Two new and one newly recorded gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) parasitising marine fishes in Iraq.

Specimens of two undescribed and one known gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) were collected in some marine fishes from off the southern coast of Iraq. Based on light and scanning electron microscopy, the following new species are described: Philometra tayeni n. sp. (males and nongravid females) from ovaries of the purple-spotted bigeye Priacanthus tayenus Richardson (Priacanthidae, Acanthuriformes), and Philometra nibeae n. sp. (males and gravid female) from the ovary of the blotched croaker Nibea maculata (Bloch et Schneider) (Sciaenidae, Acanthuriformes). Philometra tayeni is mainly characterised by a pair of postanal papillae and a V-shaped caudal mound in males and by their body lengths (2.42-2.99 mm), whereas P. nibeae differs from its gonad-infecting congeners parasitising scienids mainly based on the body length of males (2.29-2.49 mm) and their spicules (96-117 µm), absence of a pair of postanal papillae and shape of caudal mound consisting of two parts. Philometra piscaria Moravec & Justine, 2014 (males and nongravid females), a parasite of the orange-spotted grouper Epinephelus coioides (Hamilton) (Epinephelidae, Perciformes), is recorded from the Arabian (= Persian) Gulf for the first time; previously unknown females of this species are described.

Comparative Studies on The Efficiency of Neem Leaves Azadirachta indica and Flubendazole Treatment Against Diplectanum in Sea Bass Dicentrarchus labrax.

PURPOSE: Diplectanum is a life-threatening metazoan infecting the gills of Sea bass Dicentrarchus labrax causing a wide-ranging extensive economic loss in the aquaculture sector. This study has focused on verifying the most effective non-toxic dose of the Neem (Azadirachta indica) and (flubendazole) bath treatment on infested D. labrax fingerlings. METHODS: In the first phase of the experiment, a total of 180 apparently healthy fingerlings were subdivided into six groups for each treatment. The tested concentrations were 0, 50, 100, 150, 200, and 250 mg L-1 for A. indica and 0, 10, 20, 30, 40, and 50 mg L-1 for flubendazole. The second phase was conducted for one week in five groups for each treatment. The first group was untreated healthy. The remaining groups were infested and received different concentrations of 0, 50, 100, and 150 mg L-1 & 0, 10, 20, and 30 mg L-1 for A. indica and flubendazole, respectively. RESULTS: The most toxic dose exhibited high mortality rates at 200 & 250 and 40 & 50 mg L-1 for A. indica and flubendazole, respectively. In the second phase of the experiment, the most effective dose was 150 and 30 mg L-1; for A. indica and flubendazole, respectively. They demonstrated the lowest mortality rates 20.00 & 20.00 %, prevalence rates 43.33 & 23.33%, and mean parasitic intensities were 2.35 & 2.00 accompanied by the highest therapeutic efficacy value 67.85 & 74.6% for both treatments; respectively. CONCLUSION: The most effective anthelmintic efficacy has been assigned for flubendazole and A. indica at 30 and 150 mg L-1.

Bicentenariella n. g. (Monogenea: Dactylogyridae) including descriptions of three new species and two new combinations from serranid fishes (Actinopterygii: Serranidae: Anthiinae) in the South American Pacific Ocean.

