Ornamental fish mortality reveals an old parasite introduction: A case study of Koi carp and fish louse.

Koi carp are globally known for their colors and cultural significance. The introduction of these fish to new environments poses a threat to local biodiversity, in addition to releasing parasites, such as argulid ectoparasites. This study presents a record of Argulus japonicus infecting carp in an artificial lake in Southern Brazil using morphological and molecular methods, with a 100% prevalence (n = 3) and a mean intensity of 21.6 parasites per host, distributed over the body surface. The invasion history of hosts in the study locality indicates that the introduction of A. japonicus occurred decades before its first formal record in Brazil.

Genetically engineered eucalyptus expressing pesticidal proteins from Bacillus thuringiensis for insect resistance: a risk assessment evaluation perspective.

Eucalyptus covers approximately 7.5 million hectares in Brazil and serves as the primary woody species cultivated for commercial purposes. However, native insects and invasive pests pose a significant threat to eucalyptus trees, resulting in substantial economic losses and reduced forest productivity. One of the primary lepidopteran pests affecting eucalyptus is Thyrinteina arnobia (Stoll, 1782) (Lepidoptera: Geometridae), commonly referred to as the brown looper caterpillar. To address this issue, FuturaGene, the biotech division of Suzano S.A., has developed an insect-resistant (IR) eucalyptus variety, which expresses Cry pesticidal proteins (Cry1Ab, Cry1Bb, and Cry2Aa), derived from Bacillus thuringiensis (Bt). Following extensive safety assessments, including field trials across various biomes in Brazil, the Brazilian National Technical Commission of Biosafety (CTNBio) recently approved the commercialization of IR eucalyptus. The biosafety assessments involved the analysis of molecular genomics, digestibility, thermostability, non-target organism exposure, degradability in the field, and effects on soil microbial communities and arthropod communities. In addition, in silico studies were conducted to evaluate allergenicity and toxicity. Results from both laboratory and field studies indicated that Bt eucalyptus is as safe as the conventional eucalyptus clone for humans, animals, and the environment, ensuring the secure use of this insect-resistant trait in wood production.

Five-years post commercial approval monitoring of eucalyptus H421.

Eucalyptus comprises the largest planted area of cultivated production forest in Brazil. Genetic modification of eucalyptus can provide additional characteristics for increasing productivity, protecting plant yield, and potentially altering fiber for various industrial uses. With this objective, a transgenic eucalyptus variety, event H421, received regulatory approval for commercial release after 6 years of approved risk assessment studies by the Brazilian National Technical Biosafety Commission (CTNBio) in 2015, becoming the first approved genetically modified (GM) eucalyptus in the world. GM event H421 enables increased plant biomass accumulation through overexpression of the Arabidopsis 1,4-ß-endoglucanase Cel1, which remodels the xyloglucan-cellulose matrix of the cell wall during development to promote cell expansion and growth. As required, in that time, by the current normative from CTNBio, a post-commercial release monitoring plan for H421 was submitted, incorporating general surveillance for five consecutive years with the submission of annual reports. The monitoring plan was conducted on fields of H421 progenies, with conventional clones as comparators, cultivated in representative regions where eucalyptus is cultivated in the states of São Paulo, Bahia, and Maranhão, representing Southeast, Northeast, and Northern Brazil. Over the course of the five-year general surveillance monitoring plan for the approved GM eucalyptus H421, no adverse effect that could impact the biosafety of the commercially approved event was identified. Additionally, the GM eucalyptus exhibited behavior highly consistent with that of conventional commercial clones. Therefore, there was no need for an extra risk assessment study of a case-specific monitoring plan. The results show the importance of continuously updating the regulation norms of governmental agencies to align with scientific advances.

Safety Assessment of the CP4 EPSPS and NPTII Proteins in Eucalyptus.

