (o escludere) eventuali implicazioni negative di tale tecnica sullo stato di salute e benessere dell'animale. Dai risultati di tale studio si può indicare come la termografia ad infrarosso possa essere di grande importanza quale mezzo d'ausilio per la diagnosi di SARA, per il fatto che l'alterazione delle fermentazioni ruminali conduce ad una variazione di temperatura del rumine e/o di circolo superficiale in relazione ai meccanismi di termo dispersione. Inoltre, a conferma di quanto ampiamente indicato in bibliografia i risultati del presente studio confermano che la ruminocentesi rappresenta uno strumento utile e prezioso di diagnostica in medicina bovina e che tale manualità  non provoca alcun danno dal punto di vista della salute e del benessere della bovina da latte

Invasive urothelial carcinoma (UC) is the most common form of urinary bladder cancer and most dogs ultimately die of the disease. It is derived from urothelium, a unique, highly specialised epithelium which lines the lower urinary tract and functions as a physical barrier as well as a sensory structure. Canine UC is believed to be an appropriate animal model because canine UC shares many characteristics with human UC. A substantive comparative study on tumorigenic molecules expressed by canine and human UC is anticipated as an important development toward understanding the mechanism. Tyrosine kinases are proteins that phosphorylate other proteins on tyrosine residues. Evidence suggests that in both human and veterinary patients, tyrosine kinases are often abnormally activated in malignant tumours. Examples of receptor tyrosine kinases include Kit and VEGFR2, all of which are known to be dysregulated forms of cancer. Toceranib phosphate (Palladia®; Pfizer Animal Health, Madison, NJ, USA) is an oral oxindole receptor tyrosine kinase inhibitor that blocks the activity of VEGFR2, PDGFRα/β, FLT-3, KIT and CSFR1. The aims of this study were to extend the knowledge about the immunohistochemical factors involved in urothelial carcinogenesis and identify risk factors for development of UC.

The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from PDF of title page (University of Missouri--Columbia, viewed on July 14, 2010). Thesis advisor: Cristi R. Cook. "May 2010" Includes bibliographical references.

Darbo tikslas: įvertinti šunų gerklų edemų etiotropinius faktorius, taikomas diagnostikos priemones bei gydymo ypatumus. Darbo uždaviniai: 1. Apžvelgti literatūroje pateikiamus duomenis apie gerklų edemų šunims etiologiją. 2. Išanalizuoti patologinio proceso pagrindinius ir pagalbinius diagnozavimo metodus. 3. Išanalizuoti gerklų edemų gydymui taikomas priemones, pateikti patologinio proceso prevencijos būdus. Buvo renkami duomenys 2005–2012 m. pacientų, kurie sirgo ligomis, sukeliančiomis gerklų edemą. Nuo 2005–2008 m. analizuotos tik pacientų ligos istorijos, kuriems buvo diagnozuota gerklų edema (n=133), o nuo 2008–2012 m. – gyvūnai (n=163) tirti kartu su veterinarijos gydytojais, dalyvauta atliekant diagnostines procedūras bei paskiriant gydymą. Išnagrinėti gerklų edemos etiologiniai veiksniai, jų diagnozavimo ir gydymo būdai. Tyrimų rezultatai ir išvados: išanalizavus surinktus duomenis apie pacientus, sergančius ligomis, kurios iššaukia gerklų edemą, nustatyta, kad pagrindiniai etiologiniai veiksniai, sukėlę gerklų edemą šunims, buvo laringitas (54 proc. arba 159 atvejai, p>0,05), laringotracheitas (27 proc. arba 78 atvejai, p>0,05), trachėjos kolapsas (11 proc. arba 34 atvejai, p<0,05), brachicefalinis sindromas (6 proc. arba 19 atvejų, p<0,05), navikai gerklų srityje (1 proc. arba 4 atvejai, p>0,05), vabzdžių įgėlimai (0,3 proc. arba 1 atvejis, p>0,05) ir šunidžių kosulys (0,3 proc. arba 1 atvejis, p>0,05). Gerklų edema dažniau diagnozuota patinams (58 proc. arba 17... [toliau žr. visą tekstą]
The objective of the research–to understand factors about dogs laryngeal oedema, laryngeal oedema diagnostic and treatment. Task of the research: 1. To analyze information about dogs laryngeal edema in literature. 2. To identify pathological process main and supporting diagnostic procedures. 3. To analyze treatment and ways of prevention of the laryngeal edema in dogs. 2005–2012 years, the information was picked about pacients, who had diseases, which stimulate laryngeal edema, from 2005–2008 information was picked about patients who has diseases which stimulate laryngeal edema, and from 2008–2012, the pacients were researched with vet doctor near abay. Were identifyied 296 pacients and their causes and treatment of laryngeal edema. Results and conclusions: main etiology factors of laryngeal edema in dogs were: laringytis (54 percent or 159 cases, p>0,05), laringotracheitis (27 percent or 78 cases, p>0,05), collapse of trachea (11 percent. or 34 cases, p<0,05), brachicefalic syndrome (6 percent. or 19 cases, p<0,05), tumours (1 percent or 1 case, p>0,05), bite of insect (0,3 % or 1 case, p>0,05) and kennel cough (0,3 % or 1 case, p>0,05). laryngeal edema,was identified more in males (58 percent or 172 cases), than females (42 percent or 124 cases). The patient average age was from 5,2±1,17 years old. Laryngeal edema diagnosed more in pedigreed dogs (72 percent or 214 cases) than in hybrids (28 percent or 82 cases). Major pedigreed dogs had ilnesses, who were from 1 to 5... [to full text]

