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An aurantiactinomyxon type is described from the marine naidid Tubificoides pseudogaster (Dahl, 1960), collected from the lower estuary of a Northern Portuguese River. This type constitutes the first of its collective group to be reported from Portugal, and only the fourth described from a marine oligochaete worldwide. Extensive morphological comparisons of new aurantiactinomyxon isolates to all known types without available molecular data are proposed to be unnecessary, given the artificiality of the usage of morphological criteria for actinosporean differentiation and the apparent strict host specificity of the group. Recognition of naidid oligochaetes as the hosts of choice for marine types of aurantiactinomyxon and other collective groups, suggests that the family Naididae played a preponderant role in the myxosporean colonization of estuarine communities. Molecular analyses of the type in study further infer its involvement in the life cycle of Paramyxidium giardi (Cépède, 1906) Freeman and Kristmundsson, 2018, a species that infects the kidney of European eel Anguilla anguilla (Linnaeus, 1758) and that has been reported globally, including from Portuguese waters. The low intraspecific difference registered in relation to Icelandic isolates of P. giardi (0.6%) is hypothesized to result from the emergence of genotypically different subspecies due to geographic isolation.
Chagas disease (CD) is a neglected parasitic condition endemic in the Americas caused by Trypanosoma cruzi. Patients present an acute phase that may or not be symptomatic, followed by lifelong chronic stage, mostly indeterminate, or with cardiac and/or digestive progressive lesions. Benznidazole (BZ) and nifurtimox are the only drugs approved for treatment but not effective in the late chronic phase and many strains of the parasite are naturally resistant. New alternative therapy is required to address this serious public health issue. Repositioning and combination represent faster, and cheaper trial strategies encouraged for neglected diseases. The effect of imatinib (IMB), a tyrosine kinase inhibitor designed for use in neoplasias, was assessed in vitro on T. cruzi and mammalian host cells. In comparison with BZ, IMB was moderately active against different strains and forms of the parasite. The combination IMB + BZ in fixed-ratio proportions was additive. Novel 14 derivatives of IMB were screened and a 3,2-difluoro-2-phenylacetamide (3e) was as potent as BZ on T. cruzi but had low selectivity index. The results demonstrate the importance of phenotypic assays, encourage the improvement of IMB derivatives to reach selectivity and testify to the use of repurposing and combination in drug screening for CD.
Local attitudes towards carnivores often reflect the degree of damage they are perceived to cause. Consequently, understanding the interactions between people and these species is essential to conservation efforts. This study investigated local perceptions of three Cerrado canid species and current chicken management practices, to identify the potential damage they cause and how this relates to peoples’ attitudes towards these species. Results from structured interviews at 50 ranches in Goiás, Brazil, highlighted that general knowledge about Cerrado canids differed significantly by species, with interviewees unable to correctly answer questions about the hoary fox Lycalopex vetulus and crab-eating fox Cerdocyon thous in comparison to the maned wolf Chrysocyon brachyurus. Chicken coops were identified as the most effective method for preventing predation, yet only 44% of respondents employed this method. Using a perceived predation measure, interviewees reported chicken predation by all three Cerrado canids even though most of these events were stated to occur during the day, outside the species’ active periods. Reported predation events were a strong predictor of attitude. Participants who experienced predation events reported they did not like having a Cerrado canid on their property. However, 86% of the respondents agreed that Cerrado canids should nevertheless be protected. Our findings support the need to incorporate the human dimension in canid and broader carnivore conservation issues.
Intake, digestibility, nitrogen (N) balance, microbial protein synthesis, weight gain, yields of the main commercial cuts and carcass morphometric measurements were evaluated in lambs fed diets containing different levels of chitosan. Sixty Santa Inês crossbred sheep with an average body weight (BW) of 24 ± 2.2 kg were assigned to three treatments (diets containing 0, 136 or 272 mg chitosan/kg BW) in a completely randomized design. There was no effect of chitosan on dry matter (DM) intake. Ingested and retained N showed a quadratic response, with the highest values estimated at the chitosan levels of 142 and 152 mg/kg BW, respectively. Similar to N balance, microbial protein synthesis showed the same quadratic response, in which the level of 136 mg/kg BW resulted in higher synthesis when compared with the other levels. No effect of chitosan was detected on average daily gain, final weight, or carcass variables (hot carcass weight, cold carcass weight, yield of commercial cuts and morphometric measurements of the carcass). Conformation, visceral fat content and fatness of carcasses were also not altered by the use of chitosan. Chitosan improves the digestibility of DM, crude protein and neutral detergent fibre, and increases N balance and microbial protein synthesis but does not change the production performance of feedlot lambs.
