Rare earth elements in biology: From biochemical curiosity to solutions for extractive industries.

Rare earth elements (REEs) are critical for our modern lifestyles and the transition to a low-carbon economy. Recent advances in our understanding of the role of REEs in biology, particularly methylotrophy, have provided opportunities to explore biotechnological innovations to improve REE mining and recycling. In addition to bacterial accumulation and concentration of REEs, biological REE binders, including proteins (lanmodulin, lanpepsy) and small molecules (metallophores and cofactors) have been identified that enable REE concentration and separation. REE-binding proteins have also been used in several mechanistically distinct REE biosensors, which have potential application in mining and medicine. Notably, the role of REEs in biology has only been known for a decade, suggesting their considerable scope for developing new understanding and novel applications.

Non-covalent binding tags for batch and flow biocatalysis.

Enzyme immobilization offers considerable advantage for biocatalysis in batch and continuous flow reactions. However, many currently available immobilization methods require that the surface of the carrier is chemically modified to allow site specific interactions with their cognate enzymes, which requires specific processing steps and incurs associated costs. Two carriers (cellulose and silica) were investigated here, initially using fluorescent proteins as models to study binding, followed by assessment of industrially relevant enzyme performance (transaminases and an imine reductase/glucose oxidoreductase fusion). Two previously described binding tags, the 17 amino acid long silica-binding peptide from the Bacillus cereus CotB protein and the cellulose binding domain from the Clostridium thermocellum, were fused to a range of proteins without impairing their heterologous expression. When fused to a fluorescent protein both tags conferred high avidity specific binding with their respective carriers (low nanomolar Kd values). The CotB peptide (CotB1p) induced protein aggregation in the transaminase and imine reductase/glucose oxidoreductase fusions when incubated with the silica carrier. The Clostridium thermocellum cellulose binding domain (CBDclos) allowed immobilization of all the proteins tested, but immobilization led to loss of enzymatic activity in the transaminases (< 2-fold) and imine reductase/glucose oxidoreductase fusion (> 80%). A transaminase-CBDclos fusion was then successfully used to demonstrate the application of the binding tag in repetitive batch and a continuous-flow reactor.

Recuperação de larvas infectantes de Trichostrongylus colubriformis em três espécies de gramíneas contaminadas no verão / Recovery of Trichostrongylus colubriformis infective larvae from three grass species contaminated in summer

O experimento teve como objetivo avaliar a recuperação de larvas infectantes (L3) de Trichostrongylus colubriformis em Brachiaria decumbens cv. Australiana, Cynodon dactilon cv. Coast-cross e Panicum maximum cv. Aruana. Foram utilizados módulos experimentais constituídos por seis canteiros de 32,4 m² cada, perfazendo dois canteiros por espécie forrageira. Cada canteiro foi dividido em 36 parcelas, de 30 x 30 cm, de forma a permitir seis repetições por espécie e por altura da forragem em cada semana de colheita de material. A sobrevivência larval foi avaliada do meio do verão e até meados do outono, sob o efeito de duas alturas de poda das forragens: baixa, 5 cm e alta, 30 cm. A poda foi realizada imediatamente antes da deposição das fezes contaminadas com ovos de T. colubriformis, obtidos de ovinos, que ocorreu no dia 05/02/ 2004. A colheita das fezes e da forragem foi realizada uma, duas, quatro, oito, 12 e 16 semanas após a deposição das fezes nos canteiros experimentais. A altura da forragem foi medida em cada uma das subdivisões imediatamente antes da colheita. A forragem foi cortada rente ao solo, de uma área delimitada com o auxílio de um círculo de 10 cm de raio. As fezes foram recolhidas manualmente dos canteiros. O número de larvas infectantes recuperado foi muito pequeno em comparação com a quantidade de larvas produzidas nas culturas controle, mantidas no laboratório (máximo de 6,7 por cento no capim Aruana com 30 cm de altura). Arecuperação de L3 das amostras fecais foi maior quando as fezes foram depositadas em meio ao capim alto (com 30 cm - P<0,05). Porém, a recuperação de L3 das forragens foi similar em ambos os cortes. Em relação à concentração de L3 (número de L3/kg de matéria seca), não houve diferença entre o corte baixo e alto (P>0,05) em nenhuma das semanas experimentais. Dentre as espécies forrageiras, o capim aruana foi o que, no geral, apresentou maiores concentrações de L3 de T. colubriformis.

