Delivery of selenium using chitosan nanoparticles: Synthesis, characterization, and antioxidant and growth effects in Nile tilapia (Orechromis niloticus).

This study aimed to elucidate the effects of selenium-loaded chitosan nanoparticles used as a dietary supplement on Nile tilapia (Oreochromis niloticus) antioxidant and growth responses. First, chitosan-based nanoparticles containing selenium (Se) were synthesized using the ionotropic gelation method and their physicochemical characteristics, controlled release profile, and antioxidant activity properties were investigated. Thereafter, the effects on glutathione peroxidase and antioxidant activities (by radical scavenging activity), growth, and whole-body composition of Nile tilapia were evaluated when they were fed with Se-loaded chitosan nanoparticles and compared with other selenium dietary supplements. Se-loaded chitosan nanoparticles showed high entrapment efficiency (87%), spherical shape, smooth surface, and broad size distribution. The controlled release of Se consisted of an initial burst followed by a gradual release over 48 h. Se-loaded nanoparticles presented significantly higher antioxidant activity compared to free Se. A 60-day feeding trial was conducted to compare the effects of supplementing different dietary Se sources, including selenomethionine (as organic source), sodium selenite (as inorganic source), and Se-loaded chitosan nanoparticles (Se-Nano and Se-Nano x1.5) on antioxidant and growth responses of Nile tilapia. A basal diet without Se supplementation was used as the control. The dietary supplementations with different Se sources (free and encapsulated selenium) lead to significant improvements in final weight and feed efficiency of Nile tilapia fingerlings. However, dietary treatments did not affect whole-body protein and lipid content. Diets containing Se-Nano and Se-Nano x1.5 were more effective than sodium selenite and selenomethionine in preventing oxidative stress and improving antioxidant activity in Nile tilapia. Overall, Se-loaded nanoparticles presented a great potential as an efficient source for delivering dietary Se to Nile tilapia, directly affecting the growth performance, feed efficiency, oxidative stress, and antioxidant activity of this species.

Biological control of infective larvae of Ancylostoma spp. in beach sand / Control biológico de las larvas infectantes de Ancylostoma spp. en arena de playa

Background. Geohelminths are parasites that stand out for their prevalence and wide distribution, depending on the soil for their transmission. Aims. The aim of this work was to evaluate the predatory capacity of the fungal isolate of the genus Duddingtonia (CG768) on third stage larvae (L3) of Ancylostoma spp. in beach sand under laboratory conditions. Methods. In the assay A five treatment groups and 1 control group were formed. The treatment groups contained 5000, 10,000, 15,000, 20,000 or 25,000 chlamydospores of the fungal isolate and 1000 Ancylostoma spp. L3 in pots containing 30 g of sand. The control group (without fungus) contained only 1000 Ancylostoma spp. L3 and distilled water in pots with 30 g of sand. Results. Evidence of predatory activity was observed at the end of 15 days, where we observed the following percentages of reduction of L3: Group 1 (4.5%); Group 2 (24.5%); Group 3 (59.2%); Group 4 (58.8%); Group 5 (63%). However, difference was noted (p < 0.01) only at concentrations 15,000, 20,000 and 25,000 in relation to control group. In the assay B two groups were formed in Petri dishes of 9 cm in diameter containing agar water 2% medium. In the treated group, each Petri dish contained 500 Ancylostoma spp. L3 and 5 g of sand containing the isolate CG 768 at a concentration of 25,000 chlamydospores/g of sand, and the control group (without fungus) contained only 500 L3. At the end of 7 days the non-predation L3 of Petri dishes using the method of Baermann were recovered. Difference (p < 0.01) between groups on reducing the average number of Ancylostoma spp. L3 (percent reduction of 84%) was observed. Conclusions. The results of this study confirm earlier work on the efficiency of the Duddingtonia genus in the control of Ancylostoma spp. infective larvae (AU)

