RESUMEN

The cestode Dibothriocephalus nihonkaiensis (syns. Diphyllobothrium nihonkaiense and Diphyllobothrium klebanovskii), the broad fish tapeworm, is a parasitic agent of intestinal infection acquired by consumption of raw or undercooked Pacific salmon, Onchorhynchus spp. Sequencing studies conducted about a decade ago revealed the presence of two major lineages (A and B) in the broad fish tapeworm population within Asian coastal areas. However, in spite of the accumulation of sequence data on GenBank recently, no further genetic analyses of D. nihonkaiensis have been attempted. The present study assessed for the first time the global cox1 variation in D. nihonkaiensis. Novel partial cox1 sequences of 14 isolates of D. nihonkaiensis from 12 patients were generated, and a global genetic analysis was performed using the 14 novel and 79 previously published sequences for isolates from definitive and second intermediate hosts of this species was performed. A total of 48 haplotypes of three haplotype groups (Types A, B and C) were identified, and co-infections with genetically different D. nihonkaiensis were highlighted in humans and Pacific salmon.

RESUMEN

The molecular, morphological, and physiological features of 42 strains of Ochroconis collected from many limestone areas were studied. Ochroconis strains are often found in limestone areas, although they have rarely been found in other wild areas, e. g. forests. Moreover, many strains from these areas grew on alkaline media (pH 9.7) and media with soap. They were phylogenetically more variable than Ochroconis strains previously found indoors or at city parks. Thus, the Ochroconis strains are thought to have adapted to the alkaline soil, specifically found in limestones. It is assumed that some strains of Ochroconis originally grew in the limestones and immigrated into park soils with the fine dust of cement and into indoor environments. More species of Ochroconis, with the ability to use surfactants as nutrients, are distributed in limestone areas than indoors or in city parks. Moreover, these fungi were found randomly in the phylogenetic trees of Ochroconis. Although only O. humicola was often found indoors and used surfactants, this fungus was newly found in the limestone areas of Okinawa Prefecture. Ochroconis humicola originally grew outdoors and seems to have immigrated to and propagated indoors. Thus, this species may have originated from the subtropical limestone areas.

Asunto(s)

Ascomicetos/aislamiento & purificación , Carbonato de Calcio , Microbiología del Suelo , Microbiología del Aire , Contaminación del Aire Interior , Ascomicetos/clasificación , Ascomicetos/genética , Microbiología Ambiental , Humanos , Concentración de Iones de Hidrógeno , Japón , Microclima , Parques Recreativos , Filogenia

RESUMEN

Samples of diaphragm were collected from 53 sika deer from Gifu Prefecture, Japan; 220 sarcocysts were isolated, examined in wet mounts and classified according to their cyst wall protrusions. The sarcocysts were then examined molecularly in order to assign them to different species. All but 11 of the 220 sarcocysts were initially identified by means of a multiplex PCR assay targeting cox1 of five species, whereas the remaining 11 sarcocysts were identified by standard PCR and sequencing. DNA from selected sarcocysts was used for PCR amplification and sequencing of cox1 (59 sequences) and 18S rDNA (23 sequences). The 220 sarcocysts comprised seven major cox1 sequence types or species. Types 4 and 7 were assigned to the known species Sarcocystis pilosa and Sarcocystis ovalis, whereas types 1, 3 and 5 were considered to represent three new species, for which the names Sarcocystis japonica, Sarcocystis matsuoae and Sarcocystis gjerdei have been proposed. Types 2 and 6 were most similar to Sarcocystis tarandi and Sarcocystis taeniata, respectively, but could not be unequivocally assigned to these species. Sarcocysts belonging to S. japonica were macroscopic with fairly thick finger-like protrusions, whereas most sarcocysts of the six other species were microscopic. Sarcocysts of S. cf. tarandi and S. matsuoae were spindle-shaped and possessed thin finger-like cyst-wall protrusions. Sarcocysts of S. pilosa and S. gjerdei had similar hair-like protrusions, whereas those of S. cf. taeniata had a smooth surface. Sarcocysts of S. japonica, S. pilosa, S. cf. tarandi, S. gjerdei, S. matsuoae, S. cf. taeniata and S. ovalis were found in 50 (94.3%), 29 (54.7%), 22 (41.5%), 10 (18.9%), 8 (15.1%), 6 (11.3%) and 1 (1.9%) of the 53 sika deer examined, respectively. An improved multiplex PCR assay targeting cox1 was developed, through which the seven Sarcocystis spp. found in the present study could be identified.