Bicentenariella n. g. is proposed to accommodate three new species of dactylogyrid monogeneans found on the gills of the threadfin bass Pronotogrammus multifasciatus Gill (Perciformes: Serranidae) from the Southeastern Pacific Ocean off Peru: Bicentenariella claudiae n. sp. (type-species), B. sinuosa n. sp. and B. puertopizarroensis n. sp. Bicentenariella n. g. is mainly characterised by possessing a broadly X-shaped dorsal bar, which has the anterior arms longer than posterior ones and by having a ventral bar with two medial projections. Bicentenariella n. g. is also characterised by having: (i) anchors equal, each with elongate superficial root and short deep root; (ii) an haptor with bilaterally paired lobes, lacking haptoral reservoirs; (iii) hooks with protruded obtuse thumb and undilated shank; (iv) a tubular tapered-shaped male copulatory organ with basal flap bifurcated or not (MCO), lacking accessory piece; (v) a delicate membrane associated with the shaft of the MCO present or absent; (vi) a muscular trumpet-shaped vagina, vaginal aperture dextrolateral; (vii) eyespots absent, accessory chromatic granules present; and (viii) a not lobulated testis. Bicentenariella claudiae n. sp. is characterised by having a MCO with whip-shaped distal end and a rod-shaped ventral bar with hatchet-shaped lateral ends. Bicentenariella sinuosa n. sp. is typified by possessing a MCO with an irregular filamentous membrane surrounding its shaft and a dumbbell-shaped ventral bar. Bicentenariella puertopizarroensis n. sp. can be differentiated from its congeners by having a tubular-shaped MCO with twisted distal end and a narrow W-shaped ventral bar. Parancylodiscoides peruensis Cruces, Chero, Sáez & Luque, 2017 from Hemanthias peruanus (Steindachner) and P. signiferi Cruces, Chero, Sáez & Luque, 2017 from H. signifer (Garman), are transferred to Bicentenariella n. g. as B. peruensis n. comb and B. signiferi n. comb., respectively.

Ceratothoa oestroides Infection in European Sea Bass: Revealing a Long Misunderstood Relationship.

Ceratothoa oestroides (Cymothoidea, Isopoda) is a generalist crustacean parasite that negatively affects the economic sustainability of European sea bass (Dicentrarchus labrax) aquaculture in the North-East Mediterranean. While mortalities are observed in fry and fingerlings, infection in juvenile and adult fish result in approximately 20% growth delay. A transcriptomic analysis (PCR array, RNA-Seq) was performed on organs (tongue, spleen, head kidney, and liver) from infected vs. Ceratothoa-free sea bass fingerlings. Activation of local and systemic immune responses was detected, particularly in the spleen, characterized by the upregulation of cytokines (also in the tongue), a general reshaping of the immunoglobulin (Ig) response and suppression of T-cell mediated responses. Interestingly, starvation and iron transport and metabolism genes were strongly downregulated, suggesting that the parasite feeding strategy is not likely hematophagous. The regulation of genes related to growth impairment and starvation supported the growth delay observed in infected animals. Most differentially expressed (DE) transcripts were exclusive of a specific organ; however, only in the tongue, the difference between infected and uninfected fish was significant. At the attachment/feeding site, the pathways involved in muscle contraction and intercellular junction were the most upregulated, whereas the pathways involved in fibrosis (extracellular matrix organization, collagen formation, and biosynthesis) were downregulated. These results suggest that parasite-inflicted damage is successfully mitigated by the host and characterized by regenerative processes that prevail over the reparative ones.

Control of Riboscyphidia sp. (Ciliate) Infection in Asian Sea Bass (Lates calcarifer), Cultivated in the Red Sea.

BACKGROUND AND OBJECTIVE: Sessiline ciliates live as eco commensals (low numbers) and parasites (high numbers) on different hosts, like mollusks copepods, mysids and fish. Riboscyphidia ecto-protozoan is moderately pathogenic but high numbers of it on the gills can physically prevent gas exchange. The present study aimed to describe the epizoic ciliates Riboscyphidia found on the Red Sea cultured Asian sea bass and obtain more information on the Epidemiology of the parasite with special references to control and histopathological examination of naturally infected sea bass. MATERIALS AND METHODS: The occurrence of epizoic ciliates on the adult Asian Sea bass. About 100 Asian sea bass were collected by the fishing net at a private marine fish farm at Ismailia governorate and transferred to the hydrobiology laboratory at National Research Centre. A parasitological and histopathological study of epizoic sessile ciliate species was done. ANOVA test was used for Statistical analysis. RESULTS: Riboscyphidia sp. was found and isolated after parasitological examination of investigated adult's Asian sea bass. The prevalence of Riboscyphidiosis was 64%. Sessile ciliates were found on gills, skin and fins. The clinical signs of Riboscyphidiosis were respiratory distress, flashing and off food. Histopathological alterations in naturally infested Asian sea bass were investigated. CONCLUSION: The treatment of choice of Riboscyphidiosis was prolonged immersion by Copper citrate with a dose of 0.56 mg mL-1 for 7 days.