Antecedentes. Los geohelmintos son parásitos que destacan por su prevalencia y amplia distribución, puesto que su transmisión depende del suelo. Objetivos. El objetivo del presente estudio fue evaluar la capacidad predatoria de aislamientos fúngicos del género Duddingtonia (CG768) sobre las larvas de estadio 3 (L3) de Ancylostoma spp. en arena de playa, en condiciones de laboratorio. Métodos. En el ensayo A se formaron 5 grupos de tratamiento y un grupo de control. Los grupos de tratamiento contenían 5000, 10.000, 15.000, 20.000 o 25.000 clamidosporas del aislamiento fúngico y 1000 larvas L3 de Ancylostoma spp. en recipientes con 30 g de arena. Los recipientes del grupo de control (sin clamidosporas) solo contenían 1000 larvas L3 de Ancylostoma spp. y agua destilada con 30 g de arena. Resultados. Al término de 15 días, fue evidente la actividad predatoria, con los porcentajes siguientes de reducción de larvas L3: grupo 1 (4.5%); grupo 2 (24.5%); grupo 3 (59.2%); grupo 4 (58.8%), y grupo 5 (63%). Sin embargo, en relación con el grupo control, solo se identificaron diferencias significativas (p < 0.01) a las concentraciones de 15.000, 20.000 y 25.000. En el ensayo B, en placas de Petri de 9 cm de diámetro, que contenían un medio de agar agua al 2%, se formaron 2 grupos. En el grupo tratado, cada placa de Petri contenía 500 larvas L3 de Ancylostoma spp. y 5 g de arena con el aislamiento CG768 a una concentración de 25.000 clamidosporas/g de arena, y el grupo de control (sin hongo) solo contenía 500 larvas L3. Al cabo de 7 días, utilizando el método de Baermann, a partir de las placas de Petri se obtuvieron larvas L3 no sometidas a predación por el hongo. Entre los grupos se observó una diferencia significativa (p < 0.01) en la reducción del número medio de larvas L3 de Ancylostoma spp. (porcentaje de reducción del 84%). Conclusiones. Los resultados del presente estudio confirman los datos de investigaciones previas sobre la eficiencia del género Duddingtonia en el control de las larvas infectantes de Ancylostoma spp (AU)

Biological control of infective larvae of Ancylostoma spp. in beach sand.

BACKGROUND: Geohelminths are parasites that stand out for their prevalence and wide distribution, depending on the soil for their transmission. AIMS: The aim of this work was to evaluate the predatory capacity of the fungal isolate of the genus Duddingtonia (CG768) on third stage larvae (L3) of Ancylostoma spp. in beach sand under laboratory conditions. METHODS: In the assay A five treatment groups and 1 control group were formed. The treatment groups contained 5000, 10,000, 15,000, 20,000 or 25,000 chlamydospores of the fungal isolate and 1000 Ancylostoma spp. L3 in pots containing 30g of sand. The control group (without fungus) contained only 1000 Ancylostoma spp. L3 and distilled water in pots with 30g of sand. RESULTS: Evidence of predatory activity was observed at the end of 15 days, where we observed the following percentages of reduction of L3: Group 1 (4.5%); Group 2 (24.5%); Group 3 (59.2%); Group 4 (58.8%); Group 5 (63%). However, difference was noted (p<0.01) only at concentrations 15,000, 20,000 and 25,000 in relation to control group. In the assay B two groups were formed in Petri dishes of 9cm in diameter containing agar water 2% medium. In the treated group, each Petri dish contained 500 Ancylostoma spp. L3 and 5g of sand containing the isolate CG 768 at a concentration of 25,000 chlamydospores/g of sand, and the control group (without fungus) contained only 500 L3. At the end of 7 days the non-predation L3 of Petri dishes using the method of Baermann were recovered. Difference (p<0.01) between groups on reducing the average number of Ancylostoma spp. L3 (percent reduction of 84%) was observed. CONCLUSIONS: The results of this study confirm earlier work on the efficiency of the Duddingtonia genus in the control of Ancylostoma spp. infective larvae.

Biological control of Fasciola hepatica eggs with the Pochonia chlamydosporia fungus after passing through the cattle gastrointestinal tract.

