First report of a member of the family Mermithidae parasitizing the sandhopper Orchestoidea tuberculata (Amphipoda, Talitridae) in Chile.

Specimens of the sandhopper Orchestoidea tuberculata (Amphipoda; Talitridae) collected from sandy beaches in south-central Chile, were found to be parasitized by juvenile mermithids, constituting the first record of a mermithid infecting a marine amphipod in Chile. A morphological description of juveniles is provided. Sequence analyses based on mitochondrial COI and nuclear 18S rDNA of the mermithids showed extremely low genetic variation. Phylogenetic analyses indicate that the mermithid is more closely related to Hexamermis agrotis, which parasitize Coleoptera, than to Thaumamermis zealandica, which parasitizes New Zealand confamilial talitrid amphipods.

Review of the concept of Profilicollis Meyer, 1931 with a description of Profilicollis rancoensis n. sp. (Acanthocephala: Polymorphidae) from the freshwater crab, Aegla abtao Schmitt, 1942 (Decapoda: Anomura) in Chile, with a key to congeneric species.

Profilicollis rancoensis n. sp. is the tenth species of Profilicollis Meyer, 1931 which includes 9 other species mostly known from marine decapod crabs and shore birds. Cystacanths of P. rancoensis are described from the dominant freshwater crab Aegla abtao in Ranco Lake, Chile and are morphologically distinguished from cystacanths of the 9 other species based on a combination of 4 characters. These are body size, number of proboscis hook rows, number of hooks per row, and length of the largest anterior 2-4 hooks. Male and female cystacanths of P. rancoensis are 2.10-3.33 mm long having an ovoid proboscis with 14 rows of 6-7 hooks per row, with the largest anterior 2-4 hooks being 105-110 micrometers long; the anterior trunk has many small spines in 70-80 concentric rings, each with 50-60 spines around them; hook roots are simple, directed posteriorly, about as long as the blades anteriorly with unremarkable anterior manubria; the cephalic ganglion are in mid-receptacle just anterior to the level of the anterior trunk; the lemnisci are long and slender; the testes are in the anterior trunk, posterior trunk, or one in each; the primordia of 2 tubular cement glands are evident; strong bundles of fibers link the anterior and posterior trunk; and the posterior trunk has a corrugated surface cuticula. Molecular analysis (COI and 18S) sequences coincided with the morphology and support its taxonomy. The phylogenetic profile revealed that P. rancoensis n. sp. fell into the Profilicollis clade. Both sequences showed low genetic variation, and three different haplotypes were found. The new species was more closely related to P. botulus (Van Cleave, 1916) Witenberg, 1932 than to other Profilicollis species.

Title: Révision du concept de Profilicollis Meyer, 1931 avec la description de Profilicollis rancoensis n. sp. (Acanthocephala, Polymorphidae) du crabe d'eau douce Aegla abtao Schmitt, 1942 (Decapoda, Anomura) au Chili, avec une clé des espèces congénères. Abstract: Profilicollis rancoensis n. sp. est la dixième espèce de Profilicollis Meyer, 1931 qui comprend neuf autres espèces principalement connues de crabes décapodes marins et d'oiseaux de rivage. Les cystacanthes de P. rancoensis sont décrits chez le crabe d'eau douce dominant Aegla abtao dans le lac Ranco, au Chili et se distinguent morphologiquement des cystacanthes des neuf autres espèces sur la base d'une combinaison de quatre caractères. Il s'agit de la taille du corps, du nombre de rangées de crochets du proboscis, du nombre de crochets par rangée et de la longueur des 2 à 4 crochets antérieurs les plus grands. Les cystacanthes mâles et femelles de P. rancoensis mesurent de 2,10 à 3,33 mm de long et ont une trompe ovoïde avec 14 rangées de 6 à 7 crochets par rangée, les 2 à 4 crochets antérieurs les plus grands mesurant 105 à 110 micromètres de long ; le tronc antérieur a de nombreuses petites épines en 70-80 anneaux concentriques chacun avec 50-60 épines ; les racines des crochets sont simples, dirigées vers l'arrière, à peu près aussi longues que les lames vers l'avant avec une manubrie antérieure sans particularité ; les ganglions céphaliques sont au milieu du réceptacle juste en avant du niveau du tronc antérieur ; les lemnisques sont longs et minces ; les testicules sont dans le tronc antérieur, le tronc postérieur ou un dans chacun ; les ébauches des 2 glandes cémentaires tubulaires sont évidentes ; de solides faisceaux de fibres relient le tronc antérieur et postérieur ; le tronc postérieur a une cuticule à surface ondulée. Les séquences d'analyse moléculaire (COI et 18S) coïncidaient avec la morphologie et confirmaient sa taxonomie. Le profil phylogénétique a révélé que P. rancoensis n. sp. appartient au clade Profilicollis. Les deux séquences ont montré une faible variation génétique et trois haplotypes différents ont été trouvés. La nouvelle espèce était plus proche de P. botulus (Van Cleave, 1916) Witenberg, 1932 que des autres espèces de Profilicollis.

