Characterization of urban and industrial wastewaters using excitation-emission matrix (EEM) fluorescence: Searching for specific fingerprints.

Excitation-emission matrix (EEM) fluorescence spectroscopy has been applied to characterize several urban and industrial wastewaters (effluents from different types of industries: brewery, winery, dairy, biscuit, tinned fish industry, slaughterhouse, pulp mill, textile dyeing and landfill leachates), searching for specific fluorescence fingerprints. Tryptophan protein-like peaks (T1 and T2) are the predominant fluorescence in urban and food industry wastewaters (brewery, winery, dairy/milk, biscuit and fish farm industries) but no special fingerprint has been found to discriminate among them. Protein-like fluorescence also dominates the spectra of meat/fish industries (effluents from a tinned fish industry and a slaughterhouse), but in this case tyrosine protein-like peaks (B1 and B2) also appear in the spectra in addition to tryptophan-like peaks. This fact might constitute a specific feature to differentiate these wastewaters from others, since the appearance of peaks B is quite uncommon in wastewaters. The textile dyeing effluent shows a characteristic triple humic-like fluorescence (peaks A, C1 and C2) that may represent a specific fingerprint for this kind of effluent. Leachates from medium-aged and old landfills might also show a specific fingerprint in their EEM spectra: the sole presence of the humic-like peak C with very high fluorescence intensity. This feature also allows differentiating them from young landfill leachates, which show predominance of protein-like peaks. The fluorescence index (FI) does not seem to be very appropriate to characterize wastewaters and its usefulness might be limited to the study of humic substances in natural waters, although further studies are needed on this topic. However, the humification index (HIX) and the biological index (BIX) do seem to be useful for studying wastewaters, since they have provided consistent results in the present work. This study shows the potential of EEM fluorescence to identify the origin of some industrial effluents, although more research is needed to check these preliminary results.

Perception of Canine Welfare Concerns among Veterinary Students, Practitioners, and Behavior Specialists in Spain.

Veterinarians are well placed to supervise and ensure canine welfare. However, the perception of animal welfare among veterinarians may vary depending on the level of training and professional practice, including the specialization in animal behavior and welfare. The aim of this study was to survey the perception of canine welfare among veterinarians, including students, practitioners, and behavior specialists. A scale-based questionnaire including 12 issues affecting canine welfare was adapted from Yeates and Main and distributed to first-year (n=50) and fifth-year veterinary students (n=50), as well as veterinary practitioners (n=260) and specialists in behavioral medicine (n=50). For each issue, respondents were asked to rate how much they perceived each issue to affect canine welfare (on a scale of 0 to 4). A General Linear Model test was used to assess the effect of the studied group on scores. "Physical abuse or cruelty" was the highest-scoring problem in all groups and "breed-related conditions" was the lowest. In general, specialists in behavioral medicine assigned significantly higher scores to most items, particularly "behavioral problems" and "lack of sufficient company." In contrast, fifth-year students assigned significantly lower scores to most items. This study shows that situations clearly affecting canine welfare represent an important concern for veterinarians, both undergraduates and professionals. However, the level of professional experience and specialization might influence the perception of more subtle examples of poor welfare. Raising awareness regarding canine welfare, including concern for breed- or behavior-related problems, should be emphasized within university programs.

Serotonin and Dopamine Blood Levels in ADHD-Like Dogs.

As with humans, dogs can suffer from attention deficit hyperactivity disorder-like (ADHD-like) behaviors naturally and exhibit high levels of hyperactivity/impulsivity and attention deficit problems, making the domestic dog a potential animal model for ADHD. ADHD has a very complex pathophysiology in which many neurotransmitters are involved, such as serotonin and dopamine. The aim of the study was to evaluate serum serotonin and dopamine levels in dogs with ADHD-like symptomatology. Fifty-eight dogs were studied, of which, thirty-six were classified as ADHD-like after physical and behavioral assessments. Additionally, the dogs' owners performed a series of scientifically validated questionnaires which included C-BARQ, the Dog Impulsivity Assessment Scale, and the Dog-ADHD rating scale. Serum from every animal was collected after the behavioral assessments and analyzed with commercial ELISA tests for serotonin and dopamine determination. Kruskal-Wallis tests and Lasso regressions were applied to assess the relationships between both neurotransmitters and the ADHD-like behaviors (as assessed by clinical evaluation and through the different questionnaires). The dogs clinically classified as ADHD-like showed lower serotonin and dopamine concentrations. Further, serotonin and dopamine levels were also linked to aggression, hyperactivity, and impulsivity. Decreased serotonin concentrations were also related to fear, attachment, and touch sensitivity. Finally, it must be noted that our data suggested a strong relationship between serotonin and dopamine and ADHD-like behaviors.