Glyphosate herbicide treatment is essential to sustainable Eucalyptus plantation management in Brazil. Eucalyptus is highly sensitive to glyphosate, and Suzano/FuturaGene has genetically modified eucalyptus to tolerate glyphosate, with the aim of both protecting eucalyptus trees from glyphosate application damage and improving weed management. This study presents the biosafety results of the glyphosate-tolerant eucalyptus event 751K032, which expresses the selection marker neomycin phosphotransferase II (NPTII) enzyme and CP4-EPSPS, a glyphosate-tolerant variant of plant 5-enolpyruvyl-shikimate-3-phosphate synthase enzyme. The transgenic genetically modified (GM) event 751K032 behaved in the plantations like conventional non-transgenic eucalyptus clone, FGN-K, and had no effects on arthropods and soil microorganisms. The engineered NPTII and CP4 EPSPS proteins were heat-labile, readily digestible, and according to the bioinformatics analyses, unlikely to cause an allergenic or toxic reaction in humans or animals. This assessment of the biosafety of the glyphosate-tolerant eucalyptus event 751K032 concludes that it is safe to be used for wood production.

Identification and molecular characterization of digenean trematode parasites of Aylacostoma chloroticum (Gastropoda: Thiaridae) from a Neotropical Basin.

Thiarids are pantropical gastropods distributed from South America to Australia and found in lentic and lotic freshwater environments. These gastropods act as intermediate hosts for several species of digeneans. There are difficulties in the correct identification of these digeneans in certain stages of the life cycle as the larvae have a reduced size and few morphological characteristics for diagnosis. Because of this, techniques based on molecular markers have become an extremely useful tool. Our objective was to molecularly characterize the digeneans found in gastropods from the Ivaí River, Paraná, Brazil, using the mitochondrial molecular marker cytochrome c oxidase (COI) and the 28S ribosomal marker. With the molecular marker COI, gastropods showed high genetic similarity with Aylacostoma chloroticum. For the parasites, four different taxa were characterized with both markers. The 28S ribosomal marker proved to be more effective for the identification of Ivaí River parasites, being possible to suggest the identification of two species among the four groups found. Considering that there are no records in the literature of studies involving parasites and gastropods as intermediate hosts in the Ivaí River, this study is the first regarding these relationships.

The first record of Centrocestus formosanus (Trematoda: Heterophyidae) in southern Brazil supported by molecular data.

Centrocestus formosanus is a digenean parasite first described from Asia, which parasitizes Melanoides tuberculata (Gastropoda: Thiaridae) at the first intermediate stage, and different fish species as second intermediate host. C. formosanus was previously recorded in Brazilian states, but never before in the southern region of this country. Recording and identification of digenean species through morphological identification is a taxonomic challenge. In light of this, we use an integrative taxonomic approach to report the occurrence of cercariae and metacercariae of C. formosanus in molluscs and fish, respectively, in an urban park located in southern Brazil. Specimens of M. tuberculata and the fishes Poecilia reticulata and Xiphophorus sp. (Poeciliidae) were collected for screening for parasites and molecular analyses using partial fragments of the 28S rDNA gene. The identification of C. formosanus obtained from molluscs and fish specimens permitted us to partially solve the life cycle of this parasite for the first time in the studied environment, demonstrating the necessity of monitoring and controlling molluscs populations. Nevertheless, our results will support future studies aiming to elucidate the life-cycle of C. formosanus in this region, since all sampled hosts' species are invasive in this environment.

Molecular evidence of new freshwater turtle blood flukes (Digenea: Spirorchiidae) in the intermediate snail host Biomphalaria occidentalis Paraense, 1981 in an urban aquatic ecosystem in Brazil.

In this study, two potentially new species of turtle blood flukes (TBFs) (Digenea: Spirorchiidae) have been recorded from South Brazil. The spirorchiid parasites infect the vascular system of turtles, thereby compromising their health. The life cycle of these parasites is not well studied. The larval stage of cercaria is found in intermediate gastropod hosts, with some species presenting similar morphological characteristics, which can result in misinterpretations when using only morphological taxonomy for species identification. In this study, we recorded a single morphotype belonging to the family Spirorchiidae in Biomphalaria occidentalis in an urban aquatic ecosystem in Brazil. However, molecular data (28S rDNA and cytochrome oxidase subunit I) confirmed the presence of two species of Spirorchiidae in the sampled environment; both phylogenetically close to genera previously studied in freshwater turtles from the Peruvian Amazon. In this study, species characterization was possible because of molecular tools. We recommend using more than one molecular marker in future studies focusing on TBFs, which need attention about their evolutionary history and ecology to understand their distribution in South America.