La Sindrome da Asfissia Perinatale (PAS) è una delle più comuni patologie che colpiscono il puledro neonato nelle prime 72 h di vita. È una patologia difficile da diagnosticare in quanto non esistono parametri o segni clinici specifici, la sintomatologia è molto variabile in base alla durata e all’intensità dell’insulto ipossico ischemico e al tipo di organo maggiormente colpito. Lo scopo di questo studio è la ricerca e la valutazione di alcuni parametri biochimico-clinici e di alcuni biomarkers per la diagnosi precoce e il corretto trattamento dei puledri affetti da PAS. Nei puledri neonati che presentano questa patologia è stata riscontrata un’ipermagnesiemia al momento del ricovero associata a prognosi infausta, probabilmente causata da un grave danno cellulare con rilascio in circolo del magnesio intracellulare. La PAS potrebbe essere un’ulteriore causa di Euthyroid Sick Syndrome, in quanto abbiamo riscontrato una diminuzione delle concentrazioni di T3 e T4 nei puledri malati rispetto ai sani della stessa età, come avviene in altre malattie sistemiche. Lo studio del profilo proteomico ha permesso di separare le più importanti frazioni proteiche del liquido amniotico di cavalla, mettendo in evidenza similitudini e differenze qualitative e quantitative nei ferogrammi dei puledri sani e di quelli affetti da PAS ed una maggiore variabilità è stata riscontrata nei profili dei liquidi amniotici dei puledri malati. Il glutatione è risultato poco espresso nel puledro neonato, i puledri sani presentano concentrazioni più basse sia rispetto ai malati della stessa età sia agli adulti ma con una tendenza all’aumento nelle prime 24 ore di vita per i sani ed un calo nei malati. La somministrazione della terapia antiradicalica non influisce sulle concentrazioni di glutatione totale ed i puledri deceduti presentano concentrazioni più alte.
Perinatal Asphyxia Syndrome (PAS) that occurs during the perinatal period is one of the most common diseases affecting foals within the first 72 h of life. The diagnosis of PAS is very difficult and relies on an accurate history, identification of neurological signs, and exclusion of other cause of neurological deficits. No specific clinical-pathological and laboratory findings have been highly suggestive of neonatal asphyxia, many of the noted clinical signs can occur with other clinical conditions and depending on the duration and the intensity of hypoxic insult and the tissue injury. The aim of this study was to discover Perinatal Asphyxia Syndrome biomarkers in neonatal foals having an early identification and interventions of the foals at highest risk developing this syndrome. Foals affected by PAS present hypermagnesaemia at admission compared to healthy foals, may be the results of serious tissue damage with cell death and release of intracellular Mg. Higher concentrations is associated to poor outcome. Data obtained in this trial suggest that PAS may cause lower T3 and T4 concentrations in affected foals than in age-matched healthy foals, as reported for other systemic illnesses. PAS could be a cause of Euthyroid Sick Syndrome. The differences revealed between electrophoretic patterns of healthy and sick foals amniotic fluids have showed a qualitative and quantitative variability in the proteomic profile. We found a high proteomic profiling variability also among sick foals. The acute period of hypoxia-ischemia is followed by a period of reperfusion, the second tissue injury occurs in this phase. The cell damage is caused by post-ischemic release of oxygen radicals. Glutathione (endogenous antiradical) concentration (tGSH) is lower in healthy foals compared to sick foals and adults. The antiradical therapy didn’t change the tGSH concentration and no-surviving foals had higher concentration.

La Sindrome da Asfissia Perinatale (PAS) è una delle più comuni patologie che colpiscono il puledro neonato nelle prime 72 h di vita. È una patologia difficile da diagnosticare in quanto non esistono parametri o segni clinici specifici, la sintomatologia è molto variabile in base alla durata e all’intensità dell’insulto ipossico ischemico e al tipo di organo maggiormente colpito. Lo scopo di questo studio è la ricerca e la valutazione di alcuni parametri biochimico-clinici e di alcuni biomarkers per la diagnosi precoce e il corretto trattamento dei puledri affetti da PAS. Nei puledri neonati che presentano questa patologia è stata riscontrata un’ipermagnesiemia al momento del ricovero associata a prognosi infausta, probabilmente causata da un grave danno cellulare con rilascio in circolo del magnesio intracellulare. La PAS potrebbe essere un’ulteriore causa di Euthyroid Sick Syndrome, in quanto abbiamo riscontrato una diminuzione delle concentrazioni di T3 e T4 nei puledri malati rispetto ai sani della stessa età, come avviene in altre malattie sistemiche. Lo studio del profilo proteomico ha permesso di separare le più importanti frazioni proteiche del liquido amniotico di cavalla, mettendo in evidenza similitudini e differenze qualitative e quantitative nei ferogrammi dei puledri sani e di quelli affetti da PAS ed una maggiore variabilità è stata riscontrata nei profili dei liquidi amniotici dei puledri malati. Il glutatione è risultato poco espresso nel puledro neonato, i puledri sani presentano concentrazioni più basse sia rispetto ai malati della stessa età sia agli adulti ma con una tendenza all’aumento nelle prime 24 ore di vita per i sani ed un calo nei malati. La somministrazione della terapia antiradicalica non influisce sulle concentrazioni di glutatione totale ed i puledri deceduti presentano concentrazioni più alte.
Perinatal Asphyxia Syndrome (PAS) that occurs during the perinatal period is one of the most common diseases affecting foals within the first 72 h of life. The diagnosis of PAS is very difficult and relies on an accurate history, identification of neurological signs, and exclusion of other cause of neurological deficits. No specific clinical-pathological and laboratory findings have been highly suggestive of neonatal asphyxia, many of the noted clinical signs can occur with other clinical conditions and depending on the duration and the intensity of hypoxic insult and the tissue injury. The aim of this study was to discover Perinatal Asphyxia Syndrome biomarkers in neonatal foals having an early identification and interventions of the foals at highest risk developing this syndrome. Foals affected by PAS present hypermagnesaemia at admission compared to healthy foals, may be the results of serious tissue damage with cell death and release of intracellular Mg. Higher concentrations is associated to poor outcome. Data obtained in this trial suggest that PAS may cause lower T3 and T4 concentrations in affected foals than in age-matched healthy foals, as reported for other systemic illnesses. PAS could be a cause of Euthyroid Sick Syndrome. The differences revealed between electrophoretic patterns of healthy and sick foals amniotic fluids have showed a qualitative and quantitative variability in the proteomic profile. We found a high proteomic profiling variability also among sick foals. The acute period of hypoxia-ischemia is followed by a period of reperfusion, the second tissue injury occurs in this phase. The cell damage is caused by post-ischemic release of oxygen radicals. Glutathione (endogenous antiradical) concentration (tGSH) is lower in healthy foals compared to sick foals and adults. The antiradical therapy didn’t change the tGSH concentration and no-surviving foals had higher concentration.

