The Role of Melanin in the Biology and Ecology of Nematophagous Fungi.

Melanin is a heteropolymer formed by the polymerization of phenolic and indolic compounds. It occurs in organisms across all biological kingdoms and has a range different of functions, thus indicating its important evolutionary role. The presence of melanin offers several protective advantages, including against ultraviolet radiation, traumatic damage, oxidative stress, extreme temperatures, and pressure. For many species of fungi, melanin also participates directly in the process of virulence and pathogenicity. These organisms can synthesize melanin in two main ways: using a substrate of endogenous origin, involving 1,8-dihydroxynaphthalene (DHN); alternatively, in an exogenous manner with the addition of L-3, 4-dihydroxyphenylalanine (L-DOPA or levodopa). As melanin is an amorphous and complex substance, its study requires expensive and inaccessible technologies and analyses are often difficult to perform with conventional biochemical techniques. As such, details about its chemical structure are not yet fully understood, particularly for nematophagous fungi that remain poorly studied. Thus, this review presents an overview of the different types of melanin, with an emphasis on fungi, and discusses the role of melanin in the biology and ecology of nematophagous fungi.

Duddingtonia flagrans in the control of gastrointestinal nematodes of horses.

Horses can harbor a large amount of parasites that may cause serious clinical signs even death. The aim of this study was to evaluate the predatory activity of the fungus Duddingtonia flagrans against infective larvae (L3) of gastrointestinal nematodes of horses in fecal culture. The experimental design was completely randomized with three treated groups (G1, G2 and G3) and one control (CG), using eight animals/group. The treated animals received G1: 1.5 × 10(5); G2: 3 × 10(5) and G3: 6 × 10(5) chlamydospores of D. flagrans/kg body weight during 21 days. The fungi preparation was given at every other three-day interval. Faecal samples were collected during 30 days, on the same interval, to perform the fecal egg counts (EPG) and fecal culture for each horse. All groups demonstrated similar results for the EPG (P > 0.05) counts. D. flagrans significantly reduced (P < 0.05) the number of infective larvae after 72 h-interval between treatments. The G2 and G3 promoted higher results (P < 0.05) of L3 reduction compared to the CG. The biological control with the predacious fungi D. flagrans is still a promising free-living parasite regulator alternative to be use in livestock.

Kinetics of capture and infection of infective larvae of trichostrongylides and free-living nematodes Panagrellus sp. by Duddingtonia flagrans.

Duddingtonia flagrans, a nematode-trapping fungus, has been investigated as an agent for biological control against infective larvae of gastrointestinal nematode parasites of production animals. The initial process of nematode-trapping fungi infection is based on an interaction between the trap structure of the fungus and the surface of the nematode cuticle. This report investigates by light and scanning electron microscopy the kinetics of capture and infection during the interaction of D. flagrans with the infective larvae (L(3)) of trichostrongylides and the free-living nematode Panagrellus sp. D. flagrans was cultivated for 7 days in a Petri dish containing agar-water. L(3) and Panagrellus sp. were inoculated in the Petri dishes and the samples consisting of agar-L(3)-fungi and agar-Panagrellus sp.-fungi were collected after 10, 20, 30, 40, 50, 60, and 70 min and 3, 4, 5, 10, 15, 20, and 25 h of interaction. All samples were observed by light microscopy. The samples with 1, 5, 15, and 25 h of interaction were also analyzed by scanning electron microscopy. The interaction was monitored up to 25 h. An initial differentiation of predation structures was observed after 30 min of interaction. The presence of traps and of captured L(3) or Panagrellus sp. occurred after 70 min. The live captured nematodes were observed up to 3 h of interaction. However, after 4 h, all Panagrellus sp. were dead. It took 15 h of interaction for the fungus to invade the L(3), and the presence of hyphae inside the nematode near the region of penetration was evident. At this time, the hyphae had filled the whole body of Panagrellus sp. The complete occupation of the body of L(3) occurred at 20 h of interaction and with 25 h the nematode was completely damaged except for the cuticle. Although the double cuticle of L(3) slows the penetration of D. flagrans, it was possible to verify that the process of trap formation and capture occurs quickly when both nematodes were tested, suggesting that the organisms would eventually be killed once in contact with the fungi encouraging the use of the fungus as a biological control agent.

Mixed infection by Libyostrongylus douglassii and L. dentatus (Nematoda: Trichostrongylidae) in Struthio camelus (Ratites: Struthioniformes) from Brazil with further morphological characterization of adults.

