Bioestructura de la piel de la aleta caudal del salmón del atlántico, en estado smolt y sus interacciones con el estado chalimus del Caligus rogercresseyi / Biostructure of the atlantic salmon skin of the caudal fin, in the smolt stage and its interactions with the chalimus stage of Caligus rogercresseyi

RESUMEN: Caligus rogercresseyi es un copépodo que representa uno de los principales desafíos de la industria del cultivo de salmónidos en Chile, ya que afecta profusamente a la piel. Es preciso destacar que los peces en agua dulce y estuario no son afectados, a diferencia del salmón, que desde el post-smolt resulta muy parasitado cuando es trasladado al mar. Se han realizado múltiples estudios sobre el ciclo de vida del parásito y desarrollado tratamientos químicos, físicos y mecánicos para eliminarlos. Sin embargo, a la fecha, los tratamientos no han sido eficaces, lo que produce un problema permanente para el bienestar del animal. El propósito de este estudio fue el de reconocer la bioestructura de la piel de la aleta caudal del salmón del atlántico en los sitios de la interacción con chalimus. Para esto, se utilizaron 15 post-smolt infectados con Caligus y 5 post-smolt controles, sin Caligus. Los salmones fueron aportados por Fundación Chile y la experiencia se realizó en su propio centro experimental. Una vez realizada la eutanasia, mediante sobredosis del anestésico benzocaína, se obtubieron muestras de las aletas caudales, las cuales fueron fijadas en formalina al 10%, incluidas en paraplast para realizar cortes de 5 µm de espesor y teñidas con Tricrómico de Masson y PAS. Los resultados indicaron que la piel de la aleta caudal de los post-smolt afectados presentan mayor altura de la epidermis, escasa células secretoras de mucus y solución de continuidad en la epidermis. Además, la membrana basal se descontinúa y ocurre un aumento de melanomacrófagos en la dermis.

SUMMARY: Caligus rogercresseyi is a copepod that represents one of the main challenges of the salmon farming industry in Chile, since it profusely affects the skin. It should be noted that fish in freshwater and estuaries are not affected, unlike salmon, which from post-smolt is highly parasitized when transferred to the sea. Multiple studies have been carried out on the life cycle of the parasite and chemical, physical and mechanical treatments have been developed to eliminate them. However, to date, the treatments have not been effective, which produces a permanent problem for the welfare of the animal. The purpose of this study was to recognize the biostructure of Atlantic salmon caudal fin skin at sites of interaction with chalimus. For this, 15 post-smolt infected with Caligus and 5 post-smolt controls, without Caligus, were used. The salmon were provided by Fundación Chile and the experience was carried out in its own experimental center. Once the euthanasia was carried out, by means of an overdose of the anesthetic benzocaine, samples of the caudal fins were obtained, which were fixed in 10 % formalin, included in paraplast to make 5 µm-thick sections and stained with Masson's Trichrome and PAS. The results indicated that the skin of the caudal fin of the affected post-smolt presented a greater height of the epidermis, few mucus-secreting cells and a solution of continuity in the epidermis. In addition, the basement membrane is discontinued and an increase in melanomacrophages occurs in the dermis.

A new monogenean species, Gyrodactylus ajime n. sp. (Gyrodactylidae), parasitic on Niwaella delicata (Niwa), an endemic loach of Japan.

A new gyrodactylid species, Gyrodactylus ajime n. sp., is described from the skin and fins of an endemic and commercially important loach Niwaella delicata (Niwa) (Cypriniformes: Cobitoidea: Cobitidae), from the upper reach of the River Yura in Ashiu, Kyoto Prefecture, Japan with scanning electron microscopy observations and molecular data. This species can be distinguished from congenerics by characteristics of the sickle of the marginal hook: the sickle proper is composed of two straight sections and a curved section; its short point extends beyond the limit of the toe and is directed downwards; and the upper corner of heel angular is slightly raised. There are 31 species and subspecies of native loaches (Cobitoidea) known from Japan, of which 25 are red-listed nationally, and two are commercially important. I reviewed all reports of monogeneans from Japanese loaches, and found that only five fish species have been recorded as hosts of these parasites. This suggests that the parasite fauna of these fishes is poorly described. It is urgent to thoroughly describe the monogenean parasites of loaches in Japan as some of the parasite species may be threatened with co-extinction because their hosts are threatened, and to understand any negative impacts on commercially important fish species.

