Morphological and molecular characterization of brown-banded broodsacs and metacercariae of Leucochloridium (Trematoda: Leucochloridiidae) parasitizing the semi-slug Omalonyx unguis (Succineidae) in Argentina.

Trematodes of the genus Leucochloridium exhibit an unusual transmission strategy among mollusks (intermediate host). The fully developed sporocyst, housing encysted metacercariae, displays vivid coloration and rhythmic activity in the snail's tentacle, mimicking insect larvae. These strategies attract insectivorous birds, their final hosts, thereby increasing the chances of completing their life cycle. In South America, the reports of adults and larval stages of Leucochloridium are scarce. Brown-banded broodsac of Leucochloridium sp. were obtained from Omalonyx unguis collected in a shallow lake from Corrientes Province, Argentina. Here, we morphologically characterized the larval stages (broodsac and metacercaria), identified the parasite through DNA sequences from nuclear 28S-rRNA (28S) and the mitochondrial cytochrome c oxidase I (COI) genes, and explored its evolutionary affinities with the Leucochloridium species available in GenBank. The present broodsac displays brown bands, with a yellowish background in the first two-thirds and yellowish-white in the last third. Based on morphological comparisons, the broodsac and metacercaria described in this study could not be conclusively categorized under any known South American species of Leucochloridium. In relation to the phylogenetic reconstructions, Leucochloridium sp. consistently clustered with L. perturbatum, and species delimitation analyses resulted in recognized Leucochloridium sp. from Argentina as a distinct species. The DNA sequences obtained in this study constitute the first genetic data generated for sporocyst broodsacs in South America. Future studies, incorporating morphology, genetic, and biological data, will be essential for both species identification and the elucidation of leucochloridiid diversity in the region.

Scaphanocephalus spp. (Trematoda: Opisthorchiidae) in intermediate and definitive hosts of the Yucatán Peninsula, Mexico, with a re-description of Scaphanocephalus expansus.

Scaphanocephalus is a small trematode genus belonging to the family Opistorchiidae. The genus currently contains only three species associated with marine fish as intermediate hosts and fish-eating birds as definitive hosts. Here, specimens of Scaphanocephalus were collected from the Osprey, Pandion haliaetus, and the White mullet, Mugil curema in the Yucatán Peninsula, Mexico. We report for the first-time DNA sequences of adult specimens of Scaphanocephalus, particularly S. expansus, as well as a sequence of a different species sampled as metacercaria. Morphological comparisons of Scaphanocephalus expansus confirmed the identity of the adult specimens, with minor morphological variations; Scanning electron photomicrographs were included, and the species was re-described. Phylogenetic analysis based on 28S rDNA sequences showed that Scaphanocephalus is monophyletic within Opisthorchiidae and consists of three independent lineages. Sequences of adults are identical to those of S. expansus. Instead, the sequence of the metacercaria sampled from the mesentery of Mugil curema nested with specimens reported as Scaphanocephalus sp. from a labrid fish in the Mediterranean Sea, herein named it as Scaphanocephalus sp. 2.

Linking metacercariae and adults of Microphallus basodactylophallus (Digenea: Microphallidae), based on larval stages from ctenophores and adult parasites from aquatic birds found in Mexico.

Members of the genus Microphallus Ward, 1901, are endoparasites mainly of birds and mammals distributed worldwide. Unencysted metacercariae of Microphallus sp., were collected from the mesoglea of ctenophores of the genus Pleurobrachia Fleming; adult digeneans were recovered from the intestines of Eudocimus albus Linnaeus (Threskiornithidae) and Buteogallus urubitinga Gmelin (Accipitridae), in four locations from southeastern Mexico. Adult specimens were identified as M. basodactylophallus (Bridgman, 1969) based on the following features: body pyriform entirely covered by minute spines, prepharynx short, oesophagus very long, caeca short and widely divergent, testes slightly symmetrical and excretory vesicle short and V-shaped. Sequences from D1-D3 domain of the large subunit of ribosomal DNA (LSU) were generated, aligned, and compared with those of congeneric species available in GenBank. Phylogenetic analyses indicated that the metacercariae and adults formed a clade together with an isolate identified as M. basodactylophallus from Florida, USA (GenBank: AY220628). The intraspecific genetic divergence among isolates was low ranged from 0.0% to 0.6%, allowing the link between the two stages of the life cycle. We observed phenotypic plasticity in the morphological traits of M. basodactylophallus adults in definitive hosts (mammals and birds) throughout the distribution, which ranged from the USA to southeastern Mexico. Finally, the unencysted metacercariae identified as M. basodactylophallus represent the first report of a microphallid in ctenophores.

