Morphological and molecular data confirm the occurrence of Paramphistomum leydeni (Trematoda: Paramphistomidae) in ruminants from Southern Brazil.

Species belonging to the family Paramphistomidae Fischoeder, 1901, commonly known as "rumen flukes", are a group of parasites frequently related to Brazilian livestock production. They inhabit the digestive tract of ruminants and have recognized pathogenicity during the early stages of infection, which can be responsible for economic losses. These trematodes are often associated with Southern Brazil, a region heavily focused on animal farming, which also makes it ideal for the life cycle of paramphistomes. Despite their aforementioned importance, studies regarding their distribution, molecular taxonomy and biology are still scarce in the country. In the present study, rumen flukes collected from cattle (n = 22) and sheep (n = 3) from 9 batches of ruminants from the cities of Jaguarão, Pelotas and Rio Grande, state of Rio Grande do Sul, Brazil, between May and July 2022, were subjected to morphological and molecular study. The microscopic analysis of histological and manual cuts revealed diagnostical traits compatible with Paramphistomum leydeni Näsmark, 1937, including the presence of tegumental papillae, pharynx of the liorchis type and acetabulum of the leydeni type. Molecular data corroborated the morphological identification, with ITS-2 and cox-1 sequences here obtained presenting 100% and 96.8-99.8% similarity, respectively, to P. leydeni samples previously characterized in different countries from Asia, Europe, and South America. Intensity of infection ranged from 5 to 458 and 1 to3 specimens of P. leydeni in sampled cattle and sheep, respectively. The present study contributes to a better understanding of the taxonomy of the flukes involved in cattle and sheep paramphistomosis in Brazil, suggesting that P. leydeni could be the main paramphistome species found in ruminants in the studied region.

Evaluation of the ovicidal activity and fasciolicidal activity of the extract of ethyl acetate from Artemisia ludoviciana Nutt. spp. mexicana and of artemisinin against adult parasites of Fasciola hepatica.

The objective of this work was to evaluate the effect of the ethyl acetate extract from A. ludoviciana (EALM) and artemisinin against adult parasites and eggs of F. hepatica. For the ovicidal assay, cell culture plates with 24 wells were used, and 90 to 110 F. hepatica eggs were placed in each well. The eggs were exposed to concentrations of 100, 200, 300, 400, and 500 mg/L EALM and incubated for 16 days. Additionally, triclabendazole (TCBZ) was used as a reference drug at concentrations of 10 and 50 mg, and the response of artemisinin at concentrations of 10 and 20 mg was simultaneously assessed. Adult flukes were exposed to concentrations of 125, 250, 375, and 500 mg/L EALM. The results of the ovicidal action of EALM on the eggs showed that concentrations greater than 300 mg/L were significant, with ovicidal percentages greater than 60% observed on day 16 of incubation (p < 0.05). The maximum efficiency of EALM on adult flukes was reached 72 h post-exposure at a concentration of 125 mg/L (p < 0.05).

A Molecular Phylogenetic Study of the Eye Fluke Philophthalmus lacrymosus (Trematoda: Philophthalmidae) Found in Larus dominicanus (Aves: Laridae) from Brazil.

