Evolutionary affinities and morphological characterization of the enigmatic Zonocotyle bicaecata (Trematoda: Paramphistomoidea: Zonocotylidae) from the Upper Paraná River basin.

Neotropical fish amphistomes represent a highly diverse group within the Paramphistomoidea, with wide distribution across major South American hydrological drainages. However, the limited molecular characterization of these taxa has impeded a comprehensive assessment of their evolutionary relationships and the systematic relevance of morphological features in classification schemes. Our study, based on the critical evaluation of the type material of both nominal species of Zonocotyle (type genus of the monotypic Zonocotylidae), and newly collected specimens of Zonocotyle bicaecata from Steindachnerina insculpta (Curimatidae) in the Upper Paraná River basin, Brazil, presents a morphological reappraisal of Z. bicaecata and provides molecular data (28S rDNA, ITS1-5.8S-ITS2 region, and COI mtDNA) to assess its phylogenetic relationships. Our phylogenetic analyses confirm this species belongs to the Paramphistomoidea. The most comprehensive analyses (based on 28S and COI) further indicate a close relationship with other fish amphistomes from the Neotropical region. Additionally, we emphasized the necessity for a new classification within Paramphistomoidea and briefly discussed the host range of Zonocotyle among curimatid fish hosts.

Characterization and finding the origin of off-flavor compounds in Nile tilapia cultured in net cages in hydroelectric reservoirs, São Paulo State, Brazil.

An increasing demand for fish products has led to an intensive aquaculture production in Brazil, and cultivation of fish constituted 860 × 103 tons in 2022, contributing to the 87% of total fish consumption. Nile tilapia constitutes almost half of the aquaculture production, and most tilapia farms use floating net cages. One of the major constraints of intensive fish production is production of off-flavors. Release of nutrients by the fish leads to deterioration of the water quality and stimulates growth of microorganisms, also including off-flavor producing species. The objective of this study was to determine levels of taste and odor compounds (geosmin, 2-MIB and a selection of volatile compounds) and their impact on the flavor quality of Nile tilapia produced in net cages in reservoirs in São Paulo State, Brazil. GC-MS analysis of fish and water from six different farms showed concentrations of geosmin in the water from 1 to 8 ng/L, while geosmin in fish flesh ranged from 40 to 750 ng/kg. The level of 2-MIB in water was 2 to 25 ng/L, and 0 to 800 ng/kg fish. The GC-MS analysis also revealed presence of more than 100 volatile organic compounds in the fish flesh, consisting of aldehydes, alcohols, benzene derivatives, hydrocarbons, ketones and few other compounds. Geosmin and 2-MIB related flavor notes were detected in all fish by a sensory panel, and a high correlation between the chemical and sensory analyses was found. The potential impact of the volatile organic compounds on the fish flavor is discussed. Analysis of the water quality in the reservoirs indicated that levels of geosmin and 2-MIB levels were highly influenced by the nutrient levels in the water.

Diversity of helminth parasites of eight siluriform fishes from the Aguapeí River, upper Paraná basin, São Paulo state, Brazil.

Fishes of the order Siluriformes, known as catfishes, have a global distribution with more than 3,600 valid species of which 2,087 occur in the Neotropical region. Despite being highly diverse, abundant, and of economic importance as food and ornamental fishes, knowledge about the diversity and distribution of their helminth parasites is fragmentary and scarce. Eight species of catfishes (Pterodoras granulosus, Trachydoras paraguayensis, Pimelodella avanhandavae, Loricariichthys platymetopon, Pterygoplichthys ambrosettii, Rhinelepis aspera, Hemisorubim platyrhynchos, and Sorubim lima) from the Aguapeí River, Upper Paraná River basin, municipality of Castilho, São Paulo State, Brazil, were surveyed for helminth parasites. Collected fishes were weighed, measured, and examined for helminth parasites following standard methodology. Fifty helminth parasite taxa (23 monogeneans, 13 digeneans, 11 nematodes, and three cestodes) were found from a total of 405 fishes screened. The helminth taxon that showed the highest mean intensity of infection and mean abundance was the nematode Rondonia rondoni from P. granulosus, followed by the nematode Parasynodontisia petterae from R. aspera. The ecological analyses were carried out at the component community level and at the infracommunity level. Trachydoras paraguayensis had the richest helminth component community. Pterygoplichthys ambrosettii had the most diverse helminth component community and R. aspera had the lowest. Both hosts are loricariids and have similar diet. However, the high parasite diversity of P. ambrosettii is due to the number of dactylogyrids species found (10), which are directly transmitted, whereas only three dactylogyrid species were found in R. aspera. At infracommunity level, the nematode species R. rondoni and P. petterae dominated the parasite communities. This study presents 38 new host records, contributing considerably to increase the diffuse knowledge of helminth parasites of Neotropical siluriforms.