Fasciolosis is a disease caused by Fasciola hepatica responsible for causing significant losses in livestock. This study aimed to evaluate the Pochonia chlamydosporia fungus (isolate VC1) on F. hepatica eggs after passing through the cattle gastrointestinal tract. For this evaluation, 1 g pellet was given in sodium alginate matrix per kilogram live weight containing 25% of fungal mycelium from isolate VC1 per animal. Twelve animals were used, six treated and six untreated (control). Some stool samples were collected from the groups of treated and control animals, at the times of 12, 18, 24, 48, 72, and 96 h after the pellets' administration. Then, from each stool sample of treated and control groups, 2 g was placed in a Petri dish of 9 cm in diameter, containing 2% water-agar and 1,000 eggs of F. hepatica. It was observed that the fungus was effective in preying upon the eggs in the samples recovered at all of the schedules starting at 12 h. Furthermore, differences were observed (p < 0.01) in the destruction of eggs in the Petri dishes in the treated group compared with the control group. The ovicidal effect was observed after 7 days of interaction. The ovicidal P. chlamydosporia fungus was effective in destroying F. hepatica eggs; therefore, it is suggested that this fungus could be employed as agent for the control of helminth eggs.

Survival of Pochonia chlamydosporia in the gastrointestinal tract of experimentally treated dogs.

The predatory capacity of the nematophagous fungus Pochonia chlamydosporia (isolate VC4) after passage through the gastrointestinal tract of dogs was assessed in vivo against Toxocara canis eggs. Twelve dogs previously wormed were divided into two groups of six animals and caged. The treatments consisted of a fungus-treated group (VC4) and a control group without fungus. Each dog of the fungus-treated group received a single 4 g dose of mycelial mass of P. chlamydosporia (VC4). Fecal samples from animals of both groups (treated and control) were collected at five different times (6, 12, 24, 36, and 48 h) after fungal administration, and placed in Petri dishes. Each Petri dish of both groups for each studied time interval received approximately 1000 T. canis eggs. Thirty days after the fecal samples were collected, approximately one hundred eggs were removed from each Petri dish of each studied time interval and evaluated by light microscopy (LM) and scanning electron microscopy (SEM). Microscopy examination of plates inoculated with the fungus showed that the isolate VC4 was able to destroy the T. canis eggs with destruction percentages of 28.6% (6 h), 29.1% (12 h), 32.0% (24 h), 31.7% (36 h), and 37.2% (48 h). These results suggest that P. chlamydosporia can be used as a tool for the biological control of T. canis eggs in feces of contaminated dogs.

Ovicidal activity of seven Pochonia chlamydosporia fungal isolates on Ascaris suum eggs.

The ovicidal effect of the nematophagous fungus Pochonia chlamydosporia on eggs of Ascaris suum was tested under laboratory conditions. A. suum eggs were plated on 2% water-agar with seven fungal isolates (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) and control without fungus. After 5, 7, 10, 14, 15 and 21 days of incubation, approximately 100 eggs were removed from the plates and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo showing hyphal penetration and internal egg colonization. The isolates effectively destroyed A. suum eggs and all types of effects were observed during the experiment. There was no variation in ovicidal capacity (type 3 effect) among the isolates (p>0.05) throughout the experiment. After 21 days, isolate 5 showed the highest percentages of type 3 effect (58.33%). The results indicated that P. chlamydosporia (Isol. 5, Isol. 31, Isol. 1, VC1, Isol. 12, Isol. 22 and VC4) can destroy A. suum eggs and is, therefore, a potential biological control agent of nematodes.

In vitro predatory activity of nematophagous fungi and after passing through gastrointestinal tract of equine on infective larvae of Strongyloides westeri.