Mullets inhabit a wide range of habitats from tropical to temperate regions and play a critical role in their ecosystems. This commercially important fish group constitutes a significant source of food in several geographic regions, and the production of some species for consumption is an increasing trend. About 64 myxosporean species have been reported in mullets, some of which are cryptic, as is the case of Myxobolus exiguus, and M. muelleri. This paper provides, for the first time, a detailed and critical revision of the data available for myxobolids reported in mullets, determining the species that have bona fide mugiliform fish hosts, in accordance with the original species descriptions, the available molecular data and the currently accepted taxonomic and phylogenetic criteria. Phylogenetic analyses using Bayesian inference and maximum-likelihood methodologies suggest that the evolutionary history of myxobolids with bona fide mugiliform fish hosts reflects that of its vertebrate hosts, while reinforcing known evolutionary factors and old systematic issues of the clade of myxobolids. A comprehensive morphological, ultrastructural and molecular redescription is also provided for the cryptic species M. exiguus, from infections in the visceral peritoneum of the thinlip-grey mullet Chelon ramada in the River Minho, Portugal.
Studies on threatened species in highly modified and unprotected landscapes are necessary for the development of appropriate conservation policies. This is particularly important for species with large home ranges, such as the giant armadillo Priodontes maximus, whose occurrence in anthropogenic landscapes is poorly known despite its categorization as Vulnerable on the IUCN Red List. We searched and surveyed for the giant armadillo within human-modified areas in central Brazil using direct and indirect methods across a wide region dominated by diverse farming environments and scattered remnants of natural vegetation. During a 14-year period (2003–2016), we located 54 records of the species, including three road-kills and two instances of poaching. Most of the occurrence points (83%) were in native vegetation, with 17% in anthropogenic environments (pastures and roads). We confirmed the presence of the giant armadillo within a wide, intensely human-altered region. These findings indicate that Cerrado and Atlantic Forest remnants in modified landscapes in central Brazil play an important role as refuges for this armadillo species. In addition to habitat loss, road-kills and poaching persist as threats to the giant armadillo. Conservation actions are necessary to minimize human impacts and facilitate the persistence of the giant armadillo in this region. Policies that both deter illegal deforestation and strengthen incentives for the protection of natural vegetation remnants and restoration of biological corridors such as gallery forests would aid conservation of the giant armadillo in this area.
Polymorphisms in the growth hormone (GH) and IGF type-1 (IGF1) genes have been associated with the economic traits in farm animals, including BW of some sheep breeds. However, it remains unknown if these polymorphisms also affect carcass traits in sheep. Thus, we aimed to identify polymorphisms in the GH and IGF1 genes in Santa Ines sheep in order to describe their allelic and genotypic frequencies as well as to test the hypotheses that they are associated with the carcass traits. Fragments of 4550 bp (IGF1) and 1194 bp (GH) were sequenced in up to 191 lambs. In all, 18 polymorphisms were identified in the IGF1 and 21 in the GH gene. The IGF1 polymorphisms rs430457475, rs412470350, rs409110739 and rs400113576 showed an additive effect on the internal carcass length (−0.9265±0.4223), rump girth (−2.9285±1.1473), rib yield (−1.0003±0.4588) and neck weight (−0.0567±0.0278), respectively. In addition, the polymorphisms rs58957314 in the GH affected the rib weight (−0.4380±0.1272) and rib yield (−2.2680±0.6970), loin weight (−0.1893±0.0516) and loin yield (−0.9423±0.3259), palette weight (−0.2265±0.0779) and palette yield (−0.9424±0.4184), leg weight (−0.3960±0.1375), neck weight (−0.0851±0.0394) and carcass finishing score (−0.1700±0.0839). These results allow us to conclude that there are polymorphisms in the IGF1 and GH genes associated with carcass traits in Santa Ines sheep, which can provide important information for marker-assisted selection.
