El presente trabajo de tesis doctoral pretende indagar las siguientes hipótesis de trabajo: los estadios larvales de Contracaecum spp. e Hysterothylacium spp. experimentan migración hacia la musculatura estriada en las especies de peces estudiadas; los estadios larvales de Contracaecum spp. obtenidas de peces naturalmente infectados provocan lesiones en el tracto digestivo de mamíferos, provocando una respuesta inmune humoral y celular que puede caracterizarse según criterios hematológicos e histopatológicos. Para poner a prueba estas hipótesis se planteó como objetivo general determinar el rol que cumplen las larvas de Contracaecum spp. e Hysterothylacium spp. presentes en recursos pesqueros del sudoeste bonaerense, en el desarrollo de la anisakidosis humana. Los objetivos específicos del trabajo fueron: determinar el rango de hospedadores naturales de Contracaecum spp. e Hysterothylacium spp. en peces marinos y continentales, precisar el status sistemático de estos estadios, evaluar la capacidad miotrópica de los mismos en peces destinados al consumo humano y tipificar el daño tisular de los estadios larvales invasores determinando el poder infectivo en mamíferos, mediante el empleo de un modelo murino y analizando el efecto de esta parasitosis sobre los parámetros hemáticos en el mismo. Para el estudio se tomaron especies de peces de importancia comercial en la región: Cynoscion guatucupa, Odontesthes argentinensis y Odontesthes bonariensis. De un total de 316 ejemplares de peces examinados entre los años 2009-2011, 111 correspondieron a la especie C. guatucupa, 105 correspondieron a O. argentinensis y 100 a O. bonariensis. Se recolectaron larvas de Contracaecum sp. e Hysterothylacium sp. de la cavidad celómica de dichas especies. Las mismas fueron estudiadas histológica y morfológicamente. Se realizaron estudios moleculares de las larvas halladas en C. guatucupa, en los cuales se evidenció una asociación con especies de Contracaecum que parasitan mamíferos. Se tomaron muestras de filete de cada especie de pez, 3.316,85 g de C. guatucupa, 1.754,9 g de O. bonariensis y 1.327,1 g. de O. argentinensis, que fueron analizados mediante digestión enzimática. En el total de filetes analizados solo se encontraron 4 nematodes en O. bonariensis. Para tipificar el daño tisular de los estadios larvales encontrados y determinar el poder infestivo en los mamíferos se inocularon, siguiendo un protocolo de infestación, ratones de cepa Balb/cAnN de 30 g. de peso promedio. Se utilizaron técnicas histológicas de rutina y coloración de hematoxilina-eosina. Los resultados mostraron dos infestaciones positivas con larvas L3 pertenecientes a C. guatucupa. La histopatología evidenció la larva fijada al tubo digestivo, principalmente en la región fúndica del estómago, la cual penetró el epitelio de la mucosa gástrica llegando hasta la capa muscular. Se observó una compresión total del epitelio, con pérdida de la estructura glandular y adoptando un aspecto aplanado entorno a la larva. Un infiltrado leucocitario con presencia de monocitos, neutrófilos, eosinófilos y linfocitos, rodeó los tejidos próximos al sitio de penetración de la larva. En uno de los casos positivos de infestación se observó variación en la composición leucocitaria de la sangre de los ratones. Parte del material infestado fue fijado en glutaraldehído 2,5%, para la observación mediante microscopia electrónica de barrido. La identificación a nivel especifico empleando técnicas moleculares se realizó solo para las larvas de Contracaecum sp. provenientes de la pescadilla, C. guatucupa, debido a que fue la única especie de parasito de las ensayadas que produjo resultados positivos de infestación. Los análisis de las secuencias genómicas indican que éstas larvas se ubicaron en el mismo cluster junto a C. margolisi y C. ogmorhini, ambos parásitos de mamíferos marinos. Hasta tanto se disponga de mayor información a nivel genético del género Contracaecum se concluye que las larvas que produjeron lesión en la mucosa de ratones BALB /cAnN pertenecen al complejo Contracaecum ogmorhini s.l. De acuerdo con los resultados alcanzados hasta el momento, las larvas de Contracaecum ogmorhini s.l. infestivas para el modelo murino utilizado, potencialmente inducirían lesión en otros mamíferos, incluido el hombre. No obstante la exigua prevalencia e intensidad larval registrada en la carne de los pescados estudiados, sumado al hábito de cocción completa de los platos