RESUMEN

Norovirus (NoV) is a major cause of viral gastroenteritis, and GII.4 has been the predominant genotype worldwide since the mid-1990s. During the 2014 to 2015 winter, a rare genotype, NoV GII.17, emerged and became prevalent mainly in East Asia. Over the past two decades, NoV molecular surveillance in Osaka City, Japan, has revealed that NoV GII.17 was detected for the first time in February 2001 and that NoV GII.17-associated outbreaks remarkably increased during the 2014 to 2015 season, with higher incidence recorded in January to March 2015. Genetic analysis indicated that 28 GII.17 outbreak strains were closely related to the novel GII.P17-GII.17 variants represented by the Kawasaki308/2015/JP strain, similar to that in other regions. Statistical analysis showed that NoV GII.17 infections were more common in adults than GII.3 and GII.4 infections, suggesting that the affected adults most likely did not have antibodies against NoV GII.17 and the novel GII.17 variant had recently appeared. Regarding transmission, food was one of the most important factors involved in the spread of NoV GII.17 among adults; 61% of GII.17 outbreaks were foodborne, with oysters being the most common vehicle. Interplay between pathogens, hosts, and environmental factors was considered to be important in the 2014 to 2015 NoV GII.17 epidemic.

Asunto(s)

Infecciones por Caliciviridae/epidemiología , Infecciones por Caliciviridae/virología , Brotes de Enfermedades , Gastroenteritis/epidemiología , Norovirus/genética , Adulto , Animales , Anticuerpos Antivirales/sangre , Infecciones por Caliciviridae/transmisión , Niño , Ciudades/epidemiología , Enfermedades Transmitidas por los Alimentos/epidemiología , Enfermedades Transmitidas por los Alimentos/virología , Gastroenteritis/virología , Genotipo , Humanos , Incidencia , Japón/epidemiología , Ostreidae/virología , Filogenia , Estaciones del Año

RESUMEN

Several surveys of Sarcocystis infection in sika deer in Japan have shown a high prevalence, but the identification has been unclear because molecular data have been lacking or have been limited to 18S ribosomal RNA gene sequences. Thus, in our previous study based on such sequences, some Sarcocystis isolates from sika deer were not clearly separated from other species in the phylogenetic analysis. In the present study, we therefore characterized sarcocyst isolates from sika deer (Cervus nippon centralis) at the mitochondrial cytochrome c oxidase subunit I gene (cox1). Moreover, we developed a multiplex PCR based on cox1 sequences of all species found, so that we could rapidly identify sarcocysts of these species. Twenty-one sarcocysts from nine sika deer were examined. Five distinct cox1 sequence types, each with a high sequence identity (> 99%), were found, and the sarcocysts could thus be classified into five species. Based on the sequence comparisons and the phylogeny, Sarcocystis spp. of types 1, 3, and 5 are considered to represent three new species, which were most closely related to Sarcocystis silva/Sarcocystis truncata, Sarcocystis entzerothi, and Sarcocystis iberica/Sarcocystis venatoria, respectively. There was a slight uncertainly whether Sarcocystis sp. with type 2 sequences represented a new species or was identical to Sarcocystis tarandi. Type 4 sequences showed 99% identity with those of Sarcocystis pilosa from sika deer in Lithuania and have therefore been assigned to this species. In the multiplex PCR, type-specific fragments were successfully amplified for all five Sarcocystis spp., indicating that this assay may be useful for a rapid identification of sarcocysts of these species.

