An insight into the sialotranscriptome of the brown dog tick, Rhipicephalus sanguineus

Background: Rhipicephalus sanguineus, known as the brown dog tick, is a common ectoparasite of domestic dogs and can be found worldwide. R. sanguineus is recognized as the primary vector of the etiological agent of canine monocytic ehrlichiosis and canine babesiosis. Here we present the first description of a R. sanguineus salivary gland transcriptome by the production and analysis of 2,034 expressed sequence tags (EST) from two cDNA libraries, one consctructed using mRNA from dissected salivary glands from female ticks fed for 3-5 days (early to mid library, RsSGL1) and the another from ticks fed for 5 days (mid library, RsSGL2), identifying 1,024 clusters of related sequences. Results: Based on sequence similarities to nine different databases, we identified transcripts of genes that were further categorized according to function. The category of putative housekeeping genes contained similar to 56% of the sequences and had on average 2.49 ESTs per cluster, the secreted protein category contained 26.6% of the ESTs and had 2.47 EST's/clusters, while 15.3% of the ESTs, mostly singletons, were not classifiable, and were annotated as ""unknown function"". The secreted category included genes that coded for lipocalins, proteases inhibitors, disintegrins, metalloproteases, immunomodulatory and antiinflammatory proteins, as Evasins and Da-p36, as well as basic-tail and 18.3 kDa proteins, cement proteins, mucins, defensins and antimicrobial peptides. Comparison of the abundance of ESTs from similar contigs of the two salivary gland cDNA libraries allowed the identification of differentially expressed genes, such as genes coding for Evasins and a thrombin inhibitor, which were over expressed in the RsSGL1 (early to mid library) versus RsSGL2 (mid library), indicating their role in inhibition of inflammation at the tick feeding site from the very beginning of the blood meal. Conversely, sequences related to cement (64P), which function has been correlated with tick attachment, was largely expressed in the mid library. Conclusions: Our survey provided an insight into the R. sanguineus sialotranscriptome, which can assist the discovery of new targets for anti-tick vaccines, as well as help to identify pharmacologically active proteins.

Evaluation of gastrointestinal motility in awake rats: a learning exercise for undergraduate biomedical students

Souza MA, Souza MH, Palheta RC Jr, Cruz PR, Medeiros BA, Rola FH, Magalhaes PJ, Troncon LE, Santos AA. Evaluation of gastrointestinal motility in awake rats: a learning exercise for undergraduate biomedical students. Adv Physiol Educ 33: 343-348, 2009; doi: 10.1152/advan.90176.2008.-Current medical curricula devote scarce time for practical activities on digestive physiology, despite frequent misconceptions about dyspepsia and dysmotility phenomena. Thus, we designed a hands-on activity followed by a small-group discussion on gut motility. Male awake rats were randomly submitted to insulin, control, or hypertonic protocols. Insulin and control rats were gavage fed with 5% glucose solution, whereas hypertonic-fed rats were gavage fed with 50% glucose solution. Insulin treatment was performed 30 min before a meal. All meals (1.5 ml) contained an equal mass of phenol red dye. After 10, 15, or 20 min of meal gavage, rats were euthanized. Each subset consisted of six to eight rats. Dye recovery in the stomach and proximal, middle, and distal small intestine was measured by spectrophotometry, a safe and reliable method that can be performed by minimally trained students. In a separate group of rats, we used the same protocols except that the test meal contained (99m)Tc as a marker. Compared with control, the hypertonic meal delayed gastric emptying and gastrointestinal transit, whereas insulinic hypoglycemia accelerated them. The session helped engage our undergraduate students in observing and analyzing gut motor behavior. In conclusion, the fractional dye retention test can be used as a teaching tool to strengthen the understanding of basic physiopathological features of gastrointestinal motility.