Integrative analysis of new Clinostomum metacercariae (Digenea, Clinostomidae) using COI mtDNA and morphology rises the number of lineages found in South American freshwater fishes.

Clinostomidae is a diverse family of digenean parasitizing fish-eating birds as adults and fishes as metacercariae. The species composition, within the genus Clinostomum has been steadily increasing in recent years. In Argentina, four named species of Clinostomum have been documented, accompanied by four metacercariae representing distinct genetic lineages whose adults have not been identified. This study focused on examining clinostomids in three fish species - Australoheros scitulus (ASI), Cichlasoma dimerus (CDIM), and Pimelodella laticeps (PLA) - at various localities in Argentina. We conducted both morphological and molecular characterizations of the Clinostomum metacercariae collected from these fish species. Molecular phylogenetic analyses using COI mtDNA were performed to determine the placement of these metacercariae within the clinostomid phylogenetic tree. Clinostomum ASC represents a distinct lineage, morphologically distinguishable from other sequenced metacercariae due to its body shape (widest anteriorly and becoming slender towards the posterior end); this lineage was found to be closely related to C. caffarae. While Clinostomum CDIM and Clinostomum PLA exhibited morphological differences, they clustered together genetically with metacercariae reported in previous studies as Clinostomum L3 and Clinostomum CVI. This outcome, coupled with a low genetic distance (0 to 3%), suggests that they are conspecific with metacercariae found in fish across Mexico, Costa Rica, and Argentina. In light of the extensive diversity of fish species in Argentine freshwater ecosystems (over 500 species), and considering the relatively constrained extent of prior investigations, the anticipation of unearthing additional Clinostomum species or lineages is plausible.

Infection of Atlantic tripletail Lobotes surinamensis (Teleostei: Lobotidae) by brain metacercariae Cardiocephaloides medioconiger (Digenea: Strigeidae).

Three juvenile Atlantic tripletail Lobotes surinamensis caught opportunistically in Charleston Harbor (South Carolina, USA) and maintained in captivity for over three months displayed an altered swimming behavior. While no direct causation can be demonstrated herein, fish were infected in their brain by strigeid trematode larvae (metacercariae) of Cardiocephaloides medioconiger, which were identified via ITS2 and 28S ribosomal RNA gene sequencing. Histology showed nonencysted metacercariae within the brain ventricle between the optic tectum and tegmentum, causing distortion of tegmental parenchyma. Aggregates of mononuclear inflammatory cells were in the ventricle adjacent to metacercariae. Metacercarial infection by Cardiocephaloides medioconiger has been reported from the brain and eyes of only two other fish species from the northern US Atlantic coast: the grey mullet Mugil cephalus and silverside Menidia menidia, but this identification is problematic and needs molecular verification. Atlantic tripletail is a new report as a second intermediate host for C. medioconiger and South Carolina is a new locality. Cardiocephaloides species in general have a low host specificity and infection by C. medioconiger could propagate to other fishes and affect neighboring natural ecosystems.

First report of Crassiphiala sp. (Trematoda: Diplostomidae) as an etiological agent of black spot disease in commercial ornamental fish from Brazil.

