African Swine Fever Virus (ASFV) is an Asfivirus and is the only member of the family Asfarviridae. It manifests as a disease that varies from acute to sub-acute or chronic forms. A true carrier state in domestic pigs is unknown but chronically affected individuals may carry and spread the virus for extended periods. African Swine Fever (ASF) is a socio-economically important disease characterized by high morbidity and mortality affecting the livelihood of many small to big scale farmers and seriously compromising international trade. Strategic measures to control this disease are by physical containment and culling in outbreak situations. There is no vaccine available. Nevertheless, every pork producer should ideally be actively involved in having biosecurity measures in place to avoid contamination and contacting their veterinary services in case of suspicion of ASF to have appropriate samples analysed. Official veterinary services must be equipped with proper diagnostic tools in order to provide a quick response. The sensitivity of currently available diagnostic tests at the Transboundary Animal Diseases Programme, Onderstepoort Veterinary Institute was analysed in order to report the best technique available. Sensitivity to ASF virus infection and therefore diagnostic potential of cell primary cultures as bone marrow macrophages, blood macrophages and alveolar macrophages was done via comparison of titre results from inoculations of ASFV SPEC 257 as control, and ASFV MOZ 1/98. In addition, molecular detection of specific DNA fragments within the viral genome were compared using five different PCRs. Bone marrow macrophage cultures and blood macrophage cultures were the most reliable cells whereas alveolar macrophages more often showed contamination. Results show that PPA PCR and real time PCR detected the highest diluted samples, thus the lowest concentration of virus, in both trials done with ASFV MOZ 1/98 and ASFV SPEC 257. In addition, animal trials were performed by inoculating domestic pigs with four different ASFV isolates of varying pathogenicity. These viruses were all from distinct geographic origins. Non-virulent ASFV OURT 3/88 and high virulent ASFV BENIN 1/97 were previously described and used as reference viruses. ASFV MOZ 1/98, suspected of having high virulence and ASFV MKUZE, which was thought to be of low virulence were included in this study to provide further information on the pathological and clinical outcome of the disease as well as measuring viral replication in various organs and blood. The study showed that ASFV MKUZE was of intermediate virulence, whilst ASFV MOZ 1/98 was highly virulent with a high mortality rate. Results confirmed the inadequacy of ASFV MKUZE to act as vaccine opposed to ASFV OURT 3/88. Following this, a potential vaccine by use of attenuated Portuguese ASFV OURT 3/88 tested against virulent heterologous challenge with a strain now known with certainty to cause acute ASF, the isolate ASFV MOZ 1/98 collected from a diseased pig in Mozambique. Domestic commercial pigs where submitted to either one or two vaccinations before challenge. Viral load in blood and tissue samples was higher in unvaccinated animals and higher in single vaccinated than in pigs vaccinated twice. However, acute ASF afflicted all groups with severe clinical signs and post-mortem lesions. Although it did not confer total immunity it was determined that pigs vaccinated with European attenuated ASFV OURT 3/88 acquired partial protection against challenge with virulent southern Africa ASFV MOZ 1/98. / Dissertation (MSc)--University of Pretoria, 2012. / gm2014 / ab2015 / Veterinary Tropical Diseases / unrestricted

This thesis focuses on the development of an open source software capable of automatic face detection in an image captured by a thermal camera, followed by a temperature measuring. This software is supposed to aid in the COVID-19 pandemics. The developed software is independent of used thermal camera. In this thesis, I am using TIM400 thermal camera. The implementation of the face detection was achieved by an OpenCV module. The methods tested were Template Matching, Eigen Faces, and Cascade Classifier. The last-mentioned had the best results, hence was used in the final version of the software. Cascade Classifier is looking for the eyes and their surrounding area in the image, allowing the software to subsequently measure the temperature on the surface of one's forehead. One can therefore be wearing a face mask or a respirator safely. The temperature measuring works in real time and the software is able to capture several people at once. It then keeps a record of the temperature of each measured individual as well as the time of the measurement. The software as a whole is a part of an installation file compatible with the Windows operating system. The functionality of this software was tested – the video recordings are included in this thesis.