PURPOSE: Species of the genus Philophthalmus are eye flukes with a complex taxonomy, which began to be improved with the help of molecular data only recently. However, most described species have never been placed into a phylogenetic context. In this study, eye flukes previously found on kelp gulls, Larus dominicanus, from Brazil and identified as Philophthalmus lacrymosus were subjected to molecular analysis. METHODS: For the molecular analyses, we analyzed parasites found in six infected gulls (one worm per bird) collected from different municipalities of the state of Santa Catarina, Brazil. We carried out the amplification and sequencing of the partial region of the 28S and cox1 genes and the data obtained were compared with sequences available to philophthalmid species and subjected to phylogenetic analysis. RESULTS: The isolates of P. lacrymosus from Brazil grouped in well-supported clades with five other species of Philophthalmus with sequences available for comparison. Interspecific divergences of 0.1-1.6% in 28S and 8.2-14.9% in cox1 were found in relation to other isolates of Philophthalmus spp. Two cox1 haplotypes differing in one nucleotide (0.1%) were found between the six eye flukes isolates in gulls from different localities. The Brazilian isolates grouped in a subclade with parasites identified as P. lacrymosus in Portugal; however, the molecular divergences found in cox1 (8.2-8.5%) strongly suggest that these isolates belong to different species. The phylogenetic trees obtained and the intergeneric divergences to species of the genera Cloacitrema and Parorchis did not support the validity of the genus Natterophthalmus, for which P. lacrymosus was proposed as the type species in the past. CONCLUSION: As P. lacrymosus was described from Brazil, we recommend that this name be applied to the South American isolates and that the Portuguese isolates be provisionally considered as Philophthalmus sp., a probable cryptic species. Moreover, data obtained supports the previous morphology-based synonymizing between Natterophthalmus and Philophthalmus. Considering our results and most of previous reports of P. lacrymosus in South America, we suggest this species presents a marine life cycle.

Codon usage in the flatworm Schistosoma mansoni is shaped by the mutational bias towards A+T and translational selection, which increases GC-ending codons in highly expressed genes.

Schistosoma mansoni is a trematode flatworm that parasitizes humans and produces a disease called bilharzia. At the genomic level, it is characterized by a low genomic GC content and an "isochore-like" structure, where GC-richest regions, mainly placed at the extremes of the chromosomes, are interspersed with low GC-regions. Furthermore, the GC-richest regions are at the same time the gene-richest, and where the most heavily expressed genes are placed. Taking these features into account, we decided to reanalyze the codon usage of this flatworm. Our results show that a) when all genes are considered together, the strong mutational bias towards A + T leads to a predominance of A/T-ending codons, b) a multivariate analysis discriminates between highly and lowly expressed genes, c) the sequences expressed at highest levels display a significant increase in G/C-ending codons, d) when comparing the molecular distances with a closely related species the synonymous distance in highly expressed genes is significantly lower than in lowly expressed sequences. Therefore, we conclude that despite previous results, which were performed with a small sample of genes, codon usage in S. mansoni is the result of two forces that operate in opposite directions: while mutational bias leads to a predominance of A/T codons, translational selection, working at the level of speed, increment G/C ending triplets.

Morphological and molecular identification with frequency analysis of Calicophoron microbothrioides in central Chile / Identificação morfológica e molecular com análise de frequência de Calicophoron microbothrioides na região central do Chile

Abstract This research aimed to determine the presence of paramphistomids in cattle slaughtered in a slaughterhouse of the Ñuble Region of Chile, to identify flukes and to analyze the frequency of these parasites in the Maule, Ñuble, and Biobío administrative regions of Chile. Between October of 2016 and April of 2017, rumens of 494 cattle were examined for flukes in the forestomachs. Worms were identified morphologically and, in addition, molecular analysis of the internal transcriber spacer region 2 of the fluke's DNA was done and phylogenetic analyses were performed with Bayesian inference in 14 worms. The frequency was analyzed by locality (low- or highlands) and age. The overall frequency was 11.24%. The district with the highest frequency of presentation was Chillán Viejo (30.8%). Districts in the lowlands had similar frequencies to those in the mountain lands (p=0.1). The frequency of flukes was significantly higher in adult animals than in young ones (p<0.01). We obtained a 460 bp-length fragment of DNA that was identical to the sequences previously identified as Paramphistomum cervi and Calicophoron microbothrioides, and performed morphological analyses confirmed that our samples belonged to C. microbothrioides. This is the first published study of C. microbothrioides in Chile.

