Characterization and application of recombinant β-glucosidase (BglH) from <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> KCTC 1918

β-Glucosidase (β-1,4-D-glucoside glucohydrolase: EC.3.2.1.21) catalyzes the hydrolysis of β-glucosidic bonds between saccharides and aryl or alkyl groups. A gene encoding β-glucosidase from Bacillus licheniformis KCTC 1918, an anaerobic spore-forming soil bacterium, was cloned and characterized. The structural gene for the β-glucosidase consists of 1410 bp encoding 469 amino acid residues, and has a molecular weight of 53.4 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis with 12% separating gel. The enzyme activity was determined against pNPG as a substrate. The enzyme was optimally active at pH 6.0 (citrate-phosphate buffer) and 47°C. β-Glucosidase retained 100% of its original activity for 24 h. The activity of the enzyme was stimulated by glycerol and urea and was decreased by Ca²⁺, Cu²⁺, Hg²⁺, Mg²⁺, and Mn²⁺. In particular, Cu²⁺ had the strongest negative effect on β-glucosidase activity. The purified β-glucosidase was active against pNPG and cellobiose. When the β-glucosidase was tested for cellulose hydrolysis, the supplement of β-glucosidase with cellulose increased the glucose yield from pine wood powder by 139.8%.     

Dietary L-Carnitine Supplementation Protected Turbot (Scophthalmusmaximus) Against Streptococcus parauberis Infection

The protective effect of dietary L-carnitine supplementation was accessed in turbot (Scophthalmusmaximus) challenged with Streptococcus parauberis. Infected fish fed 240mg L-carnitine showed significantly (P < 0.01) higher survival (78%) than infected fish fed 40mg L-carnitine (11%). When dietary L-carnitine supplementation was suspended, a significant increase in mortality was observed in infected fish. The present report documents that dietary L-carnitine fed on a continuous basis may act as an immunostimulant in turbot.     

RETROSPECTIVE COMPARISON OF THREE‐DIMENSIONAL CONFORMAL RADIATION THERAPY VS. PREDNISOLONE ALONE IN 30 CASES OF CANINE INFRATENTORIAL BRAIN TUMORS

Infratentorial tumors are relatively infrequent in dogs and a lack of data makes it difficult to offer prognostic information. Untreated, dogs with these neoplasms have shorter survival times than those with supratentorial tumors. The role of radiation therapy (RT) in the management of infratentorial tumors is poorly defined and tumoral/peritumoral swelling in this site is a potential cause of serious acute side effects. The aim of this retrospective, cohort study was to describe cases of infratentorial tumors treated with fractionated three‐dimensional conformal RT (3D CRT) and glucocorticoids (GC), and compare outcomes and survival with dogs affected by tumors in the same location that received GC alone. Thirty patients with a MRI diagnosis of infratentorial tumors were recruited (15 received RT and GC and 15 GC alone). None had mentation changes at presentation. For both groups, MRI and medical records were reviewed; and factors associated with survival were evaluated with Kaplan–Meier product limit survival and Cox regression analysis. Overall median survival time (MST) was 294 days (95% CI 42–545). The MST in the RT group was 756 days (95% CI 209–1302) vs. 89 days (95% CI 34.7–143.3 days) for those dogs treated palliatively with GC alone. This difference was statistically significant (P = 0.001). No other factors (including neurological signs, MRI features, tumor volume and total RT dose) were statistically associated with survival in the RT group. This study suggests that 3D CRT offers a survival advantage for dogs with infratentorial tumors compared to GC alone, and significant complications are uncommon.     

Effect of provision of apiaceous flowers associated to foods on the biology of <Emphasis Type="Italic">Coleomegilla maculata</Emphasis>