Ornamental fish are becoming increasingly popular, but the lack of knowledge regarding their various diseases is a major challenge. Skin diseases commonly found in freshwater fish include black spot disease (BSD), which is characterized by melanin deposits around the metacercariae of some trematode species. Since BSD remains poorly understood, this study describes an outbreak of BSD in Etroplus maculatus raised in outdoor ponds at a Brazilian fish farm. Metacercariae samples were collected, examined, and subjected to molecular phylogenetic analysis. The parasites were conspecific to an unnamed species, Crassiphiala lineage 5, recently found in Brazilian birds (Megaceryle torquata). Sequences obtained for longifurcate cercariae of the planorbid snail Biomphalaria straminea from the same region were identical to our metacercariae of Crassiphiala sp. These results suggest that Biompahalaria snails are likely an intermediate host of this parasite on farms where E. maculatus was found to be infected. We provide the first molecular evidence that Crassiphiala are the causative agents of BSD in fish from Brazil. Combatting snails and preventing access of fish-eating birds to outdoor ponds are strategies to control this disease in ornamental fish farms.

First record of Cotylurus metacercariae (Trematoda:Strigeidae) in Biomphalaria straminea (Planorbidae) from Argentina: morphological and molecular identification.

In the study of the biology of trematode species, the knowledge of the larval stages in snail hosts is important to elucidate their complete life cycle. The goal of the present study was to describe a new tetracotyle-type metacercaria found in the freshwater mollusk Biomphalaria straminea sampled in a rice field from Corrientes province, Argentina. To this end, 1,768 snails were collected from the cultivated plots and irrigated channels during the flooding periods (from the time of sowing to soon after rice harvesting) between December 2016 and May 2017. We used morphological and molecular analysis to characterize the tetracotyle-type metacercariae. Its morphological traits and the internal transcribed spacers (ITS1 and ITS2 plus 5.8S; ~1200 pb) from nuclear ribosomal DNA (rDNA) were amplified and sequenced. From 1,768 specimens of B. straminea screened, 52 were found infected with metacercariae of tetracotyle type (2.9%) that were identified as Cotylurus genus. A total of 218 metacercariae were found encysted in the ovotestis or between the mantle and viscera of B. straminea. Bioinformatic analysis showed that the metacercarial rDNA sequences shared 94% identity with those of Cotylurus gallinulae from Mexico and 100% identity with those of Cotylurus sp. from Brazil. In this study, the morphological descriptions are supplemented with the first molecular identification of a metacercaria related to Cotylurus parasitizing planorbids from Argentina. Also, our study provides a new morphological description in B. straminea, thus broadening the geographical distribution. The life cycle of this Cotylurus metacercariae is unknown and there are no reports of adult stages parasitizing waterfowl in Argentina.

Paramonostomum deseado n. sp. (Digenea: Notocotylidae) parasitizing the South American Black Oystercatcher and their atypical life cycle from the Patagonian coast.

By way of morphological and molecular analysis we describe a new species of notocotylid, Paramonostomum deseado n. sp., parasitizing Haematopus ater from Argentina and we contribute to elucidate its life cycle. Within this genus, 4 groups can be morphologically distinguished according to body shape: 'Oval', 'Pyriform', 'Elongate', 'Overlong'. The new species belongs to the 'Elongate group', which presents a wide variation in body length (597­4500 µm). The new species, Paramonostomum caeci from Australia, Paramonostomum actitidis from the Caribbean and Paramonostomum alveoelongatum from Russia share the smallest range of body size in this group (<1130 µm). The new species more closely resembles P. actitidis but differs from it by cirrus-sac length, which is shorter in the new species (97­146 vs 280­430 µm in P. actitidis), and egg size which is larger in the new species (25­33 vs 18­20 µm in P. actitidis). Paramonostomum deseado n. sp. uses the limpet Nacella magellanica as both first and second intermediate hosts in which metacercariae encyst inside the redia. This is the first abbreviated cycle described for notocotylid species. Ribosomal RNA sequences provided for adults (ITS1, ITS2 and 28S) and metacercariae inside the rediae (ITS1) support the species identification and the correspondence among stages. Phylogenetic analysis based on 28S placed P. deseado n. sp. close to other Notocotylus spp. and Paramonostomum anatis. Molecular results demonstrate that the hosts involved in the life cycles and the habitat more than morphological differences are determining the phylogenetic relationships in members of Notocotylidae.

Metacercariae in the brain of Erythrinus cf. erythrinus (Characiformes: Erythrinidae) from Iguazú National Park (Argentina): do they belong to Dolichorchis lacombeensis (Digenea, Diplostomidae)?