Comprehensive characterization of industrial wastewaters using EEM fluorescence, FT-IR and 1H NMR techniques.

The organic matter present in six industrial wastewaters (pulp and paper mill, brewery, textile, dairy, slaughterhouse effluents and a municipal landfill leachate) has been studied in this work using three analytical techniques: excitation-emission matrix fluorescence (EEMF), proton nuclear magnetic resonance spectroscopy (1H NMR) and Fourier transform infrared spectroscopy (FTIR). The pulp and paper mill effluent shows characteristic signals of the presence of lignins, carbohydrates and carboxylic acids, as well as sulfate, carbonate and sulfonates (coming from surfactants used in the cleaning of tanks). The main constituents of the brewery effluent are peptides and proteins coming mainly from spent yeast and diatomite filters (the presence of the latter was confirmed by SiO bands in the FTIR spectrum). The municipal landfill leachate is characterized by the majority presence of humic substances (typical of an old landfill) and a residual presence of small peptides, amino acids and carboxylic acids. Additionally, several inorganic compounds were identified by FTIR, such as nitrate, sulfate, phosphate and cyanide ions. The textile effluent from a cotton-based industry contains carbohydrates, carboxylic acids and sulfonates, which can act as auxochromes in the textile industry. The dairy effluent comprises amino acids and small peptides coming from the biodegradation of milk and whey in addition to carbohydrates (lactose) and carboxylic acids (mainly lactic acid). The presence of tyrosine-like peaks B in the EEMF spectrum of the slaughterhouse effluent indicates the existence of small peptides and amino acids coming from the biodegradation of blood proteins. Additionally, residual glucose, fatty acids, phosphate and sulfate were also identified in this effluent.

Using excitation-emission matrix fluorescence to evaluate the performance of water treatment plants for dissolved organic matter removal.

This study is aimed at assessing the performance of water treatment plants (both wastewater and drinking water treatment plants) for dissolved organic matter (DOM) removal using excitation-emission matrix fluorescence (EEMF) as the monitoring technique. The influent from the wastewater treatment plant (WWTP) of Burgos (Spain) is characterized from the presence of protein-like peaks (T1 and T2) and humic-like peaks (A and C), T2 and A showing the highest fluorescence intensity. The percentages of total removal in the effluent were in the following order: peak T1 (65%) > peak A (45%) > peak C (34%) > peak T2 (26%). The humic-like peaks were the most removed at the primary sedimentation stage, whereas peak T1 was by far the most removed in the biological reactor. Protein-like peaks T1 and T2 experienced a slight increase in the final effluent in comparison to their fluorescence at the previous stage (the exit of the biological reactor), an increase that can be explained by the release of SMP (soluble microbial products) from the biomass in the secondary clarifier. A poor correlation was obtained between peak T2 fluorescence and COD, BOD (r2 = 0.34-0.38). The natural water from the Úzquiza reservoir in Burgos (Spain) is characterized by the only presence of humic substances: a majority peak A (fulvic-like) and a weak peak C (humic-like). The whole fluorescent DOM was removed by coagulation-flocculation but a low fluorescence peak T2 appeared at the final stage, coming from protein-like SMPs released by the biomass attached to the filters.

Study of Behavioural Traits in Can de Palleiro (Galician Shepherd Dog).

The Can de Palleiro (CP) is an autochthonous canine breed from Galicia (NW Spain). Interestingly, no previous research has been published about the behaviour of this breed. Thus, the aim of the present study was to obtain a deeper understanding of CP behavioural and temperamental traits and detect any potentially problematic behaviour by using the Canine Behavioural Assessment and Research Questionnaire (C-BARQ) and the Socially Acceptable Behaviour (SAB) test. Behavioural information was obtained from 377 dogs-177 CPs and 200 general population (GP) dogs-using the C-BARQ. Additionally, 32 dogs were enrolled to perform the SAB test (19 CPs and 13 GP dogs) in order to directly evaluate their temperament. Our results indicated that CP dogs had a lower tendency to show aggressiveness towards their owners (0.18 times lower, p = 0.033) and less fear of other dogs (by 0.43 times, p = 0.001), as well as higher trainability levels (2.56 times higher, p < 0.001) when compared to GP dogs. CP dogs also had increased odds of showing chasing behaviour (3.81 times higher, p < 0.001). Conversely, CPs had reduced odds of non-social fear, separation-related problems and excitability (by 0.42, 0.35 and 0.48 times, respectively; p < 0.001, p < 0.001 and p = 0.002). The current research represents a starting point for the study of the behaviour of CPs, which appear to be a working breed, with guarding and, especially, herding characteristics.