A comparative assessment of the morphology of Profilicollis altmani (Acanthocephala, Polymorphidae) from crustaceans and shore birds in Peru, with special notes on hook elemental analysis (EDXA), SEM imaging, histopathology, and molecular profile.

The morphology of cystacanths and adults of Profilicollis altmani (Perry, 1942) Van Cleave, 1947 (Polymorphidae) were studied from the Pacific mole crab Emerita analoga (Stimpson) (Crustacea, Hippidae) and Belcher's gull Larus belcheri (Vigors) (Aves, Laridae), respectively, in Peru. Comparative morphometrics with accounts of other populations of P. altmani from elsewhere off the Pacific and Atlantic coasts of North and South America revealed marked intraspecific population variations. We report scanning electron micrographs (SEM) of new features, not before noted or captured in line drawings by earlier observers. We further present microscope images that reveal internal details not previously reported or possible to see with SEM. Energy dispersive X-ray analysis (EDXA) revealed unusual patterns in the chemistry of proboscis hooks especially the high sulfur and diminished phosphorous and calcium in hook tips and low sulfur and high levels of phosphorous and calcium at mid hooks. The size and shape of all hooks of the cystacanths are reported for the first time. Histopathological studies in L. belcheri from Peru are also included. Cystacanths of P. altmani from California were also analyzed for molecular patterns and compared with other sequences reported from other locations. The molecular data and the analysis of our new sequences of cytochrome oxidase I (COI) showed that haplotypes of P. altmani had low genetic variation; the species is not geographically structured, and within its clade no monophyletic group is formed.

TITLE: Évaluation comparative de la morphologie de Profilicollis altmani (Acanthocephala, Polymorphidae) de crustacés et d'oiseaux de rivage au Pérou, en particulier l'analyse élémentaire des crochet (EDXA), l'imagerie SEM, l'histopathologie et le profil moléculaire. ABSTRACT: La morphologie des cystacanthes et des adultes de Profilicollis altmani (Perry, 1942) Van Cleave, 1947 (Polymorphidae) a été étudiée, respectivement, à partir du crustacé Emerita analoga (Stimpson) (Crustacea, Hippidae) et du Goéland Siméon Larus belcheri (Vigors) (Aves, Laridae), au Pérou. La morphométrie comparative avec des données d'autres populations de P. altmani d'autres localités au large des côtes du Pacifique et de l'Atlantique de l'Amérique du Nord et du Sud a révélé des variations intraspécifiques marquées des populations. Nous rapportons des nouvelles observations de microscopie électronique à balayage, non notées auparavant ou non capturées dans des dessins au trait par des observateurs antérieurs. Nous présentons en outre des images au microscope qui révèlent des détails internes qui n'ont pas été signalés auparavant ni possibles à voir avec MET. L'analyse aux rayons X à dispersion d'énergie (EDXA) a révélé des modèles inhabituels dans la chimie des crochets de la trompe, en particulier la teneur élevée en soufre et une diminution du phosphore et du calcium dans les pointes des crochets et des niveaux faibles en soufre et en phosphore et en calcium au milieu des crochets. La taille et la forme de tous les crochets des cystacanthes sont décrites pour la première fois. Des études histopathologiques sur L. belcheri du Pérou sont également incluses. Les cystacanthes de P. altmani de Californie ont également été analysés pour les modèles moléculaires et comparés à d'autres séquences rapportées d'autres endroits. Les données moléculaires et l'analyse de nos nouvelles séquences de cytochrome oxydase I (COI) ont montré que les haplotypes de P. altmani avaient une faible variation génétique. L'espèce n'est pas structurée géographiquement, et au sein de son clade aucun groupe monophylétique n'est formé.

