Different isolates from Leishmania braziliensis complex induce distinct histopathological features in a murine model of infection

The aim of this study was to evaluate the histopathological features in tissues of mice infected by human isolates (I, II, and III) or the reference M2903 strain of Leishmania braziliensis complex. BALB/c and C57Bl/6 mice were infected in the hind footpad with 10⁶ stationary-phase promastigotes of L. braziliensis complex. The evolution of lesions was observed for 10 weeks and the animals were then euthanized and liver, spleen and popliteal lymph nodes were collected. Tissues were stained with hematoxylin and eosin and analyzed by immunohistochemistry assay. Increased thickness of infected footpads was observed in all animals, lesions were nodular and non-ulcerated. Mice infected with isolate I presented inflammatory infiltrates consisting predominantly of mononuclear cells in all tissues examined, and also a great number of megakaryocytes, compared with other isolates. Infection with isolate II led to an infected footpad enlargement not seen in other isolates. In addition, mononuclear infiltrates in the liver and hemosiderin in spleen were noted. Conversely, mice infected with either isolate III or M2903 strain only showed an increased number of megakaryocytes in spleen. All tissues examined had detectable amastigote forms of Leishmania by immunohistochemistry in all groups. Taking together, our results showed an unforeseen behavior of different isolates of L. braziliensis complex that led to diverse pathological findings.     

Effect of nitrogen fertilization on nitrogen dynamics in oilseed rape using 15N‐labeling field experiment

In order to optimize nitrogen (N) fertilization and to reduce the environmental impact of oilseed rape without decreasing yield, a clearer understanding of N dynamics inside the plant is crucial. The present investigation therefore aimed to study the effects of different N‐application rates on the dynamics of N uptake, partitioning, and remobilization. The experiment was conducted on winter oilseed rape (Brassica napus L. cv. Capitol) under three levels of N input (0, 100, and 200 kg N ha^–1) from stem elongation to maturity using ¹⁵N‐labeling technique to distinguish between N uptake and N retranslocation in the plant. Nitrogen fertilization affected the time‐course of N uptake and also the allocation of N taken up from flowering to maturity. Most pod N came from N remobilization, and leaves accounted for the largest source of remobilized N regardless the N‐application rate. However, the contribution of leaves to the remobilized N pool increased with the N dose whereas the one of taproot decreased. Stems were the main sink for remobilized N from stem elongation to flowering. Leaves remained longer on N200 than on N0 and N100 plants, and N concentration in fallen leaves increased with the N treatment and in N100 plants along an axial gradient from the basal to the upper leaves. Overall, these results show that the timing of N supply is more crucial than the N amount to attain a high N efficiency.     

Antiproliferative Activity of Fucan Nanogel

Sulfated fucans comprise families of polydisperse natural polysaccharides based on sulfated L-fucose. Our aim was to investigate whether fucan nanogel induces cell-specific responses. To that end, a non toxic fucan extracted from Spatoglossum schröederi was chemically modified by grafting hexadecylamine to the polymer hydrophilic backbone. The resulting modified material (SNFuc) formed nanosized particles. The degree of substitution with hydrophobic chains was close to 100%, as estimated by elemental analysis. SNFfuc in aqueous media had a mean diameter of 123 nm and zeta potential of −38.3 ± 0.74 mV, as measured by dynamic light scattering. Nanoparticles conserved their size for up to 70 days. SNFuc cytotoxicity was determined using the MTT assay after culturing different cell lines for 24 h. Tumor-cell (HepG2, 786, H-S5) proliferation was inhibited by 2.0%–43.7% at nanogel concentrations of 0.05–0.5 mg/mL and rabbit aorta endothelial cells (RAEC) non-tumor cell line proliferation displayed inhibition of 8.0%–22.0%. On the other hand, nanogel improved Chinese hamster ovary (CHO) and monocyte macrophage cell (RAW) non-tumor cell line proliferation in the same concentration range. The antiproliferative effect against tumor cells was also confirmed using the BrdU test. Flow cytometric analysis revealed that the fucan nanogel inhibited 786 cell proliferation through caspase and caspase-independent mechanisms. In addition, SNFuc blocks 786 cell passages in the S and G2-M phases of the cell cycle.     

