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The expanding presence of red foxes ( Vulpes vulpes ) in urban and suburban regions could potentially lead to increased instances of human aggression towards this species. We studied 10 deceased red foxes that were submitted by law enforcement agencies in the metropolitan area of Madrid in 2014-2022 because of suspected abuse. Forensic autopsies were performed to establish the cause and manner of death. In 4 of the 10 cases, the cause of death was deemed unnatural, involving blunt-force trauma ( n  = 2), asphyxia resulting from hanging ( n  = 1), and firearm injury ( n  = 1). Among the remaining cases, most had succumbed to natural causes ( n  = 4), often marked by severe emaciation and a high burden of parasites, primarily Sarcoptes scabiei . In 2 cases, death was undetermined given the poor preservation of the carcass. The growing prevalence of wildlife species in urban areas, particularly red foxes, may require forensic veterinary investigation of deaths potentially related to abuse., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this paper.

Neurobrucellosis is a shared condition of cetaceans and humans. However, the pathogenesis and immune response in cetacean neurobrucellosis has not been extensively studied. In this multicentric investigation, 21 striped dolphin ( Stenella coeruleoalba ) neurobrucellosis ( Brucella ceti ) cases diagnosed over a 10-year period (2012-2022) were retrospectively evaluated. For each case, morphological changes were assessed by evaluating 21 histological parameters. Furthermore, the immunohistochemical expression of Brucella antigen, glial fibrillary acid protein (GFAP), and a selection of inflammatory cell (IBA-1, CD3, and CD20) and cytokine (tumor necrosis factor-alpha [TNF-α], interferon-gamma [IFN-γ], interleukin [IL]-1β, IL-2, and IL-6) markers were investigated. Inflammation of the leptomeninges, ependyma, and/or choroid plexus was lymphohistiocytic, containing macrophages/microglia (IBA-1+), T-cells (CD3+), and B-cells (CD20+) in equal proportion. B-cells occasionally formed tertiary follicles. GFAP expression showed astrocytosis in most cases. Expression of TNF-α, IFN-γ, and IL-2 indicated an intense proinflammatory response, stimulating both macrophages and T-cells. Our results showed that the inflammation and neuroinflammation in neurobrucellosis of striped dolphins mimic human neurobrucellosis and in vitro and in vivo studies in laboratory animals. Cetacean disease surveillance can be exploited to expand the knowledge of the pathogenesis and immunology of infectious diseases, particularly brucellosis, under a One Health approach., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Human-wildlife conflicts may have devastating consequences for fauna due to targeting by humans of wildlife populations suspected to have predated livestock. Suspicion of depredation of extensively raised livestock by vultures in Europe has triggered public administration-led forensic investigations intended to distinguish between predation and scavenging in order to compensate farmers for attacks on their livestock. In this study, gross and histological analyses were carried out on suspected cases of domestic animal depredation by griffon vultures (Gyps fulvus) over a 1-year period. Fifty-eight animals were affected (n = 41, suspected depredation), including domestic cattle (75.9%), sheep (22.4%) and a goat (1.7%). All the adults affected were female and most cases of suspected depredation occurred during the peripartum period (56%). Histological investigations distinguished between post-mortem (84% of diagnostic samples) and ante-mortem (16% of diagnostic samples) cases, and gross examinations revealed significant differences (P ≤ 0.002) between ante-mortem, post-mortem and non-diagnostic samples. This study highlights the need to optimize sampling protocols to increase the success of forensic studies. The forensic investigations presented here may be applied to resolve human-wildlife conflicts involving not only vultures but other endangered carnivores and contribute to human-wildlife coexistence in rural areas by protecting the interests of both the livestock sector and endangered species., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Prescribed fire (PB) is used to achieve ecological objectives and to reduce fuel hazard thus limiting detrimental impacts of wildfire and appropriate selection of prescription window is critical for these goals. Operational use of PB in the Mediterranean forest is scarce and information about its effects on soil remains incomplete. This study for the first time i) compared the immediate impact of spring and autumn PB and experimental summer fire on key properties of forest floor and mineral topsoil in Mediterranean black pine forest, and ii) assessed the capacity of PB to reduce fuel, with limited immediate impacts on soil. PB significantly reduced the 32.5 % of pre-fire forest floor depth, while summer fire consumed 88.5 % and exposed about 30 % of the mineral soil surface. Mean maximum temperature during fire at the mineral soil surface was 23 °C in PB, in contrast to 128 °C in summer fire, while soil heating at 2 cm depth was negligible in both cases. PB did not cause immediate changes in OM quality parameters, and chemical (C and N concentrations, C/N and pH) and microbiological properties (Cmic, Cmic/C, and β-glucosidase, acid phosphatase and alkaline phosphatase activities) in forest floor or mineral topsoil (0-2 cm). By contrast, summer fire greatly increased OM recalcitrance and reduced Cmic, Cmic/C and enzyme activities in forest floor immediately after fire. In the mineral topsoil, only microbial properties were significantly reduced. The maximum temperature reached during fire in forest floor and topsoil was