Zusammenfassung Katharina Regina Winkler Hämatologisch-immunologische Verlaufsuntersuchungen bei Kühen mit Gebärparese Medizinische Tierklinik der Veterinärmedizinischen Fakultät der Universität Leipzig Eingereicht im September 2014 100 Seiten, 41 Abbildungen, 13 Tabellen, 282 Literaturangaben, 17 Seiten Anhang Schlüsselwörter: Gebärparese, Stoffwechsel, Glucocorticoide, Endotoxin, Blutbild Problemstellung: Der Mineralstoffwechsel unterliegt immunologischen Einflüssen. Vitamin-D3 ist essentiell für die Antigen- und Zytokinsynthesen in den Blutzellen. Die Osteoblastenreifung wird durch Zytokine beeinflusst. Das tangiert die Gebärparese (GP) und erschließt potentiell prophylaktische sowie therapeutische Ansätze. Zielstellung: Es wurde geprüft, ob a) es Mineralstoffunterschiede bei der GP-Diagnose und im Verlauf zu früheren Studien gibt, b) die Parameter Endotoxin (ET), anti-Lipid A-IgG-Titer (ALA-IgG-Titer) und Haptoglobin (Hp) sowie Leuko- und Erythrogramme gesicherte Beziehungen zur GP sowie zu Mineralstoffen und immunologischen Parametern haben, c) die Therapie bei GP durch Glucocorticoide verbessert werden kann und d) Jungkühe mehr Geburtsstress und Belastungen des Ca-Pi-K-Stoffwechsels haben. Versuchsanordnung: Untersucht wurden 111 HF-Kühe bzw. Jungkühe: 21 GP-Kühe mit Grundbehandlung, 22 GP-Kühe mit zusätzlicher Dexamethason-21-iso-ni-cotinat-Therapie (Dexa-IN), 40 gesunde Kontrollkühe (KG) und 28 gesunde Jungkühe. Laborkontrollen erfolgten bei den GP-Kühen vor der Therapie, bei den KG 1 - 3 Tage post partum (d p. p.) sowie 1 d und 14 d nach dem Therapiedatum. Analysiert wurden neben Stoffwechselparametern Endotoxin (LAL-Test), ALA-IgG-Titer, Haptoglobin sowie das Leuko- und Erythrogramm. Ergebnisse: Die Erstbehandlung war bei 47 % ohne und 67 % mit Dexa-IN-Therapie erfolgreich; die Heilungsrate betrug 74 bzw. 82 %, d. h., die Dexa-IN-Therapie verbesserte das Behandlungsergebnis bei GP ohne Nebenwirkungen. 69,8 % der GP-Kühe hatten eine kombinierte Hypokalz- und Hypophosphatämie. 24 Stunden nach Beginn der Therapie waren beide Mineralstoffe in den GP-Gruppen wieder physiologisch. 11,6 % der GP-Kühe und 10,7 % der Jungkühe hatten eine Hypophosphatämie. Das ist offensichtlich eine Folge des Kalbestresses in diesen Gruppen. Die Mg-, Na- und Cl-Konzentrationen waren in allen Gruppen physiologisch. Mg korrelierte negativ mit Ca und Pi (p<0,01). Die K-Konzentrationen waren in den GP-Gruppen einen d p. p. signifikant niedriger als in den KG. Sie korrelierten mit Ca- und Pi in den GP-Gruppen mit 0,42 bis 0,48 (p<0,01). Auf stärkere Stresseinflüsse auf K wiesen Korrelationen zu Glucose, Bilirubin, eosinophile und basophile Granulozyten sowie Lymphozyten hin. Die Fe-Konzentrationen der GP- und KG-Kühe waren physiologisch. Fe korrelierte mit ALA-IgG-Titer gesichert negativ. Die ET-Konzentrationen ließen nur schwache Beziehungen zur GP erkennen, wie rET:Ca=-0,17 (p<0,05). ET korrelierte mit den ALA-IgG-Titern gesichert positiv (Dexa-IN-Gruppe). Die ALA-IgG-Titer differierten bei den Kühen nicht gesichert, sie korrelierten nicht mit Ca, aber mit Pi und mit der Mehrzahl der klinisch-chemischen und hämatologischen Parameter. Das zeigt die Entzündungseinflüsse auf den Pi-Stoffwechsel mit der Förderung