Asunto(s)

Ciclooxigenasa 1/genética , Complejo IV de Transporte de Electrones/genética , Sarcocystis/clasificación , Sarcocystis/genética , Sarcocistosis/veterinaria , Animales , Secuencia de Bases , Ciervos/parasitología , Genes Mitocondriales/genética , Japón , Lituania , Reacción en Cadena de la Polimerasa Multiplex/métodos , Filogenia , ARN Ribosómico 18S/genética , Análisis de Secuencia de ADN

RESUMEN

The molecular, morphological, and physiological characters of 55 Ochroconis strains collected from indoor and outdoor environments were studied. In Japan, Ochroconis species are often found in indoor detergent-rich environments, such as bathrooms and washing machines, and the predominant species have been identified as O. humicola, similar to that in other Asian and European countries. Although Ochroconis species have rarely been found in outdoor environments such as mountains, forests, and agricultural fields, in the present study, Ochroconis strains were specifically isolated from the soils of urban city parks. Phylogenetic analysis conducted using the 28S ribosomal RNA (28S rDNA) gene sequence showed that almost all of the Ochroconis strains found in indoor environments (i.e., water supply) were O. humicola. Although city parks were often surrounded by residences, more than half of the Ochroconis strains collected from the soils of city parks examined in this study were different Ochroconis species. The ability to use detergents as nutrients was found in a new genetic group (probably a new species) isolated from the soils of city parks as well as in O. humicola and O. constricta. Ochroconis humicola is assumed to adapt mostly to indoor environments and to penetrate from the outdoors, e.g., soils of urban areas. To elucidate the factors promoting indoor fungal predominance, the ability of using surfactants as nutrients was compared among these three species. Additionally, growth under alkaline and drought conditions, and heat tolerance were examined. Indoor predominance of O. humicola compared to that of the other two species was attributed to the ability of using a non-ionic surfactant as nutrient and to growth under alkaline conditions.

Asunto(s)

Microbiología Ambiental , Hongos/genética , Genoma Fúngico/genética , Ambiente , Hongos/crecimiento & desarrollo , Filogenia , Reacción en Cadena de la Polimerasa

RESUMEN

A 1-month-old brown wood owlet (Strix leptogrammica) purchased from a wholesaler and housed as a companion bird by an individual owner in Japan showed severe dehydration and anorexia following a week of vomiting and severe diarrhea. A great number of approximately 5 × 4-µm-sized Cryptosporidium oocysts were found in the feces by microscopy. The owlet was administered subcutaneous fluid and intragastric tube feeding for 2 weeks, resulting in improvement of the condition with a decreased number of oocysts in the feces. At days 51 and 119, no oocysts were found in the feces by microscope and PCR detection. These results suggested that this parasite was a possible agent of severe diarrhea in the affected bird. Molecular analysis of DNA extracted from oocysts based on the 18SrRNA loci identified C. avium; however, analysis of actin and hsp (heat shock protein) genes identified a novel genotype indicating a mixed infection with C. avium and a novel genotype.

Asunto(s)

Enfermedades de las Aves/parasitología , Criptosporidiosis/parasitología , Cryptosporidium/clasificación , Cryptosporidium/aislamiento & purificación , Estrigiformes/parasitología , Animales , Cryptosporidium/genética , Heces/parasitología , Genotipo , Japón , Oocistos/aislamiento & purificación , Reacción en Cadena de la Polimerasa , ARN Ribosómico 18S/genética

Asunto(s)

Enfermedades Transmisibles Importadas/parasitología , Teniasis/diagnóstico , Animales , Antihelmínticos/administración & dosificación , Antihelmínticos/uso terapéutico , ADN de Helmintos , ADN Mitocondrial , Femenino , Humanos , Japón , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa Multiplex , Taenia/genética , Taenia/aislamiento & purificación , Teniasis/tratamiento farmacológico