Monosodium l-glutamate (MSG)-induced obesity is a useful model for non-alcoholic fatty liver disease (NAFLD) studies. However, there is limited data on its initiation and progression. Thus, this study aimed to characterize the onset of metabolic and histopathological features of NAFLD and its progression to non-alcoholic steatohepatitis (NASH) in this model. To perform this study, Swiss mice pups were neonatally injected with MSG (4 g/kg/day, s.c.) or equiosmolar saline and followed up to 60, 120 or 180 days old. At each age, blood, liver, as well as periepididymal and retroperitoneal fat pads were collected for morphometric, biochemical and histological analyses, the later according to NAFLD activity score. MSG mice presented hypertriglyceridemia and central obesity at all ages, but peripheral insulin-resistance was verified only in 120- and 180-day-old mice. Hepatic total fat and triglycerides content were higher in MSG mice at all ages. Accordingly, histopathological analysis showed that 60-day-old MSG mice had microvesicular steatosis with occasional ballooning, which evolved into NASH from 120 days old. Retroperitoneal fat accumulation was the only variable to independently correlate with NAFLD activity total score upon multivariate analysis (R2=71.45%). There were no differences in IL-6 and TNF-α serum levels among groups. Overall, this study shows that NAFLD is a precocious outcome in MSG-obese mice, whereas the period comprised between 60 and 120 days old seems to be a crucial metabolic window for comprehending pathophysiological events involved in NAFLD-to-NASH progression in this model.
It is a general believe that the local sheep breed Churra da Terra Quente (CTQ), reared in the Northeast of Portugal is well adapted to the conditions of its production system. However, there are large gaps on our knowledge to allow understanding of such claimed adaptation. Several researchers have found differences in digestibility between sheep breeds (e.g. Givens and Moss, 1994; López et al., 2001), suggesting that those fed natural resources can have an improved ability to digest fibre. This experiment was designed to compare digestibility coefficients of a high– and a low–roughage diet measured on sheep from CTQ and Ile-de-France (IF) breeds at the same degree of maturity.
In Northeast of Portugal sheep is reared under extensive systems. These systems frequently involve expressive body composition changes due to the storing and mobilization of body reserves, mainly fat. Body condition score (BCS) is the most common way to assess these reserves and the nutritional status of ewes. However due to the subjective nature of BCS, their quality has been questioned and other alternatives has been studied. For cattle there are some studies that use the real time ultrasonography (RTU) to evaluate the BCS (Schwager-Suter et al., 2000; Broring et al., 2003), but this approach was not tested in ewes. Therefore the aim of the present study was to establish a relationship between the BCS and ultrasound subcutaneous fat (SF) and Longissimus thoracis et lumborum muscle (LM) measurements.
We studied the ontogenetic growth of goat wethers (castrated male goats) of the Saanen and Swiss Alpine breeds based on a large range of intraspecific body mass (BM). The body parts and the chemical constituents of the empty body were described by the allometric function by using BM and the empty body mass (EBM) as the predictors for morphological traits and chemical composition, respectively. We fitted the allometric scaling function by applying the SAS NLMIXED procedure, but to evaluate assumptions regarding variances in morphological and compositional traits, we combined the scaling function with homoscedastic (MOD1), and the heteroscedastic exponential (MOD2) and power-of-the-mean (MOD3) variance functions. We also predicted the ontogenetic growth by using the traditional log-log transformation and back-transformed results into the arithmetic scale (MOD4). We obtained predictions from MOD4 in the arithmetic scale by a two-step process, and evaluated MOD1, MOD2 and MOD3 by a model selection framework, and compared MOD4 with MOD1, MOD2 and MOD3 based on goodness-of-fit measures. Based on information criteria for model selection, heterogeneous variance functions were more likely to describe 10 over 36 traits with a low level of model selection uncertainty. One trait was predicted by averaging the MOD1 and MOD2 variance functions; and nine traits were better described by averaging the MOD2 and MOD3 variance functions. The predictions for other 16 traits were averaged from MOD1, MOD2 and MOD3. However, MOD4 better described 11 traits according to the goodness-of-fit measures. Depending on the variable being analyzed, the body parts and the chemical amounts exhibited the three types of allometric behavior with respect to BM and EBM, that is, positive, negative and isometric ontogenetic growth. Reference BMs, that is, 20, 27, 35 and 45 kg, were used to compute the net protein and energy requirements based on the first derivative of the scaling function, and the results were presented in reference to the EBM and EBM0.75. Both the net protein and energy requirements scaled to EBM0.75 increased from 20 to 45 kg of BM.