Canine chronic inflammatory enteropathies (CIE) are a group of diseases characterized by chronic gastrointestinal (GI) symptoms. Once other causes for GI symptoms have been ruled out, subdivision of CIEs occurs through therapeutic response which involves a unique trial. The therapeutic steps include a dietary change, subsequently the introduction of antibiotic therapy (this step has recently been discouraged), the execution of an endoscopy of the digestive tract and the introduction of an immunosuppressive therapy. Understanding the pathogenesis and the type of CIE in is fundamental for a correct diagnostic and therapeutic management. There are also several laboratory prognostic markers and clinical scores to help the clinician understand the type of CIE and the severity of the patient's clinical condition. Research in the field of dog CIE is in continuous development with the production of new therapeutic and management indications. The aim of this research was to rewire current literature about CIE, with particular focus on diagnostic and therapeutic aspects. Experimental studies, conducted during author’s PhD, were reported. Studies were focused on diagnostic markers (page 13), GI endoscopy (page 57, page 67) and therapeutic management (page 88) of canine CIE.

Šunų I tipo padidinto jautrumo reakcijos klinikinė ir laboratorinė diagnostika. Darbo apimtis 42 puslapiai, sudarytos 8 lentelės, 6 paveikslai, naudoti 62 literatūros šaltiniai. Darbo tikslas - klinikiniais ir laboratoriniais tyrimo metodais įvertinti I tipo alerginės reakcijos atvejus šunų tarpe, kurie pasitaiko nedidelės smulkių gyvūnų klinikos praktikoje. Tyrimas buvo atliekamas veterinarijos klinikoje „Pas pumą“ 2012 – 2013 m. Buvo užregistruoti 38 šunų susirgimai, susiję su I tipo padidinto jautrumo reakcijos klikiniais požymiais, iš savininkų surinkta išsami anamnezė ir atlikti hematologiniai (morfologija) tyrimai. Imunologiniam tyrimui surinkti 13 alergiškų ir 12 sveikų šunų kraujo serumo mėginiai. Imunologinis IFA (ELISA) tyrimas, nustatyti tiriamųjų šunų IgE koncentracijai kraujo serume, buvo atliktas Lietuvos sveikatos mokslų universiteto veterinarijos fakulteto anatomijos ir fiziologijos katedros imunologijos laboratorijoje. Tyrimo metu nustatyta, kad dažniausiai pasitaikanti I tipo padidinto jautrumo reakcijos forma šunų tarpe buvo atopinis dermatitas (63,2 proc.), kurios ryškiausias požymis buvo niežulys (57,9 proc.), daugiausiai apimantis ausis ir pilvo ventralinę sieną. Šunų lytis ir veislė neturėjo reikšmingos įtakos I tipo padidinto jautrumo reakcijos pasireiškimui, o amžiaus tendencija ryški – alergijos pirmieji požymiai pasireiškia 6 mėn. – 2 m. šunims (62,5 – 78,57 proc.), o šunims iki 6 metų amžiaus alergijos pasikartoja dažniau. Kraujo neutrofilų... [toliau žr. visą tekstą]
Master thesis: Clinical and laboratory diagnosis of type I hypersensitivity reaction in dogs. Master thesis volume 41 pages, consisting of 8 tables, 6 pictures, 61 references used. The main aim of the thesis is to evaluate type I hypersensitivity reaction cases in dogs in small animal veterinary practice using clinical and laboratory methods. The study was carried out in veterinary practise “Pas pumą” in 2012 – 2013. 38 cases of dogs associated with type I hypersensitivity reaction were registered and a detailed medical history was collected from these dogs owners. Haematological examination was performed. Blood samples were collected from 13 dogs with type I hypersensitivity and from 12 healthy dogs. Immunological test ELISA was done to evaluate canine serum IgE levels concentration. Testing was performed in Lithuanian University of Health Sciences , Faculty of Veterinary Anatomy and Physiology, Department of Immunology laboratory. The analysis of clinical findings has shown the most common type I hypersensitivity clinical presentation among dogs is atopic dermatitis (63.2 percent). The most common clinical features were pruritus (57.9 percent) mainly of the ears and the ventral abdominal wall. Dog gender and breed had no significant effect on type I hypersensitivity reactions. The analysis showed that first signs of allergy for dogs commonly occur from 6 months until 2 years of age (62.5 – 78.57 percent of dogs). Allergies had a higher incidence of recurrence in dogs up... [to full text]