RESUMEN

Anisakis simplex larvae are well known to cause gastrointestinal and allergic manifestations after ingestion of parasitized raw or undercooked seafood. The antibody recognition dynamics against the components of Anisakis larval antigen after primary and re-infection with Anisakis live larvae remain unclear. For this study, immunoblot analyses of serum IgG, IgE, and IgM against Anisakis larval somatic extract were performed in rats that had been orally inoculated with A. simplex live larvae. Multiple antigen fractions were recognized after primary infection. Their reaction was enhanced after re-infection. Antibody recognition was observed for 12 weeks after re-infection. The fraction of approximately 35 kDa contained a main antigen that induced strong and prolonged immunoreactions in IgG and IgE. The antibody reaction to this fraction appeared to be enhanced after inoculation of larval homogenates. This fraction was heat tolerant with boiling for 30 min. The fraction was spotted by immunoblotting after two-dimensional electrophoresis and was identified as Anisakis haemoglobin (Ani s 13) using mass spectrometry analysis. The amino acid sequences of haemoglobin mRNAs from two A. simplex sensu stricto and one Anisakis pegreffii were identified by RACE-PCR. They differed from those of two isolates of Pseudoterranova decipiens and A. pegreffii. Results of this study show that Anisakis haemoglobin, which is known to be a major allergen of A. simplex, induces strong and prolonged immunoreaction in rats. This report is the first to show the amino acid sequence variation of Anisakis haemoglobin mRNA between A. simplex sensu stricto and A. pegreffii.

Asunto(s)

Anisakiasis/inmunología , Anisakis/inmunología , Antígenos Helmínticos/inmunología , Hemoglobinas/inmunología , Alérgenos/inmunología , Secuencia de Aminoácidos , Animales , Anisakis/genética , Immunoblotting , Larva/inmunología , Masculino , Reacción en Cadena de la Polimerasa , Ratas , Ratas Wistar

Asunto(s)

Anaplasma phagocytophilum/genética , Anaplasmosis/epidemiología , Animales Salvajes/microbiología , Ciervos/microbiología , Porcinos/microbiología , Anaplasma phagocytophilum/aislamiento & purificación , Animales , ADN Bacteriano/genética , Japón/epidemiología , Filogenia , Reacción en Cadena de la Polimerasa

RESUMEN

Sapovirus (SaV) is a causative agent of gastroenteritis in humans in both sporadic cases and outbreaks. During the period from January 2005 to August 2014, SaV was detected in 30 (5.9%) of 510 gastroenteritis outbreaks in Osaka City, Japan using real-time RT-PCR. Seasonal distribution of SaV-associated outbreaks revealed an increase during the 2011-2012 season and the highest frequency of outbreaks during the 2012-2013 season. Genotyping analysis based on the capsid region demonstrated that the most common genotype was GI.2 (36.7%), in which the strains were closely related. The comparison of complete capsid gene sequences with 18 GI.2 strains (7 strains in this study and 11 from GenBank) between 1990 and 2013 showed that GI.2 strains were classified into at least three genetic clusters (1990-2000, 2004-2007, and 2008-2013) with chronologically unique amino acid residues and accumulation of mutations in the predicted P domain, suggesting the one of the causes of emergence and spread of GI.2 strains. This study will also be helpful for understanding the evolutionary mechanism of the SaV genome.