Phylogeny and Life Cycles of the Archiacanthocephala with a Note on the Validity of Mediorhynchus gallinarum.

PURPOSE: The molecular profile of specimens of Mediorhynchus gallinarum (Bhalero, 1937) collected from chickens, Gallus gallus L. in Indonesia was analysed. The aim of this study was to assess the phylogenetic position of species of Mediorhynchus within the order Giganthorhynchida. METHODS: We used one mitochondrial gene (cytochrome oxidase 1) and one nuclear gene (18S ribosomal RNA) to infer phylogenetic relationships of class Archiacanthocephala. RESULTS: The COI and 18S rDNA genes sequences showed that M. gallinarum had low genetic variation and that this species is sister to Mediorhynchus africanus Amin, Evans, Heckmann, El-Naggar, 2013. The phylogenetic relationships of the Class Archiacanthocephala showed that it is not resolved but, however, were mostly congruent using both genes. A review of host-parasite life cycles and geographic distributions of Archiacanthocephala indicates that mainly small mammals and birds are definitive hosts, while termites, cockroaches, and millipedes are intermediate hosts. CONCLUSIONS: While the intermediate hosts have wide geographic distributions, the narrow distribution of the definitive hosts limit the access of archiacanthocephalans to a wider range of prospective hosts. Additional analyses, to increase taxonomic and character sampling will improve the development of a robust phylogeny and provide more stable classification. The results presented here contribute to better understanding of the ecological and evolutionary relationships that allow the host-parasite co-existence within the class Archiacanthocephala.

GEOGRAPHICAL VARIATION OF ENTEROPARASITISM IN THE SOUTHERN SEA LION OTARIA FLAVESCENS (SHAW, 1800) OFF THE COAST OF CHILE AS EVIDENCED WITH COPROLOGICAL TESTS: IMPLICATONS FOR ZOONOTIC RISKS.

The scarce information on the helminth fauna in otariids from the Southeastern Pacific comes mainly from stranded individuals or killed for that purpose. In this study, we compared the abundance and composition of enteroparasitic assemblages of Otaria flavescens using coprological techniques. Three sampling localities from north to south spanning 2,200 km off the Chilean coast were considered (Iquique, Viña del Mar, and Talcahuano). In all, 60 fecal samples were collected, and eggs belonging to 5 taxa were found in 91.6% of the samples. They were the anisakid nematodes Contracaecum and Pseudoterranova, the cestode Adenocephalus (syn. Diphyllobothrium), the trematode Ogmogaster, and the acanthocephalan Corynosoma. Samples from southern Chile (Talcahuano) showed the highest prevalence. Adenocephalus eggs had the highest prevalence and abundance in Iquique and Talcahuano, whereas Ogmogaster was the less prevalent and abundant in all sampling localities. Corynosoma eggs had similar prevalence and abundance among sampling localities, and Pseudoterranova eggs were absent in Iquique and with median prevalence values in Viña del Mar and Talcahuano. Thus, the composition of parasite egg assemblages was different between sampling localities. These differences between sampling localities may help to explain differential records of some zoonotic parasitoses such as pseudoterranovosis and diphyllobothriosis in Peru and Chile, where consumption of raw or marinated fish (ceviche) is common. For example, the lower diversity of parasite egg assemblages in the northern Chilean coast may be due to the absence or lower abundance of first intermediate/paratenic hosts of Pseudoterranova.

