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The Rio Grande Cone is a major fanlike depositional feature in the continental slope of the Pelotas Basin, Southern Brazil. Two representative sediment cores collected in the Cone area were retrieved using a piston core device. In this work, the organic matter (OM) in the sediments was characterized for a continental vs. marine origin using chemical proxies to help constrain the origin of gas in hydrates. The main contribution of OM was from marine organic carbon based on the stable carbon isotope (δ13C-org) and total organic carbon/total nitrogen ratio (TOC:TN) analyses. In addition, the 14C data showed important information about the origin of the OM and we suggest some factors that could modify the original organic matter and therefore mask the “real” 14C ages: (1) biological activity that could modify the carbon isotopic composition of bulk terrestrial organic matter values, (2) the existence of younger sediments from mass wasting deposits unconformably overlying older sediments, and (3) the deep-sediment-sourced methane contribution due to the input of “old” (>50 ka) organic compounds from migrating fluids.
Few studies have used genomic epidemiology to understand tuberculosis (TB) transmission in rural and remote settings – regions often unique in history, geography and demographics. To improve our understanding of TB transmission dynamics in Yukon Territory (YT), a circumpolar Canadian territory, we conducted a retrospective analysis in which we combined epidemiological data collected through routine contact investigations with clinical and laboratory results. Mycobacterium tuberculosis isolates from all culture-confirmed TB cases in YT (2005–2014) were genotyped using 24-locus Mycobacterial Interspersed Repetitive Units-Variable Number of Tandem Repeats (MIRU-VNTR) and compared to each other and to those from the neighbouring province of British Columbia (BC). Whole genome sequencing (WGS) of genotypically clustered isolates revealed three sustained transmission networks within YT, two of which also involved BC isolates. While each network had distinct characteristics, all had at least one individual acting as the probable source of three or more culture-positive cases. Overall, WGS revealed that TB transmission dynamics in YT are distinct from patterns of spread in other, more remote Northern Canadian regions, and that the combination of WGS and epidemiological data can provide actionable information to local public health teams.
Expression of estrus near timed artificial insemination (TAI) is associated with greater fertility, and estrus detection could improve TAI fertility or direct TAI management, although accurate estrus detection can be difficult and time-consuming using traditional methods. The aim of this study is to evaluate influence of estrus on pregnancy (artificial insemination pregnancy rates (P/AI)) and to validate an alternative method to classify estrus/heat expression using tail chalking (HEATSC) in postpartum Bos indicus cows subjected to TAI in progesterone–estrogen-based protocols. In experiment 1 (Exp. 1), cows (5491) were subjected to visual observation of estrus after progesterone device removal, before TAI, and P/AI was evaluated according to estrus and body condition score (BCS). Cows received a progesterone device and 2 mg estradiol benzoate (EB). After 8 days, the device was removed and 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotrophin was given. Later, animals in Exp. 1 received 1 mg EB and TAI 44 to 48 h. In the Exp. 2 – 3830 cows using similar protocol, received different ovulation inducers: 1 mg EB (n=1624) or 1 mg estradiol cypionate (EC; n=2206) on day 8 (D8). Cows were then marked with chalk, and HEATSC evaluated at TAI on D10 (HEATSC1 – no chalk removal=no estrus expression; HEATSC2 – partial chalk removal=low estrus expression; HEATSC3 – near complete/complete chalk removal=high estrus expression). In Exp. 1, cows showing estrus presented greater P/AI (48.4% v. 40.2%, P<0.05). In Exp. 2, P/AI (HEATSC1 – 40.0%; HEATSC2 – 49.7%; HEATSC3 – 60.9%; P<0.001), and larger follicle timed artificial insemination (LFTAI) (<0.001) varied according to HEATSC. There was no difference in P/AI (P=0.41) or LFTAI (P=0.33) according to ovulation inducer. Cows with greater BCS showed greater P/AI in both experiments (P<0.05). Estrus presence and greater HEATSC improved P/AI, and EC v. EB used promoted differential estrus manifestation (cows showing HEATSC2 and HEATSC3: 79.5% with EB v. 69.98% with EC use, P<0.001), however, with similar P/AI. The use of HEATSC in B. indicus cows subjected to TAI is useful to identify cows with greater estrus expression and consequently improved pregnancy rates in TAI, allowing the cows with low HEATSC to be targeted for additional treatments aimed at improving P/AI.