The fatty acid profiles and energetic substrates of two Nile tilapia (Oreochromis niloticus,Linnaeus) strains,Red‐Stirling and Chitralada,and their hybrid

Protein and lipid content as well as the fatty acid (FA) composition of storage tissues were analysed in two varieties of Oreochromis niloticus (Red‐Stirling and Chitralada) and their hybrid. The animals were maintained in cages for 11 months. The samples were taken when the animals weighed 10, 50, 100, 250 and 500 g. The results showed that changes in the metabolic processes occur during an increase in body mass in both varieties of tilapia and also their hybrid, but that these differences are not found in animals collected at the commercial weight. The protein content of the fillet and liver decreased with growth and the same protein content associated with growth was found for fillet lipid content. The genetic variety did not influence the FA profile of the fillet, but different genotypes had different hepatic FA compositions. Even with the same lipid content, the hepatocytes of Chitralada accumulated higher levels of polyunsaturated fatty acids (PUFA) n6 in triglycerides and increased C22:6n3 in the hepatocyte membranes. The higher n6PUFA content was compensated by a lower fraction of saturated FA in the hepatocyte triglycerides. The skin of Chitralada also had higher n6PUFA and C22:6n3 contents, suggesting a higher ability to deposit PUFA in the skin due to alterations in the liver synthetic pathway.     

Tendonitis of branches of the superficial digital flexor tendon in Standardbred racehorses: 15 cases

This case report describes the clinical and ultrasonographic findings and performance outcome of distal tendonitis of the superficial digital flexor tendon (SDFT) in Standardbred racehorses. Cases with distal tendonitis of the SDFT in isolation (not associated with a metacarpal lesion) were reviewed retrospectively. The clinical and ultrasonographic findings and performance outcome were reviewed. The injury was located on the left forelimb in 67% (10/15) of horses and on the right forelimb in 33% (5/15). None of the horses were affected bilaterally. The lateral branch was involved in 80% of the cases (12/15) and medial branch in 20% (3/15). None of the horses were affected biaxially. Mean time between injury and first start ± s.d. was 302 ± 142 days. Returning to racing was observed in 93% of the horses (14/15), with a reinjury rate of 22% (2/9). For horses which raced after the injury, the number of starts, shows and wins was not significantly different before and after injury, regardless of the location of the lesion. Tendonitis of branches of the SDFT in isolation carries a good prognosis in Standardbred racehorses. A higher return to racing and a lower reinjury rate may be expected compared to lesions in the metacarpal region.     

Nitrate Reductase,Micronutrients and Upland Rice Development as Influenced by Soil pH and Nitrogen Sources

The average yield of upland rice under no-tillage system (NTS), a sustainable soil management, is lower than in conventional tillage (one plowing and two disking). One of the reasons given for this drop in crop grain yield would be the low-nitrate assimilation capacity of rice seedlings, due to the low activity of the nitrate reductase (NR) enzyme in the early development phase. A greenhouse experiment was conducted to evaluate the effects of the soil acidic and nitrogen source in the micronutrient concentrations, NR activity and grain yield of upland rice growing under NTS. The soil used in the experiment was an Oxisol. The experimental design was completely randomized in a factorial 3 × 4. Treatments consisted of three levels of soil acidity (high, medium, and low) combined with four nitrogen sources (nitrate, ammonium, ammonium + nitrification inhibitor, and control – without N fertilization). The reduction of soil acidity reduced the concentration of zinc and manganese in rice plants. Generally, the activity of the NR enzyme was higher in plants grown in soils with low acidity and fertilized with calcium nitrate. There was a greater response in growth and yield in rice plants grown in soils with high acidity. Under medium acidity, rice plants grown with ammonium sulfate were more productive (no differences were detected with the addition of the nitrification inhibitor).     

Toxic effects of domoic acid in the seabream Sparus aurata

Neurotoxicity induced in fish by domoic acid (DA) was assessed with respect to occurrence of neurotoxic signs, lethality, and histopathology by light microscopy. Sparus aurata were exposed to a single dose of DA by intraperitoneal (i.p.) injection of 0, 0.45, 0.9, and 9.0 mg DA kg(-1) bw. Mortality (66.67 ± 16.67%) was only observed in dose of 9.0 mg kg(-1) bw. Signs of neurological toxicity were detected for the doses of 0.9 and 9.0 mg DA kg(-1) bw. Furthermore, the mean concentrations (±SD) of DA detected by HPLC-UV in extracts of brain after exposure to 9.0 mg DA kg(-1) bw were 0.61 ± 0.01, 0.96 ± 0.00, and 0.36 ± 0.01 mg DA kg(-1) tissue at 1, 2, and 4 hours. The lack of major permanent brain damage in S. aurata, and reversibility of neurotoxic signs, suggest that lower susceptibility to DA or neuronal recovery occurs in affected individuals.     