The purpose of the experiment was to evaluate infective Trichostrongylus colubriformis larvae (L3) survival in three forage species. Experimental plots, planted with Brachiaria decumbens cv. Australian, Cynodon dactilon cv. Coast-cross, and Panicum maximum cv. Aruana, were used in the study, totaling two plots for each species. Each plot (32.4 m²) was divided into 36 subplots (30 x 30 cm) in order to allow six replicates per forage species and per herbage height in each week of material collection. Larval recovery was evaluated from middle summer to middle autumn under the effect of two forage paring heights: low, 5 cm, and high, 30 cm. The paring was carried out immediately before the fecal samples with T. colubriformis eggs, taken from sheep, were deposited on pasture in 05/Feb/2004. Feces and forage collection was performed one, two, four, eight, 12 and 16 weeks after feces deposition in the experimental plots. Forage grass height was measured in each subdivision immediately before the collections. The forage sample was cut, close to the soil, from an area delimited with a circle with a 10-cm radius. The feces were collected from the subplots. The number of infective larvae recovered from pasture was very small in compa6rison with the amount of larvae produced in cultures maintained in laboratory (maximum 6.7 percent on Aruana grass with 30 cm). L3 recovery rates from fecal samples were bigger when the feces were deposited on high grass (measuring 30 cm - P<0.05). L3 recovery from pasture and L3 concentration on herbage (L3/Kg dry matter) were similar for both cuts (P> 0.05). Among the forage species, the Aruana grass was the one that, in general, harbored the biggest concentrations of infective T. colubriformis larvae.

[Recovery of Trichostrongylus colubriformis infective larvae from three grass species contaminated in summer]. / Recuperação de larvas infectantes de Trichostrongylus colubriformis em três espécies de gramíneas contaminadas no verão.

The purpose of the experiment was to evaluate infective Trichostrongylus colubriformis larvae (L3) survival in three forage species. Experimental plots, planted with Brachiaria decumbens cv. Australian, Cynodon dactilon cv. Coast-cross, and Panicum maximum cv. Aruana, were used in the study, totaling two plots for each species. Each plot (32.4 m(2)) was divided into 36 subplots (30 x 30 cm) in order to allow six replicates per forage species and per herbage height in each week of material collection. Larval recovery was evaluated from middle summer to middle autumn under the effect of two forage paring heights: low, 5 cm, and high, 30 cm. The paring was carried out immediately before the fecal samples with T. colubriformis eggs, taken from sheep, were deposited on pasture in 05/Feb/2004. Feces and forage collection was performed one, two, four, eight, 12 and 16 weeks after feces deposition in the experimental plots. Forage grass height was measured in each subdivision immediately before the collections. The forage sample was cut, close to the soil, from an area delimited with a circle with a 10 cm radius. The feces were collected from the subplots. The number of infective larvae recovered from pasture was very small in comparison with the amount of larvae produced in cultures maintained in laboratory (maximum 6.7% on Aruana grass with 30 cm). L3 recovery rates from fecal samples were bigger when the feces were deposited on high grass (measuring 30 cm - P<0.05). L3 recovery from pasture and L3 concentration on herbage (L3/Kg dry matter) were similar for both cuts (P>0.05). Among the forage species, the Aruana grass was the one that, in general, harbored the biggest concentrations of infective T. colubriformis larvae.

[Trichostrongylus colubriformis larvae recovery from different Brachiaria decumbens and Panicum maximum strata]. / Recuperação de larvas de Trichostrongylus colubriformis EM diferentes estratos de Brachiaria decumbens E Panicum maximum.

The purpose of the experiment was to evaluate infective Trichostrongylus colubriformis larvae vertical migration in two forage grass species. Experimental modules formed by eight plots, established with Brachiaria decumbens cv. Australian and Panicum maximum cv. Aruana, were used in the study, totaling four plots for each grass species. Each plot was divided into six 30 x 30 cm subplots. Larval migration was evaluated in the four seasons of the year, in different plant strata (0-7, 7-14, 14-21, 21-28 and above 28 cm). Four feces deposits were made, one in each season of the year, in the middle of 30-cm tall forage. The feces were collected from the forage ten days after each feces deposit in the experimental subplots. Grass height was measured in each of the strata immediately before the collections. The forage of the different strata was cut from an area measuring 10-cm in radius. The feces were collected manually from the subplots. There was a grass species and grass stratum interaction in the deposit made in autumn (P<0.05). During that season, most of the larvae were recovered from the Brachiaria grass base; meanwhile, at the forage apex, the biggest average was registered in the aruana grass. Infective larvae (L3) recovery was similar among the different strata during spring. In springtime, the biggest L3 recovery occurred at the 21-28 cm stratum from both forage species. No L3 was recovered from any of the No L3 was recovered from any of the grass strata during winter and summer. Study results show that migration of T. colubriformis larvae was more influenced by weather conditions than by forage species.