The use of tail chalk and estrus/heat expression scores (HEATSC) evaluation is instrumental in identifying cows with greater estrus expression and greater artificial insemination pregnancy rates (P/AI) in cows submitted to timed artificial insemination (TAI), and cows with low or no estrus expression present lower P/AI. It was intended in this study to improve the pregnancy rates in TAI for Bos indicus beef cows, and gonadotrophin-releasing hormone (GnRH) injection was hypothesized to increase pregnancy rates in a TAI program for cows submitted to progesterone–estradiol-based protocols with low or no estrus expression, evaluated by HEATSC. Cows (n= 2284) received a progesterone device and 2 mg estradiol benzoate, after 8 days the device was removed and 1 mg estradiol cypionate, 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotropin was administered. All cows were marked with chalk and HEATSC evaluated (scales 1 to 3) at TAI performed on day 10. Animals with HEATSC1 and HEATSC2 (n= 937) received 100 μg de gonadorelin (GNRH group; n= 470), or 1 ml saline (Control group; n= 467), and cows with HEATSC3 (named HEAT group; n= 1347) received no additional treatment. The larger dominant follicle, evaluated on day 8and at TAI (day 10), was greater in HEAT group (P= 0.0145 and P <0.001, respectively). Corpus luteum (CL) area and progesterone concentration was evaluated on day 17, and CL area was larger in HEAT group, intermediary in Control and lower in GnRH group (Control= 2.68 cm2, GnRH= 2.37 cm2, HEAT group= 3.07 cm2, P <0.001). Greater progesterone concentrations were found in HEAT group than in Control and GnRH groups (Control= 4.74 ng/ml, GnRH= 4.29 ng/ml, HEAT group= 6.08 ng/ml, P<0.001). There was a difference in ovulation rate, greater in HEAT group than GnRH and Control groups (Control= 72.5%; GnRH= 81.25%; HEAT group= 90.71%; P= 0.0024). Artificial insemination pregnancy rates was greater in HEAT group (57.09% (769/1347) than in Control and GNRH groups, with positive effect of GnRH injection at the time of TAI in P/AI (Control= 36.18% (169/467), GnRH= 45.95% (216/470); P<0.0001). In conclusion, GnRH application in cows with low HEATSC (1 and 2) is a simple strategy, requiring no changes in TAI management to increase pregnancy rates in postpartum beef cows submitted to progesterone–estradiol-based TAI protocols, without reaching, however, the pregnancy rates of cows that demonstrate high estrus expression at the TAI.
This qualitative study explores the experiences of older adults participating in a creative visual arts program at a residential care facility in Victoria, British Columbia. A narrative inquiry approach was used to conduct face-to-face interviews with 10 residents and three program staff in addition to the systematic observations of program activities and an arts exhibit. The findings reveal the program fostered a sense of community among participants and enhanced their sense of self-worth as artists. A public art exhibition at a community centre underlined the value of residents’ artwork and gave meaning and purpose to their involvement in the program. Findings show the importance of arts programs in fostering creativity in later life and illustrate how people living in institutions can experience multiple dimensions of the self through artistic forms of expression. This study highlights the need to increase access to arts programs for individuals living in residential care.