Updating specific PCR primer for detection of Cryptocaryon irritans from reared Larimichthys polyactis.

Artificial breeding of small yellow croaker (Larimichthys polyactis) was recently achieved, providing a bright future for its commercial farming. In May 2019, a disease outbreak occurred among small yellow croakers in an aquaculture farm near Xiangshan Bay, charactering by white spots spotted on the surface of fish skin, gills and fins. The parasite was preliminarily identified as Cryptocaryon irritans based on morphological feature of the parasite and the symptoms on fish. However, the previously published specific primer pairs failed to confirm the existence of C. iriitans. Six nucleotides mismatches were discovered after mapping specific forward primer back to targeted gene. Therefore, an updated PCR specific primer was developed within the 9th highly variable region of 18S rRNA gene and conserved in all C. irritans sequences available in GenBank database. The specificity was verified in silico by Primer-BLAST against GenBank nucleotide. Laboratory cultured ciliates (Mesanophrys, Pseudokeronopsis and Uronema) as well as natural microbial community samples collected from sea water and river water was used as negative control to verify the specificity of the primer in situ. Besides, tank transfer method was used to evaluate the treatment of the parasite infection. By tank transfer method, 2.00 ± 0.61 out of 10 fish that already sever infected were successfully survived after 8 days treatment, meanwhile the control group died out at d 6. More loss to the treatment group during first five days was observed and may attribute to the combined effect from infection and stress the recent domesticated fish suffered during rotation. Therefore, tank transfer method was also effective to prevent small yellow croaker from further infection, however the loss of the small yellow croaker suffered from stress during rotation also needs to be carefully concerned. In conclusion, this study reported the first diagnose of C. irritans infection on small yellow croaker, provided updated specific primer to detect C. irritans infection on fish body and reported the effect of tank transfer on small yellow croaker treatment.

Modulation of local and systemic immune responses in brown trout (Salmo trutta) following exposure to Myxobolus cerebralis.

Myxobolus cerebralis, the etiological agent of Whirling Disease (WD), is a freshwater myxozoan parasite with considerable economic and ecological relevance for salmonids. There are differences in disease susceptibility between species and strains of salmonids. Recently, we have reported that the suppressor of cytokine signaling SOCS1 and SOCS3 are key in modulating rainbow trout (Oncorhynchus mykiss) immune responses and that resistant fish apparently exhibit effective Th17 cell response after exposure to M. cerebralis. It is unclear whether such molecules and pathways are also involved in the immune response of M. cerebralis infected brown trout (Salmo trutta). Hence, this study aimed to explore their role during immune modulation in infected brown trout, which is considered resistant to this parasite. Fish were exposed to the triactinomyxon (TAM) stages of M. cerebralis and quantitative real-time PCR (RT-qPCR) was carried out to examine local (caudal fin) and systemic (head kidney, spleen) immune transcriptional changes associated with WD over time in infected and control fish. All of the immune genes in the three tissues studied were differentially expressed in infected fish at multiple time points. Brown trout reduced the parasite load and demonstrated effective immune responses, likely by keeping pro-inflammatory and anti-inflammatory cytokines in balance whilst stimulating efficient Th17-mediated immunity. This study increases knowledge on the brown trout immune response to M. cerebralis and helps us to understand the underlying mechanisms of WD resistance.

Increasing the known biodiversity of cnidarian parasites of bryconid fishes from South America: two novel Myxobolus species with ultrastructure and ssrDNA-based phylogeny.

This study increases the known biodiversity of cnidarian parasites in neotropical bryconid fishes. Two novel Myxobolus species are described based on morphology, ultrastructure and small subunit ribosomal DNA (ssrDNA) sequencing: Myxobolus vetuschicanus n. sp. infecting fins of Salminus franciscanus and Myxobolus mineirus n. sp. infecting the mesentery of Brycon orthotaenia from the São Francisco River basin, Minas Gerais State, Brazil. Ultrastructural analysis of the two species revealed an asynchronous sporogenesis process, with germinative cells and young developmental stages of myxospores in the periphery of the plasmodia. In M. vetuschicanus n. sp., the plasmodia were surrounded by a layer of fibroblasts and in M. mineirus n. sp., the plasmodial membrane had direct contact with the host tissue. The phylogenetic analysis based on the ssrDNA of Henneguya/Myxobolus species showed that the two novel Myxobolus species grouped in subclades together with other parasite species of bryconid fishes.