The genus Libyostrongylus includes three species, L. douglassii, L. dentatus and L. magnus that occur as parasites in the proventriculus of Struthio camelus. We confirmed a mixed infection by L. douglassii and L. dentatus in farmed ostriches from the southeast of Brazil for the first time, and provided new information on some morphological characters that differentiate these species. Adult nematodes collected from the proventriculus of ostriches were observed by light and scanning electron microscopy. Morphologic characterization and morphometric analysis of the nematodes enabled the distinction of both species and corroborated results of prior studies. Specimens of L. dentatus have a buccal capsule with a prominent esophageal tooth. Furthermore, males and females of L. dentatus were larger (4954 and 9347 microm) than those of L. douglassii (3411 and 4229 microm), but measurements for most characters in both species were smaller then those previously reported. Besides, the cephalic structure based on scanning electron microscopy differs, and L. dentatus has thick lips with round papillae, whereas L. douglassii has fine lips with lengthened papillae. The confirmation of both species in South America strongly suggests that the mixed infection may be common in farmed ostriches.

In vitro efficacy of Duddingtonia flagrans against nematodes of sheep based on in vivo calculations

Abstract Duddingtonia flagrans has been tested as an alternative parasite control, but data from in vitro experiments based on in vivo calculations describing nematophagous fungi predation in nematodes are restricted. The objective of this work was to determine the efficacy of D. flagrans against sheep nematode larvae in vitro using in vivo calculations. Fecal samples were introduced to fungi in different concentrations: 0.0/control; 0.05; 0.1; 0.2; 0.4; 0.8; 1.6; 3.2; and 6.4 g corresponding, respectively, to 583.000; 1.166.000; 2.332.000; 4.664.000; 9.328.000; 18.656.000; 37.312.000 and 74.624.000 chlamydospores/kg of body weight. The material was incubated for 14 days, before the larvae recovery (Assay 1). Assay 2 was carried out with the doses of 0.00625; 0.0125; and 0.025 g. The results showed a negative correlation between fungal concentrations and larval numbers for both assays. The fungus demonstrated an efficacy above 89% in both assays. Thus, we consider that the data from in vitro studies based on in vivo calculations may optimize the fungi quantities for field experiments.

Resumo Duddingtonia flagrans tem sido testado como uma alternativa no controle de parasitos, entretanto, trabalhos in vitro da predação de nematoides por fungos nematófagos correlacionados com cálculos baseados para testes in vivo são restritos. O objetivo deste trabalho foi determinar a eficácia in vitro de D. flagrans contra larvas de nematoides de ovinos tendo como base cálculos in vivo. Amostras fecais receberam a adição do fungo em diferentes concentrações: 0.0/controle; 0,05; 0,1; 0,2; 0,4; 0,8; 1,6; 3,2 e 6,4 gramas correspondendo, respectivamente, às seguintes dosagens: 583.000; 1.166.000; 2.332.000; 4.664.000; 9.328.000; 18.656.000; 37.312.000 e 74.624.000 clamidósporos/Kg de peso vivo animal. O material foi incubado por 14 dias, para recuperação das larvas (Ensaio 1). O Ensaio 2 foi realizado com concentrações de 0,00625; 0,0125 e 0,025 g. Foi observada correlação negativa entre a concentração fúngica e o número de larvas, nos dois ensaios. O fungo demonstrou eficácia acima de 89% em ambos os ensaios. A partir destes dados, acreditamos que ensaios in vitro baseados em cálculos in vivo podem aprimorar as dosagens para a realização de experimentos a campo.

Helminths found in marmosets (Callithrix penicillata and Callithrixjacchus) introduced to the region of occurrence of golden lion tamarins (Leontopithecusrosalia) in Brazil.

The introduction of exotic species can increase the risk of extinction of native species through the introduction of new diseases, predation or resource competition. The marmosets Callithrix penicillata and Callithrix jacchus and hybrids of these two species have been introduced to privately owned forests in the lowland Atlantic forest of the Rio de Janeiro State in Brazil, the region of occurrence of the endangered golden lion tamarin, Leontopithecus rosalia. Because the ecology and biology of the marmosets and tamarins is similar, there is a reasonable risk that the marmosets would transmit pathogens such as endo parasites. The objective of this study was to identify the helminth fauna present in the introduced marmosets through an analysis of fecal samples of wild caught animals, and to evaluate the parasitological profile according to age, sex and geographical location. Eggs belonging to the Acanthocephala and Nematoda were found in the feces. One nematode egg type was identified as being Primasubulura jacchi. The ocurrence of nematodes was higher in males, acanthocephala were found in all age groups and sexes and P. jacchi was found with higher frequency in adult females. The geographic distribution analysis revealed that some of the forests had a higher predominance of parasites. Little is known about the pathology of parasites in free living Neotropical mammals, and this knowledge would be necessary to infer about the risk (form a parasitological standpoint) that the marmoset presence represents for the survival of the endangered golden lion tamarin.