Sugarcane is an important forage source for dairy cows in tropical countries. However, it provides limited digestible fiber and energy intake, and fat supplementation can be a way to increase energy density and decrease dietary, non-fiber carbohydrates concentrations. We aimed to evaluate the performance, digestion and metabolism of dairy cows in early lactation fed different concentrations of soybean oil (SBO) in sugarcane-based diets. Fourteen primiparous (545±17.2 kg of BW) and eight multiparous (629±26.7 kg BW) Holstein dairy cows were used according to a randomized block design. After calving, diets were randomly assigned to cows within the two parity groups. Diets were formulated with increasing concentrations of SBO (g/kg dry matter (DM)): control (0), low (LSBO; 15.7), medium (MSBO; 44.3) and high (HSBO; 73.4). The study was performed from calving until 84 days in milk, divided into three periods of 28 days each. Dry matter intake (DMI) was affected quadratically in response to SBO addition with the greatest and lowest values of 19.0 and 16.0 kg/day for LSBO and HSBO diets, respectively. The digestibility of potentially digestible NDF was quadratically affected by SBO with the greatest value of 623 g/kg for LSBO diet. Both milk and energy-corrected milk (ECM) production were quadratically affected by SBO inclusion, with greatest ECM values of 27.9 and 27.3 for LSBO and MSBO, respectively. Soybean oil inclusion linearly decreased milk fat concentration by 13.2% from control to HSBO. The CLA t10,c12-18:2 was observed in milk fat only for MSBO and HSBO diets. Soybean oil inclusion did not affect plasma glucose or serum concentrations of total proteins, globulins, albumin, urea nitrogen, beta-hydroxybutyrate, non-esterified fatty acids or insulin. Serum concentrations of total cholesterol, triglycerides and low-density lipoprotein increased with SBO supplementation. Soybean oil inclusion in sugarcane-based diets for early lactation dairy cows from 15.7 to 44.3 g/kg DM can improve energy intake and performance; however, at 44.3 g/kg DM milk fat concentration and ECM decreased. Soybean oil inclusion at 73.4 g/kg DM adversely affected energy intake, fiber digestion and performance of early lactation dairy cows and is not recommended.
The present review will present the recent published results and discuss the main effects of nutrients, mainly fatty acids, on the expression of genes involved in lipid metabolism. In this sense, the review focuses in two phases: prenatal life and finishing phase, showing how nutrients can modulate gene expression affecting marbling and fatty acid profile in meat from ruminants. Adiposity in ruminants starts to be affected by nutrients during prenatal life when maternal nutrition affects the differentiation and proliferation of adipose cells enhancing the marbling potential. Therefore, several fetal programming studies were carried out in the last two decades in order to better understand how nutrients affect long-term expression of genes involved in adipogenesis and lipogenesis. In addition, during the finishing phase, marbling becomes largely dependent on starch digestion and glucose metabolism, being important to create alternatives to increase these metabolic processes, and modulates gene expression. Different lipid sources and their fatty acids may also influence the expression of genes responsible to encode enzymes involved in fat tissue deposition, influencing meat quality. In conclusion, the knowledge shows that gene expression is a metabolic factor affecting marbling and fatty acid profile in ruminant meat and diets and their nutrients have direct effect on how these genes are expressed.
Animal evidence has suggested that maternal emotional and nutritional stress during pregnancy is associated with behavioral outcomes in offspring. The nature of the stresses applied may differ, but it is often assumed that the mother’s hippocampus–hypothalamic–pituitary–adrenal (HHPA) axis response releases higher levels of glucocorticoid hormones. The bed nucleus of the stria terminalis (BNST) is in a pivotal position to regulate the HHPA axis and the stress response, and it has been implicated in anxiety behavior. In the current study, to search whether BNST structural changes and neurochemical alterations are associated with anxiety-related behavior in adult gestational protein-restricted offspring relative to an age-matched normal protein diet (NP) rats, we conduct behavioral tests and, BNST dendritic tree analysis by Sholl analysis, associated to immunoblotting–protein quantification [11β-HSD2, GR, MR, AT1R, 5HT1A and 5HT2A, corticotrophin-releasing factor (CRH) and CRH1]. Dams were maintained either on isocaloric standard rodent chow [with NP content, 17% casein or low protein content (LP), 6% casein] chow throughout their entire pregnancy. Here, in rats subjected to gestational protein restriction, we found: (a) a significant reduction in dendritic length and impoverished dendritic arborization in BNST neurons; (b) an elevated plasmatic corticosterone levels; and (c) associated with enhanced anxiety-like behavior when compared with age-matched NP offspring. Moreover, altered protein (11β-HSD2, GR, MR and type 1 CRH receptors) expressions may underlie the increase in anxiety-like behavior in LP offspring. This work represents the first demonstration that BNST developmental plasticity by maternal protein restriction, resulting in fine structural changes and neurochemical alterations that are associated with modified behavioral states.