a base de filetes, permiten considerar de bajo riesgo epidemiológico a las especies estudiadas. / The aim of the present work is to test the following hypotheses: the larval stage of Contracaecum spp. and Hysterothylacium spp. migrate towards the musculature in the hosts fishes; the larval stage of Contracaecum spp. and Hysterothylacium spp. obtained of infected fishes caused injuries in the digestive tract of mammals, provoking humoral and cellular immune responses that can be characterized according to hematological and histophatological criteria. To test these hypotheses it was considered as a general objective to determine the role of Contracaecum spp. and Hysterothylacium spp. present in the southwest of Buenos Aires fishery resources in the development of human anisakidosis. The following specific purposes were added: to determine the range of natural hosts of Contracaecum spp. and Hysterothylacium spp. in marine and continental fishes, to confirm the systematic status of these stages, to evaluate the myotropic behavior of the helminths in fish used as human food and to assess the tissue damage of the larval stages invading the mucosa of the digestive tract and to analyzes the effect of this parasitosis on hematological parameters. Some fish species of commercial importance in the region Cynoscion guatucupa, Odontesthes argentinensis and Odontesthes bonariensis were considered to the study. A total of 316 fish individuals were analyzed between 2009 and 2011, 111 corresponded to C. guatucupa, 105 specimens to O. argentinensis and 100 to O. bonariensis. Contracaecum spp. and Hysterothylacium spp. larvae were collected from the visceral cavity of these fishes. They were studied histologically and morphologically. Fillet samples of each host were prepared and 3.316,85 grs were taken from C. guatucupa, 1.754,9 grs. From O. bonariensis and 1.327,1 grs from O. argentinensis, further analyzed by enzymatic digestion. Only four larval nematodes were found from all of the flesh examined. To assess the tissue damage and determine the infective power of the larval nematodes, they were inoculated per oss in mammals, following a protocol of infestation, using a strain of BALB / cAnN mice about 30 grs. in mean weight. There were performed histological sections stained with hematoxylin-eosin. The results showed two positive infestations with Contracaecum sp. L3 from C. guatucupa. Histopathology showed the larvae attached to the gastrointestinal tract, mainly in the fundic region of the stomach, penetrating the gastric mucosal epithelium and reaching the muscle layer. A total compression of the epithelium was observed, with loss of glandular structure and adopting a flattened appearance around the larva. A leukocyte infiltration with monocytes, neutrophils, eosinophils and lymphocytes surrounding the penetration site next larval tissues was observed. In the positive cases it was noted a variation in the blood leukocyte counts. Part of the infected material, fixed in glutaraldehide 2,5 %, was prepared for the observation under scanning electron microscopy. The identification at the specific level employing molecular technics was performed only on larvae of Contracaecum sp. proceeding from the stripped weakfish, Cynoscion guatucupa, because of these were the only that brought positive results in the experimental infections. The comparative analysis of the genomic sequences shows that these parasites cluster near the marine mammals parasites C. margolisi and C. ogmorhini. Until more information to the genetic composition of Contracaecum could be available, the larvae which provoking injuries in the mucosal tissue of BALB /cAnN mice belong to the taxonomical complex Contracaecum ogmorhini s.l. According to the results reached up to the moment using a murine model, the infective C. ogmorhini s.l. L3, could potentially induce damage to other mammals, including man. Nevertheless the exiguous prevalence and larval intensity found in the flesh, along with the habit of well done cooking food based on filet, allow consideras low epidemiological risk the anisakid species studied.