Global distribution patterns and geographic range of Cymothoa Fabricius, 1793 (Isopoda: Cymothoidae) associated with host fish.

Over recent years, fish parasites of the genus Cymothoa Fabricius, 1793, have received increased attention due to both their ecological and their economic importance to aquaculture and fishery. As the studies about Cymothoa have increased this improve our understanding on the host specificity and distribution of these parasites. The aim of this paper was to review the current global geographic distribution, distribution patterns and parasite-host interactions patterns of Cymothoa spp. associated with fish from marine and brackish water bodies around the world. A total of 144 samples were analyzed, from which 23 species of Cymothoa were found parasitizing 84 teleost fish species of 35 families and 20 orders. Most of these parasites were found in the mouth of the host fish, including in wild fish. The highest occurrence of parasites was found in host species belonging to the families Carangidae and Lutjanidae. Host specificity was an important factor in the geographic distribution of Cymothoa species as also environmental temperature. Cymothoa indica, Cymothoa exigua and Cymothoa excisa were the species with lowest specificity for host family and widest geographic distribution.

Global distribution patterns of Caligus Müller, 1785 (Copepoda: Caligidae) associated to teleost fishes, with physiological and histopathological data and description of treatment strategies.

This review surveyed information on Caligus Müller, 1785 to identify global infestation patterns and geographic distribution in teleost fishes, as well as physiological and histopathological data and description of treatment strategies. A total 990 samples of Caligus spp. (N = 212 species) obtained of 233 scientific papers on farmed and wild teleost species from 99 families and 30 orders were used, and the highest number of occurrences was on Carangidae. Caligus spp. was predominantly found in marine environments, and only Caligus lacustris and Caligus epidemicus were found in teleost fish of freshwater environments. There was a high prevalence of Caligus spp. on hosts and infestation occurred predominantly in both the tegument and the gills. Caligus species are distributed across different countries and some particularities were identified and discussed. Caligus elongatus and Caligus bonito bonito had the broadest geographic distribution. Histomorphological and hematological disorders caused by infestation by Caligus spp. were reported and discussed, as well as chemotherapeutic products used for controlling and treating the infestations. Variation in the distribution and geographic patterns of Caligus spp. were little evident in many ecosystems and due to the limited data on the infestation of these sea lice on teleost populations in different regions.

Lepeophtheirus (Copepoda: Caligidae) associated with fish: global infection patterns, parasite-host interactions and geographic range.

Lepeophtheirus Nordmann, 1832 is a genus of sea lice that have been reported to cause parasitic disease problems for fish farming and the fishery industry. This first global investigation on Lepeophtheirus species associated with fish and infestation patterns, parasite-host interactions and geographic ranges linked to these ectoparasites covered articles published from 1940 to 2022. The total of 481 samples of Lepeophtheirus spp. comprised 49 species of these ectoparasites and were found parasitizing 100 teleost fish species from 46 families and 15 orders. Globally, a total of 9 Lepeophtheirus species were found in farmed fish (1 species occurred only in farmed fish and 8 species in both farmed and wild fish) and 48 in wild fish. The highest numbers of occurrences of Lepeophtheirus were in Serranidae and Pleuronectidae. L. pectoralis and L. salmonis were the species with widest geographic distribution. Host specificity was an important factor in the geographic distribution of L. salmonis. Most of the parasite species showed specificity for host fish families, as well as specificity for geographic regions. Little is known about many Lepeophtheirus species compared to the economical important L. salmonis. This could be an obstacle to developing improved management control strategies for the parasite in the fish farming industry, in addition to the diminishing knowledge of parasite taxonomy in many regions.

Morphological and functional structure of two Ergasilidae parasites determine their microhabitat affinity on the gills of an Anostomidae fish from the Amazon.