Three isolates of predator fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), and Arthrobotrys robusta (I-31) were assessed in in vitro test regarding the capacity of prey infective larvae (L(3)) Strongyloides westeri. Compared to control, without fungus, there was a significant decrease (P < 0.01) of 80.4%, 67.9%, and 72.8% in means of infective larvae S. westeri recovered from treatments with isolates AC001, NF34, and I-31, respectively. All tested isolates were efficient in the capture of S. westeri (P > 0.01) in vitro test. Linear regression coefficients of treated and control groups were -0.21 for control, -0.32 for D. flagrans, -0.34 for M. thaumasium, and -0.22 for A. robusta. In the following, isolates AC001 and NF34 were assessed in vivo regarding the capacity of supporting the passage through equine gastrointestinal tract without loss of ability of preying infective larvae S. westeri. Fungal isolates survived the passage and were efficient in preying L(3) since the first 12 h of collection (P < 0.01) in relation to the control group (without fungus). Compared to control, there was a significant decrease (P < 0.01) of 76.4% and 76.7% (12 h), 86.4% and 85.9% (24 h), 88.3% and 87.7% (48 h), and 89.9% and 87.2% (72 h) in means of infective larvae S. westeri recovered from treatments with isolates AC001 and NF34, respectively. Linear regression coefficients of L(3) of recovered S. westeri regarding the collections due to time were 1.93 for control, -3.52 for AC001, and -2.64 for NF34. Fungi D. flagrans and M. thaumasium (NF34) have demonstrated to be promising for use in the biological control of equine parasite S. westeri.

Predatory activity of the nematophagous fungus Duddingtonia flagrans on horse cyathostomin infective larvae.

This work was performed to determine the predatory capacity in vitro of the nematophagous fungus Duddingtonia flagrans (isolate AC001) on cyathostomin infective larvae of horse (L(3)). The experimental assay was carried out on plates with 2% water-agar (2% WA). In the treated group, each plate contained 1.000 L(3) and 1.000 conidia of the fungus. The control group without fungus only contained 1.000 L(3) in the plates. Ten random fields (4 mm diameter) were examined per plate of treated and control groups, every 24 h for seven days under an optical microscope (10x and 40x objective lens) for non-predated L(3) counts. After 7 days, the non-predated L(3) were recovered from the Petri dishes using the Baermann method. The interaction there was a significant reduction (p < 0.01) of 93.64% in the cyathostomin L(3) recovered. The results showed that the D. flagrans is a potential candidate to the biological control of horse cyathostomin L(3).

Predatory activity of Pochonia chlamydosporia fungus on Toxocara (syn. Neoascaris) vitulorum eggs.

Toxocara (Neoascaris) vitulorum is a gastrointestinal nematode parasite of young ruminants, responsible for high mortality rates in parasitized cattle and buffalo calves. The objective of this work was to compare the predatory capacity under laboratory conditions of four fungal isolates of the nematophagous fungus Pochonia chlamydosporia (VC1, VC4, VC5 and VC12) on T. vitulorum eggs in 2% water-agar (2% WA). T. vitulorum eggs were plated on 2% WA Petri dishes which contained cultured fungal isolates and control plates without fungi. After 10 and 15 days one hundred eggs were removed and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo in addition to hyphal penetration and internal egg colonization. The fungal isolates were effective in the destruction of T. vitulorum eggs presenting the type 3 effect at 10 and 15 days after contact with the fungus. No nematophagous fungi were observed in the control group during the experiment. There was no variation in the predatory capacity of the fungal isolates (P > 0.01) at the intervals of 10 and 15 days. These results indicate that P. chlamydosporia (VC1, VC4, VC5 and VC12) negatively influenced the development of T. vitulorum eggs and can be considered a potential candidate for the biological control of nematodes.

[In vitro evaluation of nematode predacious fungus Duddingtonia fagrans on cyathostomes infective larvae of equines (Nematoda: Cyathostominae)]. / Avaliação in vitro do fungo predador de nematoides Duddingtonia flagrans sobre larvas infectantes de ciatostomíneos de equinos (Nematoda: Cyathostominae).