Solving a long-standing nomenclatorial controversy: designation of a neotype for the southern sea lion Otaria flavescens (Shaw, 1800).

During almost two centuries, two available specific epithets have competed for the southern sea lions of the genus Otaria Péron, 1816 (Fig. 1), which is classically regarded as monotypic (but see below). The species distributes more or less continuously along the Atlantic coast from southern Brazil south to Cape Horn and along the Pacific from northern Peru to the west South American Pacific coast . While some researchers have argued that the correct name for this species is Phoca flavescens Shaw, 1800 (e.g., Cabrera, 1940, Rice, 1977; Rodriguez Bastida, 1993; Teta et al., 2018), others have used P. byronia de Blainville, 1820 (e.g., Allen, 1905; Hamilton, 1934; King, 1978, Oliva, 1988; Berta Churchill, 2012). This controversy originates from the fact that the diagnosis of the type of flavescens, the oldest epithet and as such the one that has priority, would not match the phenotypic features of the genus Otaria. This view, which favors the usage of byronia for populations of southern sea lions, was strongly defended by Oliva (1988:768), who argued that "…(1) the total length of the holotype [of flavescens]; (2) the size of the external ear; and (3) the color and length of the specimen's fur do not correspond to any developmental stage of the species." However, Rodríguez and Bastida (1993:378) discussed these same features reaching a very different conclusion, by indicating that "Shaw's holotype, collected in the Strait of Magellan, could only have been a newborn pup of Otaria or Arctocephalus australis; its body size (circa 62 cm) could correspond to either species, but the uniform yellowish color is found exclusively in some molted pups of the Southern sea lion. Ear length, though not matching well with the described body length, lies within the recorded range of Otaria, but outside that for Arctocephalus australis."

Decreased Metabolic Rate in the Mole Crabs, Emerita analoga, Infected with the Acanthocephalan Profilicollis altmani.

The relationship between a parasite and its host will have an energy cost for the host at some point; however, this basic hypothesis has seldom been evaluated. Using Emerita analoga as a model species, we investigated the aerobic metabolism, ammonia excretion rate, and locomotor performance patterns of crabs both uninfected and infected with the acanthocephalan Profilicollis altmani. Our results show that infected Emerita had a lower metabolic rate compared to uninfected ones. Whether or not this decrease is a result of the pathology of the parasite infection or due to manipulation of the host by the parasite is still unknown. We discuss the importance of using anaerobic and aerobic metabolism and more-complex scenarios in order to understand the effects on host fitness and not only on the immediate response of the host.

Life Cycle of the Parasite Profilicollis chasmagnathi (Acanthocephala) On the Patagonian Coast of Argentina Based On Morphological and Molecular Data.

This study verifies the identity of adult specimens of the parasite Profilicollis chasmagnathi (Acanthocephala, Polymorphidae) recovered from kelp gulls Larus dominicanus (Aves, Laridae), and cystacanths found in crabs Cyrtograpsus altimanus (Crustacea, Decapoda) from the southwestern Atlantic coast. The life cycle of this parasite is elucidated in the intertidal zone of Patagonia, Argentina, based on morphological and molecular data. Preferences by size and sex of the intermediate host and seasonal variation of this parasite are provided, contributing to the knowledge of this host-parasite association.

Mesoscale spatiotemporal variability in a complex host-parasite system influenced by intermediate host body size.