associated with most of the overall changes in properties in both layers. The findings suggest that prescribed fire can significantly reduce fuel with limited initial impacts on soil. Although these findings are encouraging for operational use of prescribed burning in the ecosystem under study, long-term monitoring of repeated application of the technique on soil properties and other ecosystem components is necessary., Competing Interests: Declaration of competing interest M. Teresa Fonturbel reports financial support was provided by Spanish National Institute for Agricultural Research Department of Environment and Agronomy. Researcher reports a relationship with University of Santiago de Compostela that includes: employment., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Land use and plant-soil management influence soil organic C stocks and soil properties. This study aimed to identify the main mechanisms by which these factors alter soil organic matter (SOM) dynamics and stocks. Changes in the organic C pools and biochemical quality in different OM compartments were assessed: a) after deforestation and intensive cultivation (SOM loss) and then, b) after the conversion of cropland to grassland (SOM replenishment) in a chronosequence of recovery (1-45 years). Topsoil samples were subjected to physical fractionation to assess the distribution of free particulate OM (POM) and mineral associated OM (MAOM). SOM quality was characterized by 13 C NMR spectroscopy, thermal analysis (DSC/TG), and microbial activity was monitored by isothermal microcalorimetry. Deforestation and intensive cultivation led to the loss of 80 % of the C stored in the upper mineral soil (up to 30-35 cm). The POM was almost depleted, MAOM underwent significant losses (>40 %) and all OM compounds, including the aromatic C, were affected. The large and unexpected loss of MAOM can be attributed to the low specific surface soil area and also to the labile (biodegradable) nature of the OM in this fraction. After 45 years, conversion of cropland to grassland recovered 68 % of the C lost in the mineral soil (mainly as MAOM), at an annual rate of 1.25 Mg C ha -1 . The present findings showed that the persistence of long-term OM depends on how strongly organic compounds are adsorbed onto mineral surfaces (i.e., the specific surface area) and the biochemical nature of OM compounds. Adequate plant-soil management favoured the replenishment of the MAOM under these experimental conditions, and this fraction was an active pool in terms of C storage and biochemical quality. This study served to test current theories about changes in soil C fractions due to land use changes and soil-plant management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Animal victims of human cruelty are receiving increasing attention from the press and society. Veterinary pathologists and civic authorities have a duty not only to elucidate the cause, method and manner of death but also to address the motivation behind a case. Poultry are commonly used as offering to gods in Santería rituals (ebós). Only a few cases have been reported in scientific journals, in contrast with the number of cases described in the media. Here, a hen and a cockerel (Gallus domesticus) were submitted to the VISAVET Health Surveillance Centre for autopsy, which comprised macroscopic and histological studies and microbiological testing for Chlamydia psittaci, avian orthoavulavirus 1 (Newcastle disease virus), West Nile virus and avian influenza virus. The carcasses were in an active decay stage of decomposition, containing larvae of Lucilia eximia. Both animals had been decapitated and haemorrhages in the cutting region were observed histologically. Post-mortem burns were also observed. To the best of the authors' knowledge, this is the first post-mortem description of lesions in a domestic avian species due to religious rituals. European Union and Spanish forensic practitioners should be aware that these ritual sacrifices occur in some European countries. Moreover, future European and national legislation updates should consider this type of animal abuse., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Tauopathies are a group of neurodegenerative diseases characterized by the pathological aggregation of hyperphosphorylated tau in neurons and glia. Primary tauopathies are not uncommon in humans but exceptional in other species. We evaluate the clinical, neuropathological, and genetic alterations related to tau pathology in 16 cats aged from 1 to 21 years with different clinical backgrounds. Interestingly, a 10-year-old female cat presented a six-year progressive history of mental status and gait abnormalities. The imaging study revealed generalized cortical atrophy. Due to the poor prognosis, the cat was euthanatized at the age of ten. Neuropathological lesions were characterized by massive neuronal loss with marked spongiosis and associated moderate reactive gliosis in the parietal cortex, being less severe in other areas of the cerebral cortex, and the loss of Purkinje cells of the cerebellum. Immunohistochemical methods revealed a 4R-tauopathy with granular pre-tangles in neurons and coiled bodies in oligodendrocytes. Deposits were recognized with several phospho-site antibodies (4Rtau, tau5, AT8, PFH, tau-P Thr181, tau-P-Ser 262, tau-P Ser 422) and associated with increased granular expression of active tau kinases (p38-P Thr180/Tyr182 and SAPK/JNK-P Thr138/Thr185). The genetic study revealed well-preserved coding regions of MAPT. No similar alterations related to tau pathology were found in the other 15 cats processed in parallel. To our knowledge, this is the first case reporting a primary 4R-tauopathy with severe cerebral and Purkinje cell degeneration in an adult cat with neurological signs starting at a young age.

