Master of Veterinary Clinical Studies / Summary Canine Neural Angiostrongyliasis (CNA) is caused by the obligatory neural migration of Angiostrongylus cantonensis larvae in dogs. Characteristically, cases are juvenile dogs with progressive CNS dysfunction characterised by hyperaesthesia and often associated with eosinophilic pleocytosis of the CSF. In Australia, most cases occur between March and June. The rat lungworm, A cantonensis was first described by Chen in 1935 in Canton, China. While initially called Pulmonema cantonensis the parasite was later reclassified as A cantonensis. A disease diagnosed as eosinophilic meningoencephalitis was first described in 1944 in Taiwan. The same disease was reported in 1948 in the East Caroline Islands but it was not until 1961 that A cantonensis was confirmed as the aetiological agent when a patient in a Hawaiian mental institution, who had died of eosinophilic meningoencephalitis, had A cantonensis larvae recovered from the brain and spinal cord. The first reports of animals infected with A cantonensis were made by Mason in 1976 when he described a syndrome occurring in puppies in the Brisbane area, characterised by urinary incontinence, hind limb paresis and hyperaesthesia, often associated with eosinophilic pleocytosis of the CSF. Reports of infection in other species followed including macropods, bats, horses, primates and birds. Twenty-two cases of suspected CNA were collected prospectively to compare with those previously described, including 37 cases published by Mason in 1983, and to examine the accuracy of an ELISA used to diagnose human neural angiostrongyliasis in Australia. Samples were collected from two control populations in an attempt to validate the ELISA results. In the prospective series of cases, there was a significantly older subpopulation of dogs in addition to “classical” young dogs, suggesting that this syndrome can occur at any age and should be considered a differential in any dog with progressive neurological disease. The mortality rate in the prospective group was lower than in the published group, which is a reflection of the severity of the disease in younger animals as is the case with human patients. Definitive diagnosis of neural angiostrongyliasis in human patients has been achieved by identifying A cantonensis larvae within the CSF or aqueous humour. In dogs, the only definitive way to diagnose CNA has been via necropsy. While many cases of CNA are characteristic and presumptive diagnosis can be made based on typical history, signalment, clinical signs, CSF analysis and response to glucocorticoids, there appear to be an increasing number of cases occurring in older dogs, that displaying focal, atypical clinical signs or that develop permanent sequelae. Serology has been a useful tool in diagnosing neural angiostrongyliasis in humans. In its current form the ELISA is not sensitive or specific enough to allow a definitive diagnosis of CNA to be made using serum but is useful when applied to CSF specimens. Further refinement of the antigen or using monoclonal rather than polyclonal antibodies may improve the accuracy of the serology. Alternatively, methods such as Western Blot, Immuno-PCR or dot-blot ELISA, which have been successfully used to diagnoses angiostrongyliasis in humans, may be worthy of investigation The major differential diagnosis for CNA is neosporosis. Other differential diagnoses include idiopathic eosinophilic meningoencephalitis, parasitic infections including Toxoplasma gondii, Taenia solium, Gnathostoma spinigerum, visceral larval migrans (Toxocara canis) and schistosomiasis, fungal, bacterial, viral and rickettsial infections as well as neoplasia, trauma, drug reactions and toxicities. Treatment of CNA has been limited to glucocorticoids, however there may be adjunct therapies including anthelmintices, cyclosporine, and matrix metalloproteinase inhibitors. In Mason’s series of cases the use of anthelmintics significantly worsened the clinical outcome for patients. It does not appear, however, that the use of these agents in species other than the dog exacerbates clinical signs. Acquired immunity is short lived in rats and mice, which would suggest the same is true in dogs. Routine heartworm and intestinal parasite prophylaxis appears to have no influence on the occurrence of CNA.