Observations on Antricola ticks: Small nymphs feed on mammalian hosts and have a salivary gland structure similar to ixodid ticks

Ticks use bloodmeals as a Source of nutrients and energy to molt and survive until the next meal and to oviposit, in the case of females. However, only the larvae of some tick species are known to feed upon bats females are obligatorily autogenous, and nymphal stages are believed to not feed. We investigated the presence of blood ill a natural population of nymphal Antricola delacruzi ticks collected from bat guano; their ability to feed upon laboratory hosts: and the microscopic structure of both salivary glands and gut. DNA amplification of gut contents of freshly collected material was positive for a mammal in 4 of 11 first instar nymphs, but we were unsuccesful in the amplification of host bloodmeal DNA from late instar nymphs. All early nymphal stages (n = 10) fed oil rabbits. and host DNA was detected and sequenced from gut contents. However, all the large nymphs (n = 10) rejected feeding, and host DNA remained undetected in these ticks. All stages of A. delacruzi have salivary glands similar in morphology to the ixodid agranular Type I salivary gland acini and to granular Type II or Type B acini. All stages of A. delacruzi had a similar gut structure. consisting of digestive cells in the basal portion that contained hematin granules. Neither regenerative nor secretory cell traces were observed in the sections Of gut.

Characterization of feather-degrading bacteria from Brazilian soils

Studies on keratinolytic microorganisms have been mainly related to their biotechnological applications and association with animal pathologies. However, these organisms have an ecological relevance to recycling keratinous residues in nature. This work aimed to select and identify new culturable feather-degrading bacteria isolated from soils of Brazilian Amazon forest and Atlantic forest. Bacteria that were isolated from temperate soils and bacteria from Amazonian basin soil were tested for their capability to grow on feather meal agar (FMA). Proteolytic bacteria were tested for feather degradation and were further identified according to their morphological and biochemical characteristics. Also, molecular identification based on 165 rDNA gene sequencing was carried out. A total of 24 proteolytic and 20 feather-degrading isolates were selected; Most of the isolates were from the Bacillus genus (division Firmicutes), but one Aeromonas, two Serratia (gamma-Proteobacteria), and one Chryseobacterium (Cytophaga-Flavobacterium group). (C) 2010 Elsevier Ltd. All rights reserved.

A new approach for in vitro bioassay to measure tannin biological effects based on a gas production technique

The aim of this paper was to study a method based on gas production technique to measure the biological effects of tannins on rumen fermentation. Six feeds were used as fermentation substrates in a semi-automated gas method: feed A - aroeira (Astronium urundeuva); feed B - jurema preta (Mimosa hostilis), feed C - sorghum grains (Sorghum bicolor); feed D - Tifton-85 (Cynodon sp.); and two others prepared mixing 450 g sorghum leaves, 450 g concentrate (maize and soybean meal) and 100 g either of acacia (Acacia mearnsii) tannin extract (feed E) or quebracho (Schinopsis lorentzii) tannin extract (feed F) per kg (w:w). Three assays were carried out to standardize the bioassay for tannins. The first assay compared two binding agents (polyethylene glycol - PEG - and polyvinyl polypirrolidone - PVPP) to attenuate the tannin effects. The complex formed by PEG and tannins showed to be more stable than PVPP and tannins. Then, in the second assay, PEG was used as binding agent, and this assay was done to evaluate levels of PEG (0, 500, 750, 1000 and 1250 mg/g DM) to minimize the tannin effect. All the tested levels of PEG produced a response to evaluate tannin effects but the best response was for dose of 1000 mg/g DM. Using this dose of PEG, the final assay was carried out to test three compounds (tannic acid, quebracho extract and acacia extract) to establish a curve of biological equivalent effect of tannins. For this, five levels of each compound were added to I g of a standard feed (Lucerne hay). The equivalent effect showed not to be directly related to the chemical analysis for tannins. It was shown that different sources of tannins had different activities or reactivities. The curves of biological equivalence can provide information about tannin reactivity and its use seems to be important as an additional factor for chemical analysis. (C) 2007 Elsevier B.V. All rights reserved.