RESEARCH PAPER: Segmental dorsolumbar epidural analgesia via the caudal approach using multiple port catheters with ketamine or lidocaine or in combination in cattle

ObjectiveTo determine the analgesic and systemic effects of epidural administration of ketamine, lidocaine or a combination of ketamine/lidocaine in standing cattle.Study designProspective, randomized, experimental trial.AnimalsSix healthy male cattle weighing between 335 and 373 kg.MethodsThe animals received 0.5 mg kg^?1 of ketamine (K), 0.2 mg kg^?1 of 2% lidocaine (L) or 0.25 mg kg^?1 ketamine plus 0.1 mg kg^?1 lidocaine (KL). All the drugs were injected into the dorsolumbar epidural space via a caudal approach through a non‐styletted multiple‐port catheter. Each animal received each treatment at random. Evaluations of analgesia, sedation, ataxia, heart rate, arterial pressure, respiratory rate, skin temperature and rectal temperature were obtained at 0 (basal), 5, 10, 15, 30, 45, 60, 75, 90 minutes after epidural injection, and then at 30‐minute intervals until loss of analgesia occurred. Skin temperature was taken at these intervals up to 60 minutes. All the animals received a standard noxious stimulus; a 4‐point scale was used to score the response. A second scale was used to score ataxia and a third for sedation.ResultsThe duration of analgesia in the upper and lower flanks in cattle was 140 ± 15, 50 ± 14 and 80 ± 22 minutes (mean ± SD) after dorsolumbar epidural KL, K or L, respectively. The cardiovascular changes were within acceptable limits in these clinically healthy cattle.ConclusionsDorsolumbar epidural administration of KL to cattle resulted in longer duration of analgesia of the upper and lower flanks in standing conscious cattle, than the administration of K or L alone.Clinical relevanceFurther research is necessary to determine whether this combination using this technique provides sufficient analgesia for flank surgery in standing cattle.     

Guava decline: effect of root exudates from Meloidogyne enterolobii-parasitized plants on Fusarium solani in vitro and on growth and development of guava seedlings under controlled conditions

In guava decline, Fusarium solani-immune guava trees become susceptible to extensive root rot caused by this fungus after parasitism by Meloidogyne enterolobii. To understand the mechanisms involved in this disease, root exudates were collected from nematode-inoculated (NI) or uninoculated (UN) guava plants cultivated in sand. After filtration through a Millipore® membrane, NI and UN exudates were used: i) to prepare media to assess their effect on mycelial growth and production of propagules of F. solani isolate UENF/CF 163, and ii) to incubate macro- and microconidia to assess their effect on germination. NI exudates promoted (P?<?0.05) more mycelial growth and production of propagules than UN exudates or water. NI and UN exudates were used to water guava seedlings laid over seed germination paper inside plastic boxes. Half of the seedlings had an agar plug colonized by the fungus positioned in the collar region. Upon watering with NI exudates the fungus caused (P?<?0.05) extensive rotting of the seedlings’ rootlets. NI and UN exudates, either unlyophilized or lyophilized and re-suspended to the original concentration, were used to water guava seedlings grown in sterile sand before being inoculated (or left uninoculated) as described before. Solely upon watering with NI exudates, in its unlyophilized form or after lyophilization, the fungus caused a reduction (P?<?0.05) of shoot and root biomass associated with rotting of roots. These results suggest that M. enterolobii induces chemical changes in the root exudates of guava trees, which are necessary for root invasion causing root rotting by F. solani.     

Salix rubens and Salix triandra Species as Phytoremediators of Soil Contaminated with Petroleum-Derived Hydrocarbons

The petroleum industry activities provide potential risks to the environment because they can contaminate ecosystems with different organic compounds in the production chain. Several accidents with transport and handling of petroleum and related products occurred in urban areas with harmful effects to the quality of life and economy. In the 1990s, bioremediation and phytoremediation technologies as economically feasible alternatives to repair the environmental damage were developed. In this study, the potential of the willows Salix rubens and Salix triandra were evaluated with regard to the phytoremediation of soils contaminated with petroleum-derived hydrocarbons (total hydrocarbons and polycyclic aromatic hydrocarbons (PAHs)). The PAHs were quantified by extraction from soils and plants using dichloromethane under ultrasonication. The HPLC analysis was performed with GC/MSD equipment. The total hydrocarbons present in uncontaminated soil were quantified by the sum of animal/vegetable oils and greases and mineral oils and greases according to Standard Methods 5520 (1997). The two willows species S. rubens and S. triandra were resistant during the project development. In the contaminated soil, in which both species were planted, the total hydrocarbons concentration was reduced near 98?%. The PAHs content was remarkably reduced as well. Pyrene showed an initial concentration of 23.06???g?kg^?1, decreasing in most cases to 0.1???g?kg^?1 or to undetectable levels. Chrysene decreased from 126.27???g?kg^?1 to undetectable levels. Benzo[k]fluoranthene and benzo[a]pyrene concentrations had also showed a decrease from 28.44 and 3.82???g?kg^?1, respectively, to undetectable levels.