Redescription of the Type Species of Cardicola Short, 1953 (Digenea: Aporocotylidae) and Description of a New Congener Infecting Yellowedge Grouper, Hyporthodus flavolimbatus (Perciformes: Serranidae), from the Gulf of Mexico.

Cardicola Short, 1953 is the most speciose aporocotylid genus (35 species) and includes marine and estuarine species of fish blood flukes that infect "higher ray-finned fishes" (Euteleostei). Several clades within Cardicola are recovered in phylogenetic analyses of the large subunit ribosomal DNA (28S), but morphological synapomorphies for those nucleotide-based clades remain elusive. The type species, Cardicola cardiocola (Manter, 1947) Short, 1953, has not been recollected in 73 yr and the original description was incomplete; making a genus revision challenging because of the ambiguous systematic position of its type species. Herein, we redescribe C. cardiocola by using the holotype (USNM 1337732) and new specimens collected from the type host, jolthead porgy, Calamus bajonado (Sparidae), from nearby the type locality. It differs from its congeners by the combination of having a body that is 5 times longer than wide, an anterior sucker with concentric rows of spines, 2-6 tegumental body spines per row, an esophageal gland that is 22-43% of the esophageal length, a testis that is 3-5 times longer than wide and that fills the intercecal space, a vitelline duct connecting to the anterior aspect of the oötype, an ascending uterus that lacks any coil, a descending uterus yielding a single coil, an obvious cirrus sac separated by constriction from the seminal vesicle, a tegumental protrusion surrounding the terminal end of cirrus sac, and a male genital pore that is posterior to the remainder of the genitalia. We also describe a new congener infecting the heart of yellowedge grouper, Hyporthodus flavolimbatus (Serranidae), from the Gulf of Mexico. It differs from its congeners by the combination of having an anterior sucker that does not extend beyond the anterior body margin, 2-5 tegumental body spines per row, posterior ceca that are 9 times length of the anterior ceca and that lack any coil, a testis that is 3 times longer than wide and that does not fill the intercecal space, an ovary that is >60% of the body width, a vitelline duct that connects to the anterior aspect of the oötype, a uterus that is >10% of the body width and that extends posterior to all genitalia, and a rounded posterior body margin. It is the first species of Cardicola to be described from a grouper (Serranidae). The 28S and internal transcribed spacer 2 phylogenetic analyses recovered the new species as a distinct lineage within the clade of Cardicola spp.

Zschokkella epinepheli n. sp. (Myxosporea: Myxidiidae) infecting the gallbladder of the white grouper Epinephelus aeneus (Serranidae) from Tunisian waters.

A new coelozoic myxosporean species, Zschokkella epinepheli n. sp., collected from the gallbladder of the white grouper Epinephelus aeneus (Perciformes: Serranidae) from the bay of Bizerte, Tunisia, is described based on morphological and molecular characteristics. Myxospores and plasmodia were observed floating free in the bile. Mature plasmodia were polysporic and subspherical in shape, measuring 85.0-94.0 µm long and 70.0-82.0 µm wide. Mature myxospores were ovoid in valvular view, measuring 10.0 ± 1.7 (8.0-11.0) µm in length and 7.0 ± 0.3 (6.6-7.5) µm in width. Polar capsules were pyriform and equal in size, measuring 3.0 ± 0.2 (2.8-3.6) µm in length and 2.3 ± 0.3 (1.8-2.7) µm in width. Myxospore valves had 12-14 longitudinal striations. Based on the small subunit rDNA, the new species Z. epinepheli n. sp. differs from all other Zschokkella species for which there is a DNA sequence deposited in GenBank. Phylogenetic analysis revealed that Z. epinepheli n. sp. clustered in the marine subclade of Zschokkella species within the biliary tract IV clade. This is the first report of a Zschokkella species from the gallbladder of an epinephelin fishes.