Sequential spectrofluorimetric determination of free and total glycerol in biodiesel in a multicommuted flow system

A new procedure for spectrofluorimetric determination of free and total glycerol in biodiesel samples is presented. It is based on the oxidation of glycerol by periodate, forming formaldehyde, which reacts with acetylacetone, producing the luminescent 3,5-diacetyl-1,4-dihydrolutidine. A flow system with solenoid micro-pumps is proposed for solution handling. Free glycerol was extracted off-line from biodiesel samples with water, and total glycerol was converted to free glycerol by saponification with sodium ethylate under sonication. For free glycerol, a linear response was observed from 5 to 70 mg L(-1) with a detection limit of 0.5 mg L(-1), which corresponds to 2 mg kg(-1) in biodiesel. The coefficient of variation was 0.9% (20 mg L(-1), n = 10). For total glycerol, samples were diluted on-line, and the linear response range was 25 to 300 mg L(-1). The detection limit was 1.4 mg L(-1) (2.8 mg kg(-1) in biodiesel) with a coefficient of variation of 1.4% (200 mg L(-1), n = 10). The sampling rate was ca. 35 samples h(-1) and the procedure was applied to determination of free and total glycerol in biodiesel samples from soybean, cottonseed, and castor beans.

Breastfeeding, sleep and wake circadian rhythms show distinct temporal emerging patterns

The emerging patterns of breastfeeding, sleep and wake circadian rhythms in an infant and the breastfeeding emergence pattern of his elder sister are presented. Both children were raised under regular contact with photic and non-photic Zeitgebers. Data are related to the first four months of life of the infants, which correspond to the exclusive and ad libitum breastfeeding stage of their lives. Discrimination is made of fasting-associated-wakefulness (FAW) which is a wake state without feeding. Our data show that while FAW episodes are concentrated in the diurnal phase of the day since the first week of life, breastfeeding rhythm takes longer to show statistically significant circadian periodicity (four weeks) and to become monophasic, concentrated in the diurnal phase of the day (three/four months). This precedence of the consolidation of FAW rhythm indicates tight association between nocturnal sleep fragmentation and the drive to feed, in the first months of life of infants.

Vegetable Oil Quenchants: Calculation and Comparison of The Cooling Properties of a Series of Vegetable Oils

The compositions of canola, soybean, corn, cottonseed and sunflower oils suggest that they exhibit substantially different propensity for oxidation following the order of Canola < corn < cottonseed < sunflower approximate to soybean. These data suggest that any of the vegetable oils evaluated could be blended with minimal impact on viscosity although compositional differences would surely affect oxidative stability. Cooling curve analysis showed that similar cooling profiles were obtained for different vegetable oils. Interestingly, no film boiling or transition nucleate boiling was observed with any of the vegetable oils and heat transfer occurs only by pure nucleate boiling and convection. High-temperature cooling properties of vegetable oils are considerable faster than those observed for petroleum oil-based quenchants. (C)2010 Journal of Mechanical Engineering. All rights reserved.

VEGETABLE OIL STRUCTURE AND ANTIOXIDANTS

Five vegetable oils: canola, soybean, corn, cottonseed and sunflower oils were characterized with respect to their composition by gas chromatography and viscosity. The compositions of the vegetable oils suggest that they exhibit substantially different propensity for oxidation following the order of: canola < corn < cottonseed < sunflower approximate to soybean. Viscosities at 40 degrees C and 100 degrees C and the viscosity index (VI) values were determined for the vegetable oils and two petroleum oil quenchants: Microtemp 157 (a conventional slow oil) and Microtemp 153B (an accelerated or fast oil). The kinematic viscosities of the different vegetable and petroleum oils at 40 degrees C were similar. The VI values for the different vegetable oils were very close and varied between 209-220 and were all much higher than the VI values obtained for Microtemp 157 (96) and Microtemp 153B (121). These data indicate that the viscosity variations of these vegetable oils are substantially less sensitive to temperature variation than are the parafinic oil based Microtemp 157 and Microtemp 153B. Although these data suggest that any of the vegetable oils evaluated could be blended with minimal impact on viscosity, the oxidative stability would surely be substantially impacted. Cooling curve analysis was performed on these vegetable oils at 60 degrees C under non-agitated conditions. These results were compared with cooling curves obtained for Microtemp 157, a conventional, unaccelerated petroleum oil, and Microtemp 153B, an accelerated petroleum oil under the same conditions. The results showed that cooling profiles of the different vegetable oils were similar as expected from the VI values. However, no boiling was observed wit any of the vegetable oils and heat transfer occurs only by convection since there is no full-film boiling and nucleate boiling process as typically observed for petroleum oil quenchants, including those of this study. Therefore, high-temperature cooling is considerable faster for vegetable oils as a class. The cooling properties obtained suggest that vegetable oils would be especially suitable fur quenching low-hardenability steels such as carbon steels.