A survey of gyrodactylid parasites on the fins of Homatula variegata in central China.

In this study, two parasites on the fins of Homatula variegata were recorded from March to September 2016. A dissection mirror was used to examine the distribution and quantity of the ectoparasitic Gyrodactylus sp. and Paragyrodactylus variegatus on the host Homatula variegata in different seasons. The present study explored possible explanations for the site specificity of gyrodactylid parasites in 442 Homatula variegata infected with 4307 Gyrodactylus sp. (species identification is incomplete, only characterized to the genus level) and 1712 Paragyrodactylus variegatus. These two gyrodactylid parasites were collected from fish fins, and the fish were harvested in China's Qinling Mountains.The results indicated that the highest number of Gyrodactylus sp., which was numerically the dominant species, appeared on the fish fins in April, while the highest number of Paragyrodactylus variegatus was found on the fish fins in March. The two parasite species appeared to be partitioned spatially, with Gyrodactylus sp. occurring more frequently on pectoral and pelvic fins, and P. variegatus occurring more frequently on caudal fins. However, Gyrodactylus sp. appeared to occur on fish of all lengths, while P. variegatus tended to occur more abundantly on shorter fish rather than on longer fish. At lower Gyrodactylus sp. infection levels (<100), the pelvic and pectoral fins were the main locations of attachment, followed by the dorsal fin. For infections of more than 100 parasites, more samples of Gyrodactylus sp. were located on the pectoral fin. For a low number of Paragyrodactylus variegatus infections (<100), the pelvic and pectoral fins were the preferred locations of attachment, followed by the caudal fin. Between April and September, there were many monogenean parasites on fish fins, and the fish size was within the range of 5-10 cm. However, when a fish was longer than 10 cm long, the number of parasites on its fins greatly decreased.

Gyrodactylus ginestrae n. sp. (Monogenea: Gyrodactylidae), a parasite of the big-scale sand smelt, Atherina boyeri Risso, 1810 (Actinopterygii: Atherinidae) from the Black Sea.

We describe a new species, Gyrodactylus ginestrae n. sp., a parasite of the big-scale sand smelt (Atherina boyeri) from the Black Sea. This is the third monogenean species known from this fish host, found at 70% prevalence, but at relatively low abundance (1.9), on fish gills and fins. The new species is, both morphologically and genetically, most similar to G. salinae, which parasitizes the killifish Aphanius fasciatus (Cyprinodontidae) in the Mediterranean region. These species differ in the size of the haptoral hard parts and the number of small spines of the male copulatory organ. For molecular characterization, the internal transcribed spacer 1 (ITS1), 5.8S rRNA gene, and the internal transcribed spacer 2 (ITS2) were sequenced, completed by a fragment of the COII gene, thereby representing the first molecularly characterized gyrodactylid species from the Black Sea. Phylogenetic reconstruction based on the ITS1-5.8S-ITS2 sequence data indicated the position of G. ginestrae n. sp. in the marine "rugiensis" group of G. (Paranephrotus) and G. (Neonephrotus) subgenera which is part of the monophyletic "long ITS1" group. Taking into account the similarity of G. ginestrae n. sp. to several monogeneans from the Atlantic and Mediterranean regions, we suggest the Boreal-Atlantic origin of this species.

Distinctive site preference of the fish parasite Myxobolus cerebralis (Cnidaria, Myxozoa) during host invasion.