In Argentina, the family Diplostomidae is composed of eight genera: Austrodiplostomum Szidat & Nani; Diplostomum von Nordmann; Dolichorchis Dubois; Hysteromorpha Lutz; Neodiplostomum Railliet; Posthodiplostomum Dubois; Sphincterodiplostomum Dubois; and Tylodelphys Diesing. During a parasitological survey of fishes from the Iguazú National Park we detected diplostomid metacercariae in the brain of Erythrinus cf. erythrinus. Fish were caught using crab traps, transported alive to the field laboratory, cold-anaesthetized and euthanized by cervical dissection. Some metacercariae were heat-killed in water and fixed in 10% formalin and others were preserved in alcohol 96% for DNA extraction. They were sequenced for the partial segment of the 28S rDNA, internal transcribed spacer (ITS) rDNA and cytochrome c oxidase subunit I (COI) mtDNA genes. Phylogenetic reconstruction was carried out using Bayesian inference and the proportion (p) of absolute nucleotide sites (p-distance) was obtained. In the 28S rDNA tree, the metacercaria sequenced grouped as Dolichorchis sp. The COI mtDNA p-distance between the metacercariae with Dolichorchis lacombeensis was 0.01. There is a small number of ITS sequences for the Diplostomidae family deposited in the GenBank. The oral sucker, ventral sucker, holdfast organ and the distance between oral and ventral suckers are larger in the adult compared with the metacercariae. Additionally, hind-body length and width are larger in the adult due to the development of the genital complex. Further studies using an integrative approach will help confirm the affiliation of other species to the genus Dolichorchis.

Finding a needle in a haystack: larval stages of Didymozoidae (Trematoda: Digenea) parasitizing marine zooplankton.

Larval didymozoids (Trematoda: Digenea) were discovered parasitizing the hemocoel of the heteropod Firoloida desmarestia (redia mean intensity = 13) and the chaetognaths Flaccisagitta enflata and Flaccisagitta hexaptera (metacercaria mean intensity = 1) during a 2014-2016 systematic study of parasites of zooplankton collected in the central and southern regions of the Gulf of California, Mexico. Didymozoid infection route during the early life cycle was inferred combining morphological (light microscopy) and molecular (mitochondrial cytochrome c oxidase subunit I gene, cox1) evidence. Didymozoid rediae parasitizing F. desmarestia were observed, just after field collection of the host, containing hundredths of completely developed cystophorous cercariae, releasing them though the birth pore at approximately one cercaria every 12 s. Cercariae lost their tails developing into a 'young metacercaria' in 1 d at 22 °C without need of an intermediate host. Molecular analysis of cox1 showed that rediae found in F. desmarestia belong to two distinct didymozoid species (Didymozoidae sp. 1 and sp. 2). Metacercariae parasitizing chaetognaths were morphologically identified as Didymozoidae type Monilicaecum and cox1 sequences showed that metacercariae of chaetognaths matched with these two Didymozoidae sp. 1, and sp. 2 species found parasitizing F. desmarestia, plus a third distinct Didymozoidae sp. 3. These are the first DNA sequences of cox1 gene from didymozoid larvae for any zooplankton taxonomic group in the world. We concluded that F. desmarestia is the first intermediate host of rediae and cercariae, and the chaetognaths are the second intermediate hosts where non-encysted metacercariae were found. The definitive host is still unknown because cox1 sequences of present study did not genetically match with any available cox1 sequence of adult didymozoid. Our results demonstrate a potential overlap in the distribution of two carnivorous zooplankton taxonomic groups that are intermediate hosts of didymozoids in the pelagic habitat. The didymozoid specimens were not identified to species level because any of the cox1 sequences generated here matched with the sequences of adult didymozoids currently available in GenBank and Bold System databases. This study provides baseline information for the future morphological and molecular understanding of the Didymozoidae larvae that has been previously based on the recognition of the 12 known morphotypes.

The influence of environmental factors on Clinostomum sp. (Digenea) infection in the fish Cichlasoma paranaense (Kullander, 1983) in Central Brazil.