Coleomegilla maculata DeGeer is an omnivorous lady beetle that feeds on natural or alternative prey and artificial foods, which allows its laboratory rearing for use on augmentative biocontrol. In addition, C. maculata may supplement its diet with pollen and nectar, which helps in its conservation in agroecosystems. This study aimed to evaluate if Apiaceous flowers (Anethum graveolens L. and Coriadrum sativum L.), with and without alternative prey [eggs of Ephestia kuehniella (Zeller) or larvae of Drosophila melanogaster Meigen] or an artificial food (aqueous solution of honey), may guarantee the survival and complete development of the immature and adult stages of C. maculata in the laboratory. The immature stages developed only when Apiaceous flowers were offered with E. kuehniella eggs. The food with only one of the alternative prey (moth eggs or fly larvae) or moth eggs + honey solution resulted in fertile adults; however, the number of eggs/cluster was greater for the foods with E. kuehniella eggs + honey solution, A. graveolens flowers, or only D. melanogaster. Foods comprising only the two Apiaceous species, only the honey solution, or only water resulted in larval development up to a specific instar. Adults of C. maculata also survived on these foods, but there was no oviposition. The foods of the two Apiaceous species produced heavier adults only when associated with E. kuehniella eggs. The results indicate that the zoophytophagous habit of C. maculata should be considered in conservation biocontrol programs aimed at using this lady beetle to control crop pests.     

Epidemiological aspects of Phyllosticta citricarpa colonization and viability in Citrus sinensis

Journal of Plant Diseases and Protection - Citrus black spot (CBS) is a disease caused by the fungus Phyllosticta citricarpa that is associated with significant yield losses. The pathogen forms...     

Gastrointestinal nematode infection in beef cattle raised in silvopastoral and conventional systems in São Paulo state,Brazil

The use of silvopastoral systems (SPS) can be a good alternative to reduce the environmental impacts of livestock breeding in Brazil. Despite the advantages offered by public policies, many producers hesitate to use this system. One of the reasons is the lack of information on health and productivity of cattle raised under these conditions. The experiment reported here was designed to compare the behavior of infection by gastrointestinal nematodes and weight gain of beef cattle raised in a SPS and a conventional pasture system. We monitored the number of eggs per gram of feces, the prevalent nematode genus, data on climate, forage availability, weight gain and packed cell volume (PCV) of the animals bred in the two systems. The infection by nematodes was significantly higher in the cattle raised in the SPS (p < 0.05). The coprocultures revealed the presence of nematodes of the genera Haemonchus, Cooperia, Oesophagostomum and Trichostrongylus, in both systems, but the mean infestation rates of Haemonchus and Cooperia were higher in the SPS (p < 0.05). The average of PCV values did not differ between the cattle in the two systems. The individual weight gain and stocking rate in the period did not vary between the systems (p > 0.05). Despite the higher prevalence of nematodes in the SPS, no negative impact was detected on the animals’ weight gain and health. The results of this experiment indicate that under the conditions studied, there is no need to alter the parasite management to assure good productive performance of cattle.     

Digestion and metabolism of low and high residual feed intake Nellore bulls

Understanding the reasons why animals of similar performances have different feed requirements is important to increase profits for cattle producers and to decrease the environmental footprint of beef cattle production. This study was carried out aiming to identify the associations between residual feed intake (RFI) and animal performance, nutrient digestibility, and blood metabolites related to energy balance of young Nellore bulls during the finishing period. Animals previously classified as low (n?=?13) and high RFI (n?=?12), with average initial body weight of 398 kg and age of 503 days were used. Cattle were fed a high energy diet and were slaughtered when rib fat thickness measured by ultrasound between the 12th and 13th ribs reached the minimum of 4 mm. A completely randomized design was adopted, being data analyzed with a mixed model that included the random effect of slaughter group, the fixed effect of RFI class, and linear effect of the covariate feedlot time. No differences were found (p?>?0.10) between RFI classes for performance, dry matter, and nutrients intake. However, dry (p?=?0.0911) and organic matter (p?=?0.0876) digestibility tended to be lower, and digestibility of neutral detergent fiber corrected for ash and protein (p?=?0.0017), and total digestible nutrients (p?=?0.0657) were lower for high RFI animals, indicating lesser capacity of food utilization. Difference between low and high RFI animals was also found for blood cortisol at the end of the trial (p?=?0.0044), having low RFI animals lower cortisol concentrations. Differences in the ability to digest food can affect the efficiency of transforming feed into meat by Nellore cattle.     