The aim of this study was to investigate the microhabitat affinities of Ergasilus sp. and Therodamas longicollum on the gills of Leporinus fasciatus ("aracu-piau"), an anostomid fish from the Amazon. A total of 143 specimens of L. fasciatus were examined, of which 35% had their gills parasitized by at least one ergasilid species. A total of 159 specimens of Ergasilus sp. and 97 specimens of T. longicollum were recovered. Both ergasilids species exhibited a greater affinity for attachment in arch 1 of the gills of the host. There was a negative correlation between the abundance of Ergasilus sp. and the abundance of T. longicollum within each gill arch. Analysis of distribution along the gill arch showed a higher occurrence of Ergasilus sp. in Section 5, while T. longicollum occurred mainly in Section 3 of the gills. There was a positive correlation between the abundance of these ectoparasites and the size of the gill arch. Ergasilus sp. occurred only on the filament of the gill arch and exhibited affinity for the proximal region, while T. longicollum occurred only in the gill arch itself. The results reveal that these two parasite species compete in the gills of the host without the overlapping of their niches.

Temporal and seasonal variation of metazoan parasites in Pimelodus ornatus (Siluriformes: Pimelodidae) from the Amazon River, Brazil.

This study investigated the influence of temporal and seasonal (i.e., rainy and dry seasons) in communities and infracommunities of metazoan parasites in Pimelodus ornatus, a Siluriformes from the Amazon River in the state of Amapá, in northern Brazil. A total of 144 fish were collected during 2020 and 2021. In 2020, five species of parasites were found (1 Monogenea, 3 Nematoda, and 1 Cestoda), and in 2021, seven species of parasites were found (1 Monogenea, 3 Nematoda, 1 Cestoda, 1 Digenea, and 1 Crustacea). In 2020, Spirocamallanus inopinatus was dominant, while in 2021, Contracaecum sp. was dominant. Brillouin diversity, species richness, and evenness were higher in 2021. There were differences for some parasite infracommunities between years and between seasonal periods. Brillouin diversity and species richness were higher in the rainy season, but there were no differences in evenness and Berger-Parker dominance between the seasonal periods. Temporal and seasonal variations were mainly influenced by rainfall levels, oxygen levels, and environmental temperature and, later, by the availability of infective stages of parasites in the environment and the size of the hosts. Lastly, the influence of the temporal and seasonal variations on the structure of communities and infracommunities of parasites was weaker than expected due to the low infection rates of the majority of parasites found.

High compliance and effective treatment of fish endoparasitic infections with oral drug delivery nanobioparticles: Safety of intestinal tissue and blood parameters.

Parasite infections in fish require constant surveillance and strategies for efficient treatments which guarantee the fish health, their sale value and the non-propagation of pathogens in new environments. Fish treatments based on nanotechnology become of increasing interest since nanoparticles have been shown as efficient materials for optimizing administration of bioactives. In this study a chitosan derivative, alginate and praziquantel conjugated nanobioparticle of effective action for oral treatment of digenetic trematodes in highly infected Corydoras schwartzi was evaluated in terms of histological and hematological safety. The inherent absence of alterations in intestinal tissue and the reversible blood cells counting during a period up to 35 days showed the safety of the drug delivery nanobioparticles, which thus represent a promising strategy for effective applications in pathogens treatments by oral administration.

Integrative taxonomy of a new species of Therodamas (Ergasilidae) infecting the Amazonian freshwater fish Leporinus fasciatus (Anostomidae).

Crustaceans of the subclass Copepoda are an important component of the invertebrate aquatic fauna. They occur in all aquatic environments and include some representatives that are free-living organisms and others that have a parasitic lifestyle. The genus Therodamas comprises marine and freshwater copepods whose females are parasites of fish in their adult phase, with only seven species described so far. During a field survey of fish parasites in the Jari River, a large tributary of the Amazon River system, in Brazil, we found a new species of the genus Therodamas infecting Leporinus fasciatus. Therodamas longicollum n. sp. is the second strictly freshwater species known. Phylogenetic analysis showed that the new species is grouped in the family Ergasilidae, and divergence estimates showed that T. longicollum n. sp. diverged from its ancestor at around 66.34 Ma, in the late Upper Cretaceous. Therodamas longicollum n. sp. differs from its congeneric in that it does not have lobes and/or expansion of the anterior neck region. Besides describing a new Therodamas species, thereby increasing the diversity of the genus to eight species, this study points out the existence of a lineage of these copepods that has adapted to the freshwater environment of the Amazon. This study also corroborates the genus Therodamas as part of the family Ergasilidae.

Parasites in Leporinus macrocephalus (Anostomidae) of four fish farms from the western Amazon (Brazil).