This study analyzed the parasitism by Clinostomum sp. metacercariae of the neotropical fish Cichlasoma paranaense (Kullander 1983) and environmental variables' influence on their host-parasite relationship in Central Brazil. Fish were collected in five impoundments (I) from three towns: Itauçú (n = 2), Inhumas (n = 1), and Goiânia (n = 2), Goiás State, Brazil. Ninety-five fish were analyzed, weighted and length measured. Afterward, the presence of metacercariae in the gills, muscles, fins, and body cavities was investigated. The metacercariae were fixed in AFA solution, kept in 70% alcohol, and stained with carmine. The infection prevalence revealed that 0.64, 0.56 and 0.91% of fish was infected. The abundance was 4.14, 3.22 and 5.88, with a mean intensity of 6.44, 5.80 and 6.42 per impoundment (I 13, I 15 and I 16), respectively. The frequency of metacercariae was higher in fish collected in Goiânia. High frequency of parasites was observed in the gills, muscles, and fins. The limnological variables did not present direct interference in the parasitism. The fish' standard length influenced the number of metacercariae positively but did not differ to the relative condition coefficient.

Morphological and molecular characterization of Austrodiplostomum compactum metacercariae in the eyes and brains of fishes from the Ivaí River, Brazil.

Austrodiplostomum spp. (Platyhelminthes: Digenea) are endoparasites with a broad geographic distribution in South America. During the larval stage, they parasitize the eyes, brains, muscles, gill, kidneys and swim bladder of a wide variety of fishes. The metacercariae of Austrodiplostomum spp. have several morphological characteristics during development, but are very similar among species, which makes it necessary to use molecular tools to contribute to the elucidation during the larval stage. The objective of this study was to perform morphological and molecular analyses of Austrodiplostomum sp. found in specimens of Hypostomus sourced from the Ivaí River in the state of Paraná, Brazil. Of the 93 analyzed specimens (H. hermanni [n = 50], H. albopunctatus [n = 9], Hypostomus sp. 1 [n = 24], and Hypostomus sp. 2 [n = 10]), 60 were parasitized. A total of 577 Austrodiplostomum sp. metacercariae was collected from the infected hosts; DNA from seven of these samples was extracted, amplified, and sequenced. The morphological data associated with the genetic distance values and the relationships observed in the COI gene tree, indicate that all metacercariae were A. compactum. This is the first record of A. compactum parasitizing H. hermanni, H. albopunctatus, Hypostomus sp. 1, and Hypostomus sp. 2 in the Ivaí River.

Parasitism by metacercariae modulates the morphological, organic and mechanical responses of the shell of an intertidal bivalve to environmental drivers.

Environmental variation alters biological interactions and their ecological and evolutionary consequences. In coastal systems, trematode parasites affect their hosts by disrupting their life-history traits. However, the effects of parasitism could be variable and dependent on the prevailing environmental conditions where the host-parasite interaction occurs. This study compared the effect of a trematode parasite in the family Renicolidae (metacercariae) on the body size and the shell organic and mechanical characteristics of the intertidal mussels Perumytilus purpuratus, inhabiting two environmentally contrasting localities in northern and central Chile (ca. 1600 km apart). Congruent with the environmental gradient along the Chilean coast, higher levels of temperature, salinity and pCO2, and a lower pH characterise the northern locality compared to that of central Chile. In the north, parasitised individuals showed lower body size and shell resistance than non-parasitised individuals, while in central Chile, the opposite pattern was observed. Protein level in the organic matter of the shell was lower in the parasitised hosts than in the non-parasitised ones regardless of the locality. However, an increase in polysaccharide levels was observed in the parasitised individuals from central Chile. These results evidence that body size and shell properties of P. purpuratus vary between local populations and that they respond differently when confronting the parasitism impacts. Considering that the parasite prevalence reaches around 50% in both populations, if parasitism is not included in the analysis, the true response of the host species would be masked by the effects of the parasite, skewing our understanding of how environmental variables will affect marine species. Considering parasitism and identifying its effects on host species faced with environmental drivers is essential to understand and accurately predict the ecological consequences of climate change.