Microbial Production of the Off-Flavor Geosmin in Tilapia Production in Brazilian Water Reservoirs: Importance of Bacteria in the Intestine and Other Fish-Associated Environments.

AIM: To determine major sources of microbially produced geosmin in the commercially important aquaculture fish species tilapia. METHODS AND RESULTS: Abundance and composition of geosmin-producing bacteria in water and fish biosphere (intestine, digesta, and fins) of Nile tilapia (Oreachromis niloticus) raised in net cages in Brazilian freshwater farms were examined. By combining qPCR of the geosmin synthase geoA gene and 16S rRNA gene amplicon sequencing to identify potential geosmin-producing organisms, we observed that the proportion and composition of geosmin producers appeared to be rather similar in the water, digesta, intestinal mucous, and on skin, making up about 0.1-0.2% of the total bacterial densities. A high proportion of Cyanobacteria and other putative geosmin producers affiliated to the Actinomycetales were identified in the intestinal mucous layer. The main uptake site for geosmin in fish is traditionally assumed to be through the gill surface, but the present results suggest that uptake by the intestinal tract may represent a major source of geosmin uptake in fish. CONCLUSION: The high abundance of geosmin-producing bacteria in the intestinal mucous layer and digesta may indicate that the digestive system in fish is an important, but hitherto overlooked, source of geosmin and likely other off-flavors in fish. SIGNIFICANCE AND IMPACT OF STUDY: Tainting of fish by microbially produced off-flavors spoils fish quality and lowers consumer preferences for aquaculture-produced freshwater fish. Our results highlight the potential for the application of probiotic microorganisms for management of the intestinal microflora to improve the fish quality. HIGHLIGHTS: -Off-flavor producing bacteria are widely abundant in aquaculture.-Off-flavor producers found on skin surface of fish.-Off-flavor producing bacteria accumulate in the digestive system.-Off-flavor producers might release significant amounts of off-flavor during lysis in the gut.-Off-flavor uptake through the digestive system might be quantitatively significant.

A new genus and four new species of dactylogyrids (Monogenea), gill parasites of pimelodid catfishes (Siluriformes: Pimelodidae) in South America and the reassignment of Urocleidoides megorchis Mizelle et Kritsky, 1969.

Nanayella gen. n. is proposed to accommodate four new species of dactylogyrid monogeneans found on the gills of two species of pimelodid catfishes (Siluriformes) from the Amazon River basin in Peru and the Upper Paraná River basin in Brazil. Nanayella gen. n. is mainly characterised by the presence of modified hook pairs 5 and 6, which have a slender shank with slightly enlarged base, inconspicuous thumb, and straight point. Nanayella aculeatrium gen. n. et sp. n. (type species) from Sorubim lima (Bloch et Schneider) (type host) differs from its congeners mainly by possessing seven sclerotised spines on one side of circular ornamentation of the vaginal atrium. Nanayella fluctuatrium gen. n. et sp. n. is characterised by the presence of sclerotised undulations surrounding the circular ornamentation of the vaginal atrium. Nanayella amplofalcis gen. n. et sp. n. is typified by possessing hook pairs 1 and 2 with a robust, wide shank and a round, weakly sclerotised subunit at the base. Nanayella processusclavis gen. n. et sp. n. differs from congeners by the presence of a conspicuous posterior medial process on the dorsal bar. Urocleidoides megorchis Mizelle et Kritsky, 1969 described from the gills of S. lima in Brazil is transferred to Nanayella gen. n. as Nanayella megorchis comb. n. based on morphological and molecular evidence. Analyses of partial sequences of the 28S rRNA gene revealed all five species of the new genus to form a monophyletic clade within a bigger clade comprised of dactylogyrids of South American pimelodids.

A new species of Aphanoblastella Kritsky, Mendoza-Franco and Scholz, 2000 (Monogenea, Dactylogyridae) parasitic on heptapterid catfish (Siluriformes) in the Neotropical region.

Aphanoblastella magna n. sp. is described from the gills of the heptapterid catfish Pimelodella avanhandavae Eigenmann, 1917 in southeastern Brazil (Paraná River Basin). This new species most closely resembles Aphanoblastella chagresii Mendoza-Franco, Aguirre-Macedo and Vidal-Martínez, 2007 described from a congeneric fish host in Panama, but can be distinguished by the shape of the accessory piece and shape and size of the ventral and dorsal bars. From the other species of the genus, A. magna n. sp. differs mainly in the shape of the male copulatory organ (MCO), which is sinuous, versus spirally coiled in other species of the genus, except for A. travassosi (Price, 1938), and by the accessory piece which resembles a 'shoehorn'. Aphanoblastella magna n. sp. is the seventh species of the genus and the first dactylogyrid described from P. avanhandavae. A partial 28S rDNA gene sequence of the new species is also provided; it forms a monophyletic clade with two congeners for which molecular data are available.