Cysts and larvae of Strongyluris: A parasite of amphibians and reptiles found in an urban population of Lissachatina fulica (Férussac, 1821) in southern Brazil.

We report the finding of cysts and larvae of Strongyluris in specimens of L. fulica from an urban area of the municipality of Maringá in northern Paraná State, southern Brazil. Thirty-seven young adult snails were collected at three sites: 15 in riparian forest; 14 in a vegetable garden; and eight in a residential garden. We found a total of 16 cysts with nematode larvae in three of the 15 snails collected in riparian forest. The parasites were identified as larvae of the genus Strongyluris, which are parasites of the gastrointestinal tract of amphibians and reptiles. Lissachatina fulica is established in urban areas of 33% of the municipalities of the state of Paraná. The species has spread rapidly through the urban area of the municipality of Maringá, which may contribute to the transmission of nematode larvae of medical and veterinary interest to humans and other animals.

Integrated Stress Responses to Bacterial Pathogenesis Patterns.

Activation of an appropriate innate immune response to bacterial infection is critical to limit microbial spread and generate cytokines and chemokines to instruct appropriate adaptive immune responses. Recognition of bacteria or bacterial products by pattern recognition molecules is crucial to initiate this response. However, it is increasingly clear that the context in which this recognition occurs can dictate the quality of the response and determine the outcome of an infection. The cross talk established between host and pathogen results in profound alterations on cellular homeostasis triggering specific cellular stress responses. In particular, the highly conserved integrated stress response (ISR) has been shown to shape the host response to bacterial pathogens by sensing cellular insults resulting from infection and modulating transcription of key genes, translation of new proteins and cell autonomous antimicrobial mechanisms such as autophagy. Here, we review the growing body of evidence demonstrating a role for the ISR as an integral part of the innate immune response to bacterial pathogens.

Topological congruence between phylogenies of Anacanthorus spp. (Monogenea: Dactylogyridae) and their Characiformes (Actinopterygii) hosts: A case of host-parasite cospeciation.

Cophylogenetic studies aim at testing specific hypotheses to understand the nature of coevolving associations between sets of organisms, such as host and parasites. Monogeneans and their hosts provide and interesting platform for these studies due to their high host specificity. In this context, the objective of the present study was to establish whether the relationship between Anacanthorus spp. with their hosts from the upper Paraná River and its tributaries can be explained by means of cospeciation processes. Nine fish species and 14 monogenean species, most of them host specific, were studied. Partial DNA sequences of the genes RAG1, 16S and COI of the fish hosts and of the genes ITS2, COI and 5.8S of the parasite species were used for phylogenetic reconstruction. Maximum likelihood phylogenetic trees of the host and parasite species were built and used for analyses of topological congruence with PACo and ParaFit. The program Jane was used to estimate the nature of cospeciation events. The comparison of the two phylogenies revealed high topological congruence between them. Both PACo and ParaFit supported the hypothesis of global cospeciation. Results from Jane pointed to duplications as the most frequent coevolutionary event, followed by cospeciation, whereas duplications followed by host-switching were the least common event in Anacanthorus spp. studied. Host-sharing (spreading) was also identified but only between congeneric host species.

Molecular markers in studies on fish parasites (Platyhelminthes): Review / Marcadores moleculares aplicados a estudos de Platyhelminthes parasitos de peixes: revisão

ABSTRACT. Studies with molecular markers are currently more common for all groups of living organisms. Molecular techniques used in Platyhelminthes parasites of fishes do not merely reveal complex life cycles, but are important for species distinction and the elucidation of the phylogenetic hypothesis. Current research verified which molecular markers were mainly used phylogenetic studies on Platyhelminthes parasites of fish so that subsidies for further phylogenetic studies in Icthyoparasitology could be provided. Data base of CAPES Journals platform was employed for bibliometric analysis comprising the keywords "fish" and "phylogeny" associated with "Cestoda", "Digenea" or "Monogenea". Information retrieved was quantified and tabulated. Most studies were on Monogenea (43%), followed by Digenea (37%) and Cestoda (18%). Ribosomal molecular markers were the most used in the phylogenetic studies for fish parasites. Due to the advance of molecular biology techniques and of bioinformatics, with more robust phylogenetic analysis, the use of these techniques in other areas such as Ichytioparasitology is on the increase. In fact, molecular phylogenetics and morphological structures analysis have efficiently contributed towards the understanding of phylogenetic relationships among the groups.