Assessing the genetic diversity of the metacercariae of Posthodiplostomum minimum (Trematoda: Diplostomidae) in Middle American freshwater fishes: one species or more?

Trematode taxonomy is mainly based on the morphological traits of adults. The identification of metacercariae is challenging because such traits are not developed in larval forms, and they even may show some level of morphological variability. Studies testing the potential correspondence between morphological differences and genetic variation of parasites are still lacking. The metacercariae of Posthodiplostomum minimum are probably the diplostomids more widely distributed in North and Middle American freshwater fish, and their intraspecific morphological variability has been attributed to the effect exerted by the host. Here, we tested the hypothesis whether they represent a single species, or a species complex by assessing the genetic divergence and phylogenetic relationships of metacercariae sampled from several host species in a wide geographical range across Middle America. The internal transcribed spacers (ITS1-5.8S-ITS2), and the mitochondrial COI gene were sequenced for 124 and 55 metacercariae, respectively. Phylogenetic analysis inferred from ITS sequences uncovered six well-supported monophyletic lineages. The six lineages show no correspondence to any Posthodiplostomum species for which sequences are available thus far in GenBank. Lineages exhibit some degree of host specificity; Lineages I, II, IV and V are primarily parasites of cyprinodontiforms of the families Poeciliidae, Goodeidae, Profundulidae and Fundulidae. In poeciliids there are at least four candidate species of Posthodiplostomum, some of them occurring in sympatry; instead, Lineages II and VI are exclusively parasites of cichlids. This study contributes to our understanding of the diversity of larval forms of diplostomids and provides an opportunity to further study their life cycles.

Molecular link between the metacercariae and adults of four species Haematoloechus (Digenea: Plagiorchioidea), including scanning electron microscopy characterization.

Haematoloechus comprises a species-rich genus of trematodes commonly found in the lungs of amphibians across the globe. Of the approximately 70 species reported worldwide, Mexico stands out as a biodiversity hot spot for the genus, with 16 species described. In Los Tuxtlas tropical rainforest, in south-eastern Mexico, three species have been reported, two of them endemic of the frog species occurring in the area. Here, we added the record of a fourth species in the area, and partially elucidated the life cycle of these species using DNA sequence data from the 28S rRNA and the COI genes. Frogs, and a diverse array of species of aquatic insects were sampled in two lakes within the tropical rain forest. Adults and metacercariae of Haematoloechus were obtained. Both developmental stages were linked molecularly contributing to our understanding of their transmission pathways, and probable host-specificity patterns. Each of the four species of Haematoloechus were sampled in a particular species of anisopteran odonate, and therefore were considered as specialists towards their second intermediate host. We validated the presence of H. complexus in the region of Los Tuxtlas because their metacercariae, sampled in the anisopteran dragongly Agriogomphus tumens, matched with sequences of H. complexus from Nebraska and Illinois in USA. We further described the ultrastructure of the tegument of three of the four species of Haematoloechus, sampled from their definitive host (Rana vaillanti, Rana berlandieri), to contribute with characters that can be reliable for species delimitation of the genus.

First morphological and molecular identification of the cercaria of Stomylotrema vicarium from the endemic apple snail Pomacea americanista.

The adult fluke Stomylotrema vicarium (Stomylotrematidae, Microphalloidea) was described for the first time in Theristicus caerulescens in 1901, but the complete life cycle has remained unknown to date. Here, we found a stomylotrematid trematode in the digestive gland of the endemic apple snail Pomacea americanista. The digestive gland's tubuloacini were compressed by the trematode larvae placed on connective tissues and haemocoel spaces. Non-virgulate, stylet-bearing cercariae showed three pairs of penetration glands with a body, oral sucker and stylet morphometrically similar to those of stylet-bearing, unencysted young metacercariae of S. vicarium found in the aquatic coleopteran Megadytes glaucus, and at a lesser extent with cercariae of S. gratiosus found in the apple snail Pomacea maculata. The larvae molecular phylogeny was inferred using the markers rRNA 28S and ITS1, being these sequences grouped with the sequences of S. vicarium obtained from adult flukes. Together, these findings indicate that the life cycle of S. vicarium begins in P. americanista, thus supporting the hypothesis that the ampullariid snails act as a first intermediate host.