Improvement of Fertility in Artificially Inseminated Ewes Following Vaginal Treatment with Misoprostol Plus Terbutaline Sulphate

The effect of vaginal administration of misoprostol plus terbutaline sulphate 6 h prior to artificial insemination (AI) upon the site of AI (vaginal or cervical) and fertility was studied using a total of 87 estrous synchronized Serra da Estrela ewes (control n = 42 and treated n = 45). Artificial insemination was performed using refrigerated semen at 54–55 h after sponge removal. Lambing rate (fertility) and prolificacy were compared between control and treated ewes. The effect of the site of semen deposition on fertility was also evaluated. Prolificacy rate was not different between control (1.5) and treated (1.59) ewes. The proportion of cervical AI achieved in control (45.2%) and treated (37.8%) ewes was not significantly different. Overall, fertility was significantly lower in control than in treated ewes (42.9% vs 64.4%; p < 0.04). Fertility following vaginal AI was significantly lower for control for than treated ewes (30.4% vs 60.7%; p < 0.03) but the difference was smaller and not significant for cervical AI (control 57.9% vs 70.6%). It was concluded that vaginal administration of misoprostol plus terbutaline sulphate 6 h prior to artificial insemination did not affect the proportion of cervical inseminations but significantly improved the fertility of treated ewes. Although needing confirmation, it was hypothesized that drugs might have induced local secretory modifications leading to an increase of cervical ability to retain more viable spermatozoa for fertilization.     

Nutritional evaluation of “Braquiarão” grass in association with “Aroeira” trees in a silvopastoral system

This study aimed at evaluating the influence of a silvopastoral system (SPS) composed of “Aroeira” trees (Myracrodruon urundeuva) on the nutritional value of Brachiaria brizantha cv. Marandu forage grass over the rainy season. The experiment was conducted in the SPS located in the “Cerrado” biome in Lagoa Santa County, Minas Gerais state, Brazil, in the agricultural year of 2004–2005. It was evaluated the influence of “Aroeira”, whose density in the area was 140 trees/ha, on the following parameters of the B. brizantha cv. Marandu forage grass: forage mass harvest above stubble height, morphogenic attributes, bromatological composition and nutritional value. The effects of “Aroeira” trees on the micro-climatic conditions were also evaluated. The experiment adopted a randomized block design with five blocks (cutting seasons), two treatments (a shaded pasture—B. brizantha cv. Marandu shaded by M. urundeuva—and a pasture in full sun—B. brizantha cv. Marandu in full sun), and three replicates. The tree component, composed of the tree species M. urundeuva in the experimental density adopted, increased dry matter and crude protein of the forage mass harvest above stubble height, without affecting the concentration of neutral detergent fibre or acid detergent fibre, thus causing dry matter and organic matter in vitro degradability as well as effective degradability of the forage grass to remain the same as those obtained in full sun. Therefore, the results of this study show the possibility of using trees and grass to increase both animal production and environmental services.     

Direct penetration of Rhizopus stolonifer into stone fruits causing rhizopus rot

Rhizopus rot, caused by Rhizopus stolonifer, is a major postharvest disease of stone fruits. The disease is related to the occurrence of mechanical and physical damage; however, observations at a Brazilian wholesale market suggest that direct penetration can occur. Therefore, the penetration mechanisms of R. stolonifer in stone fruits were evaluated. To identify the production of enzymes that help with direct penetration by the pathogen, esterase activity, both in mycelial discs and in spore suspensions of the fungus in water and in modified Van Etten nutrient solution, was measured. Assays were also conducted to evaluate the growth of R. stolonifer on glucose or cutin as a sole carbon source. The pathogen grew on both media, and higher esterase activity was observed in the cutin medium. Wounded and unwounded peaches and nectarines were inoculated with R. stolonifer spore suspensions in water or in modified Van Etten nutrient solution. Wounded fruit inoculated with either of the R. stolonifer spore suspensions developed rhizopus rot, whereas unwounded fruit developed the rot only in the presence of spores in the modified Van Etten nutrient solution. Scanning electron and light microscopic examination showed the fungus can directly penetrate the nectarine cuticle. Diisopropyl fluorophosphate, a serine hydrolase inhibitor, prevented rot development in peaches. The results provide valuable evidence for the ability of R. stolonifer to directly penetrate unwounded stone fruits, probably due to the production of esterase enzymes.