A new species of Wallinia Pearse, 1920 (Digenea: Allocreadiidae) collected from Astyanax fasciatus (Cuvier, 1819) and A. lacustris Lucena and Soares, 2016 (Characiformes: Characidae) in Brazil based on morphology and DNA sequences.

Wallinia brasiliensis n. sp. is described from the intestine of two species of tetras-Astyanax fasciatus (Cuvier, 1819) and Astyanax lacustris Lucena and Soares, 2016-collected from the Batalha River in São Paulo State, Brazil. The new species can be clearly distinguished from the other three congeneric species by its vitelline follicles extending from the genital pore to the end of the caeca, eggs lacking operculum, a larger egg size with a consequently lower number relative to the other three species, and the ovary located opposite the anterior testis. The validity of the new species was confirmed through a phylogenetic analysis of the 28S rRNA gene which showed that the new species is the sister taxon to Wallinia mexicana Pérez-Ponce de León, Razo-Mendivil, Mendoza-Garfía, Rubio-Godoy and Choudhury, 2015, a species infecting Astyanax mexicanus (De Filippi, 1853) in Mexico.

A new Synthesium species (Digenea: Brachycladiidae) from the bottlenose dolphin Tursiops truncatus (Cetacea: Delphinidae) in Southwestern Atlantic waters.

A new species of Synthesium from the bottlenose dolphin Tursiops truncatus in South Brazilian waters is described. Morphological and molecular identification was performed, and phylogenetic analyses were carried out using the ribosomal small subunit and internal transcribed spacer 1 and the mitochondrial NDH dehydrogenase subunit 3 and cytochrome c oxidase subunit 1 genes. The main characteristics of the new species are the subterminal round-shaped oral sucker, the anterior distribution of vitellaria reaching the level of the ovary and the oval-shaped testes. The results obtained with the molecular markers supported the inclusion of the specimens into the genus Synthesium. The nucleotide divergence detected for the mitochondrial genes among the new species and others of the same genus supported the erection of a new species. This is the ninth species assigned to the genus and the third Synthesium species recorded in the South Atlantic Ocean.

A new species of Cosmocercoides (Nematoda: Cosmocercidae) from a gymnophthalmid lizard of western Brazil.

A Cosmocercoides sauria n. sp., an intestinal parasite of the gymnophthalmid lizard Iphisa elegans from western Brazil, is described. Of the 18 species previously described, C. sauria represents the 19th species, and is the third to be reported in the Neotropical region. Moreover, C. sauria is the second species reported from a reptilian host and is distinguished by a smaller number of rosette papillae (4 pairs). Additionally, the new species can be distinguished from the other Neotropical species (Cosmocercoides lilloi Ramallo, Bursey and Goldberg, 2007 and C. variabilis (Harwood, 1930)) by possessing smaller spicule size and presence of both gubernaculum and lateral alae in Cosmocercoides lilloi .

First report of Strongyloides sp. (Nematoda, Strongyloididae) in Leopardus tigrinus (Carnivora: Felidae) in the municipality of Botucatu, State of São Paulo, Brazil.

The present study reports the first case of infection by Strongyloides sp. in Leopardus tigrinus in the municipality of Botucatu, State of São Paulo, Brazil. Feces of the infected L. tigrinus specimen were cultivated in sterilized equine feces and a cat (Felis catus domesticus) was experimentally infected with three thousand infective L3 subcutaneous route, in order to identify the Strongyloides species involved in the parasitism. Parthenogenetic females recovered from the experimental animals were analyzed but comparison between the biometric data found and the data in the literature did not enable identification of the species. This is the first report on the occurrence of Strongyloides sp. in L. tigrinus.

A new species of Africana (Nematoda: Heterakidae) from lizards of Southern Amazon, Brazil.

Africana dardanelosi n. sp. is described from the intestine of the lizard Hoplocercus spinosus Fitzinger, 1843 collected from southern Amazon, Brazil. Africana dardanelosi is the third species of the genus occurring in the Neotropical realm and is the tenth species assigned to the genus. From other species of Africana from Africa, and especially from the Neotropical species (A. telfordi Bursey and Goldberg, 2002 and A. chabaudi Baker, 1981), A. dardanelosi is easily distinguished by spicule size (2,157-2,403 microm versus 366-458 microm and 644-869 microm, respectively) and by the number of caudal papillae (13 versus 17 and 10, respectively).