Master of Veterinary Clinical Studies / Summary Canine Neural Angiostrongyliasis (CNA) is caused by the obligatory neural migration of Angiostrongylus cantonensis larvae in dogs. Characteristically, cases are juvenile dogs with progressive CNS dysfunction characterised by hyperaesthesia and often associated with eosinophilic pleocytosis of the CSF. In Australia, most cases occur between March and June. The rat lungworm, A cantonensis was first described by Chen in 1935 in Canton, China. While initially called Pulmonema cantonensis the parasite was later reclassified as A cantonensis. A disease diagnosed as eosinophilic meningoencephalitis was first described in 1944 in Taiwan. The same disease was reported in 1948 in the East Caroline Islands but it was not until 1961 that A cantonensis was confirmed as the aetiological agent when a patient in a Hawaiian mental institution, who had died of eosinophilic meningoencephalitis, had A cantonensis larvae recovered from the brain and spinal cord. The first reports of animals infected with A cantonensis were made by Mason in 1976 when he described a syndrome occurring in puppies in the Brisbane area, characterised by urinary incontinence, hind limb paresis and hyperaesthesia, often associated with eosinophilic pleocytosis of the CSF. Reports of infection in other species followed including macropods, bats, horses, primates and birds. Twenty-two cases of suspected CNA were collected prospectively to compare with those previously described, including 37 cases published by Mason in 1983, and to examine the accuracy of an ELISA used to diagnose human neural angiostrongyliasis in Australia. Samples were collected from two control populations in an attempt to validate the ELISA results. In the prospective series of cases, there was a significantly older subpopulation of dogs in addition to “classical” young dogs, suggesting that this syndrome can occur at any age and should be considered a differential in any dog with progressive neurological disease. The mortality rate in the prospective group was lower than in the published group, which is a reflection of the severity of the disease in younger animals as is the case with human patients. Definitive diagnosis of neural angiostrongyliasis in human patients has been achieved by identifying A cantonensis larvae within the CSF or aqueous humour. In dogs, the only definitive way to diagnose CNA has been via necropsy. While many cases of CNA are characteristic and presumptive diagnosis can be made based on typical history, signalment, clinical signs, CSF analysis and response to glucocorticoids, there appear to be an increasing number of cases occurring in older dogs, that displaying focal, atypical clinical signs or that develop permanent sequelae. Serology has been a useful tool in diagnosing neural angiostrongyliasis in humans. In its current form the ELISA is not sensitive or specific enough to allow a definitive diagnosis of CNA to be made using serum but is useful when applied to CSF specimens. Further refinement of the antigen or using monoclonal rather than polyclonal antibodies may improve the accuracy of the serology. Alternatively, methods such as Western Blot, Immuno-PCR or dot-blot ELISA, which have been successfully used to diagnoses angiostrongyliasis in humans, may be worthy of investigation The major differential diagnosis for CNA is neosporosis. Other differential diagnoses include idiopathic eosinophilic meningoencephalitis, parasitic infections including Toxoplasma gondii, Taenia solium, Gnathostoma spinigerum, visceral larval migrans (Toxocara canis) and schistosomiasis, fungal, bacterial, viral and rickettsial infections as well as neoplasia, trauma, drug reactions and toxicities. Treatment of CNA has been limited to glucocorticoids, however there may be adjunct therapies including anthelmintices, cyclosporine, and matrix metalloproteinase inhibitors. In Mason’s series of cases the use of anthelmintics significantly worsened the clinical outcome for patients. It does not appear, however, that the use of these agents in species other than the dog exacerbates clinical signs. Acquired immunity is short lived in rats and mice, which would suggest the same is true in dogs. Routine heartworm and intestinal parasite prophylaxis appears to have no influence on the occurrence of CNA.