Here, we experimentally studied the site preference of Myxobolus cerebralis, one of the most pathogenic myxozoan (Cnidaria, Myxozoa) fish parasites, which causes whirling disease in salmonids. Parasite invasion was examined in three fish species with various susceptibility levels: the type host brown trout, the highly susceptible rainbow trout, and the non-susceptible gibel carp, in which parasite spores do not develop. We investigated the first two hours of fish invasion, and measured the site preference of triactinomyxons (TAMs) during attachment and penetration of fish in three body parts (gills, fins, skin). Infection prevalence and intensity were estimated using a species-specific nested PCR, optimised in the present study. The highest infection prevalence was detected in the most susceptible fish species, rainbow trout. Interestingly, higher prevalence was observed in gibel carp than in the type host, brown trout (95.2% vs. 85.7%). Considering body locations, remarkable differences were detected in infection intensities. The highest intensity was observed in fins, whereas skin was the least infected body part in every fish species examined. Infection prevalence and intensity did not differ significantly among fish species. Thus, we confirmed that M. cerebralis TAMs cannot discern fish species. Furthermore, we proved experimentally that fish fin is significantly more attractive to fish-invading parasite TAMs than gills or skin.

Description of Myxobolus xiantaoensis n. sp. from the fins of yellow catfish in China: a species previously attributed to Myxobolus physophilus Reuss, 1906 in Chinese records.

Myxozoans are economically important cnidarian endoparasites. Members of this group have been traditionally characterized by a morphology-based taxonomic system. Because myxozoans possess few morphological characters, these data are routinely accompanied by biological traits (host/organ/tissue specificity) and molecular data when describing or identifying myxozoan species. In the present study, a species of Myxobolus was collected from the fins of yellow catfish Tachysurus fulvidraco Richardson, 1846, which was consistent in spore morphology and host/organ specificity with Chinese records of Myxobolus physophilus Reuss, 1906. However, these earlier records and our own findings are inconsistent with the original description of M. physophilus from Russia. Specifically, there are differences in spore morphology (shape, intercapsular appendix, and polar capsule size), the infection site (air bladder vs. fins), and the host affinity (common rudd vs. yellow catfish). The inconsistencies allow us to conclude that both the present Myxobolus species and Chinese records of M. physophilus are distinct from the original description of M. physophilus and represent a new Myxobolus species, which we named Myxobolus xiantaoensis n. sp.

Molecular confirmation of Henneguya adiposa (Cnidaria: Myxozoa) and associated histologic changes in adipose fins of channel catfish, Ictalurus punctatus (Teleost).

Henneguya adiposa is one of ten known, closely related myxozoan species that parasitize a variety of tissue sites in the channel catfish, Ictalurus punctatus. Reported to specifically target the adipose fin, H. adiposa is not associated with morbidity or mortality, although detailed descriptions of its associated histologic pathology are lacking. The objective of this work was to confirm the presence of H. adiposa within fin lesions of affected channel catfish using DNA sequenced from histologic sections obtained by laser capture microdissection, as well as to describe pathologic changes induced by infection. The parasite formed large, white, elongate, nodular plasmodia that caused localized tissue damage and incited a granulomatous inflammatory response within a deep connective tissue layer at the base of the adipose fin. Myxospores released from ruptured plasmodia into adjacent tissue were observed to migrate superficially in tracts through the skin, indicating a portal of exit for environmental dispersal. Defects in the connective tissue layer created by ruptured plasmodia were infiltrated by granulomatous inflammation and fibroplasia, suggesting lesion resolution by scar formation over time. Sequencing of the 18S rRNA gene amplified from excised myxospores confirmed the myxozoan's identity as H. adiposa, with 100% similarity to the reference sequence from previous published work.

Morphological and molecular characterization of Myxobolus dibombensis sp. n. (Myxozoa: Myxobolidae), a parasite of the African carp Labeobarbus batesii (Teleostei: Cyprinidae) from Dibombe River, Cameroon.

Myxobolus dibombensis sp. n. (Cnidaria: Myxosporea: Bivalvulida) is described from the fins of the African carp, Labeobarbus batesii, based on morphological and molecular data. Prevalence of infection was 51.9% (67/129). Ovoid to spherical cyst-like plasmodia were found in the intrasegmental region and among the fin rays. No pathological changes were found in the fish host tissue surrounding the cyst-like plasmodia. Mature myxospores were ovoid in frontal view and lenticular in lateral view, with slightly truncated anterior and rounded posterior ends. Myxospores measured 16.8 (15.8-18.0) µm long and 11.4 (10.0-13.0) µm wide. There was a triangular intercapsular appendix measuring 3.8 (2.6-4.5) µm long. Polar capsules were ovoid and slightly unequal in size, occupying approximately one-third of the myxospore length. The larger polar capsule measured 7 (6-8) µm long and 3.6 (3-4) µm wide, while the smaller one measured 5.8 (4.8-7.0) µm long and 3 (2-4) µm wide. The larger polar capsule contained nine to 11 filament coils, whereas the smaller one contained seven to nine coils. SSU rDNA gene sequence of M. dibombensis sp. n. did not match any sequences available in the GenBank. The similarity with available Myxobolus spp. sequences ranged from 65 to 81%. The novel species clustered with M. algonquinensis, which infects the cyprinid Luxilus cornutus from Canada.