Caligus tunisiensis n. sp. (Copepoda: Caligidae) parasitic on the painted comber Serranus scriba (L.) (Perciformes: Serranidae) from the Mediterranean Sea, off the Tunisian coast.

A new species of parasitic copepod, Caligus tunisiensis n. sp. (Caligidae), is described based on two female specimens collected from the gills of the painted comber, Serranus scriba (L.), caught in the Mediterranean Sea, off the Tunisian coasts. The new species belongs to the Caligus productus-species group established by Boxshall & Gurney (1980) as it shares the following set of character states: (i) antenna with well-developed posterior process on proximal segment; (ii) posterior margin of distal exopodal segment of leg 1 lacking typical plumose setae, or retaining single vestigial seta; and (iii) 2-segmented exopod of leg 4 armed with IV spines on compound distal exopodal segment. Detailed morphological comparisons between the new species and the core members of the C. productus-species group revealed that the new species closely resembles with C. productus Dana, 1852 and C. temnodontis Brian, 1924. However, the new species can be distinguished from its congeners in having: (i) a female maxilliped bearing a prominent bi-lobate myxal process, opposing tip of the claw; (ii) leg 4 protopod ornamented with a patch of spinules on the posterolateral surface; and (iii) an abdomen ornamented with two rows of minute spinules at the posterolateral corners.

Transcriptome Analysis of Amyloodinium ocellatum Tomonts Revealed Basic Information on the Major Potential Virulence Factors.

The ectoparasite protozoan Amyloodinium ocellatum (AO) is the etiological agent of amyloodiniosis in European seabass (Dicentrarchus labrax) (ESB). There is a lack of information about basic molecular data on AO biology and its interaction with the host. Therefore, de novo transcriptome sequencing of AO tomonts was performed. AO trophonts were detached from infested ESB gills, and quickly becoming early tomonts were purified by Percoll® density gradient. Tomont total RNA was processed and quality was assessed immediately. cDNA libraries were constructed using TruSeq® Stranded mRNA kit and sequenced using Illumina sequencer. CLC assembly was used to generate the Transcriptome assembly of AO tomonts. Out of 48,188 contigs, 56.12% belong to dinophyceae wherein Symbiodinium microadriaticum had 94.61% similarity among dinophyceae. Functional annotations of contigs indicated that 12,677 had associated GO term, 9005 with KEGG term. The contigs belonging to dinophyceae resulted in the detection of several peptidases. A BLAST search for known virulent factors from the virulence database resulted in hits to Rab proteins, AP120, Ribosomal phosphoprotein, Heat-shock protein70, Casein kinases, Plasmepsin IV, and Brucipain. Hsp70 and casein kinase II alpha were characterized in-silico. Altogether, these results provide a reference database in understanding AO molecular biology, aiding to the development of novel diagnostics and future vaccines.

Temperature is a key factor influencing the invasion and proliferation of Toxoplasma gondii in fish cells.