von Hypophosphatämien. Die Hp-Konzentrationen streuten stark und waren in allen Gruppen am Diagnose- und noch mehr am Folgetag erhöht (p>0,05). Bei Jungkühen wies der höhere Anstieg auf stärkeren Kalbestress hin. Die Leukozytenzahl war am GP-Diagnosetag erhöht (Leukozytose; p>0,05) und sank zum Folgetag in den Normbereich ab. In der Dexa-IN-Gruppe war der Abfall am stärksten (p<0,05). Die Leukozytenzahl korrelierte gesichert negativ mit den ALA-IgG-Titer sowie in den GP-Gruppen mit den Pi- sowie Ca-Konzentrationen, ebenso die neutrophilen Granulozyten. Eosinophile, basophile Granulozyten sowie Lymphozyten sanken p. p. ab (p<0,05) und korrelierten gesichert mit Ca und Pi. Die GP-Kühe hatten am Diagnosetag eine Monozytose (p<0,05). Die Monozyten korrelierten mit den ALA-IgG-Titern, mit dem Pi und dem Ca gesichert negativ. Sie hatten die engsten Beziehungen zum Entzündungsgeschehen sowie den Pi- und Ca-Konzentrationen. Die CK-Aktivitäten waren am Diagnose- und am Folgetag gegenüber der KG signifikant erhöht. Mit Ca korrelierte die CK in allen Kuh-Gruppen gesichert negativ, mit Pi nur in der GP-Gruppe ohne Dexa-IN. Die CK stand in enger gesicherter Beziehung zu Entzündungsindikatoren. Hämoglobin war in den GP-Gruppen am Diagnosetag signifikant, der Hämatokrit und die Erythrozytenzahlen tendenziell gesteigert. Schlussfolgerungen: Die Studie zeigt bei GP-Kühen vielfältige Beziehungen des Ca-Pi-K-Stoffwechsels zu Entzündungsindikatoren und legt ursächliche Einflüsse nahe, besonders zu ALA-IgG-Titern und Monozyten. Sie können Ursachen für Hypophosphat- und Hypokalämie sein. Zusätzliche Dexa-IN-Therapie verbessert das Behandlungsergebnis.

The group A beta-haemolytic streptococcus is known to be the aetiologic agent in acute rheumatic fever, but the exact pathogenesis remains obscure. A review of the histopathology of the Aschoff body suggests that the cardiac pathology is a granulomatous hypersensitivity reaction. However the streptococcus has not been found in the lesions, and the agent responsible for the granuloma has not yet been identified. Circulating immune complexes have previously been measured in some children with acute rheumatic fever. The normal or raised complement components measured by some workers in acute rheumatic fever suggests that the immune complexes may not be complement fixing. Considering that the usual assays for measuring immune complexes depend on complement fixation, the failure of the immune complexes to fix complement might produce false negative results. A physical, non-complement fixing assay (polyethylene glycol precipitation - PEG), was therefore used to measure circulating immune complexes. Results were expressed as total IgG precipitated (g/L), or as a percentage of serum IgG. Immune complexes were also measured by two complement dependent assays, a Clq binding assay (ClqBA), and conglutinin binding assay (CBA). Complexes were assayed in 15 children with acute rheumatic carditis (ARC), 11 with non-active, chronic rheumatic heart disease (CRHD), 13 with acute poststreptococcal glomerulonephritis (APSGN), and 15 normal children and adults (NORMAL). Total haemolytic complement, complement components as well as the complement breakdown product C3d, were measured.