Estudos com marcadores moleculares são cada vez mais comuns em todos os grupos de seres vivos. Para os platelmintes parasitos de peixes, as técnicas moleculares possibilitam desvendar ciclos de vida complexos, sendo importantes também na distinção de espécies e na elucidação de hipóteses filogenéticas. Neste sentido, este trabalho teve como objetivo verificar quais são os principais marcadores moleculares utilizados nos estudos de platelmintos parasitos de peixes, visando fornecer subsídios para futuros estudos filogenéticos na Ictioparasitologia. Para a análise bibliométrica foi utilizado o banco de dados dos Periódicos da CAPES, tendo como palavras-chave "fish" e "phylogeny" associadas a "Cestoda", "Digenea" e "Monogenea". As informações obtidas nos trabalhos foram tabuladas e quantificadas. Dos 143 trabalhos obtidos 43% foram com monogenéticos, 37% com digenéticos e 18% com cestoides. Os marcadores moleculares ribossomais foram os mais utilizados nos estudos filogenéticos com estes parasitos de peixes. Com o avanço das técnicas de biologia molecular e da bioinformática, com análises filogenéticas mais robustas, é crescente a utilização destas técnicas na Ictioparasitologia. A filogenética molecular, juntamente com a análise de estruturas morfológicas tem contribuído de maneira mais eficiente para o entendimento das relações de parentesco, entre estes grupos de parasitos de peixes.

Parasitism by larval tapeworms genus Spirometra in South American amphibians and reptiles: new records from Brazil and Uruguay, and a review of current knowledge in the region.

Spargana are plerocercoid larvae of cestode tapeworms of the genus Spirometra, Family Diphyllobothriidae, parasitic to frogs, reptiles, birds and mammals. This parasitic disease in humans can be transmitted through the use and consumption of amphibians and reptiles. The available knowledge about Spirometra in South America is scarce, and there are only a few reports on the occurrence of sparganum in amphibians and reptiles, many of them published in old papers not easily available to researchers. In this work we present a review on this topic, provide new records in two species of amphibians and 7 species of reptiles from Brazil and Uruguay respectively. We also summarize current knowledge of Spirometra in the continent, along with an updated of host taxonomy. We could gather from the literature a total of 15 studies about amphibian and reptile hosts, published between 1850 and 2016, corresponding to 43 case reports, mostly from Brazil (29) and Uruguay (8), Argentina (3), Peru (2), and Venezuela (1); the majority of them related to reptiles (five lizards and 26 snake species), and 14 corresponded to amphibians (9 anurans). Plerocercoid larvae were located in different organs of the hosts, such as subcutaneous tissue, coelomic cavity, peritoneum, and musculature. The importance of amphibians and reptiles in the transmission of the disease to humans in South America is discussed. Relevant issues to be studied in the near future are the taxonomic characterization of Spirometra in the region and the biological risk of reptile meat for aboriginal and other rural communities.

Rust disease of eucalypts, caused by Puccinia psidii, did not originate via host jump from guava in Brazil.

The rust fungus, Puccinia psidii, is a devastating pathogen of introduced eucalypts (Eucalyptus spp.) in Brazil where it was first observed in 1912. This pathogen is hypothesized to be endemic to South and Central America and to have first infected eucalypts via a host jump from native guava (Psidium guajava). Ten microsatellite markers were used to genotype 148 P. psidii samples from eucalypts and guava plus five additional myrtaceous hosts across a wide geographic range of south-eastern Brazil and Uruguay. Principal coordinates analysis, a Bayesian clustering analysis and a minimum-spanning network revealed two major genetic clusters among the sampled isolates, one associated with guava and another associated with eucalypts and three additional hosts. Multilocus genotypes infecting guava differed by multiple mutational steps at eight loci compared with those infecting eucalypts. Approximate Bayesian computation revealed that evolutionary scenarios involving a coalescence event between guava- and eucalypt-associated pathogen populations within the past 1000 years are highly unlikely. None of the analyses supported the hypothesis that eucalypt-infecting P. psidii in Brazil originated via host jump from guava following the introduction of eucalypts to Brazil approximately 185 years ago. The existence of host-associated biotypes of P. psidii in Brazil indicates that this diversity must be considered when assessing the invasive threat posed by this pathogen to myrtaceous hosts worldwide.

