In vitro and ex vivo anti-Pythium insidiosum potential of ozonated sunflower oil.

This study sought to evaluate the in vitro and ex vivo susceptibility of Pythium insidiosum to ozonized sunflower oil (OSO) and verify the morphological alterations of OSO-exposed hyphae. Susceptibility assays were performed according to the broth microdilution protocol M38-A2/CLSI, and the minimal inhibitory (MIC) and minimal oomicidal (MOC) concentrations were also determined. Non-ozonated sunflower oil (SO) was used as the oil control. Additionally, kunkers from equine pythiosis were exposed to OSO. Damages caused by OSO and SO on P. insidiosum hyphae ultrastructure were verified using scanning electron microscopy. The MIC range for OSO was 7000 to 437.5 mg/mL, and the values for SO were higher, ranging from 56000 to 14000 mg/mL. The MOC was equal to MIC for both oil formulations. The OSO fully inhibited the oomycete growth from kunkers, although there was P. insidiosum growth in the kunker control in 24 h of incubation. The SEM analyses showed that both OSO and SO caused morphological alterations in P. insidiosum hyphae, highlighting the presence of cavitation along the hyphae with loss of continuity of the cell wall, which was more evident in the OSO-treated hyphae. The OSO had the best oomicidal activity, leading us to believe that our findings may support future research containing this formulation to be applied in integrative medicine protocols to control pythiosis in animals and humans.

Melaleuca alternifolia formulations in the treatment of experimental pythiosis.

Essential oils (EO) are aromatic compounds from the plant secondary metabolism. Melaleuca alternifolia EO is well known for its medicinal properties and promising use as an antimicrobial agent. Pythiosis is a difficult-to-treat and emerging disease caused by the oomycete Pythium insidiosum. This study evaluated a nanoemulsion formulation of M. alternifolia (NEMA) in topical and intralesional application to treat experimental pythiosis. Dermal toxicity tests were performed on M. alternifolia EO in Wistar rats. Pythiosis was reproduced in rabbits (n = 9) that were divided into groups: group 1 (control), cutaneous lesions with daily topical application of a non-ionizable gel-based formulation and intralesional application of sterile distilled water every 48 h; group 2 (topical formulation), lesions treated daily with topical application of a non-ionizable gel-based formulation containing 5 mg/ml of NEMA; and group 3 (intralesional formulation), lesions treated with NEMA at 5 mg/ml in aqueous solution applied intralesionally/48 h. The animals were treated for 45 days, and the subcutaneous lesion areas were measured every 5 days. M. alternifolia EO showed no dermal toxicity. The lesion areas treated with intralesional NEMA reduced at the end of treatment, differing from groups 1 and 2 (P < 0.05). In the topically treated group, the lesion areas did not differ from the control group, although the number of hyphae significantly reduced (P < 0.05). Under the experimental conditions of this study, the NEMA formulations presented a favorable safety profile. However, further studies are required to evaluate if this safety applies to higher concentrations of NEMA and to validate its use in clinical pythiosis.

Biogenic silver nanoparticles in the treatment of experimental pythiosis Bio-AgNP in pythiosis therapy.

Pythiosis is a rapidly progressing disease that can be lethal to affected individuals due to resistance to available therapeutic protocols. The disease affects mammals, with the largest number of reports in horses and humans. The present study investigated the activity of biogenic silver nanoparticles (bioAgNP) in the treatment of experimental pythiosis. The disease was reproduced in nine female 90-day-old New Zealand rabbits. Animals were divided into three groups: group1 (control, n = 3) daily and topically treated with a nonionized gel-based formulation and 1 ml of sterile distilled water intralesion administered every 48 hours; group 2 (n = 3), daily and topically treated with gel-based formulation containing 1 µg/ml bio-AgNP; group 3 (n = 3), treated with 1 ml bio-AgNP in 1 µg/ml aqueous solution intralesion administered every 48 hours. Animals were treated for 45 days, and the area of subcutaneous lesions was measured every 5 days. Results showed that groups 2 and 3 differed from control group (P < .05) in the lesion area, as well as the amount of hyphae within the lesions. It was observed that lesions of treated animals (groups 2 and 3) did not differ from each other, showing that the application route did not influence the regression of lesions. However, it was observed that one animal from group 2 reached clinical cure at 35 days of treatment. This research is pioneer in the application of nanocomposites for the treatment of experimental pythiosis and showed that bio-AgNP can be powerful allies of integrative medicine and can be included in pythiosis therapeutic protocols.