Host cell attachment by pathogenic bacteria can play very different roles in the course of infection. The pathogenic spirochetes Borrelia hermsii and Borrelia burgdorferi sensu lato which cause relapsing fever and Lyme disease, respectively, are transmitted by the bite of infected ticks. After transmission, these spirochetes can cause systemic infection. Relapsing fever spirochetes remain largely in the bloodstream causing febrile episodes, while Lyme disease will often colonize a variety of tissues, such as the heart, joint and nervous system, resulting in a chronic multisystemic disorder. Borrelia species have the ability to bind to various cell types, a process which plays a crucial role in pathogenesis and may influence spirochetal clearance from the bloodstream. Colonization of multiple tissues and cell types is likely promoted by the ability to bind to components found in target tissues, and many B. burgdorferi adhesins have been shown to promote attachment to a wide variety of cells and extracellular matrix components. Different Lyme disease strains have been shown to preferentially colonize certain tissues, although the basis of this tissue tropism is not well understood. In this study we found that among different Lyme disease strains, allelic variation of the adhesin DbpA contributes to variation in its in vitro binding activities raising the possibility that this variation contributes to tissue tropism in vivo. In studying B. hermsii infection, we found evidence by both histological and fluorescence in situ hybridization (FISH) analysis of tissues that indicated that red blood cells were removed by tissue resident macrophages in infected mice. Spirochetes in the spleen and liver were often visualized associated with RBCs, lending support to the hypothesis that direct interaction of B. hermsii spirochetes with RBCs leads to clearance of bacteria from the bloodstream. Our findings indicate that host cell attachment play a key role in the establishment of Lyme disease infection, and in contrast contributes to the clearance of relapsing fever infection.

L’importance et l’expansion des arboviroses comme la dengue ou le virus Zika nécessite des modèles pour mieux comprendre et prédire leurs dynamiques. La propagation vectorielle de ces maladies est influencée par de multiples facteurs humains et environnementaux qui rendent complexe la construction de modèles épidémiologiques parcimonieux. Parallèlement, de nombreux outils théoriques et computationnels existent désormais pour confronter ces modèles aux données observées. L’objectif de ce travail de thèse est donc d’apporter l’éclairage des données sur les modèles de propagation des arboviroses. Dans un premier temps, il s’agit d’identifier les éléments les plus importants à incorporer pour modéliser les dynamiques de la dengue en milieu rural, dans la région de Kampong Cham (Cambodge). Différents modèles sont comparés, complexifiant à la fois le détail de l’histoire de la maladie et la prise en compte des formes de stochasticité. Dans le cadre déterministe, on a pu souligner l’importance des interactions entre sérotypes, et le faible intérêt pour la représentation explicite des moustiques vecteurs et des individus asymptomatiques. Par ailleurs, la prise en compte des incertitudes indique qu’une large part de la dynamique est capturée seulement par la stochasticité et non par les éléments du squelette déterministe du modèle. Aussi étudie-t-on dans un second temps d’autres aspects de la transmission de la dengue, comme la saisonnalité et la structure spatiale, grâce à des données d’épidémies à Rio de Janeiro (Brésil). Dans un dernier temps, ces méthodes et modèles sont appliqués à l’étude d’un arbovirus émergent, le virus Zika. A partir de données d’épidémies survenues dans le Pacifique, les paramètres-clé de la propagation du virus sont estimés dans le cadre stochastique, et leur variabilité est envisagée à la fois en termes de contexte géographique et de modèle épidémiologique, par la comparaison de quatre îles et de deux modèles à transmission vectorielle. Par ailleurs, la question des interactions potentielles du virus Zika avec celui de la dengue est explorée. / Arboviruses such as the dengue and Zika viruses are expanding worldwide and mo- deling their dynamics can help to better understand and predict their propagation, as well as experiment control scenarios. These mosquito-borne diseases are influenced by a multiplicity of human and environmental factors that are complex to include in parsimonious epidemiological models. In parallel, statistical and computational tools are nowadays available to confront theore- tical models to the observed data. The objective of this PhD work is therefore to study arbovirus propagation models in the light of data. Firstly, in order to identify the most important elements to incorporate in models for dengue dynamics in a rural setting, several dengue models are com- pared using data from the Kampong Cham region in Cambodia. Models incorporate increasing complexity both in the details of disease life history and in the account for several forms of sto- chasticity. In the deterministic framework, including serotype interactions proved decisive, whereas explicit modeling of mosquito vectors and asymptomatic infections had limited added value, when seasonality and underreporting are already accounted for. Moreover, including several forms of un- certainties highlighted that a large part of the disease dynamics is only captured by stochasticity and not by the elements of the deterministic skeleton. Therefore, secondly, we explore other aspects of transmission, such as seasonality and spatial structure, in the case of dengue epidemics in Rio de Janeiro (Brazil). Finally, the models and estimation methods are applied to study an emerging arbovirus, the Zika virus. Using data from epidemics in the Pacific, we estimate the key parameters of disease propagation in the stochastic framework and explore their variability in terms of geogra- phic setting and model formulation by comparing four islands and two models with vector-borne transmission. In addition, potential interactions with the dengue virus are explored.