Glyphosate reduces urediniospore development and Puccinia psidii disease severity on Eucalyptus grandis.

BACKGROUND: Recent studies have shown the effects of glyphosate drift on decreasing rust intensity on Eucalyptus grandis plants. However, the effects of the herbicide on Puccinia psidii initial development are unknown. In this study the systemic action of glyphosate on rust severity was evaluated on Eucalyptus plants maintained under greenhouse conditions. Urediniospore germination and apressorium formation on detached leaves and on water agar medium, previously treated with glyphosate, were also evaluated. RESULTS: Rust severity and the number of urediniospores per leaf area were significantly reduced with increasing glyphosate doses, even on branches not directly treated with the herbicide, indicating a systemic effect of glyphosate on pathogen development. Similarly, higher glyphosate doses also reduced germination and apressorium formation on detached Eucalyptus leaves, regardless of the direct application of the product on the leaf limb or on the petiole base. Puccinia psidii urediniospore germination in water agar medium also decreased with increasing herbicide doses. CONCLUSIONS: Reductions in germination and apressorium formation of P. psidii urediniospores with increasing glyphosate dose indicate that a lower severity and intensity of the disease may perhaps be due to blockage of the shikimic acid pathway in the fungal metabolic system.

Occurrence and ecological aspects of the metazoan fish parasites from Ingá lake, Maringá, Paraná State - DOI: 10.4025/actascibiolsci.v29i3.507 / Ocorrência e aspectos ecológicos de metazoários parasitos de peixes do Lago do Parque do Ingá, Maringá, Estado do Paraná - DOI: 10.4025/actascibiolsci.v29i3.507

Ingá lake was dammed in 1970 and, since then, no study on fish and parasites had been done, but it has suffered anthropic influences. This study analyzed some ecological aspects of the metazoan parasites form this lake. From analyzed fish, 69.5% had at least one species of parasite. Among them, 1.372 parasites were found, prevailing Nematoda (larvae) and Platyhelminthes (Monogenea and Cestoda). The discovery of cestode larvae in Tilapia rendalli and Oreochromis nitolicus confirms the importance of these species as participants of intermediate trophic levels. The great number of Tilapia rendalli and Oreochromis nitolicus in the lake can be a determining factor for the presence of parasites, because they take advantage of host species that are plentiful to guarantee the development of their life cycle and also because they are more susceptible to predator birds, which makes it a place of great importance for the continuation of these parasites life cycle. The low diversity of endoparasites collected can be justified by the presence of heavy metal Cu and Pb on the sediment, which must be interfering in the invertebrate development, a possible intermediate host.

Nenhum estudo de peixes e parasitos foi realizado, até o momento, no Lago do Parque do Ingá que sofre influências antrópicas desde seu represamento, em 1970. Este trabalho analisou alguns aspectos ecológicos dos parasitos metazoários dos peixes deste lago. Dos peixes analisados, (69,5%) estavam parasitados por pelo menos uma espécie de parasito. Foram encontrados 1.372 parasitos pertencentes à Nematoda (larvas) e Platyhelminthes (Monogenea e Cestoda), sendo estes os mais prevalentes e numerosos. O encontro de larvas de cestóides em Tilapia rendalli e Oreochromis niloticus confirma a importância destas espécies como participantes de níveis tróficos intermediários. A maior abundância de T. rendalli e O. niloticus, no local, pode ser um fator determinante no recrutamento de parasitos. O parasito aproveita-se das espécies de hospedeiros mais abundantes para garantir o desenvolvimento de seu ciclo de vida e também por serem os mais disponíveis à predação pelas aves piscívoras ali presentes, o que torna o local de grande importância para a sustentação do ciclo de vida desses parasitos. A baixa diversidade de endoparasitos coletados pode ser justificada pela presença de metais pesados Cu e Pb no sedimento, o que deve estar interferindo no desenvolvimento dos invertebrados, possíveis hospedeiros intermediários.