Oxidative stress and nitric oxide (NO) appear to represent important links between obesity and cardiovascular, metabolic and/or renal disease. We investigated whether oxidative stress and NO production/metabolism are increased in overweight and obese prepubertal children and correlate with cardiometabolic risk and renal function. We performed a cross-sectional evaluation of 313 children aged 8–9 years. Anthropometrics, 24-h ambulatory blood pressure, pulse wave velocity (PWV), insulin resistance (homoeostasis model assessment index (HOMA-IR)), inflammatory/metabolic biomarkers, estimated glomerular filtration rate (eGFR), plasma total antioxidant status (TAS), plasma and urinary isoprostanes (P-Isop, U-Isop), urinary hydrogen peroxide (U-H2O2), and plasma and urinary nitrates and nitrites (P-NOx, U-NOx) were compared among normal weight, overweight and obese groups, according to WHO BMI z-score reference. U-Isop were increased in the obese group, whereas U-NOx were increased in both overweight and obese children. U-Isop were positively correlated with U-H2O2, myeloperoxidase (MPO), high-sensitivity C-reactive protein, HOMA-IR and TAG. TAS correlated negatively with U-Isop and MPO and positively with PWV. HOMA-IR and U-H2O2 were associated with higher U-Isop, independently of BMI and eGFR, and total cholesterol and U-H2O2 were associated with U-NOx, independently of BMI, eGFR values and P-NOx concentration. In overweight and obese children, eGFR decreased across P-NOx tertiles (median: 139·3 (25th, 75th percentile 128·0, 146·5), 128·0 (25th, 75th percentile 121·5, 140·4), 129·5 (25th, 75th percentile 119·4, 138·3), Pfor linear trend=0·003). We conclude that oxidant status and NO are increased in relation to fat accumulation and, even in young children, they translate into higher values of cardiometabolic risk markers and affect renal function.
Information-flow control mechanisms are difficult both to design and to prove correct. To reduce the time wasted on doomed proof attempts due to broken definitions, we advocate modern random-testing techniques for finding counterexamples during the design process. We show how to use QuickCheck, a property-based random-testing tool, to guide the design of increasingly complex information-flow abstract machines, leading up to a sophisticated register machine with a novel and highly permissive flow-sensitive dynamic enforcement mechanism that is sound in the presence of first-class public labels. We find that both sophisticated strategies for generating well-distributed random programs and readily falsifiable formulations of noninterference properties are critically important for efficient testing. We propose several approaches and evaluate their effectiveness on a collection of injected bugs of varying subtlety. We also present an effective technique for shrinking large counterexamples to minimal, easily comprehensible ones. Taken together, our best methods enable us to quickly and automatically generate simple counterexamples for more than 45 bugs. Moreover, we show how testing guides the discovery of the sophisticated invariants needed for the noninterference proof of our most complex machine.
In tropical regions, protein supplementation is a common practice in dairy and beef farming. However, the effect of highly degradable protein in ruminal fermentation and microbial community composition has not yet been investigated in a systematic manner. In this work, we aimed to investigate the impact of casein supplementation on volatile fatty acids (VFA) production, specific activity of deamination (SAD), ammonia concentration and bacterial and archaeal community composition. The experimental design was a 4×4 Latin square balanced for residual effects, with four animals (average initial weight of 280±10 kg) and four experimental periods, each with duration of 29 days. The diet comprised Tifton 85 (Cynodon sp.) hay with an average CP content of 9.8%, on a dry matter basis. Animals received basal forage (control) or infusions of pure casein (230 g) administered direct into the rumen, abomasum or divided (50 : 50 ratio) in the rumen/abomasum. There was no differences (P>0.05) in ruminal pH and microbial protein concentration between supplemented v. non-supplemented animals. However, in steers receiving ruminal infusion of casein the SAD and ruminal ammonia concentration increased 33% and 76%, respectively, compared with the control. The total concentration of VFA increased (P<0.05) in steers receiving rumen infusion of casein. SAD and the microbial protein concentration did not vary significantly among treatments during the feeding cycle, but mean SAD values were greater in steers supplemented in the rumen and rumen/abomasum. Ruminal ammonia concentration was positively correlated with SAD in animals receiving ruminal infusion of casein. Polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE) analysis revealed low similarity between treatments, animals and time of sample collection. Richness analysis and determination of the Shannon–Wiener index indicated no differences (P>0.05) in species richness and diversity of γ-proteobacteria, firmicutes and archaea between non-supplemented Nellore steers and steers receiving casein supplementation in the rumen. However, species richness and the Shannon–Wiener index were lower (P<0.05) for the phylum bacteroidetes in steers supplemented with casein in the rumen compared with non-supplemented animals. Venn diagrams indicated that the number of unique bands varied considerably among individual animals and was usually higher in number for non-supplemented steers compared with supplemented animals. These results add new knowledge about the effects of ruminal and postruminal protein supplementation on metabolic activities of rumen microbes and the composition of bacterial and archaeal communities in the rumen of steers.