[pt] Os antibióticos eritromicina e canamicina, pertencentes às classes dos macrolídeos e aminoglicosídeos respectivamente, têm sua importância como agentes preservativos no processo de preparação e durante o processo de utilização da vacina contra a febre amarela - apresentação de cinco doses - produzida no Instituto de Tecnologia em Imunobiológicos, Bio-Manguinhos, FIOCRUZ, RJ. Este trabalho propõe apresentar duas novas abordagens para a determinação espectrofluorimétrica desses antibióticos, considerando o desempenho, inclusive com relação à seletividade, na análise dessa matriz específica (vacina). Um estudo preliminar do comportamento luminescente (fosforescência e fluorescência) dessas substâncias foi realizado em diferentes condições experimentais a fim de encontrar as que permitissem o desenvolvimento dos métodos analíticos. Para a eritromicina, que não apresentou luminescência nativa, foi utilizado um procedimento de derivação fotoquímica de soluções do analito preparadas em meio contendo ácido sulfúrico. Como resultado obteve-se um foto-produto que apresentou fluorescência em 412/465 nm. As condições experimentais foram otimizadas visando a maximização desse sinal fluorescente. Nesse caso, foram estudados o tempo de irradiação com UV, o tipo e a concentração do ácido utilizado, o tempo e a temperatura de aquecimento. O método foi parcialmente validado apresentando limites de detecção e de quantificação iguais a 0,25 ug mL-1 e 0,85 ug mL-1 respectivamente. A faixa linear do método estendeu-se até 200 ug mL-1 e os parâmetros de precisão e robustez foram bastante satisfatórios. O método foi aplicado na análise de uma simulação de vacina contra a febre amarela e na análise de medicamentos comerciais com um percentual de recuperação entre 98,2 e 105,2 %. A metodologia desenvolvida para a canamicina, que também não apresentou fluorescência natural, baseou-se na reação de oxirredução deste composto com o Ce (IV) produzindo o Ce (III), que é uma espécie fortemente fluorescente (255/360 nm). As intensidades dos sinais fluorescentes medidos no par excitação/emissão do Ce (III) foram diretamente proporcionais às concentrações de canamicina. As condições para a realização da reação foram estudadas, avaliando a quantidade de Ce (IV), a concentração e tipo de ácido usado no meio reacional e o tempo e temperatura de aquecimento. A ampla faixa linear (até 1000 ug mL-1) foi obtida, com limites de detecção e quantificação de 1,22 ug mL-1 e 4,08 ug mL-1, respectivamente. O percentual de recuperação obtido na análise da vacina diluída foi de 109,8 % e em amostra diluída de urina enriquecida com o analito foi de 103,4 %. / [en] The antibiotics erythromycin and kanamycin, members of the macrolide and aminoglycoside classes respectively, have their importance as preservative agents for the preparation process and during the using of the yellow fever vaccine, five doses presentation, produced in Technology on Immunobiologicals Institute, Bio- Manguinhos, FIOCRUZ, RJ. In this work two new approaches for the spectrofluorimetric determination of these antibiotics are presented, taking into consideration their selectivity performance for this specific matrix (vaccine). A preliminary study was performed to evaluate the luminescent behavior (phosphorescence and fluorescence) of these substances in different experimental conditions aiming to find the best ones that would lead to the development of the analytical methods. For erythromycin, which does not present native luminescence, a photochemical derivatization was employed using analite solutions prepared in sulfuric acid. As a result, a photo- product was obtained, which presented fluorescence at 412/465 nm. The experimental conditions were optimized aiming the maximization of the fluorescent signal. In this case, the studied parameters were the UV irradiation time, the type and concentration of the acid utilized, and the time and temperature used for the heating step. The method was partially validated, indicating limits of detection and quantification of 0.25 ug mL-1 and 0.85 ug mL-1, respectively. The linear dynamic range of the method extended up to 200 ug mL-1 and the parameters related to precision and robustness were very satisfactory. The method was applied in the analysis of a simulated yellow fever vaccine and in commercial pharmaceutical formulations. Recovery percent between 98.2 and 105.2 % were achieved. The methodology developed for kanamycin, which also do not presented natural fluorescence, was based on the oxirreduction reaction of this compound with Ce (IV), producing Ce (III), a strongly fluorescent species (255/360 nm). The fluorescence intensities, measured in the excitation/emission pair of Ce (III), was directly proportional to the kanamycin concentration. The reaction conditions were studied by the evaluation of the amount of Ce (IV), the type and concentration of the acid utilized in reactional medium and heating time and temperature. Large linear range (up to 1000 ug mL-1) was obtained, with detection and quantification limits of 1.22 ug mL-1 and 4.08 ug mL-1, respectively. The recovery percent obtained in the analysis of a diluted vaccine was 109.8 % while the recovery achieved for diluted spiked urine was 103.4 %.