Relationship of intestinal histology with the resistance to Trichostrongylus colubriformis infection in three breeds of sheep

The study was carried out to evaluate the relationship of inflammatory intestinal cells with the resistance to Trichostrongylus colubriformis infections in three breeds of sheep (Santa Ines, Suffolk and Ile de France), naturally infected. Mast cells, eosinophils, and globule leucocytes were enumerated in intestinal mucosa. Histamine concentration was estimated in intestinal tissue samples and the length of male and female specimens were determined. The three breeds of sheep showed similar cellular response in the small intestine mucosa (P>0.05). There was extensive variation among sheep in the parasitological and inflammatory cell variables, even in lambs of the same breed. In general, animals presenting less inflammatory cells had a larger worm burden, higher fecal egg counts, and larger T. colubriformis worms. The inflammatory cells possibly impaired the parasite's establishment, development, and survival.

Avaliaram-se a associação entre o número de células inflamatórias no intestino delgado e a resistência à infecção por Trichostrongylus colubriformis em ovinos de três raças (Santa Inês, Suffolk e Ile de France), naturalmente infectados. Mastócitos, eosinófilos e leucócitos globulares foram quantificados na mucosa intestinal. A concentração de histamina foi estimada em amostras teciduais do intestino, bem como foi determinado o comprimento de machos e fêmeas de T. colubriformis. A resposta celular foi similar na mucosa intestinal das três raças ovinas (P>0,05). Houve grande variação entre os ovinos em relação aos resultados parasitológicos e celulares, mesmo nos animais de mesma raça. Em geral, os animais que apresentaram número menor de células inflamatórias tiveram cargas parasitárias maiores, contagens de ovos por grama de fezes mais altas e exemplares de T. colubriformis maiores. Os resultados indicaram que mastócitos, eosinófilos e leucócitos globulares prejudicaram o estabelecimento, o desenvolvimento e a sobrevivência dos parasitas.

Efficacy of homeopathic treatment against natural infection of sheep by gastrointestinal nematodes.

The efficacy of the homeopathic treatment with the Fator Vermes, administered according to the manufacturer's recommendations, was evaluated against gastrointestinal nematodes infections in sheep. The experiment was divided into two phases: in the first phase (January/06/2004 to April/30/2004), the animals of the treated (n=10) and control (n=10) groups were treated individually with conventional anthelmintics to avoid deaths. In the second phase (April/30/2004 to July/06/2004), the sheep from the group that received the Fator Vermes were treated as they had been in the previous phase, while the control group animals were treated with conventional anthelmintics at 14 day intervals. In the first phase of the experiment, there was no significant difference (P>0.05) between group means regarding egg counts in feces (EPG), weight gain, or packed cell volume (PCV). Meanwhile, in the second phase, the control group sheep had a significantly higher weight gain, higher PCV values, and lower EPG counts. Infective larvae of Haemonchus spp., Trichostrongylus spp., Cooperia spp., and Oesophagostomum spp. were identified in the fecal cultures. After six months of daily treatment with the Fator Vermes, it was not possible to substantiate the product's benefits in both sheep health and productivity or in the prophylaxis of gastrointestinal nematode infections.

Resistance of Santa Inês and Ile de France suckling lambs to gastrointestinal nematode infections.

A trial was carried out to determine the resistance to natural infection by gastrointestinal nematodes in 12 Santa Inês and nine Ile de France lambs before weaning. Faecal samples were obtained for faecal nematode egg counts (FEC). Blood samples were collected to determine packed cell volume (PCV), total plasma protein levels and peripheral eosinophil counts. Most Ile de France lambs (77.8%) were treated with an anthelmintic at 43 days of age, while 50% off Santa Inês lambs were treated at weaning, 57 days of age. The mean PCV values were normal in Santa Inês lambs, while in Ile de France lambs showed lower values reaching 22.3% at 43 days of age. The lowest mean plasma protein values were observed in Ile de France lambs (4.13 g/dl) at 43 days of age and in Santa Inês lambs (5.0 g/dl) at 57 days of age. Before weaning, Santa Inês lambs were susceptible to natural infections by gastrointestinal nematodes but with a greater capacity to stand the adverse effects of parasitism compared to Ile de France lambs.