Toxoplasma gondii has long been considered a ubiquitous parasite possessing the capacity of infecting virtually all warm-blooded animals globally. Occasionally, this parasite can also infect cold-blooded animals such as fish if their body temperature reaches 37 °C. However, we are currently lacking an understanding of key details such as the minimum temperature required for T. gondii invasion and proliferation in these cold-blooded animals and their cells. Here, we performed in vitro T. gondii infection experiments with rat embryo fibroblasts (REF cells), grouper (Epinephelus coioides) splenocytes (GS cells) and zebra fish (Danio rerio) hepatocytes (ZFL cells), at 27 °C, 30 °C, 32 °C, 35 °C and 37 °C, respectively. We found that T. gondii tachyzoites could penetrate REF, GS nd ZFL cells at 27 °C but clear inhibition of multiplication was observed. Intriguingly, the intracellular tachyzoites retained the ability to infect mice after 12 days of incubation in GS cells cultured at 27 °C as demonstrated by bioassay. At 30 °C, 32 °C and 35 °C, we observed that the mammalian cells (REF cells) and fish cells (GS and ZFL cells) could support T. gondii invasion and replication, which showed a temperature-dependent relationship in infection and proliferation rates. Our data demonstrated that the minimum temperature for T. gondii invasion and replication was 27 °C and 30 °C respectively, which indicated that temperature should be a key factor for T. gondii invasion and proliferation in host cells. This suggests that temperature-dependent infection determines the differences in the capability of T. gondii to infect cold- and warm-blooded vertebrates.

Prevalence, intensity and histopathology of Zeylanicobdella arugamensis infestation on groupers reared on different aquaculture systems.

The marine leech, Zeylanicobdella arugamensis, is a major threat to aquaculture in grouper-producing countries including Indonesia. This study aimed at investigating prevalence, intensity and histopathology of the ectoparasite in humpback and hybrid groupers cultured in different rearing systems. A total of 260 groupers (60 humpback groupers and 200 hybrid groupers) were used for samples. The marine leech was observed on skin, fins, gills and mouth, followed by histopathological assay on the skin tissue. The results showed that prevalence of the leech in both groupers was higher when they were cultured in the floating net cages compared with the hatchery, p < .05. Furthermore, humpback grouper had a higher prevalence than hybrid grouper when they were cultured in a similar system, p < .05. Meanwhile, there was no significant difference in intensity between the two groupers, p > .05. Within the hybrid groupers, the highest prevalence was obtained from hybrid groupers reared in the earthen ponds. Histopathological studies showed that the infected groupers exhibited inflammation, congestion and erosion of the epidermis layer. Hybrid grouper had more severe histopathological lesions in the skin tissues. These results suggested that species and type of aquaculture system had significantly determined the prevalence, intensity and severity of lesion in Z. arugamensis infestation.

Ceratomyxa mennani n. sp. (Myxosporea: Bivalvulida) parasitizing the gallbladder of the dusky grouper Epinephelus marginatus (Serranidae) from Tunisian waters.

Ceratomyxa mennani n. sp. is a new coelozoic Ceratomyxa species found in the gallbladder of Epinephelus marginatus from the Gulf of Tunis, Tunisia. Mature plasmodia were disporic, ovoid in shape measuring 9-12 µm in width and 11-14 µm in length. Mature myxospores were slightly crescent-shaped with almost straight posterior margin, measuring 5.8 ± 0.2 (5.4-6.1) µm in length and 12.7 ± 0.3 (11.9-13.0) µm in thickness. The two valves were unequal with rounded ends. Polar capsules were spherical, equal in size with 2.1 ± 0.2 (1.9-2.6) µm in diameter. The binucleated sporoplasm filled the entire cavity of the myxospore. Molecular analysis of SSU rDNA sequences indicated that C. mennani n. sp. was distinct from all other Ceratomyxa sequences in GenBank. Phylogenetic analysis revealed that C. mennani n. sp. clustered with Ceratomyxa species infecting Epinephelinae fishes. Seasonal prevalence of infection over one year was significantly higher in winter and the lowest in autumn. This is the third report of Ceratomyxa species infecting the gallbladder of Epinephelus marginatus from Tunisia and the first study to include molecular data.

A new genus and three new species of dactylogyrids (Monogenea), gill parasites of the threadfin bass, Pronotogrammus multifasciatus Gill (Perciformes: Serranidae) in the Southeastern Pacific Ocean off Peru.