La sindrome cardiorenale (CRS) può essere definita come un disordine fisiopatologico di cuore e reni in cui la disfunzione acuta o cronica di un organo può indurre una disfunzione acuta o cronica dell'altro. La cardiopatia acquisita più comune che colpisce i cani anziani e che porta a insufficienza cardiaca congestizia (CHF) è la degenerazione mixomatosa della valvola mitrale (MMVD). Il peggioramento della performance cardiaca e la riduzione della perfusione renale contribuiscono allo sviluppo della malattia renale cronica (CKD) (CRS tipo 2). Nei gatti la più frequente malattia cardiaca è la cardiomiopatia ipertrofica (HCM), un disturbo miocardico primario caratterizzato da un aumento della massa cardiaca ipertrofia ventricolare sinistra. L'anemia è spesso associata a insufficienza cardiaca e insufficienza renale in una triade sfavorevole denominata “complesso sindrome cardiorenale - anemia". La carenza di ferro può essere presente, da sola o in combinazione con l’anemia, con peggioramento della qualità della vita e riduzione della sopravvivenza. Lo scopo della tesi era di descrivere la sindrome cardiorenale nella medicina dei piccoli animali attraverso lo studio delle comorbidità e degli strumenti diagnostici innovativi per migliorare la diagnosi rapida e l'approccio medico alle malattie cardiovascolari. In primo luogo, è stato valutato lo stato di ferro nei cani con MMVD; sebbene non così frequente come negli esseri umani, la carenza di ferro è stata riconosciuta in quasi il 20% dei cani con MMVD, con un rischio di 6,3 maggiore di essere incluso in una classe ACVIM più alta per cani con basse concentrazioni di ferro. La dimetilarginina simmetrica, un nuovo biomarker di velocità di filtrazione glomerulare, è stata valutata su sieri di cani con MMVD e si è dimostrata non influenzata dalla presenza della malattia cardiaca, potendo quindi essere considerata un biomarker di funzionalità renale affidabile in questi cani. La creatin-chinasi è stata misurata in sieri di gatti sani e gatti con diverse forme di cardiomiopatia per evidenziare il suo eventuale ruolo nella diagnosi precoce della malattia; i risultati suggeriscono che la macro-CK1 felina può avere una struttura diversa rispetto a quella di altre specie e che, fra le frazioni espresse, CK-MB potrebbe avere un ruolo nella diagnosi delle cardiomiopatie feline. La precisione e l'affidabilità di D-Heart, il primo elettrocardiografo portatile a più derivate utilizzabile nel paziente canino, sono state valutate, per la diagnosi rapida delle aritmie. Il dispositivo ha dimostrato una registrazione efficace e accurata dell'ECG nei cani inclusi nello studio. Infine, il kit ELISA selezionato per rilevare l'aldosterone su campioni di urina di cane è risultato essere accurato ed efficace, costituendo un'alternativa semplice, sicura ed economica al metodo RIA. Il confronto tra cani sani e cani con MMVD (B1) non ha mostrato differenze statisticamente significative nel rapporto aldosterone urinario:creatinina (UAldo:C). I valori medi del rapporto UAldo: C nel presente studio erano maggiori di quelli riportati in letteratura, suggerendo la necessità di uno studio approfondito per rivalutare la soglia di normalità stabilita dalla letteratura (1.0 μg/g), che non sembra essere valida nella popolazione del presente studio.
Cardiorenal syndrome (CRS) can be defined as a pathophysiologic disorder of the heart and kidneys whereby acute or chronic dysfunction of one organ may induce acute or chronic dysfunction of the other. The most common acquired heart disease affecting old dogs and leading to congestive heart failure (CHF) is myxomatous mitral valve disease (MMVD). The worsening of cardiac performances and reduction of renal perfusion contribute to the development of chronic kidney disease (CKD) (CRS type 2). In cats the most common cardiac disease is hypertrophic cardiomyopathy (HCM), a primary myocardial disorder characterized by increased cardiac mass and a hypertrophied, not dilated, left ventricle. Anemia is often associated with heart failure and renal insufficiency in an unfavorable triad called “CRS-anemia” complex. Iron deficiency can be present, alone or in combination with anemia, worsening quality of life and shortening survival. The aim of the thesis was to describe cardiorenal syndrome in small animal medicine trough the identification of general comorbidities and particular novel diagnostics tools to improve fast diagnosis and the medical approach to the cardiovascular diseases in small animal medicine. Firstly, iron status in dogs with MMVD was evaluated; although not as frequent as in humans, iron deficiency was recognized in almost the 20% of dogs with MMVD, with a 6.3 higher risk of being included in a higher ACVIM class. Symmetric dimethylarginine, a novel biomarker of glomerular filtration rate, was evaluated on sera of dogs with MMVD and proved to be not influenced by the presence of the heart condition and can thus be considered a reliable biomarker in these dogs. Creatine-kinase was measured in sera of healthy cats and cats with different forms of cardiomyopathy to highlight its role in the early diagnosis of the disease; the results suggests that feline macro-CK1 may have a different structure compared with other species and a potential role of CK-MB in the evaluation of feline cardiomyopathies. Accuracy and reliability of D-Heart, the first portable, multiple lead smartphone electrocardiograph in the canine patient, was evaluated, for rapid diagnosis of arrhythmias. The device proved effective and accurate recording of ECG in the canine patient. Finally, the selected ELISA kit to detect aldosterone in canine urine was found to be accurate and effective, constituting a simple, safe and economical alternative to the radioimmunoassay method. Comparison between healthy dogs and dogs with MMVD (B1) showed no statistically significant difference in urinary aldosterone:creatinine (UAldo:C) ratio. The mean values of UAldo:C ratio in the present study were greater than those reported in literature, suggesting the need of a deep study to re-evaluate the normality threshold set by the literature (1.0 μg/g), which does not seem to be valid in the population of the present study.