Antitumoural effect of an L-amino acid oxidase isolated from Bothrops jararaca snake venom.

An L-amino acid oxidase (BjarLAAO-I) from Bothrops jararaca snake venom was highly purified using a stepwise sequential chromatography on Sephadex G-75, Benzamidine Sepharose and Phenyl Sepharose. Purified BjarLAAO-I showed a molecular weight around 60,000 under reducing conditions and about 125,000 in the native form, when analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, respectively. BjarLAAO-I is a homodimeric acidic glycoprotein, pI approximately 5.0, and N-terminal sequence showing close structural homology with other snake venom LAAOs. The purified enzyme catalysed the oxidative deamination of L-amino acids, the most specific substrate being L-Phe. Five amino acids, L-Ser, L-Pro, L-Gly, L-Thr and L-Cys were not oxidized, clearly indicating a significant specificity. BjarLAAO-I significantly inhibited Ehrlich ascites tumour growth and induced an influx of polymorphonuclear cells, as well as spontaneous liberation of H(2)O(2) from peritoneal macrophages. Later, BjarLAAO-I induced mononuclear influx and peritoneal macrophage spreading. Animals treated with BjarLAAO-I showed higher survival time.

Cytokine profile of Ehrlich ascites tumor treated with Bothrops jararaca venom.

We previously demonstrated that Bothrops jararaca venom (BjV) has an antitumor effect on Ehrlich ascites tumor (EAT) cells and induces an increase of polymorphonuclear leukocytes in early stages of tumor growth. It has been reported that this venom presents an important inflammatory effect when inoculated in animal models and in human snakebites, and that cytokine levels have been detected in these cases. To evaluate whether the cytokines can be involved with the suppression of the tumoral growth, we evaluate the cytokine profile in the peritoneal cavity of mice inoculated with EAT cells and treated with BjV. Swiss mice were inoculated with EAT cells by the intraperitoneal route and treated with BjV venom (0.4 mg/kg, intraperitoneally), on the 1st, 4th, 7th, 10th, and 13th day. Mice were evaluated for cytokine levels on the 2nd, 5th, 8th, 11th and 14th day. Analysis was performed using an enzyme-linked immunosorbent assay for interleukin (IL)-1alpha, IL-2, IL4, IL-6, IL-10, IL-13, and tumor necrosis factor-alpha (TNF-alpha) levels in the peritoneal washing supernatant. Results were analyzed statistically by the Kruskal-Wallis and Dunn's tests at the 5% level of significance. We observed that EAT implantation induces IL-6 production on the 11th and 14th days of tumor growth, IL-10 on the 11th day and TNF-alpha on the 14th day. The treatment with BjV suppresses production of these cytokines. In addition, IL-13 was produced by animals that were inoculated only with venom on the 11th and 14th days, and by the group inoculated with EAT cells and treated with venom on the 2nd and 14th days. Furthermore, we suggest that the IL-6 detected in the present study is produced by the EAT cells and the suppression of its production could be associated with the antitumor effect of BjV.

Antitumor effect of Bothrops jararaca venom.

Many experimental studies have been carried out using snake venoms for the treatment of animal tumors, with controversial results. While some authors have reported an antitumor effect of treatment with specific snake venom fractions, others have reported no effects after this treatment. The aim of this study was to evaluate the effect of Bothrops jararaca venom (BjV) on Ehrlich ascites tumor (EAT) cells in vivo and in vitro. In the in vivo study, Swiss mice were inoculated with EAT cells by the intraperitoneal (i.p.) route and treated with BjV venom (0.4 mg/kg, i.p.), on the 1st, 4th, 7th, 10th, and 13th days. Mice were evaluated for total and differential cells number on the 2nd, 5th, 8th, 11th and 14th days. The survival time was also evaluated after 60 days of tumor growth. In the in vitro study, EAT and normal peritoneal cells were cultivated in the presence of different BjV concentrations (2.5, 5.0, 10.0, 20.0, 40.0, and 80 microg) and viability was verified after 3, 6, 12 and 24 h of cultivation. Results were analyzed statistically by the Kruskal-Wallis and Tukey tests at the 5% level of significance. It was observed that in vivo treatment with BjV induced tumor growth inhibition, increased animal survival time, decreased mortality, increased the influx of polymorphonuclear leukocytes on the early stages of tumor growth, and did not affect the mononuclear cells number. In vitro treatment with BjV produced a dose-dependent toxic effect on EAT and peritoneal cells, with higher effects against peritoneal cells. Taken together, our results demonstrate that BjV has an important antitumor effect. This is the first report showing this in vivo effect for this venom.