Pronotogrammella n. g. is erected to accommodate Pronotogrammella boegeri n. sp. (type-species), Pr. scholzi n. sp. and Pr. multifasciatus n. sp. (Monogenea: Dactylogyridae). The new species are gill parasites of the threadfin bass Pronotogrammus multifasciatus Gill (Perciformes: Serranidae), a demersal teleost collected from off the coastal zone of Puerto Pizarro, Tumbes, Peru. Pronotogrammella n. g. is mainly characterised by having broadly fork-shaped dorsal anchors, which have an accessory anchor sclerite articulated to the tip of the superficial roots. Pronotogrammella n. g. is also characterised by having: (i) a tubular tapered-shaped male copulatory organ (MCO), filamentous distally, with a clockwise coil at distal end or not, lacking accessory piece; (ii) a dorsal bar with an anteromedial delicate umbelliform membrane supported by two processes; (iii) hooks with upright blunt thumb and uniform shank; (iv) a vaginal aperture dextrolateral; (v) a subquadrangular haptor, with inconspicuous lateral flaps and lacking haptoral reservoirs; and (vi) eye-spot or chromatic granules absent. Pronotogrammella boegeri n. sp. is characterised by its crosier-shaped MCO having a clockwise coil at distal end and by its dorsal bar with a straight anteromedial processes. Pronotogrammella scholzi n. sp. is typified by possessing of a dorsal bar with the anteromedial processes like cow horns, hoof-shaped deep roots of the dorsal anchors and a broader shaft of the MCO. Pronotogrammella multifasciatus n. sp. differs from all congeners by having a tubular MCO with twisted shaft and a base with a short and broad arm and by having an almost dumbbell-shaped ventral bar.

Pseudorhabdosynochus kasetsartensis n. sp. (Monogenea: Diplectanidae) from the cloudy grouper Epinephelus erythrurus (Valenciennes) (Perciformes: Serranidae) in the lower Gulf of Thailand.

Pseudorhabdosynochus kasetsartensis n. sp. is described from the gills of the cloudy grouper Epinephelus erythrurus (Valenciennes) caught in the lower Gulf of Thailand. The new species is distinguished from other species assigned to the genus by the structure of its sclerotised vagina which has a wide and prominent sclerotised trumpet, long, thin, coiled or curved primary canal, short secondary canal, and primary and secondary chambers that are blind extremities of the primary and secondary canals, respectively. This is the first species of Pseudorhabdosynochus Yamaguti, 1958 described from E. erythrurus and the first record of a species of Pseudorhabdosynochus in Thailand.

Saprolegnia molecular phylogeny among farmed teleosts in Nova Scotia, Canada.

To identify the pathogens causing saprolegniosis among farmed fish in Nova Scotia, 172 infected tissues and 23 water samples were collected from six species of teleosts: Atlantic salmon (Salmo salar), brown trout (Salmo trutta), Arctic charr (Salvelinus alpinus), brook trout (Salvelinus fontinalis), striped bass (Morone saxatilis) and rainbow trout (Oncorhynchus mykiss) at nine facilities over a 600 km range. Following laboratory culture, 132 isolates were recovered. Six species of oomycetes were identified from analysis of the internal transcribed spacer (ITS) sequence of the nrDNA: Saprolegnia parasitica, Saprolegnia ferax, Saprolegnia diclina, Saprolegnia aenigmatica, Saprolegnia torulosa, Saprolegnia sp. and Pythiopsis cymosa. Further phylogenetic analyses of the ITS and cytochrome c oxidase subunit 1 (Cox1) regions revealed four strains of Saprolegnia parasitica (named here as S1, S2, S3 and S4), of which S1 and S2 were common (37% and 42% of the isolates), and two strains of S. ferax. Among S. parasitica, S2 and S3 are more closely related to each other than to S1 based on the phylogenetic analyses and predicted RNA secondary structure of the ITS region. Sexual structures with a similar morphology were formed by S1 and S3 in vitro, but were not formed by S2.