Aeromonas salmonicida is one of the major threats to world aquaculture, causing fish furunculosis and high mortality rates in cultured fish, particularly salmonids. Although Aeromonas spp. is a thoroughly studied pathogen, little is known regarding aeromoniasis in sturgeons. After a mortality outbreak, four juvenile sturgeons ( Acipenser baerii ) were submitted for autopsy and tissue samples were collected for histopathological and microbiological studies. The external examination revealed size heterogenicity, skin hyperpigmentation and reduced body condition of sturgeons. Within the abdominal cavity, mild hepatomegaly and splenomegaly were observed, as well as generalized organic congestion. Histology revealed severe multifocal haemorrhagic and ulcerative dermatitis, mainly localized in the dorsal and latero-ventral areas of fish. The histological study also showed moderate to severe inflammation of gills and organic lesions compatible with septicaemia. Bacterial isolates were identified as Aeromonas salmonicida subsp. salmonicida using MALDI-TOF MS and PCR. Overall, the lesions first described here are consistent with those previously reported in other cultured fish species and contribute to a better understanding of the pathogenesis of Aeromonas salmonicida subsp. salmonicida in the Siberian sturgeon, aside from providing new diagnostic tools for bacterial diseases impacting the fast-growing industry of caviar.

Flavobacterium psychrophilum affects many cultured fish species and is considered one of the most important bacterial pathogens causing substantial economic losses in salmonid aquaculture worldwide. Here, F. psychrophilum was identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and nested PCR as the aetiological agent causing mortality in diseased juvenile Siberian sturgeons (Acipenser baerii) reared on a freshwater fish farm. Diseased sturgeons were lethargic and displayed dark skin pigmentation, increased mucus production and the presence of skin ulcerations and haemorrhages specially on the ventral side and the base of fins. The histological examination of fish revealed proliferative branchitis, ulcerative and necrotizing dermatitis and myositis, lymphoid tissue atrophy, liver and kidney degeneration and thrombosis. To the best of our knowledge, this is the first report describing the infection of Siberian sturgeons by F. psychrophilum. The detection of F. psychrophilum in diseased Siberian sturgeons and the description of the pathological findings observed during the outbreak may contribute to a better understanding of the bacterium pathogenicity and the range of fish species susceptible to infection., (© 2023 The Authors. Journal of Fish Diseases published by John Wiley & Sons Ltd.)