This study evaluated the presence of metazoan parasites in Leporinus macrocephalus from four fish farms from the western Amazon (Brazil). In 160 fish examined, prevalence was 61.9%, and parasites found were: Urocleidoides paradoxus, Urocleidoides eremitus, Tereancistrum parvus, Jainus leporini, Procamallanus (Spirocamallanus) inopinatus, Rhabdochona (Rhabdochona) acuminata, Dolops discoidalis and Ergasilus sp., but U. paradoxus was the dominant parasite. Jainus leporini and Ergasilus sp. occurred only in L. macrocephalus from one fish farm, while U. paradoxus, U. eremitus and T. parvus were found in fish from three fish farms. Dolops discoidalis, P. (S.) inopinatus and R. (R.) acuminata occurred only in L. macrocephalus from two fish farms. Higher infection levels were caused by U. paradoxus, U. eremitus and P. (S.) inopinatus, which had an aggregated dispersion. There was positive correlation between abundance of parasites and the length of hosts. No difference in the condition factor of parasitized and non-parasitized fish were found. Such differences between fish farms were attributed to differences in management and quality of cultivation environments, and data indicate the need to adopt prophylactic measures in the fish farms to prevent diseases in the future. This was the first report of D. discoidalis and Ergasilus sp. for L. macrocephalus.

Can a biopesticide based on Bacillus thuringiensis affect the physiology and histomorphology of Arapaima gigas?

The use of biopesticides based on Bacillus thuringiensis (Berliner 1915) in agriculture has been considered harmless for non-target organisms such as fish. The present study aimed to investigate the effects of a biopesticide based on B. thuringiensis on the physiology and histology of the liver, kidney and intestine of Arapaima gigas (Schinz, 1822), via exposure to water (0.13 g/L) and in the diet (0.13 g), after 24 and 48 h. Fish subjected to B. thuringiensis in the water of their breeding and feeding tanks exhibited mortality due to changes in erythrogram (hematocrit, hemoglobin, erythrocytes), thrombogram and leucogram blood parameters, and plasma (sodium, chloride, potassium, cholesterol, glucose, triglycerides, cortisol and total proteins) and enzymatic (Aspartate Amino Transferase and Alanine Amino Transferase) biochemistry. Histopathological alterations in the liver and kidney ranged from mild to severe and were characterized by the presence of cytoplasmic vacuolization, nuclear hypertrophy and atrophy, melanomacrophage centers and necrosis, and in the intestine by changes to the number of villi and goblet cells. Therefore, these physiological and histopathological alterations indicate that care should be taken with the dispersion of biopesticides based on agricultural B. thuringiensis in fish farming.

Nanoemulsions with oleoresin of Copaifera reticulata (Leguminosae) improve anthelmintic efficacy in the control of monogenean parasites when compared to oleoresin without nanoformulation.

This study compared the in vitro anthelmintic activity of Copaifera reticulata oleoresin (200, 400, 600, 800 and 1,000 mg/L) and of nanoemulsions prepared with this oleoresin (50, 100, 150, 200 and 250 mg/L) against monogeneans on the gills of Colossoma macropomum. The major compounds present in the oleoresin of C. reticulata were γ-macrocarpene (14.2%), α-bergamotene (13.6%), ß-selinene (13.4%) and ß-caryophyllene (11.7%). All concentrations of the nanoemulsion and the oleoresin without nanoformulation showed anthelmintic efficacy against monogeneans, and higher concentrations led to more rapid parasite mortality. Structural damages to the tegument of the parasites exposed to C. reticulata oleoresin were observed with scanning electron microscopy. At two hours of exposure, fish showed 100% tolerance to all nanoemulsion concentrations used in the in vitro assays, whereas 100% mortality was shown in the fish exposed to the oleoresin without nanoformulation after one hour. The results of this study suggest that nanoemulsions with oleoresin of C. reticulata have advantages in the control and treatment of monogenean infections in C. macropomum when compared to the oleoresin without nanoformulation. In addition, since nanoemulsions with the C. reticulata oleoresin are safe to control monogeneans, the efficacy of these nanoformulations may be assayed in therapeutic baths to treat C. macropomum infected by monogeneans.

Lethal concentration of Cymbopogon citratus (Poaceae) essential oil for Dolops discoidalis and Argulus sp. (Crustacea: Argulidae).