First description of the male and subgravid female of Philometra serranellicabrillae Janiszewska, 1949 (Nematoda: Philometridae), a gonad-infecting parasite of the comber Serranus cabrilla (Linnaeus) (Serranidae) off Tunisia.

The male and subgravid female of Philometra serranellicabrillae Janiszewska, 1949 (Philometridae) collected from the gonads of Serranus cabrilla (Linnaeus) (Serranidae) off Tunisia are described for the first time based on light and scanning electron microscopical studies. The male of this nematode can be separated from other congeneric gonad-infecting nominal species in the structure and shape of the gubernaculum (e.g. absence of a dorsal protuberance and a median smooth field). The shape of the male posterior region is unique in that it bears a pair of big circular papillae posterior to the cloacal opening, which is also present in other Philometra spp. from serranids, i.e. P. indica Moravec & Manoharan, 2014, P. inexpectata Moravec, Chaabane, Justine & Neifar, 2016 and P. jordanoi (López-Neyra, 1951) Yamaguti, 1961. Moreover, P. serranellicabrillae differs from its congeners in other fish families from the Mediterranean Sea, in the length of spicules and gubernaculum.

First report of trypanosomiasis in farmed largemouth bass (Micropterus salmoides) from China: pathological evaluation and taxonomic status.

The aim of this study was to evaluate the effect of trypanosomes on cultured largemouth bass (Micropterus salmoides) and describe the taxonomic identification of the parasite. The effects of the parasite on M. salmoides were examined based on clinical symptoms, hemograms, histopathology, and serum biochemistry. Diseased fish showed typical clinical symptoms of trypanosomiasis, which included lethargy, anorexia, and histopathological lesions in the liver, head kidney, and spleen. The serum of diseased fish had significantly lower concentrations of glucose, triglyceride, and low-density lipoprotein, and significantly higher alanine transaminase (ALT), aspartate transaminase (AST), and lactate dehydrogenase (LDH) activities. The morphology of the trypanosomes was also analyzed using light microscopy, and their 18S rDNA sequence was analyzed to establish genetic relationships with other known strains. We found that the trypomastigote form of the trypanosomes from M. salmoides was similar to those isolated from Pelteobagrus fulvidraco. The trypanosomes had a slender and narrow body with a relatively long free flagellum, not well-developed undulating membrane, and an oval kinetoplast located near the subterminal posterior end of the body. The 18S rDNA sequences of the trypanosome from M. salmoides had the highest similarity (99.8%) with that of P. fulvidraco, suggesting they are identical species. Based on the differences in morphological characteristics and 18S rDNA sequence compared to trypanosomes isolated from other freshwater fish, it is considered as a new species and we propose the name Trypanosoma micropteri n. sp.

Newly Described Coccidia Goussia bayae from White Perch Morone americana: Morphology and Phylogenetics Support Emerging Taxonomy of Goussia within Piscine Hosts.

In March and April 2016, 150 white perch ( Morone americana) were collected from various localities in Chesapeake Bay and examined for coccidia. A previously undescribed species of coccidia was observed in the hepatic bile ducts and gallbladder of all white perch (100%) examined. We describe this species using morphological characteristics, histology, and gene sequences of the small-subunit ribosomal DNA ( rDNA), large-subunit rDNA, and mitochondrial genes cytochrome oxidase 1 ( COI), cytochrome oxidase b ( Cytb), and cytochrome oxidase 3 ( COIII). Oocysts of Goussia bayae n. sp. were subspherical with a single-layered smooth wall and measured (length [L] × width [W]) 26.2 × 21.8 µm, with a L/W ratio of 1.2. A micropyle was present but a micropyle cap, polar granules, and oocyst residuum were absent. Each oocyst contained 4 sporocysts that were ellipsoidal and measured (L × W) 12.6 × 7.8 µm, with a L/W ratio of 1.6. A pair of sporozoites was present, but sporocysts lacked a Stieda body and residuua. Meronts and gamonts were epicellular in biliary epithelial cells and oocysts were coelozoic in hepatic and common bile ducts and gallbladder. This is the first report of Goussia spp. from white perch and the first mitochondrial DNA sequence reported from a Goussia species. Phylogenetic analysis indicates basal placement of G. bayae to Eimeriidae, Choleoeimeria, and Sarcocystidae.