Non-native parasitic copepod Neoergasilus japonicus (Harada, 1930) utilizes non-native fish host Lepomis gibbosus (L.) in the floodplain of the River Dyje (Danube basin).

The parasitic copepod Neoergasilus japonicus (Harada, 1930) (Ergasilidae), native to east Asia, is widely distributed in Asia, Europe, and North and Central America. Recently, this species appeared in lentic water bodies of the River Dyje floodplain (Danube basin, Czech Republic). It was first recorded in 2015 and in 2 years it reached a 100% prevalence in recently expanding non-native fish host, Lepomis gibbosus (Linnaeus, 1758) (Centrarchidae, native to North America) at two borrow pits. Abundance of N. japonicus increased with fish length, with maximum intensity of infection reaching 99 parasites per fish. The parasite was most frequently found attached to the dorsal and anal fins of fish, while preference for the dorsal fin was more evident with lower infection intensities. Utilization of expanding fish hosts in water bodies that are regularly interconnected via natural or managed flooding may support the rapid dispersal of this non-native parasite.

A new species of Peniculus (Copepoda: Siphonostomatoida) parasitizing mesopelagic myctophid fish: first discovery of colonization of the genus in deep water.

Peniculus hokutoae n. sp. is described on the basis of an ovigerous adult female parasitizing the caudal fin of the myctophid fish Symbolophorus evermanni (Gilbert, 1905), collected from Suruga Bay, Japan. This is the first record of parasitism by this genus on mesopelagic myctophid fish. The new species is easily distinguished from other congeners in: (1) the presence of a conical process anterior to the rostrum; (2) the secondary elongation of the first pedigerous somite; (3) the incorporation of the third and fourth pedigerous somites into the trunk; (4) the unilobate maxillule bearing two unequal apical setae; (5) the lack of any processes on the first segment of the maxilla. Four morphological patterns of the cephalothorax, neck and anterior parts of the trunk can be found in the genus. We infer that initial colonization of a mesopelagic myctophid fish as host is likely to have occurred when the diurnally-migrating myctophid host was feeding in near-surface waters at night and was exposed to infective stages of Peniculus.

Protocol identification and preservation of myxozoan parasites for microscopy with silver nitrate (Klein's dry) staining technique.

The present study, discusses a first report of staining techniques with silver nitrate (AgNO3 ) stain for the preservation and identification of myxozoans globally. The silver nitrate stain was used to prepare permanent slide preparation of myxozoans with some adaptations made in our laboratory. Fresh air dried smear were stained with silver nitrate stain ensuing dark brown color polar capsules and light brown color of spore-wall. The stain is everlasting for years differing to other stains like Geimsa, iron-heamotoxylin Zeihl-Neelsen Blue, and trichome stains. RESEARCH HIGHLIGHTS: Stains usually fade with time and fail to disclose the morphological characters of the specimen. Present staining method helps to detect less infection in the tissue locating the myxospores. The Klein dry method (1958) is useful and suitable for long term preservation of the myxozoan slides and morphological description.

Association between adaptive immunity and neutrophil dynamics in zebrafish (Danio rerio) infected by a parasitic ciliate.

The protective immune response in zebrafish (Danio rerio) against the parasitic ciliate Ichthyophthirius multifiliis, targeting host skin, fins and gills, comprises an accelerated and manifold elevated immunoglobulin gene expression as well as a significantly elevated number of neutrophils at infected sites. Experimental fish were subjected to a primary I. multifiliis infection followed by a series of secondary exposures before they were challenged by a high dosage of infective theronts. Immunized fish responded immediately with a protective response suggesting existence of immunological memory whereas fish exposed to the parasite for the first time obtained a marked infection. The primary response to infection was dominated by expression of genes encoding acute phase reactants and inflammatory cytokines as well as recruitment of neutrophils at infected locations. Immunized fish showed a significantly upregulated immunoglobulin gene expression following challenge, which indicates existence of a secondary response effected by antibodies. Both responses induced a significantly elevated expression of the Th2 signature cytokine Il13. The increased presence of neutrophils in immunized fish suggests that innate cell mediated immunity supplements or influence the protective response against the parasite.