Neprilysin is poorly expressed in the prefrontal cortex of aged dogs with cognitive dysfunction syndrome.

Neprilysin (NEP) is the principal amyloid ß (A ß ) degrading peptidase; this activity may protect against Alzheimer's disease (AD), the most important age-related neurodegenerative process. The aim of this work was to analyze NEP mRNA expression in the frontal cortex of dogs with and without canine cognitive dysfunction syndrome (CDS), which is considered a natural model for AD. Expression of canine cerebral NEP mRNA was assessed by RT-PCR followed by qPCR in young, aged-cognitively unimpaired (CU), and aged-cognitively impaired (CI) dogs. On average, aged-CI dogs showed 80% (P < 0.01) lower expression levels of NEP mRNA than their aged-CU counterparts. Furthermore, the standard deviation of the qPCR measurements was more than 6 times higher in the cognitively healthy animals (young and aged-CU) than in the aged-CI group. Another interesting find is the determination of a positive correlation between NEP expression and the number of cholinergic neurons in basal telencephalon, indicating a probable connection between both events in these types of neurodegeneration processes. These results suggest that high expression levels of NEP might be a protective factor for canine CDS and, most likely, for other A ß -associated neurodegenerative diseases, such as AD.

Identification of ß-amyloid species in canine cerebrospinal fluid by mass spectrometry.

It is well known that several Aß species, including Aß40 and Aß42, are present in cerebrospinal fluid (CSF). Experimental results suggest that these species could play a role in Alzheimer's disease and might also have diagnostic significance. In the present work, the canine CSF ß-amyloid species profile has been identified by matrix-assisted laser desorption and ionization-time-of-flight/time of flight mass spectrometry (MALDI-TOF/TOF) analysis after immunoprecipitation with different Aß-specific antibodies. The results show that species arising from combined ß- and γ-secretase activities in humans, such as Aß1-33, Aß1-34, Aß1-37, Aß1-38, Aß1-39, Aß1-40, and Aß1-42, are also present in dogs. Species arising from combined α- and ß-secretase activities, as well as other Aß-degrading enzymes, are also present in both human and canine CSF, with the exception of Aß1-13, Aß1-14, and Aß1-18, which are not detected in dogs. A large number of species truncated at Glu-3 and Glu-11 have also been detected. To our knowledge, this work describes a most complete Aß species profile from canine CSF. The similarities between the canine and human CSF Aß profile reinforce the dog as a highly appropriate animal model for research in Alzheimer's disease.

Expression of p75(NTR), a marker for basal forebrain cholinergic neurons, in young and aged dogs with or without cognitive dysfunction syndrome.

The canine cognitive dysfunction syndrome (CDS) has been identified as a natural model for Alzheimer's disease (AD). We have used unbiased stereology to estimate the total number of basal forebrain cholinergic neurons expressing the nerve growth factor p75(NTR) receptor in young, aged cognitively-unimpaired (CU) and aged cognitively-impaired (CI) dogs. Aged-CI dogs showed a ∼20% decrement (p = 0.009) in p75(NTR) neurons compared to both the young and the aged-CU animals. These results suggest that the basal forebrain cholinergic system is affected in dogs with CDS and provide additional support for the use this canine syndrome as a model for AD research.

Plasma ß-amyloid peptides in canine aging and cognitive dysfunction as a model of Alzheimer's disease.

Aging dogs naturally demonstrate cognitive impairment and neuropathology that model early Alzheimer's disease (AD). In particular, there is evidence that canine cognitive dysfunction syndrome (CDS) in aged dogs is accompanied by cortical deposition of Aß peptides and neurodegeneration. Plasma Aß levels have been examined in humans as putative biomarkers for AD, but to date, no similar studies have been conducted for canine dementia. The aim of the present study was to assess plasma Aß1-42 and Aß1-40 levels in a blind study using pet dogs that were either successfully aging or exhibiting CDS. The severity of cognitive impairment was assessed using an owner-based questionnaire. On average, young dogs presented significantly higher plasma levels of Aß1-42 and Aß1-40 than aged, cognitively unimpaired dogs. Notably, among aged dogs, the levels of Aß1-42 and the Aß42/40 ratio were significantly higher in those showing mild cognitive impairment than in either cognitively unimpaired or severely affected dogs. These results suggest that increased plasma Aß1-42 levels and Aß42/40 ratio could be a biomarker for canine cognitive dysfunction, which is considered an excellent natural model of early AD.