This study investigated the acute toxicity (LC50-24 hr ) effects of the essential oil of Cymbopogon citratus for adult Argulus sp. and Dolops discoidalis, before and during oviposition. In vitro acute toxicity (LC50-24 hr ) was tested using 20, 40, 60, 80, 100, 120, 140 and 160 µg/L of C. citratus essential oil, and two control groups (one with cultivation tank water and one with cultivation tank water + alcohol) were used. Specimens of Argulus sp. and D. discoidalis submitted to acute toxicity were evaluated using histological procedures. The major chemical compounds of C. citratus essential oil were geranial (47.5%), neral (35.6%) and myrcene (6.7%). The LC50-24 hr for Argulus sp. adults was 67.97 µg/L, while for D. discoidalis it was 59.55 µg/L. In the oviposition of both species of argulids, maximum mortality began with treatments of 140 µg/L, while the LC50-24 hr for Argulus sp. and D. discoidalis was 83.98 µg/L and 82.48 µg/L, respectively. In both argulid species exposed to C. citratus essential oil, morphological alterations were observed only in the eyes, and they occurred in the ommatidium and rhabdomeres and were dependent on the concentration of C. citratus essential oil and the parasite species.

Morphometric Characterization of Trypanosoma spp. and blood parameters in Pterygoplichthys pardalis (Pisces: Loricariidae) from the Brazilian Amazon.

The present study describes Trypanosoma spp. infection and blood parameters in Pterygoplichthys pardalis from the Tapajós River basin in eastern Amazon (Brazil). Of 32 fish examined, 40.6% were infected by Trypanosoma spp., while a total of 112 trypomastigotes were found. Two Trypanosoma morphotypes were characterized and compared with species described in literature infecting other Loricariidae, and a similarity of 94% was found with one species described for another host. The plasma glucose and aspartate aminotransferase levels, hematocrit, number of total erythrocytes, mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) in the infected and uninfected fish were similar, but hemoglobin was lower in fish infected with Trypanosoma spp. Hemoglobin levels declined with the abundance of the hemoparasites, but the condition factor was similar among fish infected and uninfected by Trypanosoma spp. This is the first study on the hemoparasitism by Trypanosoma spp. and blood parameters in P. pardalis.

Diet composition of the electric eel Electrophorus voltai (Pisces: Gymnotidae) in the Brazilian Amazon region.

The diet composition of the electric eel Electrophorus voltai was studied in specimens collected from the River Jari, state of Amapá, eastern Amazon region, Brazil. Analysis on their stomach contents revealed that fish, especially Megalechis thoracata, were the most frequent prey item, whereas arthropods and plant material were the least frequent intakes. This is the first stomach content analysis on E. voltai, and it corroborates that electric eel species are piscivorous.

Growth, physiological and immune responses of Arapaima gigas (Arapaimidae) to Aeromonas hydrophila challenge and handling stress following feeding with immunostimulant supplemented diets.

The current study tested the efficacy of a dietary immunostimulant additive (Aquate Fish™®) on the growth performance, and on the physiological and immune responses of Arapaima gigas. Two trials were carried out: a feeding trial for 30 days with the experimental diets and a challenge trial for 7 days, in which fish were bacterial challenge (Aeromonas hydrophila) following by 60 s handling stress. During the feeding trial, fingerlings were fed diets supplemented with 0 (control), 6, 9 and 12 g Aquate Fish™®/kg diet. Dietary supplementation did not influence feed intake, feed conversion and condition factor, but increased the final biomass, number of erythrocytes, thrombocytes, leukocytes, lymphocytes, monocytes, hemoglobin, glucose, globulins and plasma triglycerides in fish fed at a concentration of 12 g/kg diet. After bacterial infection, mortality occurred only in fish fed control treatment, whereas respiratory burst of leukocytes, number of leukocytes and lymphocytes increased in fish that received 12 g of dietary supplementation. The results indicated that dietary supplementation with 12 g of Aquate Fish™® improved biomass and immunity performance of A. gigas fingerlings, without negatively affecting blood biochemical parameters.

Toxicological, hematological and immunological effects of levamisole and ivermectin diet supplementation on Colossoma macropomum (Serrasalmidae).