Prevalence and risk factors of parasites in tambaqui Colossoma macropomum fingerling fish farming from São Francisco region AL/SE / [Prevalência e fatores de risco de parasitos na piscicultura de alevinos de tambaqui Colossoma macropomum da região do baixo São Francisco AL/SE]

This study identified the risk factors associated with parasite infestation in tambaqui fingerlings Colossoma macropomum from São Francisco region SE/AL. The fingerlings fish farms were and characterized about water quality, handling, feeding management and sanitary aspects. Parasitological indexes (prevalence, mean intensity and abundance) were obtained from 380 fish and correlated to the aspects of fish farms as well as biotic and abiotic factors to determine the risk factors. The fish rearing in earthen ponds increased the parasite infestations in tambaqui fingerlings, and the mainly parasites founded were monogenean, Myxobolus sp., Ichthyophthirius multifiliis and digenean metacercaria. The main risk factors identified were associated to irregular water supply and consequently low water quality (total ammonia); lack of biometric procedure which promoted an inadequate feeding rate; and absence of disinfection of tools and ponds. Thus, improvements on these specific aspects in fish farms from São Francisco region must be carried out to avoid disease outbreaks, dissemination, mortalities and economic losses.

Este estudo identificou os fatores de risco associados à ocorrência de parasitas em alevinos de tambaqui Colossoma macropomum da região do baixo São Francisco, SE/AL. As pisciculturas foram caracterizadas segundo parâmetros de qualidade da água, manejo, alimentação e dos aspectos sanitários. Os índices parasitológicos (prevalência, intensidade média e abundância) foram obtidos mediante a análise de 380 peixes e correlacionados com as características das pisciculturas, bem como com os fatores bióticos e abióticos, a fim de determinar os fatores de risco. A fase de crescimento dos alevinos realizada em tanques de terra aumentou as infestações de parasitas, e os principais parasitas encontrados foram monogenéticos, Myxobolus sp., Ichthyophthirius multifiliis e metacercária de digenético. Os principais fatores de risco identificados foram: abastecimento irregular de água e consequente baixa qualidade da água (amônia total); falta de procedimento biométrico, que promoveu uma taxa de alimentação inadequada; e ausência de desinfecção dos tanques e das ferramentas. Pelo exposto, melhorias, especificamente nesses aspectos, nas pisciculturas de alevinos da região do São Francisco, devem ser realizadas para evitar surtos e disseminação de doenças, assim como mortalidades e perdas econômicas.

Morphological and molecular characterization of Austrodiplostomum compactum metacercariae in the eyes and brains of fishes from the Ivaí River, Brazil / Caracterização morfológica e molecular de metacercárias de Austrodiplostomum compactum em olhos e cérebros de peixes do Rio Ivaí, Brasil

Abstract Austrodiplostomum spp. (Platyhelminthes: Digenea) are endoparasites with a broad geographic distribution in South America. During the larval stage, they parasitize the eyes, brains, muscles, gill, kidneys and swim bladder of a wide variety of fishes. The metacercariae of Austrodiplostomum spp. have several morphological characteristics during development, but are very similar among species, which makes it necessary to use molecular tools to contribute to the elucidation during the larval stage. The objective of this study was to perform morphological and molecular analyses of Austrodiplostomum sp. found in specimens of Hypostomus sourced from the Ivaí River in the state of Paraná, Brazil. Of the 93 analyzed specimens (H. hermanni [n = 50], H. albopunctatus [n = 9], Hypostomus sp. 1 [n = 24], and Hypostomus sp. 2 [n = 10]), 60 were parasitized. A total of 577 Austrodiplostomum sp. metacercariae was collected from the infected hosts; DNA from seven of these samples was extracted, amplified, and sequenced. The morphological data associated with the genetic distance values and the relationships observed in the COI gene tree, indicate that all metacercariae were A. compactum. This is the first record of A. compactum parasitizing H. hermanni, H. albopunctatus, Hypostomus sp. 1, and Hypostomus sp. 2 in the Ivaí River.