Engineered live bacteria could provide a new modality for treating lung infections, a major cause of mortality worldwide. In the present study, we engineered a genome-reduced human lung bacterium, Mycoplasma pneumoniae, to treat ventilator-associated pneumonia, a disease with high hospital mortality when associated with Pseudomonas aeruginosa biofilms. After validating the biosafety of an attenuated M. pneumoniae chassis in mice, we introduced four transgenes into the chromosome by transposition to implement bactericidal and biofilm degradation activities. We show that this engineered strain has high efficacy against an acute P. aeruginosa lung infection in a mouse model. In addition, we demonstrated that the engineered strain could dissolve biofilms formed in endotracheal tubes of patients with ventilator-associated pneumonia and be combined with antibiotics targeting the peptidoglycan layer to increase efficacy against Gram-positive and Gram-negative bacteria. We expect our M. pneumoniae-engineered strain to be able to treat biofilm-associated infections in the respiratory tract., (© 2023. The Author(s).)

Ovine brucellosis is an infectious disease that causes alterations in the reproductive tract in ram and abortion in ewes. Their negative economic impact in ovine production warrants a thorough understanding the interactions between B. ovis and the host. Here, epididymis lesions of rams infected by B. ovis were histopathologically staged into early and advanced. Expression by immunohistochemistry of Brucella antigens, inflammatory cell markers (CD3, CD79αcy) and cytokines (IFN-γ, TNF-α, TGF-β1) was assessed in both stages. Early lesions were characterized by epithelial changes, interstitial inflammation, and mild fibrosis; whereas advanced lesions displayed caseous granulomas containing numerous macrophages, multinucleated giant cells, lymphocytes, and plasma cells. Expression of Brucella antigens were observed in both stages. The cellular response in B. ovis lesions were predominantly of T-cells (CD3+) whereas low numbers of B-cells and plasma cells (CD79αcy+) were present in both early and advanced lesions. IFN-γ was expressed by lymphocytes in early lesions suggesting that the adaptive immune response against B. ovis is initiated by Th1 cells, this response was also preserved in advanced stages. Expression of TNF-α was observed in neutrophils of epithelial microabscesses and intraepithelial T-cells of early lesions suggesting a promotion of neutrophil phagocytosis triggered by TNF-α. On the other hand, advanced lesions showed a reduction of TNF-α expression which may permit B. ovis persistence in granulomas. Lastly, TGF-β1 expression (fibroblast, macrophages and less in lymphocytes) were increased with time, suggesting that B. ovis promotes TGF-β1 secretion promoting chronicity of the lesions., Competing Interests: Declaration of Competing Interest The authors report no conflicts of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Porcine brucellosis, which is caused by Brucella suis biovar (bv) 2, is a re-emerging disease that causes reproductive problems in pigs in Europe. The pathogenesis and lesions of B. suis intrauterine infection are poorly characterized; characterization could facilitate the diagnosis and investigation of porcine brucellosis. We collected samples of placentas and fetuses for histologic and microbiologic studies during an outbreak of abortions on a pig-breeding farm in Spain. Brucella was cultured from the vaginal swabs obtained from sows that had aborted, some placentas, and fetal tissues (spleen, liver, lung, gastric content); molecular testing confirmed B. suis bv 2 infection. Histologically, there was necrotizing and hemorrhagic placentitis; suppurative hepatitis; lymphoid depletion and sinusoidal histiocytosis in the spleen, lymph nodes, and thymus; and bronchointerstitial pneumonia. Hemorrhages were observed in the umbilical cord, heart, kidneys, and brain. We detected Brucella by immunohistochemistry (IHC) in all of the placentas and fetal organs studied, specifically in the trophoblasts of the chorionic epithelium, in the cytoplasm of macrophages in the chorionic stroma, and extracellularly in necrotic debris. Furthermore, we assessed the lymphocyte population in the placentas through the use of IHC (anti-CD3, anti- Pax5 antibodies), revealing that the lymphocytic response was composed of T cells but not B cells.