Economic viability of anthelmintic treatment in naturally infected beef cattle under different nutritional strategies after weaning.

The aim of the present study was to evaluate the effects of treatment with different anthelmintic compounds on the productivity of naturally infected calves and the economic viability of these treatments within extensive breeding systems employing different nutritional strategies after weaning. For this purpose, 4 farms with 42-60 calves naturally infected with gastrointestinal nematodes were selected. The calves were distributed into 6 groups (7-10 animals each) per farm and treated with ivermectin 1%, ivermectin 3.15%, eprinomectin 5%, levamisole 7.5%, albendazole 15%, and control group (no treatment). These animals were evaluated over an experimental period of 150 days. Levamisole 7.5% presented the best capacity for the reduction of eggs per gram (EPG) of feces in all herds evaluated, followed by albendazole 15% and eprinomectin 5%. Parasite resistance to multiple drugs was found in all herds, especially those of Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. For farm 1, differences in weight gain and EPG reduction percentages led to a difference of US$285.06 between the levamisole and ivermectin 3.15% groups. Similar findings were noted for the levamisole and ivermectin 1% groups of farm 3, with a difference of US$399.37 because of the final weight gain in these groups. For farms 2 and 4, the ivermectin 3.15% and control groups, respectively, were the most profitable; these unexpected results were possibly influenced by variables not measured during the experimental period. This study suggested that anthelmintic treatments should always precede an efficacy test, once they are demonstrated to be most profitable under adequate breeding conditions, to ensure adequate control of gastrointestinal nematode infection.

Ingestive behavior and forage intake of heifers receiving corn grain with or without crude glycerin on italian ryegrass pasture / Comportamento ingestivo e ingestão de forragem por bezerras recebendo grão de milho com ou sem glicerina bruta em pastagem de azevém

The objective was to analyze the ingestive behavior, forage intake and pasture characteristics when beef heifers are kept exclusively on ryegrass pasture (Lolium multiflorum Lam.) or receiving ground corn grain as supplement (0.9% of body weight (BW )) with or without addition of crude glycerin (0.2% BW). Experimental animals were Angus heifers with initial age and body weight of eight months and 166.2 ± 9.5 kg, respectively. The grazing method was continuous with variable number of animals. The experimental design was completely randomized with repeated measures. Dry matter intake was estimated using chromium oxide as an indicator of fecal output. Heifers ingested a similar amount of dry matter and neutral detergent fiber. Forage intake was 19.0% lower when heifers receiving ground corn grain as supplement with or without addition of crude glycerin and these reduction in dry matter intake of forage provided increase of 38.2% in the stocking rate. Heifers that received crude glycerin as a supplement spent more time in the trough than heifers that received only ground corn grain.

Objetivou-se avalizar o comportamento ingestivo, ingestão de forragem e as características da pastagem quando bezerras de corte são mantidas exclusivamente em pastagem de azevém (Lolium multiflorum Lam.) ou recebendo grão de milho quebrado como suplemento (0,9% do peso corporal (PC)) com ou sem adição de glicerina bruta (0,2% PC). Os animais experimentais foram bezerras Angus com idade e peso inicial de oito meses e 166,2 ± 9,5 kg, respectivamente. O método de pastejo foi o contínuo com número variável de animais. O delineamento experimental foi o inteiramente casualizado com medidas repetidas no tempo. A ingestão de matéria seca foi estimada usando óxido de cromo como indicador de produção fecal. As bezerras ingeriram similares quantidades de matéria seca e fibra em detergente neutro. A ingestão de forragem foi 19.0% menor quando as bezerras recebem grão de milho quebrado como suplemento com ou sem adição de glicerina bruta e essa redução na ingestão de matéria seca de forragem promove aumento de 38,2% na taxa de lotação. Bezerras que recebem glicerina bruta como suplemento permanecem mais tempo no cocho do que bezerras que recebem somente grão de milho quebrado.