Thesis (Ph.D.) - University of Glasgow, 2008.
Ph.D. thesis submitted to the Faculty of Veterinary Medicine, University of Glasgow, 2008. Includes bibliographical references. Print version also available.

Doctor of Philosophy
Department of Diagnostic Medicine/Pathobiology
Daniel U. Thomson
The widespread use of beta adrenergic agonists in beef cattle production has been adopted by the beef industry in recent years to improve weight gain and feed efficiency at the end of the feeding period. During this feeding period, anecdotal reports of increased mortality during the timeframe in which beta agonists were being fed to cattle was also reported, and confirmed in epidemiologic studies. Additionally, adverse animal welfare events at abattoirs in cattle fed beta adrenergic agonists were reported in August 2013. The objectives of this dissertation were to investigate physiologic and management factors that may be associated with adverse effects of the use of beta adrenergic agonists in cattle. Two studies were conducted, one to establish normal Holter monitor registration values and evaluate the electrocardiographic effects of zilpaterol and ractopamine hydrochloride on finishing steers, and one to develop a model to investigate the physiologic effects of forced exercises in finished cattle, which was hypothesized to be a possible factor in reported adverse cattle welfare events in August 2013. Thirty steers were enrolled to evaluate the effect of ractopamine, zilpaterol or negative control on arrythmia and mean heart rate at 4 different time periods during a 28 day feeding period. Cattle fed ractopamine and zilpaterol had increased heart rate (P < 0.05) but no differences in arrythmia rates were found. Forty steers were enrolled in a study at a commercial feeding facility to develop a model for fatigue in cattle forced to run 1,540 m compared to control cattle walked 1,540 m. Blood lactate, cortisol, rectal temperature, heart rate was increased (P < 0.05), blood pH decreased (P < 0.01) and to have reduced locomotion, as measured by pedometers, during the 48 hour period following handling compared to controls. Additionally cattle that were fatter and forced to run had increased lactate (P =0.057) and lower blood pH (P < 0.01) than thinner cohorts. Cattle handling method is a factor in the health and welfare of cattle and the continued adoption of low stress handling methods throughout the beef industry should be pursued.

La sindrome di Cushing (SC) è una delle endocrinopatie più comuni nel cane. La diagnosi richiede l'integrazione di anamnesi, segnalamento, segni clinici, esami ematobiochimici, test endocrini specifici e diagnostica per immagini . Nel corso degli anni diversi sono i principi attivi testati per la terapia della SC del cane. In passato, il mitotane è stato il farmaco più utilizzato, sebbene il suo uso risulti complicato e non privo di potenziali effetti collaterali. Recentemente, il trilostano ha dimostrato di essere un trattamento efficace per il controllo dei sintomi ed è stato approvato per tale uso nel cane. Al fine di testare metodiche non invasive per la diagnosi di SC nel cane abbiamo valutato le concentrazioni di cortisolo nel pelo (HCC) .Queste risultavano significativamente più elevate nei cani con SC rispetto ai cani sani e malati. Questo test può essere quindi considerato una procedura diagnostica non invasiva in cani con un elevato sospetto di SC. Inoltre, a causa della difficile reperibilità dell’ACTH esogeno sono state valutate le concentrazioni di cortisolo basale come strumento di monitoraggio in cani con SC trattati con trilostano. Tuttavia la singola valutazione del cortisolo basale non rappresenta un parametro efficace ed accurato per il monitoraggio della terapia con trilostano. Sono stati inoltre valutati i fattori prognostici in cani con SC alla diagnosi. L' iperfosfatemia è risultata un riscontro comune nei cani SC, rappresentando un fattore prognostico negativo. La terapia chirurgica non è una procedura di routine nella SC del cane, tuttavia abbiamo descritto l'approccio di ipofisectomia transsfenoidale in un Galgo spagnolo di 8 anni con SC . Il cane è stato sottoposto per due volte ad ipofisectomia transsfenoidale che ha permesso di rimuovere completamente il macroadenoma ipofisario. In conclusione, questi studi ci hanno permesso di indagare alcuni aspetti patogenetici, clinici e diagnostici della SC del cane.
The Cushing Syndrome is one of the most common canine endocrinopathies. The diagnosis requires the integration of anamnesis, signalment, clinical signs, blood work, specific endocrine tests and the diagnostic imaging. Over the years several drugs were tested for the treatment of canine Cushing's syndrome (CS). In the past, mitotane has been the most widely used drug, although its use is complicated and with many potential side effects. Recently, trilostane has been proven to be an effective treatment for the control of symptoms and it is approved for this use in dog. In order to test noninvasive techniques as a diagnostic tool for dogs with CS we measured the hair cortisol concentrations (HCC). HCC were significantly higher in CS dogs compared with healthy dogs and sick control dogs.Because of the lack of specificity, this test could be considered a noninvasive procedure only in dogs with a high suspicion of HC. Moreover due to the difficult supplying of exogenous ACTH we evaluated the baseline cortisol concentration as a monitoring tool in dogs with CS treated with trilostane. It is evident from our study that the ACTH stimulation test cannot be replaced by basal cortisol concentration. We investigated the prognostic factors of dogs with newly diagnosed hypercortisolism and this study the hyperphosphatemia is a common finding in newly diagnosed CS dogs and represents a negative prognostic factor. The surgical therapy it is not a routine procedure in the canine CS however we described the transsphenoidal hypophysectomy approach in 8-year-old, female Spanish Galgo dog with CS. The dog was underwent for two time at transsphenoidal hypophysectomy that have removed the completely the pituitary macroadenoma. In conclusion, these studies allowed us to investigate some aspects of pathogenetic, clinical and diagnostic of the SC of the dog.