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-03-22T13:55:27Z No. of bitstreams: 1 2016 - Mariana Gomes Lima.pdf: 3236185 bytes, checksum: 3517b88022c7943663ae0274085cc96a (MD5) / Made available in DSpace on 2017-03-22T13:55:27Z (GMT). No. of bitstreams: 1 2016 - Mariana Gomes Lima.pdf: 3236185 bytes, checksum: 3517b88022c7943663ae0274085cc96a (MD5) Previous issue date: 2016-02-23 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / The nematode Angiostrongylus cantonensis has gained notoriety under the public health point of view out of its endemic area in Asia, since in recent years cases of eosinophilic meningoencephalitis have been reported in other continents, such as in the Americas. In Brazil over thirty cases of this zoonotic disease have been recorded and there are reports of the presence of this nematode in 11 states. In its life cycle A. cantonensis has, along their development, need for intermediate and final host, involving predominantly molluscs and rodents, respectively, and different paratenic hosts, humans are accidental hosts. They can also infect a rich variety of terrestrial and aquatic molluscs and in this study, there were used specimens of neotropical planorbids transmitters of schistosomiasis, Biomphalaria straminea and Biomphalaria tenagophila, born and reared in the laboratoryfor experimental infection by A. cantonensis. In order to characterize the resulting physiological profile that host-parasite relationship, the host molluscs were separated into groups of infected and uninfected containing thirty molluscs per group, all made in triplicates. The investigation lasted 21 days, after the pre-patent period molluscs were dissected to collect hemolymph, removal of tissues and use the shell for calcium concentration check. The reproductive biology of both species was observed on the viability of the eggs, as well as (A) Number of eggs/ eggs mass; (B) Number of eggs/mollusc; (C) Number of eggs mass/mollusc and (D) Number of hatched molluscs/mollusc. In the hemolymph there were quantified total protein, glucose, uric acid, urea, the activity of transaminases AST and ALT, lactate dehydrogenase - LDH, and organic acids (oxalic, pyruvic, succinic and lactic). In the cephalopedal mass and digestive gland the glycogen content was measured and in the albumen gland, it was measured the galactogen. The results showed that the snails infected, of both species showed significant reduction in the viability of eggs, 50% for B. straminea and 10% for B. tenagophila. The metabolic status was also strongly affected, with mobilization of glycogen and hemolymph glucose reduction of 68% to B. straminea and 6.48% for B. tenagophila, followed by anincrease of LDH activity of 518.25% to 320.14% B. straminea and to B. tenagophila as well as the concentration of lactic acid, 5,7mM to B. straminea and 0.34 mM for B. tenagophila, and reduction in the concentration of pyruvic and succinic acids. The intense release of gluconeogenic amino acids from protein catabolism reversed the excretory pattern from uricotelic to ureotelic in both B. tenagophila and B. straminea. The tissue injury showed increase of 69% in the concentration of total proteins in the hemolymph of B. straminea and the intense activity of aminotransferases, ALT and AST, an increase of 241.26% and 360%, respectively, compared to the control group. The infection reduced by 51.57% the concentration of total circulating proteins to B. tenagophila, and increased ALT and AST activity at 310% and 280% compared to uninfected snails. The results of this study indicate that the energetic and structural metabolism of both species is strongly affected by the infection, showing the activation of an anaerobic to compensate for the depletion of glicidics stores as well as the change in the excretory pattern depending on the use of substrates protein for the production of ATP. Although studies on clinical, pathological and epidemiological neural angiostrongyliasis have been extensively carried xiii out, data on the metabolic and reproductive profile of snails infected by A. cantonensis are scarce. This pioneering study focused on species of host-parasite relationship certainly contribute to the knowledge of this interaction, as well as to the epidemiology of transmission A. cantonensis in addition to providing new subsidies which can be used