BACKGROUND: Parasites are essential components of natural communities, but the factors that generate skewed distributions of parasite occurrences and abundances across host populations are not well understood. METHODS: Here, we analyse at a seascape scale the spatiotemporal relationships of parasite exposure and host body-size with the proportion of infected hosts (i.e., prevalence) and aggregation of parasite burden across ca. 150 km of the coast and over 22 months. We predicted that the effects of parasite exposure on prevalence and aggregation are dependent on host body-sizes. We used an indirect host-parasite interaction in which migratory seagulls, sandy-shore molecrabs, and an acanthocephalan worm constitute the definitive hosts, intermediate hosts, and endoparasite, respectively. In such complex systems, increments in the abundance of definitive hosts imply increments in intermediate hosts' exposure to the parasite's dispersive stages. RESULTS: Linear mixed-effects models showed a significant, albeit highly variable, positive relationship between seagull density and prevalence. This relationship was stronger for small (cephalothorax length >15 mm) than large molecrabs (<15 mm). Independently of seagull density, large molecrabs carried significantly more parasites than small molecrabs. The analysis of the variance-to-mean ratio of per capita parasite burden showed no relationship between seagull density and mean parasite aggregation across host populations. However, the amount of unexplained variability in aggregation was strikingly higher in larger than smaller intermediate hosts. This unexplained variability was driven by a decrease in the mean-variance scaling in heavily infected large molecrabs. CONCLUSIONS: These results show complex interdependencies between extrinsic and intrinsic population attributes on the structure of host-parasite interactions. We suggest that parasite accumulation-a characteristic of indirect host-parasite interactions-and subsequent increasing mortality rates over ontogeny underpin size-dependent host-parasite dynamics.

Morphological and molecular evidence on the existence of a single estuarine and rocky intertidal acanthocephalan species of Profilicollis Meyer, 1931 (Acanthocephala: Polymorphidae) along the Atlantic and Pacific coasts of southern South America.

Profilicollis chasmagnathi Holcman-Spector, Mañé-Garzón & Dei-Cas, 1977 (Acanthocephala: Polymorphidae) has been reported to parasitise different grapsid species as intermediate hosts along the South Atlantic shores, i.e. Cyrtograpsus angulatus (Dana) and Neohelice granulata (Dana) in Uruguay and Cyrtograpsus altimanus (Rathbun) in Argentina. Larvae of a similar acanthocephalan described as Profilicollis antarcticus Zdzitowiecki, 1985 were recorded in the crab Hemigrapsus crenulatus (Milne-Edwards) from an estuarine habitat on the Southeast Pacific shore in Chile. Earlier studies have questioned the specific assignation of the Chilean estuarine populations of Profilicollis Meyer, 1931. The aim of this study was to re-examine the identification of these acanthocephalans by means of morphological and molecular analyses of cystacanths of Profilicollis spp. gathered from C. angulatus, N. granulata, C. altimanus and H. crenulatus. Our analyses showed that a single species of Profilicollis, P. chasmagnathi, parasitises these four crab species. The assessment of specimens from the South Shetlands Islands, the type-locality of P. antarcticus, is needed before formally proposing that P. antarcticus is a junior subjective synonym of P. chasmagnathi.

Serine proteinases with gelatinolytic activity in an Aspergillus fumigatus allergenic extract.