The Asian small-clawed otter ( Aonyx cinereus ) is an endangered species that is common in zoologic collections. A 17-y-old female Asian small-clawed otter under human care, with a clinical history of chronic renal disease, was euthanized because of deteriorating health. Histologically, the jejunal wall was infiltrated by a monomorphic population of small neoplastic lymphocytes that expanded the lamina propria of the villi and crypts, and on rare occasions invaded the submucosa. The tumor was composed of T cells (CD3+) with a proliferation index of 16%. Based on the World Health Organization (WHO) Classification of Hematopoietic Neoplasms in Domestic Animals, this lymphoma was classified as an enteropathy-associated T-cell lymphoma (EATL) type 2. We also present here a review of the literature on intestinal lymphoma in the subfamily Lutrinae (otters).

The lesion resulting from the interaction between Mycobacterium and the host immune response is the tuberculous granuloma. Tuberculous granulomas, except in incipient stages, are partially or totally encapsulated by connective tissue. The aim of this study was to assess the immunoexpression of the extracellular matrix proteins fibronectin, collagen III, and collagen I in granulomas caused by Mycobacterium caprae in goats (Capra aegagrus hircus) to understand capsule development at different granuloma stages. For this purpose, a retrospective study of 56 samples of tuberculous granulomas in lung (n = 30) and mediastinal lymph node (n = 26) from 17 goats naturally infected with M. caprae in stages I (n = 15), II (n = 14) and III (n = 27) was carried out. Fibronectin immunoreaction was extracellular, fibrillar-reticular in the center of stage I, II and III granulomas and peripheral in stages II and III granulomas. Collagen III immunoexpression was extracellular and fibrillar in the center of stages I, II and III tuberculous granulomas in lung and mediastinal lymph node, and progressive expression was observed in the periphery of stages II and III granulomas. Finally, collagen I immunoexpression was extracellular and fibrillar, showing a progressive loss of central expression and an increase in peripheral expression in stage III granulomas compared to stage I granulomas. Immunoexpression of these extracellular matrix proteins could help understand fibrogenesis and dating in tuberculous granuloma in both animal models and humans., (© 2022. The Author(s).)

Porcine brucellosis, caused by Brucella suis ( B. suis ), is a notifiable disease causing significant economic losses in production systems. Most infected pigs may act as carriers and shed B. suis even if asymptomatic. This can contribute to environmental persistence, thus hindering control efforts. Here, the environment and the offspring were investigated during and after a B. suis outbreak at a sow breeding farm. The diagnosis of B. suis in sows ( n = 1,140) was performed by culture and polymerase chain reaction (PCR) from vaginal swabs, indirect enzyme-linked immunosorbent assay (I-ELISA) from sera, and brucellin skin test (BST). B. suis diagnosis in post-weaning pigs ( n = 899) was performed by I-ELISA in sera and BST. The environmental surveillance programme was implemented by placing gauze sponges ( n = 175) pre-hydrated in a surfactant and inactivating liquid for Brucella DNA detection by PCR in different farm areas. Our results showed that the offspring of infected sows reacted to in vivo techniques for B. suis . Furthermore, the offspring born during the outbreak displayed higher seropositivity (I-ELISA) and reactivity (BST) than those pigs born after. Brucella DNA was detected in pregnant sow areas, boxes, boots, and post-weaning pig areas. In addition, Brucella DNA environmental detection was higher during the B. suis outbreak than the post B. suis outbreak. The environmental approach has proven to be a simple, practical, valuable, and safe method to detect and monitor B. suis . These results suggest a role of the environment and the offspring that should be considered in porcine brucellosis surveillance and control programmes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Rebollada-Merino, Pérez-Sancho, Rodríguez-Bertos, García, Martínez, Navarro, Domínguez and García-Seco.)