Since September 2020, Germany has experienced the first ever outbreak of African swine fever (ASF). The first known cases occurred exclusively in wild boar in forest areas in Brandenburg and Saxony; in July 2021, infected domestic pigs were also confirmed for the first time. As wild boar are considered the main reservoir for the virus in the European region, an effective interruption of this infection chain is essential. In particular, the removal and safe disposal of infected carcasses and the direct disinfection of contaminated, unpaved ground are priorities in this regard. For the disinfection, highly potent as well as environmentally compatible disinfectants must be used, which are neither influenced in their effectiveness by the soil condition nor by increased organic contamination. Thus, in this study, slaked lime, milk of lime and quicklime (1% to 10% solutions) were selected for efficacy testing against the test virus recommended by the German Veterinary Society (DVG), Modified Vaccinia Ankara virus (MVAV), and ASF virus (ASFV) in conjunction with six different forest soils from Saxony in two different soil layers (top soil and mineral soil) each. In summary, 10% of any tested lime type is able to inactivate both MVAV and ASFV under conditions of high organic load and independent of the water content of the soil. At least a 4 log reduction of the virus titer in all tested forest soil types and layers and by all applied lime types was observed. In conclusion, the high efficacy and suitability of all tested lime products against both viruses and in the presence of high organic load in forest soil can be confirmed and will help to control ASF spread.

Arbovirus infections are widespread, and their disease burden has increased in the past decade. In Africa, arbovirus infections and fever with unknown etiology are common. Due to the lack of well-established epidemiologic surveillance systems and accurate differential diagnosis in most African countries, little is known about the prevalence of human arbovirus infections in Africa. The aim of this review is to summarize the available epidemiological data and diagnostic laboratory tools of infections with dengue, yellow fever, Zika, and chikungunya viruses, all transmitted by Aedes mosquitoes. Studies indicate that these arboviral infections are endemic in most of Africa. Surveillance of the incidence and prevalence of the infections would enable medical doctors to improve the diagnostic accuracy in patients with typical symptoms. If possible, arboviral diagnostic tests should be added to the routine healthcare systems. Healthcare providers should be informed about the prevalent arboviral diseases to identify possible cases.

Behavioral fever is an essential coping mechanism across ectothermic phyla to aid in combating pathogenic threats. Ectotherms lack internal temperature regulation associated with fever in endotherms; thus, ectotherms can exhibit a behavioral fever response when immunocompromised to thermoregulate by moving to warmer locations. The salamander order Caudata, tend to be keystone species in their resident ecosystems through their role as secondary consumers of invertebrates to maintain the food chain. With growing interest about ecology and conservation of salamanders as species diversity declines, this study was designed to determine if salamanders use their environment to take advantage of behavioral fever. The lungless salamander, Plethodon glutinosus, was used to investigate behavioral fever through exposure to the synthetic viral pyrogen polyinosinic:polycytidylic acid (Poly (I:C)) at doses of 15 µg/g by live wet weight. After 24 hours, the induced fever specimens were placed in a linear temperature gradient with a controlled humidity of 95% throughout. Average temperature preferences were then monitored over a 12-hour period and resulted in control animals preferring cold and moderate temperatures between 15-19 °C while all poly (I:C) injected animals preferred higher temperatures from 19-21°C (p<<0.0001). This result supports that P. glutinous responds to immunocompromising threats such as presented by synthetic viral pyrogen Poly (I:C) through use of behavioral fever.