Resumo Este trabalho teve como objetivo determinar a presença de paramphistomídeos em bovinos abatidos em um matadouro da Região do Ñuble do Chile, para identificar parasitas e analisar a frequência desses parasitos nas regiões administrativas de Maule, Ñuble e Biobío, no Chile. Entre outubro de 2016 e abril de 2017, rúmens de 494 bovinos foram examinados à procura de vermes no pré-estômago. Os vermes foram identificados morfologicamente e, além disso, a análise molecular da região interna do espaçador do transcritor 2 do DNA e análises filogenéticas foram realizadas com inferência bayesiana em 14 vermes. A frequência foi analisada pela altitude da localidade (baixa ou alta) e idade. A frequência geral foi de 11,24%. O distrito com as maiores frequências de parasitismo foi Chillán Viejo (30,8%). Os distritos das terras baixas tinham frequências semelhantes às encontradas nas terras das montanhas (p=0,17). A frequência foi significativamente maior em animais adultos do que em jovens (p<0.01). Obtivemos um fragmento de DNA de 460 pb que era idêntico às sequências anteriores identificadas como Paramphistomum cervi e Calicophoron microbothrioides, e realizamos análises morfológicas que permitiram confirmar que nossas amostras pertenciam a C. microbothrioides. Este é o primeiro estudo publicado sobre C. microbothrioides no Chile.

Morphological and molecular identification with frequency analysis of Calicophoron microbothrioides in central Chile / Identificação morfológica e molecular com análise de frequência de Calicophoron microbothrioides na região central do Chile

This research aimed to determine the presence of paramphistomids in cattle slaughtered in a slaughterhouse of the Ñuble Region of Chile, to identify flukes and to analyze the frequency of these parasites in the Maule, Ñuble, and Biobío administrative regions of Chile. Between October of 2016 and April of 2017, rumens of 494 cattle were examined for flukes in the forestomachs. Worms were identified morphologically and, in addition, molecular analysis of the internal transcriber spacer region 2 of the flukes DNA was done and phylogenetic analyses were performed with Bayesian inference in 14 worms. The frequency was analyzed by locality (low- or highlands) and age. The overall frequency was 11.24%. The district with the highest frequency of presentation was Chillán Viejo (30.8%). Districts in the lowlands had similar frequencies to those in the mountain lands (p=0.1). The frequency of flukes was significantly higher in adult animals than in young ones (p 0.01). We obtained a 460 bp-length fragment of DNA that was identical to the sequences previously identified as Paramphistomum cervi and Calicophoron microbothrioides, and performed morphological analyses confirmed that our samples belonged to C. microbothrioides. This is the first published study of C. microbothrioides in Chile.(AU)

Este trabalho teve como objetivo determinar a presença de paramphistomídeos em bovinos abatidos em um matadouro da Região do Ñuble do Chile, para identificar parasitas e analisar a frequência desses parasitos nas regiões administrativas de Maule, Ñuble e Biobío, no Chile. Entre outubro de 2016 e abril de 2017, rúmens de 494 bovinos foram examinados à procura de vermes no pré-estômago. Os vermes foram identificados morfologicamente e, além disso, a análise molecular da região interna do espaçador do transcritor 2 do DNA e análises filogenéticas foram realizadas com inferência bayesiana em 14 vermes. A frequência foi analisada pela altitude da localidade (baixa ou alta) e idade. A frequência geral foi de 11,24%. O distrito com as maiores frequências de parasitismo foi Chillán Viejo (30,8%). Os distritos das terras baixas tinham frequências semelhantes às encontradas nas terras das montanhas (p=0,17). A frequência foi significativamente maior em animais adultos do que em jovens (p 0.01). Obtivemos um fragmento de DNA de 460 pb que era idêntico às sequências anteriores identificadas como Paramphistomum cervi e Calicophoron microbothrioides, e realizamos análises morfológicas que permitiram confirmar que nossas amostras pertenciam a C. microbothrioides. Este é o primeiro estudo publicado sobre C. microbothrioides no Chile.(AU)

Two new species of Elopicola (Digenea: Aporocotylidae) from Hawaiian ladyfish, Elops hawaiensis (Eastern Sea) and Atlantic tarpon, Megalops atlanticus (Gulf of Mexico) with a comment on monophyly of elopomorph blood flukes.