Asunto(s)

Infecciones por Caliciviridae/epidemiología , Brotes de Enfermedades/estadística & datos numéricos , Epidemias , Gastroenteritis/epidemiología , Gastroenteritis/virología , Sapovirus/genética , Adolescente , Adulto , Anciano , Infecciones por Caliciviridae/virología , Proteínas de la Cápside/genética , Niño , Preescolar , Heces/virología , Genoma Viral , Genotipo , Humanos , Vigilancia Inmunológica , Lactante , Recién Nacido , Japón/epidemiología , Persona de Mediana Edad , Mutación , Filogenia , ARN Viral/genética , Estaciones del Año , Análisis de Secuencia de ADN , Adulto Joven

RESUMEN

This report describes Ascaridia nymphii n. sp., a new species isolated from the alimentary tract of cockatiel Nymphicus hollandicus in Japan. More than 63 nematodes were found in the formalin-fixed small intestine, ventriculus, proventriculus and crop of a 48-day-old young cockatiel that died after exhibiting severe emaciation. No nematode eggs were observed in the faecal examination performed while the cockatiel was alive, but Cryptosporidium oocysts were found. The intestinal mucosa was damaged considerably. Male worms had two alate spicules, well-developed precloacal sucker and a tail with ventrolateral caudal alae and mainly 11 pairs of papillae. Nuclear partial (813 bp) 18S ribosomal RNA gene (18S rDNA) sequences obtained from two female samples were mutually identical. They respectively showed 99.1 and 98.6% identities to those from Ascaridia numidae and Ascaridia galli. Phylogenetic analysis using this locus indicated the present nematode as Ascaridia species. The mitochondrial NADH dehydrogenase subunit 2 gene (nad2) sequences obtained for four samples were mutually identical. They respectively showed 98.7, 85.7 and 82.2% identities with those from Ascaridia columbae, Ascaridia sp. and A. galli. Combining the morphological and sequencing data from two loci, the present nematode was identified as A. nymphii n. sp., which is closely related with A. columbae. This report is the first of a study examining the distribution of Ascaridia species in captive parrots in Japan. This study also identified the trachea and cloaca, like Cryptosporidium baileyi, as the possible location of Cryptosporidium avian genotype V in avian hosts.

Asunto(s)

Ascaridia/aislamiento & purificación , Ascaridiasis/veterinaria , Enfermedades de las Aves/parasitología , Cacatúas/parasitología , Tracto Gastrointestinal/parasitología , Animales , Ascaridia/clasificación , Ascaridia/genética , Ascaridiasis/parasitología , Enfermedades de las Aves/mortalidad , Cryptosporidium/clasificación , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Femenino , Japón , Masculino , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 18S/genética

RESUMEN

Didymozoids found in the muscles of marine fish are almost always damaged because they are usually found after being sliced. Therefore, identifying muscle-parasitizing didymozoids is difficult because of the difficulty in collecting non-damaged worms and observing their organs as key points for morphological identification. Moreover, muscle-parasitizing didymozoids are not easily found because they parasitize at the trunk muscles. Therefore, muscle-parasitizing didymozoid classification has not progressed because there are few opportunities to detect them. Our recent report was the first to describe the usefulness of sequencing analysis for discrimination among muscle-parasitizing didymozoids. Recently, we found a didymozoid in the trunk muscle of a chub mackerel Scomber japonicus. The present study genetically compares the present isolate with other muscle-parasitizing didymozoids. The present isolate differs markedly from the previously unidentified didymozoid from an Atlantic mackerel S. scombrus by phylogenetic analysis of 18S rDNA. It also differs from other muscle-parasitizing didymozoids from other host species based on phylogenetic analyses of 18S, 28S rDNAs, and coxI loci. These results suggest that sequencing analysis is useful for the discrimination of muscle-parasitizing didymozoids. Combining the present data with earlier data for sequencing analysis, muscle-parasitizing didymozoids from seven marine fish species were classified as seven species. We proposed appellations for six distinct muscle-parasitizing didymozoids for future analysis: sweetlips fish type from Diagramma pictum and Plectorhinchus cinctus, red sea bream type from Pagrus major, flying fish type from Cypselurus heterurus, Atlantic mackerel type from Scomber scombrus, chub mackerel type from S. japonicus, and purple rockcod type from Epinephelus cyanopodus.