Expression of infection-related immune response in European sea bass (Dicentrarchus labrax) during a natural outbreak from a unique dinoflagellate Amyloodinium ocellatum.

In the Mediterranean area, amyloodiniosis represents a major hindrance for marine aquaculture, causing high mortalities in lagoon-type based rearing sites during warm seasons. Amyloodinium ocellatum (AO) is the most common and important dinoflagellate parasitizing fish, and is one of the few fish parasites that can infest several fish species living within its ecological range. In the present study, A. ocellatum was recorded and collected from infected European sea bass (Dicentrarchus labrax) during a summer 2017 outbreak in north east Italy. Histological observation of infected ESB gill samples emphasized the presence of round or pear-shaped trophonts anchored to the oro-pharingeal cavity. Molecular analysis for small subunit (SSU) rDNA of A. ocellatum from gill genomic DNA amplified consistently and yielded 248 bp specific amplicon of A. ocellatum, that was also confirmed using sequencing and NCBI Blast analysis. Histological sections of ESB gill samples were addressed to immunohistochemical procedure for the labelling of ESB igm, inos, tlr2, tlr4, pcna and cytokeratin. Infected gills resulted positive for igm, inos, pcna and cytokeratin but negative to tlr-2 and tlr-4. Furthermore, ESB immune related gene response (innate immunity, adaptive immunity, and stress) in the course of A. ocellatum infection using quantitative polymerase chain reaction (qpcr) for infected gills and head kidney was analysed. Among the twenty three immune related gene molecules tested, cc1, il-8, il-10, hep, cox-2, cla, cat, casp9, and igt were significantly expressed in diseased fish. Altogether, these data on parasite identification and expression of host immune-related genes will allow for a better understanding of immune response in European sea bass against A. ocellatum and could promote the development of effective control measures.

Functional ultrastructure and cytochemistry of vitellogenesis and mature vitellocytes of the digenean Cainocreadium labracis (Dujardin, 1845), parasite of Dicentrarchus labrax (L., 1758).

Vitellogenesis and vitellocytes of Cainocreadium labracis were studied by transmission electron microscopy (TEM) and TEM cytochemistry. Four developmental stages were distinguished during vitellogenesis: (I) stem cell of high nucleo-cytoplasmic ratio; (II) early differentiation with chief activity focused on the beginning of protein synthesis and shell globule formation; (III) advanced differentiation with rapid intensification of protein synthesis, progressive fusion of single shell globules into large globule clusters, and formation of unsaturated lipid droplets surrounded by ß-glycogen particles; and (IV) mature vitellocyte. Early vitellogenesis with vitellocyte maturation consists of: (1) increase in cell volume; (2) increased development of large, parallel cisternae of GER with production of proteinaceous granules; (3) development of small Golgi complexes that package granules; and (4) within vacuoles, progressive enlargement of proteinaceous granules into shell globule clusters formed during vitellogenesis. Three types of inclusions accumulate in large amounts in mature vitelline cells: (1) shell globule clusters, important component in the formation of egg shell; (2) numerous unsaturated lipid droplets. Though fewer, there are also diphasic droplets consisting of saturated and unsaturated lipids in the same droplet, and (3) a relatively small amount of ß-glycogen particles, usually surround a few groups of lipid droplets. The ß-glycogen and lipid droplets are nutritive reserves for embryogenesis. General pattern and functional ultrastructure of vitellogenesis greatly resemble those observed in some lower cestodes, such as bothriocephalideans and diphyllobothrideans. Variations and differences in the amount of lipids and of glycogen during vitellogenesis in lower cestodes and other trematodes are compared and discussed.