Feline gastrointestinal eosinophilic sclerosing fibroplasia (FGESF) has been described as an inflammatory disorder with an eosinophilic component with etiopathogenesis that is still unknown. Sixteen intestinal samples from two veterinary diagnostic services (2014-2017) were included in the study. A histopathological criterion classified the cases into three grades (mild, moderate, and severe) according to the distribution of the lesions and the course. An immunohistochemical study of collagen I, collagen III, fibronectin, and transforming growth factor β1 (TGF-β1) was performed in each case. An immunohistochemical study of mild grades shows greater collagen III immunoexpression, compared to collagen I and fibronectin, which suggests an "early" stage of fibrosis. In more intense grades, an increased immunoexpression of collagen I, compared to collagen III, suggests a "late" stage of fibrosis. Otherwise, the highest expression of TGF-β1 was observed in the moderate phase, due to the high proliferation of reactive fibroblast and intense inflammation. The results suggest that the inflammatory infiltrate is the trigger for the elevation in TGF-β1, altering the collagen type III:I ratio. In conclusion, immunohistochemical studies can be a very useful method in diagnosing cases of FGESF of mild grades and could help to apply a differential diagnosis regarding feline eosinophilic chronic enteritis (CEE) in the context of inflammatory bowel disease (IBD).

Lymphoma is the most common malignant hematopoietic neoplasm in domestic felines. Twenty-two cases of feline epitheliotropic duodenal T-cell lymphoma were characterized morphologically and immunohistochemically (CD3, Pax5, Ki-67), and Bcl-2 immunoexpression was established. Most cases were in domestic shorthair cats (88.2%), with a mean age of 11.2 years. All lymphomas were CD3+, with a low-to-moderate expression of Ki-67 (<30%). A correlation between the tumoral pattern of infiltration in the lamina propria and the intraepithelial distribution of the neoplastic lymphocytes was established (p = 0.0155). Intraepithelial nests of neoplastic lymphocytes were predominantly observed in lymphomas with a patchy distribution in the lamina propria, whereas intraepithelial plaques were seen in lymphomas with an obliteration pattern. Bcl-2 was expressed in neoplastic cells in all cases, and a higher expression was associated with increased villous stunting (p = 0.0221), and tended to be present in those cases with increased epithelial damage. The expression of Bcl-2 and the degree of epitheliotropism were correlated with neoplastic progression in epitheliotropic intestinal T-cell lymphomas; those displaying high Bcl-2 immunoexpression showed increased villous stunting and epithelial damage, suggesting that Bcl-2 is overexpressed in advanced tumor stages, and may be used as a predictor of tumoral behavior in feline epitheliotropic intestinal T-cell lymphomas. This entity showed many similarities with human MEITL, so the latter entity should be considered in further lymphoma classifications of domestic animals.

Colistin has a long story of safe use in animals for the treatment and prevention of certain bacterial diseases. Nevertheless, the first description of the mcr-1 gene showed that colistin resistance can spread by horizontal gene transfer and changed the landscape. This study aimed to assess the effect of colistin administration on the dispersion of resistance in the microbiota of day-old broiler chicks and how the presence of mcr-1 genes influences the spread of colistin resistance determinants. In this study, 100 one-day-old chicks were divided into four groups of 25 animals (G1, G2, G3, and G4). Animals from G3/G4 were challenged with mcr-1 -carrying Salmonella (day 7), while colistin (600 mg/L) was administered daily to G2/G4 animals through drinking water (from day 8 to day 15). Two quantitative PCR assays were performed to compare the amount of Salmonella and mcr-1 that were present in the caecal samples. We observed that levels of mcr-1 were higher in G3/G4 animals, especially G4, due to the spread of mcr-1 -carrying Salmonella . On day 21, Salmonella levels decreased in G4, reaching similar values as those for G3, but mcr-1 levels remained significantly higher, suggesting that colistin may accelerate the spreading process when mcr-1 -carrying bacteria reach the gut.