in prevention and control of eosinophilic meningitis, zoonosis considered emerging in Brazil. / O nemat?deo Angiostrongylus cantonensis tem ganhado notoriedade sob o ponto de vista da sa?de p?blica fora de sua ?rea end?mica na ?sia, uma vez que nos ?ltimos anos casos de meningoencefal?te eosinof?lica v?m sendo reportados em outros continentes, como nas Am?ricas. No Brasil mais de trinta casos dessa zoonose j? foram registrados e h? relatos da presen?a desse nemat?deo em 11 estados. Em seu ciclo de vida heteroxeno A. cantonensis tem, ao longo do seu desenvolvimento, a necessidade de hospedeiro intermedi?rio e definitivo, envolvendo predominantemente, moluscos e roedores, respectivamente, al?m de v?rios hospedeiros parat?nicos, sendo o homem um hospedeiro acidental. Pode infectar uma rica variedade de moluscos terrestres e aqu?ticos e neste estudo, foram utilizadas para infec??o experimental popula??es nascidas e criadas em laborat?rio de duas esp?cies de planorb?deos neotropicais transmissores da esquistossomose mans?nica, Biomphalaria straminea e Biomphalaria tenagophila. Com a finalidade de caracterizar o perfil fisiol?gico resultante dessa rela??o hospedeiro-parasito, os moluscos hospedeiros foram separados em grupos de infectados e n?o infectados, contendo trinta moluscos por grupo, todos feitos em triplicatas. A investiga??o durou 21 dias, ao fim do per?odo pr?-patente os moluscos foram dissecados para a coleta de hemolinfa, retirada de tecidos e utiliza??o da concha para verifica??o de concentra??o de c?lcio. A oviposi??o dos moluscos de ambas as esp?cies foi observada quanto ? viabilidade dos ovos, assim como (A) N?mero de massas ov?geras/molusco; (B) N?mero de ovos/molusco; (C) N?mero de ovos/massa ov?gera e (D) N?mero de moluscos eclodidos/molusco. Na hemolinfa foram quantificadas as prote?nas totais, glicose, ?cido ?rico, ureia, a atividade das transaminases ALT e AST, lactato desidrogenase - LDH, e os ?cidos org?nicos (ox?lico, pir?vico, succ?nico e l?tico). Na massa cefalopediosa e gl?ndula digestiva, foram mensurados os conte?dos de glicog?nio e na gl?ndula de alb?men, foi mensurado o galactog?nio. Os resultados obtidos mostraram que os moluscos infectados, de ambas as esp?cies, apresentaram redu??o significativa na viabilidade dos ovos, de 50% para B. straminea e de 10% para B. tenagophila. O estado energ?tico tamb?m foi vigorosamente afetado, com mobiliza??o das reservas de glicog?nio, e queda da glicemia de 68% para B. straminea e de 6,48% para B. tenagophila, seguida de uma eleva??o na atividade da LDH de 518,25% para B. straminea e 320,14% para B. tenagophila, bem como na concentra??o do ?cido l?tico, 5,7mM para B. straminea e 0,34 mM para B. tenagophila, e redu??o na concentra??o dos ?cidos pir?vico e succ?nico. A intensa libera??o de amino?cidos gliconeog?nicos a partir do catabolismo proteico inverteu o padr?o excretor de uricot?lico para ureot?lico tanto em B. tenagophila quanto em B. straminea. A les?o tecidual mostrou eleva??o de 69% na concentra??o das prote?nas totais na hemolinfa de B. straminea, bem como a intensa atividade das aminotransferases, AST e ALT, com aumento de 241,26% e 360%, respectivamente, em rela??o ao grupo controle. A infec??o reduziu em 51,57% a concentra??o de prote?nas totais circulantes para B. tenagophila, e elevou a atividade de ALT e AST em 310% e xi 280% em compara??o com os moluscos n?o infectados. Os resultados observados neste estudo indicam que o metabolismo energ?tico e estrutural de ambas as esp?cies foi vigorosamente afetado pela infec??o, mostrando a ativa??o de uma via anaer?bia para compensar a exaust?o das reservas glic?dicas, assim como a mudan?a no padr?o excretor em fun??o da utiliza??o de substratos proteicos para produ??o de ATP. Embora, estudos sobre aspectos cl?nicos, patol?gicos e epidemiol?gicos da angiostrongil?ase neural t?m sido extensivamente realizados, dados acerca do perfil metab?lico e reprodutivo de moluscos infectados por A. cantonensis s?o escassos. Este estudo pioneiro na rela??o parasito-hospedeiro das esp?cies focadas seguramente contribuir? tanto para o conhecimento dessa intera??o, quanto da epidemiologia da transmiss?o de A. cantonensis, al?m de fornecer novos subs?dios que poder?o ser utilizados em medidas de preven??o e controle da meningite eosinof?lica, zoonose considerada emergente no Brasil