La sindrome di Cushing (SC) è una delle endocrinopatie più comuni nel cane. La diagnosi richiede l'integrazione di anamnesi, segnalamento, segni clinici, esami ematobiochimici, test endocrini specifici e diagnostica per immagini . Nel corso degli anni diversi sono i principi attivi testati per la terapia della SC del cane. In passato, il mitotane è stato il farmaco più utilizzato, sebbene il suo uso risulti complicato e non privo di potenziali effetti collaterali. Recentemente, il trilostano ha dimostrato di essere un trattamento efficace per il controllo dei sintomi ed è stato approvato per tale uso nel cane. Al fine di testare metodiche non invasive per la diagnosi di SC nel cane abbiamo valutato le concentrazioni di cortisolo nel pelo (HCC) .Queste risultavano significativamente più elevate nei cani con SC rispetto ai cani sani e malati. Questo test può essere quindi considerato una procedura diagnostica non invasiva in cani con un elevato sospetto di SC. Inoltre, a causa della difficile reperibilità dell’ACTH esogeno sono state valutate le concentrazioni di cortisolo basale come strumento di monitoraggio in cani con SC trattati con trilostano. Tuttavia la singola valutazione del cortisolo basale non rappresenta un parametro efficace ed accurato per il monitoraggio della terapia con trilostano. Sono stati inoltre valutati i fattori prognostici in cani con SC alla diagnosi. L' iperfosfatemia è risultata un riscontro comune nei cani SC, rappresentando un fattore prognostico negativo. La terapia chirurgica non è una procedura di routine nella SC del cane, tuttavia abbiamo descritto l'approccio di ipofisectomia transsfenoidale in un Galgo spagnolo di 8 anni con SC . Il cane è stato sottoposto per due volte ad ipofisectomia transsfenoidale che ha permesso di rimuovere completamente il macroadenoma ipofisario. In conclusione, questi studi ci hanno permesso di indagare alcuni aspetti patogenetici, clinici e diagnostici della SC del cane.
The Cushing Syndrome is one of the most common canine endocrinopathies. The diagnosis requires the integration of anamnesis, signalment, clinical signs, blood work, specific endocrine tests and the diagnostic imaging. Over the years several drugs were tested for the treatment of canine Cushing's syndrome (CS). In the past, mitotane has been the most widely used drug, although its use is complicated and with many potential side effects. Recently, trilostane has been proven to be an effective treatment for the control of symptoms and it is approved for this use in dog. In order to test noninvasive techniques as a diagnostic tool for dogs with CS we measured the hair cortisol concentrations (HCC). HCC were significantly higher in CS dogs compared with healthy dogs and sick control dogs.Because of the lack of specificity, this test could be considered a noninvasive procedure only in dogs with a high suspicion of HC. Moreover due to the difficult supplying of exogenous ACTH we evaluated the baseline cortisol concentration as a monitoring tool in dogs with CS treated with trilostane. It is evident from our study that the ACTH stimulation test cannot be replaced by basal cortisol concentration. We investigated the prognostic factors of dogs with newly diagnosed hypercortisolism and this study the hyperphosphatemia is a common finding in newly diagnosed CS dogs and represents a negative prognostic factor. The surgical therapy it is not a routine procedure in the canine CS however we described the transsphenoidal hypophysectomy approach in 8-year-old, female Spanish Galgo dog with CS. The dog was underwent for two time at transsphenoidal hypophysectomy that have removed the completely the pituitary macroadenoma. In conclusion, these studies allowed us to investigate some aspects of pathogenetic, clinical and diagnostic of the SC of the dog.

Abstract Objectives: To retrospectively compare and contrast the clinical presentation, diagnostic findings and survival in Boxer dogs with cardiomyopathy, with or without left ventricular (LV) systolic failure. Animals, materials and methods: Medical records of Boxers referred between 1993 and 2008 in which a diagnosis of ventricular arrhythmias and/or cardiomyopathy was made, were reviewed. Dogs were divided into two groups according to their left ventricular (LV) systolic diameter, groupAnormal (20 dogs) or group Bdilated (59 dogs). Results: Dogs in group A had a better outcome than dogs in group B (median survival time of 124 and 17 weeks respectively, p < 0.001). In group B, dogs with a history of collapse had a worse outcome (median survival time of 10 weeks) compared with dogs not showing collapse (median survival time 24 weeks) (p ¼ 0.031). Conclusions: The majority of dogs, in this UK study, presented with the myocardial dysfunction form of the disease, with LV dilation and congestive heart failure signs. The prognosis was worse in dogs with LV dilation compared to dogs with a normal LV and ventricular arrhythmias. In the Boxers with LV dilation, dogs with collapse had a worse prognosis than those without.