In this study, the effects of co-administration with biscuit flour and fermented defatted "alperujo" (FDA) on gut health were evaluated in a batch of laying hens (Hy-Line 2015) on a commercial farm. Animals were divided into two groups: control group and treatment group; and histological and morphometric analyses of all sections of the intestine (duodenum, jejunum, ileum, cecum and rectum) were performed at 10, 18, 25, 50 and 75 weeks of age. During the whole productive period, a decrease in the mortality rate ( p = 0.01) was observed in treated hens, as well as an increase in the number of eggs produced ( p < 0.001), their size ( p < 0.025), and weight ( p < 0.024). In the early and late stages of production (10, 18 and 50 weeks), a significant increase ( p < 0.001) in the height and depth of the intestinal villi was observed in the treatment group. Villi height was also significantly higher ( p < 0.001) in the treatment group up to week 50 in the cecum, and at weeks 18 and 50 in the rectum. We concluded that an economical and sustainable feeding system with less environmental impact, such as co-supplementation with biscuit flour and FDA, could maintain gut health without negatively impacting laying hens' productive performance.

The highland forests of tropical regions are highly vulnerable to climate change because changes in soil organic quality due to the increased soil water deficit conditions through rising temperatures. Several authors have reported that labile molecules dominate soil organic matter at higher elevations, and it is therefore more vulnerable to the rising temperatures associated with climate change. The objective of the present study was to analyze the effect of interaction between the chemical composition of organic matter derived from the dominant plant species and the metabolism of microbial community along an elevational gradient in a highland forest in Central Mexico. The study compared three vegetation-soil systems that represent three different elevational levels: Alnus-system (3100 m.a.s.l.), Abies-system (3500 m.a.s.l.) and Pinus-system (3700 m.a.s.l.). The SOM produced in the lowest site is more recalcitrant (i.e., higher Alkyl:O-Alkyl ratio) as a result of the lower water availability than in the highest site. The results of Threshold Elemental Ratio C:N (TER C:N ) and TER C:P for the organic layer were lower than their C:N and C:P ratios in the organic layer, supporting that the microbial community of the organic layer in the site of lowest elevation must be limited by the carbon source, rather than by N and P. However, these results were not found in the mineral soil, suggesting that the drivers of organic matter decomposition differ between the organic layer and the mineral soil. As a conclusion, our results suggest that the chemical recalcitrance of organic matter (at the lowest site) and temperature (at the highest site) reduce the microbial metabolic activity in the forest floor. Integrated study of plant-derived organic material and the microbial metabolism of the forest floor is therefore required to achieve a full understanding of the vulnerability of tropical mountain ecosystems to climate change., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Companion animal abuse is an issue that concerns not only veterinarians and law enforcement agencies but also society in general. Animals that die under suspicious or violent circumstances should be submitted to reference laboratories for a postmortem examination by veterinary forensic pathologists trained to recognize animal abuse. Nevertheless, the low notification rate of such cases in Spain may explain the country's lag in the development of veterinary forensics and the limited information available on animal abuse epidemiology in comparison with other countries. By sharing information among the entities involved in recognizing animal abuse, we can better address these issues, thus improving veterinary forensics in Spain (and elsewhere). In this study, we analysed the cause and manner of death of 96 dog and cat carcasses suspected of animal abuse. These cases were submitted to our diagnostic laboratory for forensic postmortem examination by public agencies and animal protection centres. To our knowledge, this is the first study to focus on the postmortem analysis of forensic cases of suspected dog and cat abuse in Spain. On the basis of gross and histopathological findings, we distinguished between natural and non-natural abuse-related deaths, classifying the latter. We confirmed that most of the dog deaths were related with abuse, though the suspected abuse and the cause of death did not always coincide. In contrast, cause of death was determined to be natural in many of the cat suspected abuse cases. The most frequent non-natural abuse-related cause of death in dogs was blunt force trauma (n=24, 43.64%), followed by firearm injuries (n=10, 18.18%), asphyxia (n=5, 9.09%), heatstroke (n=3, 5.45%), starvation (n=2, 3.64%), bite injury (n=1, 1.82%), and sharp force trauma (n=1, 1.82%). In cats, the most common cause of death was blunt force trauma (n=9, 21.95%), followed by firearm injuries (n=3, 7.32%) and bite injury (n=2, 4.88%). The main goal of our study is to share our results with the scientific community to advance the field of veterinary forensics in Spain, which will lead to more successful prosecutions by law enforcement agencies. Finally, we highlight that veterinarians who can accurately recognize signs of animal abuse may be able to better respond to cases of companion animal cruelty, which in turn, may prevent possible escalation to interpersonal violence., Competing Interests: Declaration of Competing Interest The author report no declarations of interest., (Published by Elsevier B.V.)