In Vitro Activity of Melaleuca alternifolia (Tea Tree) in Its Free Oil and Nanoemulsion Formulations Against Pythium insidiosum.

Pythium insidiosum is an important aquatic oomycete which can cause pythiosis in both animals and humans. This microorganism shows low susceptibility to antifungal drugs available. This study analyzed the in vitro antimicrobial activity of Melaleuca alternifolia in its free oil (FO) and nanoemulsion (NE) formulations against Brazilian P. insidiosum isolates. The antimicrobial activity evaluation was performed by the broth microdilution method according to CSLI M38-A2 document adapted to phytopharmaceuticals. Twenty-six P. insidiosum isolates were evaluated, and the minimum inhibitory concentration was determined at 100 % growth inhibition. Melaleuca alternifolia essential oil or FO was obtained commercially. The NE containing 1 % M. alternifolia essential oil was prepared by the spontaneous emulsification method. All P. insidiosum isolates evaluated showed minimum inhibitory concentrations (MIC) ranging from 531.5 to 2125 µg/mL for the FO formulation; MIC50 and MIC90 showed values between 1062.5 and 2125 µg/mL, respectively. When the NE formulation was evaluated, MIC values ranged from 132.7 to 2125 µg/mL and both MIC50 and MIC90 corresponded to 1062.5 µg/mL. FO and NE formulations of M. alternifolia showed antimicrobial activity against P. insidiosum. This study demonstrated that M. alternifolia oil can be an additional therapy in pythiosis treatment; however, further studies are needed to evaluate the applicability of the plant essential oils in the treatment of clinical pythiosis.

Treatment of experimental pythiosis with essential oils of Origanum vulgare and Mentha piperita singly, in association and in combination with immunotherapy.

This study investigated the in vivo antimicrobial activity of the essential oils of Origanum vulgare and Mentha piperita both singly, associated and in combination with immunotherapy to treat experimental pythiosis. The disease was reproduced in 18 rabbits divided into six groups (n=3): group 1, control; group 2, treated with essential oil of Mentha piperita; group 3, treated with essential oil of Origanum vulgare; group 4, treated with commercial immunotherapic; group 5, treated with a association of oils of M. piperita and O. vulgare and group 6, treated with a combination of both oils plus immunotherapy. Essential oils were added in a topical cream base formula, and lesions were treated daily for 45 days. The animals in groups 4 and 6 received a dose of immunotherapeutic agent every 14 days. The results revealed that the evolution of lesions in groups 5 and 6 did not differ from one another but differed from the other groups. The lesions of group 5 increased 3.16 times every measurement, while those of group 6 increased 1.83 times, indicating that the smallest growth of the lesions occurred when the combination of therapies were used. A rabbit from group 5 showed clinical cure at day 20 of treatment. This research is the pioneer in the treatment of experimental pythiosis using essential oils from medicinal plants and a combination of therapies. This study demonstrated that the use of essential oils can be a viable alternative treatment to cutaneous pythiosis, particularly when used in association or combination with immunotherapy.