Asunto(s)

Músculos/parasitología , Perciformes/parasitología , Trematodos/clasificación , Trematodos/aislamiento & purificación , Animales , Análisis por Conglomerados , ADN Ribosómico/química , ADN Ribosómico/genética , Complejo IV de Transporte de Electrones/genética , Japón , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 18S/genética , ARN Ribosómico 28S/genética , Análisis de Secuencia de ADN , Trematodos/genética

RESUMEN

Sarcocystis nesbitti, using snakes as the definitive host, is a causative agent of acute human muscular sarcocystosis in Malaysia. Therefore, it is important to explore the distribution and prevalence of S. nesbitti in snakes. Nevertheless, epizootiological information of S. nesbitti in snakes remains insufficient because few surveys have assessed Sarcocystis infection in snakes in endemic countries. In Japan, snakes are popular exotic pet animals that are imported from overseas, but the degree of Sarcocystis infection in them remains unclear. The possibility exists that muscular sarcocystosis by S. nesbitti occurs in contact with captive snakes in non-endemic countries. For a total of 125 snake faecal samples from 67 snake species collected at animal hospitals, pet shops and a zoo, this study investigated the presence of Sarcocystis using polymerase chain reaction (PCR) for the 18S ribosomal RNA gene (18S rDNA). Four (3.2%) faecal samples were positive by PCR. Phylogenetic analysis of the 18S rDNA sequences obtained from four amplification products revealed one isolate from a beauty snake (Elaphe taeniura), Sarcocystis zuoi, which uses rat snakes as the definitive host. The isolate from a Macklot's python (Liasis mackloti) was closely related with unidentified Sarcocystis sp. from reticulated pythons in Malaysia. The remaining two isolates from tree boas (Corallus spp.) were closely related with Sarcocystis lacertae, Sarcocystis gallotiae and unidentified Sarcocystis sp. from smooth snakes, Tenerife lizards and European shrews, respectively. This report is the first of a study examining the distribution of Sarcocystis species in captive snakes in Japan.

Asunto(s)

Sarcocystis/genética , Sarcocistosis/veterinaria , Serpientes/parasitología , Animales , Animales de Zoológico , Secuencia de Bases , ADN Ribosómico/genética , Heces/parasitología , Humanos , Japón/epidemiología , Mascotas , Filogenia , Reacción en Cadena de la Polimerasa , ARN Ribosómico 18S/genética , Sarcocistosis/epidemiología , Sarcocistosis/parasitología

RESUMEN

Parasitic Entamoeba spp. are found in many vertebrate species including humans, as well as many livestock including pigs. In pigs, three Entamoeba spp., E. suis, and E. polecki and E. histolytica as zoonotic species, have been identified, but their pathogenicity has not been fully characterized. Here, we report the bacteriological, virological, and histopathological examination of three piglets with chronic diarrhea. Two animals appeared to be additionally infected with Lawsonia intracellularis, which caused a characteristic proliferative ileitis. In the piglet infected with Entamoeba spp., the trophozoites (approximately 10-15 µm with one nucleus in their cytoplasm) invaded into the lamina propria and the disease was worsened by the formation of ulcers and pseudomembranes. Genetic analysis identified the parasite as E. polecki (99.5% identity). Although E. polecki in humans or animals might be less pathogenic in the case of a single infection, coinfections with other pathogens including L. intracellularis may increase the severity of the disease.

Asunto(s)

Infecciones por Desulfovibrionaceae/veterinaria , Entamoeba/aislamiento & purificación , Entamebiasis/veterinaria , Ileítis/veterinaria , Lawsonia (Bacteria) , Enfermedades de los Porcinos/parasitología , Animales , Coinfección , Infecciones por Desulfovibrionaceae/complicaciones , Diarrea/microbiología , Diarrea/parasitología , Diarrea/veterinaria , Entamoeba/genética , Entamebiasis/complicaciones , Entamebiasis/epidemiología , Entamebiasis/parasitología , Heces/parasitología , Humanos , Ileítis/microbiología , Ileítis/parasitología , Ileítis/patología , Japón/epidemiología , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/microbiología