Resumo Austrodiplostomum spp. (Platyhelminthes: Digenea) são endoparasitos com uma ampla distribuição geográfica na América do Sul. Durante a fase larval, parasitam os olhos, cérebros, músculos, brânquias, rins e bexiga natatória de uma grande variedade de peixes. As metacercárias de Austrodiplostomum spp. apresentam várias características morfológicas durante o desenvolvimento, as quais são muito semelhantes entre as espécies, o que torna necessário o uso de ferramentas moleculares para contribuir para a elucidação durante a fase larval. O objetivo deste estudo foi realizar análises morfológicas e moleculares de Austrodiplostomum sp. encontradas em espécimes de Hypostomus provenientes do rio Ivaí, no Paraná, Brasil. Dos 93 espécimes analisados (H. hermanni [n = 50], H. albopunctatus [n = 9], Hypostomus sp. 1 [n = 24], e Hypostomus sp. 2 [n = 10]), 60 foram parasitados. Um total de 577 metacercárias de Austrodiplostomum foram coletadas dos hospedeiros infectados; o DNA de sete dessas amostras foi extraído, amplificado e sequenciado. Os dados morfológicos, associados aos valores de distância genética e as relações observadas na árvore gênica do COI, indicam que todas as metacercárias são A. compactum. Este é o primeiro registo de A. compactum parasitando H. hermanni, H. albopunctatus, Hypostomus sp. 1, e Hypostomus sp. 2 no rio Ivaí.

Distribution of cysts holding Ascocotyle (Phagicola) longa metacercariae in tissues and organs of mugilid.

Ascocotyle (Phagicola) longa is an etiological agent of human phagicolosis. Mugilids are the second intermediate host, the first being Heleobia australis, and mugilids predatory birds and mammals are its definitive hosts. The occurrence of cysts holding A. longa metacercariae is described in mugilids with a prevalence of up to 100%. The wide geographical distribution of A. longa and its intermediate hosts coupled with the rise in the consumption of raw or poorly cooked fish may elevate the risk of human infection. Therefore, in this study, we aimed to verify the distribution pattern of cysts holding A. longa in mugilids. The tissue and organ samples of these fish were processed in a domestic blender and examined under a stereoscopic microscope to identify the cysts holding the digenetic metacercariae. Of the 24 (100%) fish samples that were analyzed, 12 of Mugil curema and 12 of Mugil liza possessed cysts holding A. longa metacercariae. Digenetic cysts were identified to be present in the gills, heart, stomach, liver, intestines, mesentery, and muscular tissues collected from M. curema and M. liza. Conclusively, in M. curema, the cysts holding A. longa metacercariae were found to be distributed randomly throughout the fish body in almost every tissue and organ that was examined.

Morphomolecular and pathological study of Scaphanocephalus sp. in new host Siganus argenteus in the Arabian Gulf.

In this study, Scaphanocephalus parasite (Platyhelminthes: Heterophyidae) metacercariae were found in Siganus argenteus (forktail rabbitfish or streamlined spinefoot) in the Arabian Gulf of Jubail province, Saudi Arabia. The findings may constitute new host and locality records for this parasite. Based on the number of black spots containing parasite cysts per fish, our study indicated that Siganus argenteus had high infection intensities of encysted metacercariae belonging to the genus Scaphanocephalus. Of the 3500 S. argenteus specimens examined, 800 (22.9%) showed multiple black cysts over the entire body surface, including the membranous parts of fins, while none were seen on the internal organs. The prevalence of infection was highest in summer (June-August) (8.8%). The excysted metacercariae differed morphologically from previously identified Scaphanocephalus spp. Molecular analysis of rDNA showed 100% identity with an unnamed Scaphanocephalus sp. reported in Caribbean fish. Therefore, our findings may indicate a new species of this previously rarely recorded fish parasite. The histopathological examination revealed that the encysted parasites were restricted to the dermal layer of the skin and surrounded by melanophores and a fibrous connective tissue capsule, with focal myositis and Zenker's necrosis in the underlying muscle tissue. The characteristically 'winged' parasite was clearly observed within the cysts. The high prevalence of Scaphanocephalus infection in siganid fish we detected requires further epidemiological study.