Consumo de forragem e desempenho de novilhas de corte recebendo suplementos em pastagem de azevém / Herbage intake and animal performance of beef heifers receiving supplements in Italian ryegrass pasture

Foram estudados o desempenho, comportamento e a ingestão de forragem por novilhas de corte em dois estádios de desenvolvimento do azevém (Lolium multiflorum Lam.): pré-florescimento e florescimento. As novilhas permaneceram exclusivamente em pastejo ou em pastejo e recebendo grão de milho ou suplemento extrusado com gordura. O delineamento experimental foi o inteiramente casualizado, em arranjo fatorial 3x2 (três sistemas alimentares x dois estádios fenológicos do azevém). A ingestão de forragem foi estimada usando o óxido de cromo como indicador da produção fecal. Os valores médios de massa de forragem e oferta de lâminas foliares foram de 1636kg ha-1 de matéria seca (MS) e 2,3kg de MS 100kg-1 de peso corporal (PC), respectivamente. Novilhas que não receberam suplemento consumiram mais pasto sem diferir das que receberam gordura. Animais exclusivamente a pasto realizaram maior número de bocados diários. As ingestões de MS do pasto e total, que foram de 2,74 e 3,01% do PC, respectivamente, são similares no pré-florescimento e florescimento do azevém. A ingestão de matéria seca total e os tempos de pastejo, ruminação e ócio de bezerras de corte recebendo suplementos são semelhantes ao uso exclusivo da pastagem de azevém, resultando em desempenho individual semelhante.

The performance, behavior and forage intake of grazing beef heifers at two phenological stages of Italian ryegrass (Lolium multiflorum Lam.): pre-flowering and flowering, were studied. The heifers remained exclusively on pasture or receiving corn grain or extruded supplement plus fat. The experimental design was completely randomized in a factorial 3x2 arrangement (three feeding systems x two phenological stages of Italian ryegrass). The forage intake was evaluated using chromic oxide as a faecal production indicator. Mean values of herbage mass and leaf allowance were 1636kg ha-1 of dry matter (DM) and 2.3kg of DM 100kg-1 live weight (LW), respectively. Heifers kept exclusively on pasture consumed more grass without difference of those receiving fat. Heifers exclusively on pasture presented greater daily bite rate. The forage and total DM intake, which averaged 2.74 and 3.01% of LW, respectively, are similar at pre-flowering and flowering of ryegrass. The total forage DM intake and the grazing, rumination and idling times of beef heifers receiving supplements are similar to the use of the ryegrass exclusively, resulting in similar individual performance of heifers.

Desenvolvimento de novilhas de corte sob alternativas de mineralização em pastagem de azevém / Development of beef heifers under mineralization alternatives, on Italian ryegrass pasture

Foi avaliado o desenvolvimento corporal de bezerras de corte em pastagem de azevém (Lolium multiflorum Lam.), recebendo diferentes alternativas de suplementação mineral, associadas ou não à adição de ionóforo. Os tratamentos testados foram: "Sal comum": mineralização com cloreto de sódio; "Sal 40P": mineralização com sal mineral contendo 40g de fósforo por kg de produto; "Sal 40P + ionóforo": mineralização com 40g de fósforo + 2.000mg de lasalocida sódica por kg de produto. Os valores de proteína bruta, fibra em detergente neutro, Ca e P na forragem aparentemente consumida, consumo de sal, escore de condição corporal, relação peso: altura e GMD foram semelhantes (P>0,05) entre animais que receberam "Sal comum", "Sal 40P" e "Sal 40P + ionóforo". Em pastagem de azevém, sem restrição ao consumo voluntário, a lasalocida sódica adicionada ao sal mineral proporciona maior peso vivo de bezerras aos 12 meses de idade e um retorno financeiro positivo.

Body development of beef heifers grazing Italian ryegrass pasture (Lolium multiflorum Lam.) was evaluated. The animals received different alternatives of mineral supplement: associated or not to ionophore addition and the treatments were: 'Common salt': mineralization with sodium chloride; '40P salt': mineralization with mineral salt plus 40g of phosphorus per kilo of product; 40P salt + ionophore: mineralization with 40g of phosphorus plus 2000mg of lasalocid per kg of product. The values of crude protein, neutral detergent fiber and Ca and P of the apparently consumed forage, salt intake, body condition and body weight:height relation were similar (P>0.05) between animals receiving 'common salt', '40P salt' and '40P salt+ionophor'. For beef heifers grazing ryegrass pasture without intake restriction, the lasalocid associated to mineral salt provides a greater body weight at twelve months age and a positive financial.