RESUMEN

The Cryptosporidium horse genotype, a zoonotic protozoan parasite first found in a Prezewalski wild horse, has not been found in any other mammal but calves, horses, and humans. Hedgehogs, popular exotic pet animals in Japan, are a reservoir of two zoonotic Cryptosporidum: C. parvum and C. erinacei (previously known as the hedgehog genotype). Recently, after finding Cryptosporidium infection in a four-toed hedgehog (Atelerix albiventris), we identified the isolate genetically as the Cryptosporidium horse genotype. Its subtype (VIbA13) was the same as that of an isolate from a pet shop employee with severe clinical symptoms, as reported previously from sequencing analysis of the partial Cryptosporidum 60kDa glycoprotein gene sequence. The occurrence of this genotype in hedgehog indicates that the horse genotype has broad host specificity. This report is the first of a study identifying isolates from pet reptiles genetically in Japan. The study identified a new host (Teratoscincus scincus) in C. serpentis lizard genotype by sequencing analysis of partial SSU rRNA and actin genes.

Asunto(s)

Animales Exóticos , Criptosporidiosis/parasitología , Cryptosporidium/clasificación , Erizos , Mascotas , Animales , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/parasitología , Caballos , Japón/epidemiología

RESUMEN

Human pinworms, Enterobius vermicularis, are normally recognized as minor pathogens. However, a fatal case of human pinworm infection has been reported in a nonhuman primate, a zoo reared chimpanzee. Here, we histopathologically examined the lesions in tissues from the deceased chimpanzee and genetically characterized the isolated worms to investigate the pathogenicity and determine the phylogeny. We identified ulcers deep in the submucosa where many parasites were found to have invaded the lamina propria mucosa or submucous tissue. An inflammatory reaction consisting mainly of neutrophils and lymphocytes but not eosinophils was observed around the parasites, and intense hemorrhage in the lamina propria was confirmed. The parasites were morphologically similar to E. vermicularis based on the shape of the copulatory spicules. Mitochondrial cytochrome c oxidase subunit 1 gene products were amplified from worm DNA by PCR and were genetically identified as E. vermicularis based on >98.7% similarity of partial sequences. Phylogenetic analysis revealed that the sequences clustered together with other chimpanzee E. vermicularis isolates in a group which has been referred to as type C and which differs from human isolates (type A). The samples were negative for bacterial pathogens and Entamoeba histolytica indicating that E. vermicularis could be pathogenic in chimpanzees. Phylogenetic clustering of the isolates indicated that the parasite may be host specific.

Asunto(s)

Colitis/parasitología , Enterobiasis/veterinaria , Enterobius/genética , Pan troglodytes/parasitología , Animales , Colitis/patología , Colon/parasitología , Colon/patología , ADN de Helmintos/genética , Enterobiasis/parasitología , Enterobius/aislamiento & purificación , Femenino , Filogenia , Reacción en Cadena de la Polimerasa

RESUMEN

Classification and identification of muscle-parasitizing didymozoids found in marine fish is difficult because of their novel parasitism and morphology. Recent sequence analysis has helped, but only seven sequences are available. Therefore, the usefulness of molecular methods for differentiation of muscle-parasitizing didymozoids, as well as genetic differences between the muscle and the other site-parasitizing didymozoids are quite unclear. In the present study, six unidentified didymozoid isolates from the trunk muscles of four marine fish species (Diagramma pictum, Plectorhinchus cinctus, Pagrus major and Cypselurus heterurus) were examined genetically using sequence analysis (18S rDNA, 28S rDNA, ITS-2 and coxI). All isolates were placed phylogenetically as a lineage independent of other site-parasitizing didymozoids at 18S rDNA, ITS-2 and coxI. They were grouped into three distinct lineages. The present and the previous unidentified or identified didymozoids from trunk muscles were found to differ clearly for every host species by sequence analysis, suggesting that muscle-parasitizing didymozoids are host-specific. This report is the first describing the molecular characteristics of muscle-parasitizing didymozoids by sequence analysis targeting the nuclear and mitochondrial DNA loci, which is proposed as a superior method for didymozoid differentiation.