Comparison of Sampling and Detection Methods for Chinook Salmon and Steelhead Naturally Infected with Myxobolus cerebralis.

Myxobolus cerebralis (Mc) is a myxozoan parasite causing whirling disease in hatchery- and natural-origin salmonids. To minimize spread of this parasite and the incidence of its associated disease, fish health professionals routinely screen fish for Mc before stocking or moving the fish to Mc-free waters. Sample collection for Mc traditionally entails lethal sampling of cranial tissue followed by pepsin-trypsin digest (PTD) and screening of the sample for mature myxobolid myxospores (PTD method); however, nonlethal sampling methods would be advantageous in some circumstances, such as when dealing with rare or otherwise valuable fish. Accordingly, we compared Mc detections in cranial cartilage by using the PTD method with PCR assays of fin biopsies collected from juvenile Chinook Salmon Oncorhynchus tshawytscha and adult steelhead O. mykiss. Cranial samples were also analyzed using PCR methods for comparative purposes. Results indicated that Mc could be detected by PCR in fin clips, but the results generated by this approach differed significantly from those associated with PTD- and/or PCR-based analysis of cranial cartilage samples. Polymerase chain reaction-based analysis-of individual head samples and head digest pools in both species as well as fins in steelhead-yielded more positive detections than PTD analysis alone. The PCR-based analysis of head and fin tissues yielded different Mc detection rates in both species, but the nature of the detection disparity varied depending on the species and/or life stage of the fish. We conclude that for lethal cranial samples, neither PTD nor PCR should be used alone, but using these techniques in concert may provide the most complete and accurate estimation of Mc presence in a group of salmonids. If imperiled or highly valuable fish are in question, nonlethal fin samples may be used to generate some information regarding Mc status, with the understanding that parasite DNA detections do not necessarily signify mature infections or disease.

Morphological and ssrDNA sequence based molecular characterization of a novel Thelohanellus species (Myxosporea: Myxobolidae) infecting the fins of Goldfish, Carassius auratus L. with special reference to its histopathological alteration.

A new species of myxozoan, Thelohanellus goldi n. sp. is described using morphological and molecular data, parasitizing the fin filaments from 18 of 25 host specimens (72.5%) of Carassius auratus collected from different ornamental fish farms of India. Mature spore of the new species were oval to spherical in frontal view having rounded posterior ends and tapering anterior end measures 8.7-10.26 (9.50) × 4.10-7.89 (5.84) µm. The single large polar capsule, round to oval in shape but slightly pointed at the anterior end measuring 4.91-7.63 (5.60) × 2.3-3.1 (2.96) µm and located just below the anterior end of the spore. Polar filament only at distal end with 5-6 loose coils. The most differentiating feature from closely related species was carried out by morpho-taxonomic affinities with previously described species which are tremendously supported by molecular taxonomy by partial sequencing of the 18S rDNA gene resulted in a total of 2124 bp fragment of newly obtained small subunit ribosomal RNA gene sequence of the new species which Exhibit 93-95% homogeneity with other closely related species available in GenBank. The BLAST search and high genetic diversity of distance matrix of Myxobolus sp. did not properly match with any available sequences in GenBank and make sister clade with Thelohanellus caudatus and Thelohanellus habibpuri in the Thelohanellus clade including most of Thelohanellus spp. The study of evolutionary history enables us to understand the evolution of modern species and supports some uncertain topologies which are being presented regarding the morphometric analysis. The severity of myxozoan infection has been assessed in this article by observing the histopathological changes of fins of the C. auratus along with the diversity, distribution and taxonomic description of the new Thelohanellus species with their new host and locality records.

Molecular data reveal high diversity of Uvulifer (Trematoda: Diplostomidae) in Middle America, with the description of a new species.