Globally, the prevalence of tuberculosis (TB) disease is higher in males. This study examined the effect of sex and age on Mycobacterium tuberculosis (Mtb) infection. Demographic and exposure data were collected on household contacts of sputum smear-positive pulmonary TB patients in Brazil. Contacts with tuberculin skin test induration ⩾10 mm at baseline or 12 weeks were considered Mtb infected. The study enrolled 917 household contacts from 160 households; 508 (55.4%) were female, median age was 21.0 years (range 0.30–87.0) and 609 (66.4%) had Mtb infection. The proportion infected increased with age from 63.3% in girls <5 years to 75.4% in women ⩾40 years and from 44.9% in boys <5 years to 73.6% in men ⩾40 years. Multivariable modelling showed the odds of infection increased between age 5 and 14 years among female contacts (OR 1.5 per 5-year age increase; 95% CI 1.1–2.2; P = 0.02) and between ages 0–4 and 15–39 years among male contacts (OR 2.7, 95% CI 0.83–8.9 and 1.1, 95% CI 0.99–1.3 per 5-year age increase; P = 0.10, 0.07, respectively). The study suggests that the age at which Mtb infection increases most is different in females compared with males. Studies are needed to explore whether these findings are due to differences in host susceptibility, exposure outside the household or other factors.
Background: Heterozygous loss-of-function mutations in the synaptic scaffolding gene SHANK2 are strongly associated with autism spectrum disorder (ASD). However, their impact on the function of human neurons is unknown. Derivation of induced pluripotent stem cells (iPSC) from affected individuals permits generation of live neurons to answer this question. Methods: We generated iPSCs by reprogramming dermal fibroblasts of neurotypic and ASD-affected donors. To isolate the effect of SHANK2, we used CRISPR/Cas9 to knock out SHANK2 in control iPSCs and correct a heterozygous nonsense mutation in ASD-affected donor iPSCs. We then derived cortical neurons from SOX1+ neural precursor cells differentiated from these iPSCs. Using a novel assay that overcomes line-to-line variability, we compared neuronal morphology, total synapse number, and electrophysiological properties between SHANK2 mutants and controls. Results: Relative to controls, SHANK2 mutant neurons have increased dendrite complexity, dendrite length, total synapse number (1.5-2-fold), and spontaneous excitatory postsynaptic current (sEPSC) frequency (3-7.6-fold). Conclusions: ASD-associated heterozygous loss-of-function mutations in SHANK2 increase synaptic connectivity among human neurons by increasing synapse number and sEPSC frequency. This is partially supported by increased dendrite length and complexity, providing evidence that SHANK2 functions as a suppressor of dendrite branching during neurodevelopment.
Phosphorus and calcium deficiency in horses represents an important factor responsible for the low equine production in Brazil. The basic mechanisms of P and Ca metabolism differ substantially among species. Regulation of P and Ca metabolism is less well understood in horses than in others species. With the use of the isotopic dilution technique is possible to evaluate the metabolism for this mineral. The aim of the present experiment was to study the effect of different Ca levels in the diet on P and Ca metabolism in horses.
In the recent years it has been generally accepted that seismic parameters add an important observational constraint for the study of stellar populations and galaxy evolution. Padova-Trieste (PARSEC) evolutionary tracks are widely used to characterise stellar objects and stellar populations. Stellar models at the base of these studies suffer from uncertainties and, more important, degeneracy among different input parameters: stellar mass, chemical composition, central chemical mixing, age, etc. Adding seismic properties to the classic parameters for stars at different evolutionary states, from the H main-sequence to the asymptotic giant branch, is a powerful tool to calibrate physical processes in stellar models, and hence to improve our interpretation of Galactic and extra-Galactic observations.