A generalized multi-period mean-variance portfolio optimization with Markov switching parameters

In this paper, we deal with a generalized multi-period mean-variance portfolio selection problem with market parameters Subject to Markov random regime switchings. Problems of this kind have been recently considered in the literature for control over bankruptcy, for cases in which there are no jumps in market parameters (see [Zhu, S. S., Li, D., & Wang, S. Y. (2004). Risk control over bankruptcy in dynamic portfolio selection: A generalized mean variance formulation. IEEE Transactions on Automatic Control, 49, 447-457]). We present necessary and Sufficient conditions for obtaining an optimal control policy for this Markovian generalized multi-period meal-variance problem, based on a set of interconnected Riccati difference equations, and oil a set of other recursive equations. Some closed formulas are also derived for two special cases, extending some previous results in the literature. We apply the results to a numerical example with real data for Fisk control over bankruptcy Ill a dynamic portfolio selection problem with Markov jumps selection problem. (C) 2008 Elsevier Ltd. All rights reserved.

Apparent digestibility coefficients of selected feed ingredients for dourado Salminus brasiliensis

The excellence of its flesh and fast growth makes the dourado, Salminus brasiliensis, a carnivorous fish native to the Prata basin, a potential candidate for intensive fish farming. This study evaluated the apparent digestibility coefficients (ADC) of energy and nutrients of animal and plant protein sources for the carnivorous Characin dourado S. brasiliensis. Fish (19.5 +/- 5.0 g) were stocked in plastic cages (80-L) and fed pelleted test diets containing 30% of the test ingredient [fish meal (FM), poultry by-product meal (PBM), soybean meal (SBM), and corn gluten meal (CGM) plus 70% of a reference diet (481.4 g kg(-1) of crude protein and 18.63 kJ of gross energy per gram]. After the last daily meal, cages were transferred to cylindrical, conical-bottomed aquarium (200-L) where faeces were collected by sedimentation in a refrigerated container. Except for ADC of protein and energy, all other ADC of nutrients showed significant differences (P < 0.01). ADC values were: 94.3%, 91.3%, 93.1%, and 93.5% for crude protein; 91.0%, 90.3%, 87.8%, and 88.8% for gross energy; 92.1%, 84.5%, 80.6%, and 79.3% for ash; 83.9%, 80.3%, 84.3%, and 84.6% for dry matter; 97.4%, 96.7%, 93.3%, and 91.5% for lipid for FM, PBM, SBM, and CGM, respectively. The average amino acid ADC was: 93.6%, 90.0%, 92.1%, and 92.5% of FM, PBM, SBM, and CGM, respectively. All test ingredients were efficiently used and can partially replace FM in diets for carnivorous dourado.

Fatty acid profile and qualitative characteristics of meat from zebu steers fed with different oilseeds