La poliradicoloneurite acuta idiopatica (ACIP) è una patologia infiammatoria che interessa le radici di più nervi spinali, descritta soprattutto nel cane, più raramente nel gatto, caratterizzata da insorgenza acuta di paresi/paralisi flaccida. L’ACIP mostra notevoli similitudini con la sindrome di Guillan-Barrè dell’uomo (GBS), in cui la patogenesi è su base autoimmunitaria ed è stata correlata con la presenza di alcuni fattori scatenanti (trigger). Lo scopo di questo lavoro è stato quello di caratterizzare l’ACIP in 26 cani, descrivendone la sintomatologia, l’evoluzione clinica, i risultati degli esami diagnostici. La diagnosi si è basata sui riscontri dell’anamnesi, della visita neurologica e del decorso confermata, quando possibile, dai rilievi elettrodiagnostici. Su tutti i cani è stata valutata l’esposizione a specifici agenti infettivi (Toxoplasma gondii, Neospora canunim, Ehrlichia canis, Leishmania infantum), o altri fattori (come vaccinazioni) che potrebbero aver agito da “trigger” per l’instaurarsi della patologia; sull’intera popolazione e su 19 cani non neurologici (gruppo di controllo), si è proceduto alla ricerca degli anticorpi anti-gangliosidi. La sintomatologia di più frequente riscontro (25/26) ha coinvolto la funzione motoria (paresi/plegia) con prevalente interessamento dei 4 arti (24/25) . Sei cani hanno ricevuto una terapia farmacologica, che non ne ha influenzato il decorso, favorevole in 24/26 casi. In 9 pazienti è stata rilevata una precedente esposizione a potenziali trigger; in 10 casi si è riscontrato un titolo anticorpale positivo ad almeno un agente infettivo testato. In 17/26 cani si è ottenuto un titolo anticorpale anti-GM2 e anti-GA1; nella popolazione di controllo solo un caso è risultato positivo. Questi risultati hanno contribuito a consolidare le conoscenze di questa patologia, validando l’utilità della ricerca anticorpale anti-gangliosidica per la diagnosi di ACIP e facendo intravedere la possibilità che l’ACIP possa essere assimilate alla GBS anche dal punto di vista patogenetico, per la quale potrebbe essere considerata come modello animale spontaneo.
Acute canine idiopathic polyradiculoneuritis (ACIP) is an immune-mediated disorders affecting peripheral myelin and axons. It is described mainly in dogs, rarely in cats, and is characterized by an acute onset of flaccid paraparesis, which usually progresses rapidly to tetraparesis/tetraplegia. ACIP is considered to be the canine equivalent of the human peripheral nerve disorder Guillain-Barré syndrome (GBS) but an aetiological relationship remains to be demonstrated. In human medicine, various antecedent events have been associated with GBS, including bacterial or viral infections, thought to trigger the immune system. The aim of this work was to characterize the ACIP in 26 dogs, describing clinical signs, outcome and results of diagnostic tests. The ACIP diagnoses was based on history, neurological examination and outcome, confirmed by electrophysiological (21/26) features. It was also evaluated a potential exposure to specific infectious agents (Toxoplasma gondii, Neospora caninum, Ehrlichia canis, Leishmania infantum), or other factors such us vaccinations, as a trigger for the onset of the disease. Furthermore, the sera of all ACP dogs and 19 non-neurological control dogs were screened for IgG Antibodies to 10 glycolipids and their heteromeric complexes. The majority of dogs (25/26) showed gait abnormalities, which involved the four limbs in most cases (24/25). Only 6 dogs received drug treatment, that did not affected the outcome, which was favorable in 24/26. Nine dogs showed a previous exposure to triggers and in ten was detected a positive antibodies title against an infectious agent. The antibodies anti-GM2 and GA1 were detected in 17/26 of ACP dogs, whilst all controls except for one were negative for anti-glycolipid Antibodies. It suggested that, as in humans, anti-ganglioside antibodies could be an effective biomarker in confirming ACP diagnosis. This also might suggest that ACP share a similar pathophysiology with GBS, for which it could be considered a naturally occurring animal model