Elopicola bristowi sp. n. infects the blood vascular system of Hawaiian ladyfish, Elops hawaiensis, in the Eastern Sea. It differs from the only nominal congener Elopicola nolancribbi by the combination of having rows of ventrolateral tegumental spines, a proportionally long oesophagus, anterior caeca, vasa efferentia coalescing ventral to the posterodextral margin of the testis, a post-testicular metraterm, a dextral common genital pore lateral to the oötype, and genitalia that are enantiomorphic relative to those of E. nolancribbi. Elopicola franksi sp. n. infects the heart and blood vascular system of Atlantic tarpon, Megalops atlanticus, in the Gulf of Mexico. It differs from its congeners by the combination of lacking ventrolateral tegumental spines and having an elongate body (6× longer than wide), a proportionally long oesophagus, a compact testis at level of the distal ends of the posterior caeca, and a post-testicular common genital pore at level of the oötype. Phylogenetic analyses based on the small subunit ribosomal DNA (18S), large subunit ribosomal DNA (28S), and internal transcribed spacer 2 (ITS2) genes revealed considerable genetic differences between these taxa. The 18S+28S tree showed a monophyletic Elopicola sister to all aporocotylids infecting fishes of Euteleosteomorpha. The ITS2 tree showed Paracardicoloides yamagutii as the sister taxon to Elopicola spp.

Novas ocorrências de metacercária de Austrodiplostomum compactum (Lutz, 1928) (Platyhelminthes: Digenea) parasito de olhos de peixes da Bacia do Rio Paraná / New occurrences of metacercariae of Austrodiplostomum compactum (Lutz, 1928) (Platyhelminthes: Digenea) eye flukes of fish from the Paraná Basin

Austrodiplostomum compactum (Platyhelminthes: Digenea) parasita os olhos de várias de espécies de peixes. A presença deste parasito, em casos extremos, pode causar exoftalmia, deslocamento da retina, opacidade do cristalino e cegueira ou até a morte. O presente estudo registra novas ocorrências desta metacercária infectando os olhos de quatro novos hospedeiros de peixes, Serrasalmus maculatus coletado no reservatório de Rosana no rio Paranapanema e Hypostomus regani, Schizodon borellii e Auchenipterus osteomystax coletados na planície de inundação do alto rio Paraná.

Austrodiplostomum compactum (Platyhelminthes, Digenea) eye flukes of several species of fishes. The presence of this parasite, in extreme cases, can cause swelling of the eyelids, displacement of the retina, opacity of the crystalline lens and blindness or even death. The present study it registers new occurrences of this metacercariae infecting the eyes of four new hosts of fish, Serrasalmus maculatus collected in the Rosana reservoir in the Paranapanema river and Hypostomus regani, Schizodon borellii and Auchenipterus osteomystax collected in the the Upper Paraná River floodplain.

Novas ocorrências de metacercária de Austrodiplostomum compactum (Lutz, 1928) (Platyhelminthes: Digenea) parasito de olhos de peixes da Bacia do Rio Paraná

Austrodiplostomum compactum (Platyhelminthes, Digenea) eye flukes of several species of fishes. The presence of this parasite, in extreme cases, can cause swelling of the eyelids, displacement of the retina, opacity of the crystalline lens and blindness or even death. The present study it registers new occurrences of this metacercariae infecting the eyes of four new hosts of fish, Serrasalmus maculatus collected in the Rosana reservoir in the Paranapanema river and Hypostomus regani, Schizodon borellii and Auchenipterus osteomystax collected in the the Upper Paraná River floodplain.

Austrodiplostomum compactum (Platyhelminthes: Digenea) parasita os olhos de várias de espécies de peixes. A presença deste parasito, em casos extremos, pode causar exoftalmia, deslocamento da retina, opacidade do cristalino e cegueira ou até a morte. O presente estudo registra novas ocorrências desta metacercária infectando os olhos de quatro novos hospedeiros de peixes, Serrasalmus maculatus coletado no reservatório de Rosana no rio Paranapanema e Hypostomus regani, Schizodon borellii e Auchenipterus osteomystax coletados na planície de inundação do alto rio Paraná.