Members of the genus Uvulifer are distributed worldwide and infect aquatic snails and freshwater fishes as first and second intermediate hosts, respectively, and fish-eating birds (kingfishers) as definitive hosts. Metacercariae of Uvulifer spp. were collected from the fins and skin of 20 species of freshwater fishes in Mexico, Guatemala, Honduras, Nicaragua and Costa Rica, and the adults were recovered from the intestine of kingfishers in four localities of Mexico. The genetic divergence among 76 samples (64 metacercariae and 12 adults) was estimated by sequencing the 28S and 5.8S nuclear genes, as well as the internal transcribed spacers ITS1 and ITS2, and one mitochondrial gene (cox1). Maximum likelihood and Bayesian inference analyses inferred with each dataset showed a high genetic diversity within the genus Uvulifer across Middle America, revealing the existence of four genetic lineages that exhibit some level of host specificity to their second intermediate hosts. The metacercariae of lineage 1 were associated with characids and cyprinids in central and northern Mexico. Metacercariae of lineages 2 and 3 were associated with cichlids distributed widely across Middle America. The lack of adults of these lineages in kingfishers, in lineages 2 and 3, or the fact that just a few adult specimens were recovered, as in lineage 1, prevented a formal description of these species. The metacercariae of lineage 4 were found in poeciliids, across a distribution range comprising Mexico, Guatemala, Honduras and Nicaragua, and the adult was found in the green kingfisher in Mexico. The number of specimens sampled for lineage 4, for both gravid adults and metacercariae, allowed us to describe a new species, Uvulifer spinatus n. sp. We describe the new species herein and we discuss briefly the genetic diversity in Uvulifer spp. and the importance of using DNA sequences to properly characterize parasite diversity.

Studies on histopathological alteration of three major organs of the goldfish, Carassius auratus L., of India due to myxozoan infection with special reference to scanning electron microscopic observation.

Severe infections by the protozoan parasite belonging to the genera Myxozoa are causing serious problems in ornamental fish reared in breeding farms. Histopathological study is being used for diagnosis of the severity of diseases. Myxozoan infections result in large scale histopathological damages in different fish tissues. No information is available regarding the histopathological changes of goldfish due to infection with myxozoans. The present study deals with the histopathological changes of the gill, fin, and skin of goldfish, infected with myxozoan parasites along with an ultrastructural study using scanning electron microscope. Several histological alterations have been observed in goldfish, like tissue damage, epithelial hyperplasia, necrosis, anoxia, localized lymphocytic infiltration, excess mucus, cellular necrosis, and epithelial proliferation. The present study revealed the invasion intensified by the occurrence of morphological lesions in the gill, skin, and fin exposed to Myxosporidia, which may lead to respiratory insufficiency in fish and even cause mass mortality.

Ultrastructural, ssrDNA sequencing of Myxobolus prochilodus and Myxobolus porofilus and details of the interaction with the host Prochilodus lineatus.

Myxobolus prochilodus and Myxobolus porofilus are parasites of Prochilodus lineatus, an economically important South American fish found in La Plata and Paraiba do Sul river basins. This study focusing on parasite-host interaction provides an ultrastructural and phylogenetic analysis, the latter based on ssrDNA sequencing of these parasites respectively infecting the gill filaments and fins of P. lineatus taken from the Mogi Guaçu River, São Paulo, Brazil. A total of 13 adult specimens were examined in this study. The prevalence of infection was 7.69 % for M. prochilodus and 15.38 % for M. porofilus. Phylogenetic analysis showed M. prochilodus and M. porofilus clustered in a subclade composed of parasites of the Prochilodontidae family. In M. prochilodus infecting gill filaments, where cellular degeneration in the epithelium was observed, the plasmodia were surrounded by a capsule composed of layers of fibrocyte-like cells, with cellular projections joined to the projections of other fibrocyte-like cells by desmosomes, and more externally typical fibroblast layers. Some granular leukocytes were seen interspersed among these layers. In M. porofilus infecting the fins, the capsule of connective tissue was represented only by loosely arranged collagen fibers, and no granular leucocytes were observed. Finally, several unusual vacuoles with filamentous content and some characteristics usually described as degenerative alterations, as myelin figure, were noted in plasmodia and pansporoblasts of both myxosporean species. The possible influence of inflammatory response and xenobiotics was considered to be the explanation for the alterations observed in Myxobolus species and its host.