This study investigated the toxicological, hematological and immunological effects of diets supplemented with levamisole and ivermectin on Colossoma macropomum. Fish were fed for 24, 96 and 240 h with diets containing 300, 600, 900 and 1200 mg of levamisole kg-1 of feed. None of the levamisole concentrations caused either mortality or behavioral alterations among the fish over 10 d of feeding. In comparison, a single feeding of ivermectin at 4500, 9000, 13500 or 18000 mg kg-1 caused 100% mortality among the fish within 10 h. There were no alterations in erythrocyte parameters or albumin in any treatments with levamisole after 24, 96 and 240 h of feeding. At concentrations of 900 and 1200 mg kg-1, levamisole caused decreases in the albumin-to-globulin ratio compared to the control group and the 300 and 600 mg kg-1 diet treatment groups. Levels of glucose and total plasma protein were higher in the fish fed with 600, 900 and 1200 mg kg-1 than in the controls and the 300 mg kg-1 diet treatment group. After 10 d of receiving feed supplemented with levamisole, globulin levels were higher in the 600 mg kg-1 group than in the controls. Respiratory burst activity of leukocytes also increased in the fish supplemented with the 900 and 1200 mg kg-1 diets compared to the controls and other treatments. This is the first study to investigate how diets supplemented with these drugs affect C. macropomum. Our research indicates that all the levamisole concentrations tested can be used in the diet of C. macropomum for antiparasitic treatments against helminth species, and that dietary treatments with levamisole can stimulate components of the innate immune system.

Albendazole, levamisole and ivermectin are effective against monogeneans of Colossoma macropomum (Pisces: Serrasalmidae).

This study evaluated the efficacy of albendazole, ivermectin, levamisole, mebendazole and praziquantel on monogeneans of Colossoma macropomum, based on in vitro and in vivo assays. In vitro assays indicated that albendazole (500, 100, 1,500 and 2,000 mg/L), ivermectin (200, 250, 300 and 350 mg/L) and levamisole (50, 75, 100 and 125 mg/L) were 100% effective against Anacanthorus spatulatus, Notozothecium janauachensis, Mymarothecium boegeri and Linguadactyloides brinkmanni, while mebendazole (125, 150, 175 and 200 mg/L) and praziquantel (5, 10, 15 and 20 mg/L) were ineffective. Fish mortality in 24 hr therapeutic baths with 500 mg/L of albendazole was 6.6%, but the behaviour of the animals remained unchanged, while 200 mg/L of ivermectin caused lethargy, signs of hypoxia and 100% mortality within 2 hr, and 125 mg/L of levamisole caused no mortality. The efficacy of 500 mg/L of albendazole was 48.6% in the 24 hr baths, while that of 125 mg/L levamisole was 88.2%. Although ivermectin showed in vitro efficacy, the lowest concentration used in baths was highly toxic to fish. Therefore, we recommend the use of 125 mg/L of levamisole to control and treat monogenean infestations on C. macropomum in fish farming.

Long-term temporal variation in the parasite community structure of metazoans of Pimelodus blochii (Pimelodidae), a catfish from the Brazilian Amazon.

The Amazon represents one of the most complex biomes in the world; however, the temporal variations in parasite community structure of fishes inhabiting this region remain poorly understood. Therefore, processes generating such variations are still unknown. The present study evaluated the long-term temporal variation of community structure of metazoan parasites of Pimelodus blochii collected in Iaco River, State of Acre (Southwestern Brazilian Amazon). A total of 196 parasites were collected over a 6-year period (2012-2017). Twenty-four different taxa of parasites were found, of which 5 Monogenea, 11 Nematoda, 3 Digenea, 1 Acanthocephala, 1 Cestoda, and 3 Crustacea. The overall species richness ranged from 4 in 2012 to 17 in 2016, in which nematodes (larvae and adults) showed higher numerical dominance, diversity, and species richness. However, the annual species richness was similar between the study years, except in 2016, where it showed a distinctly higher value. The overall parasite diversity was also different in 2012 and 2016, whereas the overall abundance differed in 2013 and 2017. The prevalence and abundance of some infracommunities of parasites varied over time. The temporal changes in the parasite community structure of P. blochii are probably related to variations in host-related features, i.e., body size and shift in diet composition as well as to the occurrence of parasites with distinct life history and biology (mainly monogeneans, digeneans, and nematodes). This is the first evaluation of a long-term temporal variation in the structure of the parasite community in fish from the Amazon.