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-10-21T10:29:56Z No. of bitstreams: 1 2010 - Caroline Duarte Ribeiro Saad.pdf: 2978488 bytes, checksum: 8e2ece3be63cb39291f012f0bb36a1dc (MD5) / Made available in DSpace on 2016-10-21T10:29:56Z (GMT). No. of bitstreams: 1 2010 - Caroline Duarte Ribeiro Saad.pdf: 2978488 bytes, checksum: 8e2ece3be63cb39291f012f0bb36a1dc (MD5) Previous issue date: 2010-12-20 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / In order to investigate and identify anisakid larvae present in the viscera, the general cavity, mesentery and muscles of marine fish of commercial importance from the coast of Rio de Janeiro, Brazil, were examined 36 specimens of Lophius gastrophysus, 34 specimens Pagrus pagrus and four specimens of Trichiurus lepturus. The vast majority of fish used was obtained from fish warehouses located in Cabo Frio coast of Rio de Janeiro (23? S and 42 ? W), 36 specimens of L. gastrophysus and 31 P. pagrus. We also obtained three specimens of P. pagrus and four T. lepturus at Mercado S?o Braz, located in Campo Grande, west of the city of Rio de Janeiro, commum fish fauna from the coast of Rio de Janeiro, (21- 23 ? S and 41-45 ? W). Six anisakid species of nematode larvae were found in the mesentery, the general cavity, liver and muscles. Those species are: Anisakis simplex, Terranova sp. Contracaecum sp. Hysterothylacium sp.1, sp.2 and Raphidascaris sp.. Anisakis simplex larvae were found in L. gastrophysus with prevalence (P) of 22.2% and mean abundance (MA) 0.53 for T. lepturus P = 50%; Terranova sp. From L. gastrophysus P = 8.3% and AM = 0.08, T. lepturus P = 25% and P. pagrus P = 5.8%; Contracaecum sp. From L.gastrophysus P = 8.3% and AM = 0.08; Hysterothylacium sp.1 from T. lepturus P = 75% and P. pagrus from P = 100%; Hysterothylacium sp.2 from L. gastrophysus P = 25% and AM = 0.47 and T. lepturus with P = 25%; Raphidascaris sp. From L. gastrophysus from P = 11.1% and AM = 0.42 and P. pagrus from P = 55.8%. These nematodes are described and illustrated herein. A revision of available literature, concerning nematodes belonging to these genera, was made. This is the first record of occurrence of Anisakis simplex, Terranova sp. e Contracaeum sp. parasitizing L. gastrophysus the coast of the state of Rio de Janeiro. It was concluded that infection in fish of high consumption and commercial importance, make the consumption of these fish, especially without prior adequate thermal processing, a potential risk from the point of view public health / Com o objetivo de pesquisar e identificar as larvas de anisak?deos presentes nas v?sceras, cavidade geral, mesent?rio e musculatura de peixes marinhos de import?ncia comercial provenientes do litoral do estado do Rio de Janeiro, Brasil, foram examinados 36 esp?cimes de Lophius gastrophysus, 34 esp?cimes de Pagrus pagrus e 4 esp?cimes de Trichiurus lepturus. A grande maioria dos peixes utilizados foi obtida de entrepostos de pescados localizados em Cabo Frio, litoral do estado do Rio de Janeiro (23? S e 42? W), sendo 36 exemplares da esp?cie L. gastrophysus e 31 de P. pagrus. Foram obtidos ainda 3 exemplares de P. pagrus e 4 de T. lepturus no Mercado S?o Braz, localizado em Campo Grande, zona oeste da cidade do Rio de Janeiro, peixes estes oriundos da costa mar?tima do estado do Rio de Janeiro, (21-23? S e 41-45? W). Seis esp?cies de larvas de nemat?ides anisak?deos foram encontradas, Anisakis simplex, Terranova sp., Contracaecum sp., Hysterothylacium sp.1, Hysterothylacium sp.2 e Raphidascaris sp., provenientes do mesent?rio, cavidade geral, f?gado e musculatura. Larvas de Anisakis simplex foram encontradas em L. gastrophysus com preval?ncia (P) de 22,2% e abund?ncia m?dia (AM) 0,53 e em T. lepturus com P=50%; Terranova sp. em L. gastrophysus com P=8,3% e AM=0,08, em T. lepturus com P=25% e P. pagrus com P=5,8%; Contracaecum sp. em L. gastrophysus com P=8,3% e AM=0,08; Hysterothylacium sp.1 em T. lepturus com P=75% e em P. pagrus com P=100%; Hysterothylacium sp.2 em L. gastrophysus com P=25% e AM=0,47 e em T. lepturus com P=25%; Raphidascaris sp. em L. gastrophysus com P=11,1% e AM=0,42 e em P. pagrus com P=55,8%. Estes nemat?ides foram descritos e ilustrados, junto ? revis?o da literatura existente a respeito dos nemat?ides pertencentes aos presentes g?neros. Este ? o primeiro registro da ocorr?ncia de Anisakis simplex, Terranova sp. e Contracaeum sp., parasitando L. gastrophysus do litoral do estado do Rio de Janeiro. Concluiu-se que a presen?a de larvas de anisak?deos dos presentes g?neros com preval?ncias de infec??o consider?veis em peixes de elevado consumo e import?ncia comercial, torna o consumo destes peixes, principalmente sem pr?vio e adequado processamento t?rmico, um risco em potencial sob o ponto de vista de sa?de p?blica.