The presence of proteolytic activity in Aspergillus fumigatus (Af) extract was analyzed in polyacrylamide gelatine containing gels at different pHs. The gelatinolytic pattern showed three major activities (180, 70, and 34 kDa bands) over a broad pH range (pH 4-8). All bands showed maximum activity at pH 8, and the activity declined at lower pH, showing no activity at pH 2. The susceptibility of the proteolytic activity to protease inhibitors confirmed that the fungal extract contained serine proteinases, and one of them, the 34 kDa band, showed a chymotrypsin-like proteinase. The immunochemical fungal extract properties were studied by western blot, and the samples were run in the presence or absence of denaturing and reducing (DTT) conditions. Two protein bands located at approximately 180 and 84 kDa showed immunoreactivity with allergenic patients' anti-Af serum, when the samples were run under denaturing and reducing conditions. In the native form an additional third immunoreactive band of about 70 kDa was found. There were no immunoreactive bands toward IgE about 34 kDa. Periodate treatment caused the decrease or disappearance of the immunoreactivity in the bands, demonstrating that epitopes recognized by serum IgE specific to Af are partially composed by carbohydrates (180 and 70 kDa bands) while others (84 kDa) are carbohydrates. Although further investigations are needed, these preliminary results suggest alkaline serine proteinases from Aspergillus fumigatus as one of the factors responsible for the fungal allergenicity.

Anticuerpos contra Aspergillus fumigatus en asmáticos atópicos / Antibody against Aspergillus fumigatus in atopic asthamtics

La observación de pruebas cutáneas inmediatas positivas a los extractos de Aspergillus fumigatus (Af.) en pacientes asmáticos atópicos libres de aspergiliosis broncopulmonar alérgica instó a estudiar más profundamente la dinámica inmunológica de estos pacientes en relación al antígeno mencionado. De cien asmáticos extrínsecos habitantes de Buenos Aires y sus alrededores con una IgE-PRIST basal promedio de 485,1 KU/L, 26 de ellos evidenciaron una prueba cutánea positiva a dicho alergeno preparado según técnicas convencionales. El RAST-IgE-anti-Af. fue positivo en 21 de esos 26 asmáticos. Por otro lado, las técnicas de inmunoprecipitación fueron negativas para los asmáticos y los controles; en estos últimos el RAST también fue negativo. Se destaca la importancia de la ausencia de precipitinas anti-Af. y la proclividad a practicar inmunoterapia con hongos anemófilos que incluyen al Af. en los atópicos portadores de alergia respiratoria. Se propone un seguimiento con técnicas específicas de inmunoprecipitación para detectar un efecto indeseable de dicha terapia con consecuencias imprevisibles

Anticuerpos contra Aspergillus fumigatus en asmáticos atópicos / Antibody against Aspergillus fumigatus in atopic asthamtics

La observación de pruebas cutáneas inmediatas positivas a los extractos de Aspergillus fumigatus (Af.) en pacientes asmáticos atópicos libres de aspergiliosis broncopulmonar alérgica instó a estudiar más profundamente la dinámica inmunológica de estos pacientes en relación al antígeno mencionado. De cien asmáticos extrínsecos habitantes de Buenos Aires y sus alrededores con una IgE-PRIST basal promedio de 485,1 KU/L, 26 de ellos evidenciaron una prueba cutánea positiva a dicho alergeno preparado según técnicas convencionales. El RAST-IgE-anti-Af. fue positivo en 21 de esos 26 asmáticos. Por otro lado, las técnicas de inmunoprecipitación fueron negativas para los asmáticos y los controles; en estos últimos el RAST también fue negativo. Se destaca la importancia de la ausencia de precipitinas anti-Af. y la proclividad a practicar inmunoterapia con hongos anemófilos que incluyen al Af. en los atópicos portadores de alergia respiratoria. Se propone un seguimiento con técnicas específicas de inmunoprecipitación para detectar un efecto indeseable de dicha terapia con consecuencias imprevisibles (AU)

Asma polínica: estudio químico e inmunológico del polen de la gramínea Lolium perenne / Pollen asthma: chemical and immunologic study of the gramineous Lollium perenne pollen