La poliradicoloneurite acuta idiopatica (ACIP) è una patologia infiammatoria che interessa le radici di più nervi spinali, descritta soprattutto nel cane, più raramente nel gatto, caratterizzata da insorgenza acuta di paresi/paralisi flaccida. L’ACIP mostra notevoli similitudini con la sindrome di Guillan-Barrè dell’uomo (GBS), in cui la patogenesi è su base autoimmunitaria ed è stata correlata con la presenza di alcuni fattori scatenanti (trigger). Lo scopo di questo lavoro è stato quello di caratterizzare l’ACIP in 26 cani, descrivendone la sintomatologia, l’evoluzione clinica, i risultati degli esami diagnostici. La diagnosi si è basata sui riscontri dell’anamnesi, della visita neurologica e del decorso confermata, quando possibile, dai rilievi elettrodiagnostici. Su tutti i cani è stata valutata l’esposizione a specifici agenti infettivi (Toxoplasma gondii, Neospora canunim, Ehrlichia canis, Leishmania infantum), o altri fattori (come vaccinazioni) che potrebbero aver agito da “trigger” per l’instaurarsi della patologia; sull’intera popolazione e su 19 cani non neurologici (gruppo di controllo), si è proceduto alla ricerca degli anticorpi anti-gangliosidi. La sintomatologia di più frequente riscontro (25/26) ha coinvolto la funzione motoria (paresi/plegia) con prevalente interessamento dei 4 arti (24/25) . Sei cani hanno ricevuto una terapia farmacologica, che non ne ha influenzato il decorso, favorevole in 24/26 casi. In 9 pazienti è stata rilevata una precedente esposizione a potenziali trigger; in 10 casi si è riscontrato un titolo anticorpale positivo ad almeno un agente infettivo testato. In 17/26 cani si è ottenuto un titolo anticorpale anti-GM2 e anti-GA1; nella popolazione di controllo solo un caso è risultato positivo. Questi risultati hanno contribuito a consolidare le conoscenze di questa patologia, validando l’utilità della ricerca anticorpale anti-gangliosidica per la diagnosi di ACIP e facendo intravedere la possibilità che l’ACIP possa essere assimilate alla GBS anche dal punto di vista patogenetico, per la quale potrebbe essere considerata come modello animale spontaneo.
Acute canine idiopathic polyradiculoneuritis (ACIP) is an immune-mediated disorders affecting peripheral myelin and axons. It is described mainly in dogs, rarely in cats, and is characterized by an acute onset of flaccid paraparesis, which usually progresses rapidly to tetraparesis/tetraplegia. ACIP is considered to be the canine equivalent of the human peripheral nerve disorder Guillain-Barré syndrome (GBS) but an aetiological relationship remains to be demonstrated. In human medicine, various antecedent events have been associated with GBS, including bacterial or viral infections, thought to trigger the immune system. The aim of this work was to characterize the ACIP in 26 dogs, describing clinical signs, outcome and results of diagnostic tests. The ACIP diagnoses was based on history, neurological examination and outcome, confirmed by electrophysiological (21/26) features. It was also evaluated a potential exposure to specific infectious agents (Toxoplasma gondii, Neospora caninum, Ehrlichia canis, Leishmania infantum), or other factors such us vaccinations, as a trigger for the onset of the disease. Furthermore, the sera of all ACP dogs and 19 non-neurological control dogs were screened for IgG Antibodies to 10 glycolipids and their heteromeric complexes. The majority of dogs (25/26) showed gait abnormalities, which involved the four limbs in most cases (24/25). Only 6 dogs received drug treatment, that did not affected the outcome, which was favorable in 24/26. Nine dogs showed a previous exposure to triggers and in ten was detected a positive antibodies title against an infectious agent. The antibodies anti-GM2 and GA1 were detected in 17/26 of ACP dogs, whilst all controls except for one were negative for anti-glycolipid Antibodies. It suggested that, as in humans, anti-ganglioside antibodies could be an effective biomarker in confirming ACP diagnosis. This also might suggest that ACP share a similar pathophysiology with GBS, for which it could be considered a naturally occurring animal model

Resumo: Com o objetivo de estabelecer valores de referência para os parâmetros hematotógicos de cutias (Dasyprocta azarae Lichtenstein, 1823), foram realizadas análises de hemograma de 42 exemplares, 29 machos e 13 fêmeas com idades variando entre 1 e 37 meses. Para fins de classificação, os animais foram divididos em duas faixas etárias: jovens quando tinham 8 meses ou menos, e adultos quando tinham mais de oito meses de idade. Para a colheita das amostras os animais foram contidos através de injeção da associação de cloridrato de cetamina, cloridrato de xilazina e de sulfato de atropina. Os resultados foram comparados quanto às variáveis sexo e idade, sendo que diferenças significativas foram observadas somente quanto a idade e não quanto ao sexo dos animais. Os parâmetros número total de eritrócitos, concentração de hemoglobina e concentração de hemoglobina globular média foram mais baixos nos animais jovens, enquanto o número total de leucócitos, número total de linfócitos e número total de monócitos foram mais baixos nos animais adultos. Quanto aos demais parâmetros avaliados, conclui-se que os animais jovens e adultos podem ser agrupados em intervalos de valores (média ± erro padrão) iguais, cujos resultados foram: volume globular: 50,52 ± 0,95 %; volume globular médio: 69,42 ± 1,37 fl; proteínas plasmáticas totais: 6,11 ± 0,11 g/dl; número total de neutrófilos segmentados: 2198,95 ± 244,91 /mm3; número total de neutrófilos bastonetes: 48,05 ± 8,58 /mm3; número total de eosinófilos: 61,28 ± 9,76 /mm3; número total de basófilos: 13,05 ± 2,50 /mm3. Nos parâmetros, onde a diferença estatística quanto a idade foi observada estabeleceu-se um intervalo de valor (média ± erro padrão) para cada faixa etária correspondente, com os seguintes resultados: número total de eritrócitos: 7,23 ± 0,18 x106 /mm3 para animais jovens e 7,91 ± 0,26 x106 /mm3 para animais adultos; concentração de hemoglobina: 14,22 ± 0,46 g/dl para animais jovens e 16,44 ± 0,28 g/dl para animais adultos; concentração de hemoglobina globular média: 28,55 ± 0,55 % para animais jovens e 31,16 ± 0,48 % para animais adultos; número total de leucócitos: 4,97 ± 0,32 x103 /mm3 para animais jovens e 3,14 ± 0,145 x103 /mm3 para animais adultos; número total de linfócitos: 2424,56 ± 130,00 /mm3 para animais jovens e 1238,76 ± 159,33 /mm3 para animais adultos; número total de monócitos: 81,09 ± 9,54 /mm3 para animais jovens e 43,59 ± 10,45 /mm3 para animais adultos. Palavras-chave: Dasyprocta azarae, hemograma; roedores.