Haemogregarina daviesensis sp. nov. (Apicomplexa: Haemogregarinidae) from South American lungfish Lepidosiren paradoxa (Sarcopterygii: Lepidosirenidae) in the eastern Amazon region.

Based on morphology and morphometry of gametocytes in blood and molecular phylogenetic analysis, we described a new species of hemoparasite from the genus Haemogregarina isolated from Lepidosiren paradoxa in the eastern Amazon region. Haemogregarina daviesensis sp. nov. is characterized by monomorphic gametocytes of varying maturity stage and their dimensions were 16 ± 0.12 µm (range 13-18) in length and 6 ± 0.97 µm (range 5-8) in width. The morphological and morphometric data were not identical with other haemogregarine species from fish. All specimens of L. paradoxa analyzed were infected by H. daviesensis sp. nov. and the parasitemia level was moderate (1-28/2000 blood erythrocytes). Two sequences were obtained from L. paradoxa, and these constituted a monophyletic sister clade to the Haemogregarina species. In addition, H. daviesensis sp. nov. detected here grouped with Haemogregarina sp. sequences isolated from chelonian Macrochelys temminckii, with 99% bootstrap support. This study provides the first data on the molecular phylogeny of an intraerythrocytic haemogregarine of freshwater fish and highlights the importance of obtaining additional information on aspects of the general biology of these hemoparasites in fish populations, in order to achieve correct taxonomic classification.

Community ecology of parasites in four species of Corydoras (Callichthyidae), ornamental fish endemic to the eastern Amazon (Brazil).

This study compared the parasites community in Corydoras ephippifer, Corydoras melanistius, Corydoras amapaensis and Corydoras spilurus from tributaries from the Amapari River in State of Amapá (Brazil). A total of 151 fish of these four ornamental species were examined, of which 66.2% were parasitized by one or more species, and a total of 732 parasites were collected. Corydoras ephippifer (91.2%) and C. spilurus (98.8%) were the most parasitized hosts, while C. amapaensis (9.6%) was the least parasitized. A high similarity (≅ 75%) of parasite communities was found in the host species. Hosts were parasitized by Procamallanus (Spirocamallanus) inopinatus, Camallanus sp. and metacercariae of digeneans. The parasites had an aggregated dispersion pattern, but in C. ephippifer a random dispersion of P. (S.) inopinatus was found. The parasite community was characterized by a low species richness (1-4 parasites per host), low diversity and low evenness, and consisted mainly of endoparasites with high prevalence and low abundance. An important component in the determination of the helminth parasite community composition was the dominance of species that use these fish species as secondary intermediate or paratenic hosts. This was the first study on the parasite community for these four hosts. A checklist of parasite species in wild populations of Corydoras spp. from different localities was also reported here.

Physiological parameters of Piaractus mesopotamicus (Osteichthyes: Characidae) exposed to a biopesticide based on Bacillus thuringiensis.

This study investigated the physiological effects of a pesticide based on Bacillus thuringiensis (Dipel-WP®) added in the water and diet of Piaractus mesopotamicus during 24 and 48 h. It was added 0.13 g of de B. thuringiensis per kg of commercial feed; and for the fish subjected to the biopesticide in the water of the tanks, it was added 0.13 g/L of the biopesticide. Plasma levels of sodium, chloride, potassium, cholesterol, glucose, triglycerides, cortisol, total protein, alanine aminotransferase (ALT), aspartate aminotransferase (AST), hematocrit, hemoglobin, erythrocytes number, mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC), number of thrombocytes and leukocytes were determined. Cortisol, lactate, glucose, total protein, cholesterol, triglycerides, ALT, AST, sodium, potassium, hematocrit, hemoglobin, MCV, number of erythrocytes, leukocytes, lymphocytes, eosinophils and PAS-positive granular leukocytes suffered alterations derived from the addition of B. thuringiensis in water and diet of the fish. The toxicity of the concentrations of biopesticide in short-term exposure in water and diet of the fish led to blood alterations (increase or decrease). Therefore, care must be taken to avoid a possible prolonged contamination in the tanks of fish farming by agricultural pollution based on B. thuringiensis.