The objective of this study was to evaluate the effects of dietary addition of ground oilseed sources on the quality, fatty acid profile, and CLA content of meat from zebu steers. Thirty-one zebu steers with an initial average age of 23 mo and an initial BW of 365 kg were used in this study. The experimental period was 84 d, which was preceded by an adaption period of 28 d. The diet was provided ad libitum with a forage: concentrate ratio of 40:60. Corn silage was used as the forage source. Four different concentrates were formulated for each treatment: without additional lipids (control) or with ground soybeans (SB), ground cottonseed (CS), or ground linseed (LS). The SB, CS, and LS diets were formulated to have 6.5% ether extract on a total dietary DM basis. The experiment was set up as a completely randomized design. After slaughter, samples were taken from the longissimus thoracis muscle for the measurement of fatty acid concentration and the evaluation of meat quality. The luminosity index was greater in the control and LS diets (P < 0.01). The greatest percentages of myristic acid (C14:0), palmitic acid (C16:0), trans octadecenoic acid (C18:1 trans-10, trans-11, or trans-12), and SFA in the subcutaneous fat were observed in the CS treatment (P < 0.01). Moreover, the least percentages of oleic acid (C18:1 cis-9) and total unsaturated fatty acids in the subcutaneous fat were observed in the CS diet (P < 0.01). The meat linoleic acid and a-linolenic acid percentages were greatest in the SB and LS treatments, respectively (P < 0.001). The unsaturated fatty acid: SFA ratio was smallest for the CS diet (P < 0.01). A gradual increase in oxidation was observed as a function of storage time; however, the diets did not affect the rancidity of the meat (P > 0.05). The fatty acid profile of subcutaneous fat was impaired by the addition of CS. Supplying ground oilseeds did not increase the content of CLA in the meat.

Digestibility of plant protein-based diets by largemouth bass Micropterus salmoides

Commercial farming of carnivorous fish demands the reduction of environmental impact of feeds; that requires minimal use of dietary animal protein. This study investigated the digestibility of diets formulated exclusively out of plant protein, added feed attractants, by the carnivore largemouth bass, Micropterus salmoides. Juvenile largemouth bass (14.0 +/- 1.0 cm) conditioned to accept artificial, dry feed were confined in polypropylene cages and fed ad libitum in three daily meals, seven experimental diets containing varying levels of vegetable and animal protein sources, added of different feed stimulants. After last daily meal, cages were transferred to cylindrical-conical-bottomed, 200-L aquaria, where faeces were collected by sedimentation into refrigerated containers, preserved and later analysed for chemical composition. Soybean meal can be used as partial substitute of animal protein in diets for largemouth bass; the poultry by-product meal shows as a good option as animal protein source in these rations. Control treatment - 50PP : 50AP - yielded best performances; the need for the use of fish meal in the formulation for carnivorous diets is, at least, questionable. Results of the digestibility trials demonstrated the importance of determining the diet digestibility, if precision in the formulation of least-cost feeds for carnivorous fish is the ultimate goal.

Apparent digestibility coefficient of protein and amino acids of acid, biological and enzymatic silage for Nile tilapia (Oreochromis niloticus)

Using the fish silage to partially replace proteic feedstuff in aquafeeds is an alternative to mitigate sanitary and environmental problems caused by the lack of adequate destination for fisheries residues. It would also lower feed costs, consequently improving fish culture profitability. However, using fish silages in aquafeeds depends on determination of its apparent digestibility coefficients (ADC). This work aimed to determining the ADC of crude protein and amino acids of acid silage (AS), biological silage (BS) and enzymatic silage (ES) for juvenile Nile tilapia (94.5 +/- 12.7 g). The ADC(CP) was: 92.0%, 89.1% and 93.7% for AS, BS and SE respectively. The average ADC of amino acids was: 91.8%, 90.8% and 94.6% for AS, BS and ES respectively. Results encourage the use of AS, BS and ES to partially replace protein sources in balanced diets for neotropical fish.

Prerequisite Programs at Schools: Diagnosis and Economic Evaluation

In this study, 20 Brazilian public schools have been assessed regarding good manufacturing practices and standard sanitation operating procedures implementation. We used a checklist comprised of 10 parts ( facilities and installations, water supply, equipments and tools, pest control, waste management, personal hygiene, sanitation, storage, documentation, and training), making a total of 69 questions. The implementing modification cost to the found nonconformities was also determined so that it could work with technical data as a based decision-making prioritization. The average nonconformity percentage at schools concerning to prerequisite program was 36%, from which 66% of them own inadequate installations, 65% waste management, 44% regarding documentation, and 35% water supply and sanitation. The initial estimated cost for changing has been U.S.$24,438 and monthly investments of 1.55% on the currently needed invested values. This would result in U.S.$0.015 increase on each served meal cost over the investment replacement within a year. Thus, we have concluded that such modifications are economically feasible and will be considered on technical requirements when prerequisite program implementation priorities are established.