An experiment was carried to evaluate the energy and protein requirements for the growth and maintenance of lambs of different sex classes. In all, 38 hair lambs (13.0±1.49 kg initial BW and 2 months old) were allocated in a factorial design with diet restriction levels (ad libitum, 30% and 60% feed restriction) and sex classes (castrated and non-castrated males). Four animals from each sex class were slaughtered at the beginning of the trial as a reference group to estimate the initial empty BW and body composition. The remaining lambs were weighed weekly to calculate BW gain (BWG), and when the animals fed ad libitum reached an average BW of 30 kg, all of the experimental animals were slaughtered. Before slaughter, fasted BW (FBW) was determined after 18 h without feed and water. Feed restriction induced reductions in body fat and energy concentration, whereas water restriction showed the opposite effect, and the protein concentration was not affected. The increase in BW promoted increases in body fat and energy content, and these increases were greater in castrated lambs, whereas the protein content was similar between classes tending to stabilize. The net energy required for gain (NEg) and the net protein required for gain (NPg) were not affected by sex class; therefore, an equation was generated for the combined results of both castrated and non-castrated lambs. The NEg varied from 1.13 to 2.01 MJ/day for lambs with BW of 15 and 30 kg and BWG of 200 g. The NPg varied from 24.57 to 16.33 g/day for lambs with BW of 15 and 30 kg and BWG of 200 g. The metabolizable energy efficiency for gain (kg) was 0.37, and the metabolizable protein efficiency for gain (kpg) was 0.28. The net energy required for maintenance (NEm) and the net requirement of protein for maintenance (NPm) did not differ between castrated and non-castrated lambs, with values of 0.241 MJ/kg FBW0.75 per day and 1.30 g/kg FBW0.75 per day, respectively. The metabolizable energy efficiency for maintenance (km) was 0.60, and the efficiency of metabolizable protein use for maintenance (kpm) was 0.57. Nutritional requirements for growth and maintenance did not differ between castrated and non-castrated lambs. This study emphasizes the importance of updating the tables of international committees and of including data obtained from studies with sheep breeds raised in tropical conditions, with the purpose of improving the productive efficiency of the animals
The thermomechanical processing of NiTi shape memory alloys usually involves several steps of hot and/or cold deformation. The present work presents the structural characterization of a Ni-rich NiTi alloy bar, produced by vacuum-induced melting and thermomechanical processing in laboratory scale, aiming at massive production in the future. This study focused on the first step of hot working at 800 °C during rotary forging. Microstructural characterization was performed using differential scanning calorimetry, high- and low-temperature X-ray diffraction (XRD) using a laboratory source and synchrotron XRD. Thus, it was possible to obtain the phase transformation characteristics of the material: the transformation temperatures and the transformation sequence. Proposed thermomechanical processing is intended for production of bars and wires that will be subsequently drawn to get thin wires, for different applications, including orthodontic arch wires.
Previous research has indicated that biological older brothers increase the odds of androphilia in males. This finding has been termed the fraternal birth order effect. The maternal immune hypothesis suggests that this effect reflects the progressive immunization of some mothers to male-specific antigens involved in fetal male brain masculinization. Exposure to these antigens, as a result of carrying earlier-born sons, is hypothesized to produce maternal immune responses towards later-born sons, thus leading to female-typical neural development of brain regions underlying sexual orientation. Because this hypothesis posits mechanisms that have the potential to be active in any situation where a mother gestates repeated male fetuses, a key prediction is that the fraternal birth order effect should be observable in diverse populations. The present study assessed the association between sexual orientation and birth order in androphilic male-to-female transsexuals in Brazil, a previously unexamined population. Male-to-female transsexuals who reported attraction to males were recruited from a specialty gender identity service in southern Brazil (n=118) and a comparison group of gynephilic non-transsexual men (n=143) was recruited at the same hospital. Logistic regression showed that the transsexual group had significantly more older brothers and other siblings. These effects were independent of one another and consistent with previous studies of birth order and male sexual orientation. The presence of the fraternal birth order effect in the present sample provides further evidence of the ubiquity of this effect and, therefore, lends support to the maternal immune hypothesis as an explanation of androphilic sexual orientation in some male-to-female transsexuals.