The predatory capacity of one isolate of nematode-trapping fungus Duddingtonia flagrans (AC001) on infective larvae of cyathostomes was evaluated in laboratorial conditions in medium water-agar 2% (WA 2%). There was significant reduction (p<0.01) of 93.64% in the average of infective larvae of cyathostomes recovered of medium WA 2% at seven day. These results show that the isolated AC001 could be used in the biological control of cyathostomes of horses.

Avaliação in vitro do fungo predador de nematoides Duddingtonia flagrans sobre larvas infectantes de ciatostomíneos de equinos (Nematoda: Cyathostominae) / In vitro evaluation of nematode predacious fungus Duddingtonia flagrans on cyathostomes infective larvae of equines (Nematoda: Cyathostominae)

A capacidade predatória de um isolado de fungo predador de nematoides Duddingtonia flagrans (AC001) sobre larvas infectantes de ciatostomíneos foi avaliada em condições laboratoriais em ensaio experimental em meio ágar-água 2% (AA 2%). Houve redução significativa (p < 0,01) de 93,64% na média de larvas infectantes de ciatostomíneos recuperadas do meio AA2%, ao final de sete dias. Os resultados desse ensaio evidenciam que o isolado fúngico AC001 poderia ser utilizado no controle biológico de ciatostomíneos de equinos.

The predatory capacity of one isolate of nematode-trapping fungus Duddingtonia flagrans (AC001) on infective larvae of cyathostomes was evaluated in laboratorial conditions in medium water-agar 2% (WA 2%). There was significant reduction (p < 0. 01) of 93.64% in the average of infective larvae of cyathostomes recovered of medium WA 2% at seven day. These results show that the isolated AC001 could be used in the biological control of cyathostomes of horses.

Interaction and ovicidal activity of nematophagous fungus Pochonia chlamydosporia on Taenia saginata eggs.

The ovicidal activity of the nematophagous fungi Pochonia chlamydosporia (isolates VC1 and VC4), Duddingtonia flagrans (isolate AC001) and Monacrosporium thaumasium (isolate NF34) on Taenia saginata eggs was evaluated under laboratory conditions. T. saginata eggs were plated on 2% water-agar with fungal isolates and controls without fungus and examined after 5, 10 and 15 days. At the end of the experiment P. chlamydosporia showed ovicidal activity against T. saginata eggs (p<0.05), mainly for internal egg colonization with results of 12.8% (VC1) and 2.2% (VC4); 18.1% (VC1) and 7.0% (VC4); 9.76% (VC1) and 8.0% (VC4) at 5, 10 and 15 days, respectively. The other fungi showed only lytic effect without morphological damage to the eggshell. Results demonstrated that P. chlamydosporia was effective in vitro against T. saginata eggs unlike the other fungi.

[Evaluation of the efficacy of antihelmintics compounds on parasitics gastrointestinal nematodes (Strongyloidea) of goats]. / Avaliação da eficácia de compostos anti-helmínticos sobre nematóides parasitos gastrintestinais (Strongyloidea) de caprinos

The present study was performed in order to evaluate the action of anthelmintics compounds on gastrointestinal parasite nematodes of 27 Alpine and Saanen adult goats. The animals were divided into three groups. The animals of groups 1 and 2 had been dealt with two different associations of antihelminthics in day zero. The goats in group 1 were treated with closantel (75 mg/mL), albendazol (38 mg/mL) and ivermectin B1a (2 mg/mL) orally (1 ml/ 10 Kg body weight); animals in group 2 were treated with closantel (100 mg/mL), albendazol (50 mg/mL), levamisol (64 mg/mL), ivermectin B1a (2 mg/mL), selenium (1 mg/mL) and cobalt (4.4 mg/mL) orally (1 ml/10 Kg of body weight) and the animals in the group 3 (control) received distilled water. Eggs per gram counts on faeces (EPG) and coprocultures of all animals were made at intervals of days 0, 3, 5, 7, 14, 21 and 28. The haematocrit, global counting and differential white blood cells, total protein and the Famacha test were determined at intervals of days 0, 14 and 28. Six animals of each group had suffered euthanasia and slaughters on the 28th day. The results showed that only the combination used in the animals of group 2 was effective.