Asunto(s)

Enfermedades de los Peces/parasitología , Músculos/parasitología , Trematodos/clasificación , Trematodos/aislamiento & purificación , Infecciones por Trematodos/veterinaria , Animales , Organismos Acuáticos/parasitología , Análisis por Conglomerados , ADN de Helmintos/química , ADN de Helmintos/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Complejo IV de Transporte de Electrones/genética , Peces , Microscopía , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 18S/genética , ARN Ribosómico 28S/genética , Análisis de Secuencia de ADN , Trematodos/genética , Infecciones por Trematodos/parasitología

RESUMEN

Unicapsula seriolae (Myxozoa; Multivalvulida) was found in the trunk muscle of Malabar grouper Epinephelus malabaricus caught off Wakayama Prefecture, Japan. Numerous filamentous or sesamoid brown to black lesions were observed in the skeletal muscle. Histopathological observation indicated that the lesions were myxosporean plasmodia encapsulated by a fibrous layer, accompanied by melanin deposition. Spores having one large and two rudimentary polar capsules were subspherical in shape and 6.6 × 6.9 µm in size. Scanning electron microscopy revealed that spores were composed of three spore valves. Morphological characteristics were consistent with U. seriolae, which is reported to cause myoliquefaction in yellowtail kingfish Seriola lalandi in Australia. Molecular analysis of the SSU and LSU rDNA supported identification of the species as U. seriolae. This is the first report of Unicapsula in Japan.

Asunto(s)

Lubina/parasitología , Enfermedades de los Peces/parasitología , Músculo Esquelético/parasitología , Myxozoa/aislamiento & purificación , Enfermedades Parasitarias en Animales/parasitología , Alimentos Marinos/parasitología , Animales , Japón , Microscopía Electrónica de Rastreo , Datos de Secuencia Molecular , Myxozoa/clasificación , Myxozoa/genética , Filogenia , Reacción en Cadena de la Polimerasa , ARN Ribosómico 18S/genética , ARN Ribosómico 28S/genética , Análisis de Secuencia de ADN

RESUMEN

Live Anisakis simplex third-stage larvae (L3) penetrate gastrointestinal mucosa after they are ingested in raw or undercooked seafood, thereafter causing gastrointestinal manifestations and allergic manifestations such as urticaria and anaphylaxis. These allergic reactions are mediated by specific IgE to L3 allergens, especially excretory-secretory (ES) allergens. Recent evidences suggest that only live larvae can cause allergic reactions, although cases attributable to ingestion of cooked, frozen seafood have been reported. Therefore the risk of Anisakis-associated hypersensitivity by ingestion of properly cooked and frozen fish remains controversial. No prior report describes the kinetics of antibody production in experimental animals after oral inoculation with dead L3. This study used ELISA to assess antibody production in rats inoculated orally with dead L3. Positive absorbance value in IgG, IgM, and IgE specific to ES antigen from L3 were found in rats inoculated with live L3 but not with dead L3 (frozen, heated, cut, or homogenized). At one week post re-inoculation with live or frozen L3 to the initially sensitized rats, the absorbance value of the specific IgM and IgE to ES antigen elevated quickly and highly in rats that had been re-inoculated with live L3, but they decreased slightly or did not change in rats inoculated with frozen L3. These results suggest that only ingestion of live L3 can produce the specific antibody and induce initial and secondary sensitizations to L3.

Asunto(s)

Anisakis/inmunología , Anticuerpos Antihelmínticos/inmunología , Animales , Anticuerpos Antihelmínticos/sangre , Especificidad de Anticuerpos , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Larva/inmunología , Ratas