This experiment compared insulin sensitivity parameters, milk production and reproductive outcomes in lactating dairy cows consuming excessive energy, and receiving in a 2×2 factorial arrangement design: (1) concentrate based on ground corn (CRN; n=13) or citrus pulp (PLP; n=13), and (2) supplemented (n=14) or not (n=12) with 2.5 g/day of chromium (Cr)-propionate. During the experiment (day 0 to 182), 26 multiparous, non-pregnant, lactating Gir×Holstein cows (initial days in milk=80±2) were offered corn silage for ad libitum consumption, and individually received concentrate formulated to allow diets to provide 160% of their daily requirements of net energy for lactation. Cow BW and body condition score (BCS) were recorded weekly. Milk production was recorded daily and milk samples collected weekly. Blood samples were collected weekly before the morning concentrate feeding. Glucose tolerance tests (GTT; 0.5 g of glucose/kg of BW) were performed on days −3, 60, 120 and 180. Follicle aspiration for in vitro embryo production was performed via transvaginal ovum pick-up on days −1, 82 and 162. No treatment differences were detected (P⩾0.25) for BW and BCS change during the experiment. Within weekly blood samples, concentrations of serum insulin and glucose, as well as insulin : glucose ratio were similar among treatments (P⩾0.19), whereas CRN had less (P<0.01) non-esterified fatty acid concentrations compared with PLP (0.177 v. 0.215 mmol/l; SEM=0.009). During the GTT, no treatment differences were detected (P⩾0.16) for serum glucose concentration, glucose clearance rate, glucose half-life and insulin : glucose ratio. Serum insulin concentrations were less (P=0.04) in CRN supplemented with Cr-propionate compared with non-supplemented CRN (8.2 v. 13.5 µIU/ml, respectively; SEM=1.7), whereas Cr-propionate supplementation did not impact (P=0.70) serum insulin within PLP cows. Milk production, milk fat and solid concentrations were similar (P⩾0.48) between treatments. However, CRN had greater (P<0.01) milk protein concentration compared with PLP (3.54% v. 3.14%, respectively; SEM=0.08). No treatment differences were detected (P⩾0.35) on number of viable oocytes collected and embryos produced within each aspiration. In summary, feeding a citrus pulp-based concentrate to lactating dairy cows consuming excessive energy did not improve insulin sensitivity, milk production and reproductive outcomes, whereas Cr-propionate supplementation only enhanced insulin sensitivity in cows receiving a corn-based concentrate during a GTT.
Leptospirosis is a zoonosis caused by bacteria of the genus Leptospira. The disease is globally distributed and a major public health concern. The Norway rat (Rattus norvegicus) is the main reservoir of the pathogen in urban slums of developing and developed countries. The potential routes of intra-specific leptospire transmission in rats are largely unknown. Herein, we identified pathogenic Leptospira spp. in breast tissue and milk of naturally infected rats. We examined kidney, breast tissue and milk from 24 lactating rats for the presence of leptospires using immunofluorescence, immunohistochemistry, polymerase chain reaction (PCR) and scanning electronic microscopy. All 24 rats had evidence for Leptospira in the kidneys, indicating chronic carriage. The majority of kidney-positive rats had detectable leptospires in milk (18, 75%) and breast tissue (16, 67%), as evidenced by immunofluorescence assay and immunohistochemistry. Four (17%) milk samples and two (8%) breast tissue samples were positive by quantitative real-time PCR. Scanning electron microscopy confirmed the presence of leptospires in breast tissue. No major pathological changes in breast tissue were found. This study, for the first time, identified leptospires in the milk and breast tissue of wild Norway rats, suggesting the possibility of milk-borne transmission of leptospirosis to neonates.
The Brazilian Amazon rain forest has lost c. 17% of its originally forested portion, due to deforestation and selective logging. Forest degradation caused by logging contributes to loss of animal species that require specialized habitats to survive, such as woodcreepers that inhabit understorey areas. Habitat associations of woodcreeper species can be important for identifying species that have restricted distribution and/or habitat specialization. Our study investigates the effects of spatial variation in forest structure and some landscape features (canopy openness, altitude, distance to stream and exploited basal area) on the abundance and composition of woodcreeper assemblage in selectively logged tropical forests in Southern Amazonia. We used mist-nets and points count to quantify the composition and abundance of woodcreepers in 32 plots in three sites. Plots were spatially arranged in PPBio LTER sites (long-term ecological research plots, systematically spaced at 1-km intervals) in Southern Amazonia. A total of 240 individuals (captured, observed and/or heard) belonging to 11 woodcreeper species were detected. Mantel tests showed that there is no spatial autocorrelation among woodcreeper assemblage and distance between plots. Altitude and canopy openness were significantly associated with the composition of the woodcreeper assemblage. Altitude was negatively associated with species richness, and the abundance of the two dominant species (Glyphorynchus spirurus and Xiphorhynchus elegans). The negative relationship with canopy openness suggests that woodcreeper assemblages that inhabit understorey are likely to be indirectly affected by selective logging which reduces canopy cover. The selective logging indirectly changes bird species assemblages, and depending on the intensity, may result in the local extinction of some insectivorous species. Short- and long-term studies addressing different intensities of selective logging are needed to determine the impacts on the bird species and forest structure.