La polinosis es una entidad clínica claramente establecida y su tratamiento hiposensibilizante provoca gratificantes resultados desde 1911 hasta la actualidad. Sin embargo, los autores han tendido siempre a emplear antígenos polínicos preparados con la mejor tecnología y estandarización, con el propósito de evitar substancias contaminantes cuyo destino se desconoce. Esta presentación aporta nuevas evidencias acerca de la composición química y la reactividad inmunológica del polen de Lolium perenne (Lp) frente a un antisuero de conejo específico y en humanos riníticos y asmáticos sensibilizados. Un extracto del polen de Lp con 10.000 UNP/nl fue pasado por columnas de Sephadex G-50 y de DEAE-celulosa. Los contenidos en proteínas y hexosas fueron registrados en un espectrofotómetro LKB Uvicord a 280 nm y 470 nm, respectivamente. Si bien se analizaron 400 tubos en cada fraccionamiento, se emplearon en las experiencias aquellos con los contenidos más altos en proteínas y en hexosas. Los pesos moleculares deteminados por columna, y comparados con marcadores especiales, fueron de 18.400, 25.500 y 68.000 daltons para las fracciones proteicas Nro. 33, 28 y 13 respectivamente. La determinación cuantitativa de proteínas arrojó valores significativos para las fracciones 33 (350 mcg/ml), 38 (250 mcg/ml) y 55 (200 mcg/ml). Por otro lado, conejos adultos fueron inmunizados con Lp más adyuvante de Freund completo, de acuerdo con el esquema clásico, durante 10 semanas. Este antisuero fue testificado con Lp y cada una de sus fracciones mediante el Ouchterlony y el Boyden. Cien pacientes afectados de rinitis estacional y asma con un IgE sérica total elevada y alta reactividad cutánea a Lp, fueron testificados con las 14 fracciones más significativas (7 proteínas y 7 hexosas) mediante pruebas cutáneas de lectura inmediata, destacándose los resultados encontrados con las fracciones proteicas Nro. 33, 38 y 359 y las hexosas Nro. 56, 192 y 360...

Asma polínica: estudio químico e inmunológico del polen de la gramínea Lolium perenne / Pollen asthma: chemical and immunologic study of the gramineous Lollium perenne pollen

La polinosis es una entidad clínica claramente establecida y su tratamiento hiposensibilizante provoca gratificantes resultados desde 1911 hasta la actualidad. Sin embargo, los autores han tendido siempre a emplear antígenos polínicos preparados con la mejor tecnología y estandarización, con el propósito de evitar substancias contaminantes cuyo destino se desconoce. Esta presentación aporta nuevas evidencias acerca de la composición química y la reactividad inmunológica del polen de Lolium perenne (Lp) frente a un antisuero de conejo específico y en humanos riníticos y asmáticos sensibilizados. Un extracto del polen de Lp con 10.000 UNP/nl fue pasado por columnas de Sephadex G-50 y de DEAE-celulosa. Los contenidos en proteínas y hexosas fueron registrados en un espectrofotómetro LKB Uvicord a 280 nm y 470 nm, respectivamente. Si bien se analizaron 400 tubos en cada fraccionamiento, se emplearon en las experiencias aquellos con los contenidos más altos en proteínas y en hexosas. Los pesos moleculares deteminados por columna, y comparados con marcadores especiales, fueron de 18.400, 25.500 y 68.000 daltons para las fracciones proteicas Nro. 33, 28 y 13 respectivamente. La determinación cuantitativa de proteínas arrojó valores significativos para las fracciones 33 (350 mcg/ml), 38 (250 mcg/ml) y 55 (200 mcg/ml). Por otro lado, conejos adultos fueron inmunizados con Lp más adyuvante de Freund completo, de acuerdo con el esquema clásico, durante 10 semanas. Este antisuero fue testificado con Lp y cada una de sus fracciones mediante el Ouchterlony y el Boyden. Cien pacientes afectados de rinitis estacional y asma con un IgE sérica total elevada y alta reactividad cutánea a Lp, fueron testificados con las 14 fracciones más significativas (7 proteínas y 7 hexosas) mediante pruebas cutáneas de lectura inmediata, destacándose los resultados encontrados con las fracciones proteicas Nro. 33, 38 y 359 y las hexosas Nro. 56, 192 y 360...(AU)