Qualitative samples of ‘opisthobranchs’ were collected from different beaches and estuaries in Ceará State, North-east Brazil. Further species distributions from the literature were used to infer patterns of distribution and to analyse faunal similarity among Atlantic biogeographic areas, Brazilian zones and the Ceará State. A total of 35 new records are reported for the Ceará State, including 19 new records from the North-east Brazilian Shelf (NBS), 12 new from the Brazilian Province and two new records from the South Atlantic. More than half of the species in this area are common between the Caribbean and Brazilian provinces, which demonstrates their capacity for crossing the Amazon–Orinoco barrier. The high percentage of occurrence in the NBS and the South Brazilian Bight, when compared to neighbouring zones, may be attributed to the poor knowledge of the Brazilian ‘opisthobranch’ fauna.
The precocious interruption of lactation is a prime factor for developmental plasticity. Here we analyzed whether flour or flaxseed oil treatment contributes to body and brain mass in male rats subjected to early weaning. Pups were weaned for separation from their mother at 14 (early weaning, EW) and 21 days (control, C). At 21 days, some of the pups were evaluated (C21 v. EW21). After 21 days, control pups (C60) were fed a control diet. EW pups were divided into those fed a control diet (EWC60), those given flaxseed flour (EWFF60), and those given flaxseed oil (EWFO60) until 60 days. EW21 showed lower body and absolute brain mass and higher relative brain mass. At 60 days, EWC60 and EWFO60 had lower body mass. With regard to relative brain mass, EWC60 was heavier; EWFO60 had lower values compared with EWC60 and higher values compared with C60 and EWFF60. These results indicated that flaxseed flour, in comparison with flaxseed oil, contributes to brain development after EW.
Different dietary interventions have been identified as potential modifiers of adiponectin concentrations, and they may be influenced by lipid intake. We identified studies investigating the effect of dietary lipids (type/amount) on adiponectin concentrations in a systematic review with meta-analysis. A literature search was conducted until July 2013 using databases such as Medline, Embase and Scopus (MeSH terms: ‘adiponectin’, ‘dietary lipid’, ‘randomized controlled trials (RCT)’). Inclusion criteria were RCT in adults analysing adiponectin concentrations with modification of dietary lipids. Among the 4930 studies retrieved, fifty-three fulfilled the inclusion criteria and were grouped as follows: (1) total dietary lipid intake; (2) dietary/supplementary n-3 PUFA; (3) conjugated linoleic acid (CLA) supplementation; (4) other dietary lipid interventions. Diets with a low fat content in comparison to diets with a high-fat content were not associated with positive changes in adiponectin concentrations (twelve studies; pooled estimate of the difference in means: − 0·04 (95 % CI − 0·82, 0·74) μg/ml). A modest increase in adiponectin concentrations with n-3 PUFA supplementation was observed (thirteen studies; 0·27 (95 % CI 0·07, 0·47) μg/ml). Publication bias was found by using Egger's test (P= 0·01) and funnel plot asymmetry. In contrast, CLA supplementation reduced the circulating concentrations of adiponectin compared with unsaturated fat supplementation (seven studies; − 0·74 (95 % CI − 1·38, − 0·10) μg/ml). However, important sources of heterogeneity were found as revealed by the meta-regression analyses of both n-3 PUFA and CLA supplementation. Results of new RCT would be necessary to confirm these findings.