Nutraceuticals are not only nutritionally beneficial for animals but also their use as feed supplements may reduce environmental contamination. The effect of fermented defatted "alperujo," an olive oil by-product, supplementation on the intestinal health of broiler chickens was assessed by analyzing the intestinal mucosal morphology of the duodenum and the cecum. The microbiota of the cecum was also characterized by analyzing the V3-V4 region of the 16S rRNA gene on days 7, 14, 21, 28, 35, and 42. Supplemented broilers from 14 to 35 D of age showed an increase in villus height in the duodenum. This increase likely improved digestibility and absorption capacity during growth, leading to the observed increase in BW at day 35 of life. A progressive increase in crypt depth in both the duodenum and the cecum was also observed. This modification likely enhanced epithelial renewal, thus safeguarding the turnover capacity of the intestinal mucosa. Our molecular analysis of cecal microbiota suggests that this dietary supplement may favor the growth of certain bacteria and may control the spread of pathogenic bacteria by means of competitive exclusion., (Copyright © 2020. Published by Elsevier Inc.)

Salmonella spp. contaminates egg and poultry meat leading to foodborne infections in humans. The emergence of antimicrobial-resistant strains has limited the use of antimicrobials. We aimed to determine the effects of the food supplement, fermented defatted 'alperujo' (FDA), a modified olive oil by-product, on Salmonella Typhimurium colonisation in broilers. One hundred and twenty 1-day-old broilers were divided into four experimental groups-two control groups and two treated groups, and challenged with S . Typhimurium at day 7 or 21. On days 7, 14, 21, 28, 35, and 42 of life, duodenum and cecum tissue samples were collected for histopathological and histomorphometric studies. Additionally, cecum content was collected for Salmonella spp. detection by culture and qPCR, and for metagenomic analysis. Our results showed a significant reduction of Salmonella spp. in the cecum of 42-day-old broilers, suggesting that fermented defatted 'alperujo' limits Salmonella Typhimurium colonization in that cecum and may contribute to diminishing the risk of carcass contamination at the time of slaughter. The improvement of the mucosal integrity, observed histologically and morphometrically, may contribute to enhancing intestinal health and to limiting Salmonella spp. colonisation in the host, mitigating production losses. These results could provide evidence that FDA would contribute to prophylactic and therapeutic measures to reduce salmonellosis prevalence in poultry farms.

Nasopharyngeal hairy polyps are rare benign tumors. We present a newborn case with a hairy polyp mass causing cyanosis and respiratory distress due to obstruction of the upper airway during the first 24 hours of life. The patient required resuscitation and endotracheal intubation. Diagnostic studies confirmed the presence of a mass in the lateral pharyngeal wall. Surgical treatment and removal of the mass was performed with satisfactory evolution of the patient., Competing Interests: None, (Sociedad Argentina de Pediatría.)

African swine fever (ASF) is a notifiable disease that in recent years has spread remarkably in Europe and Asia. Eurasian wild boar ( Sus scrofa ) plays a key role in the maintenance and spread of the pathogen. Here we examined gross pathology of infection in wild boar with a highly virulent, hemadsorbing genotype II ASF virus (ASFV) strain. To this end, six wild boars were intramuscularly inoculated with the 10 HAD50 Arm07 ASFV strain, and 11 wild boars were allowed to come into direct contact with the inoculated animals. No animals survived the infection. Clinical course, gross pathological findings and viral genome quantification by PCR in tissues did not differ between intramuscularly inoculated or contact-infected animals. Postmortem analysis showed enlargement of liver and spleen; serosanguinous effusion in body cavities; and multiple hemorrhages in lungs, endocardium, brain, kidneys, urinary bladder, pancreas, and alimentary system. These results provide detailed insights into the gross pathology of wild boar infected with a highly virulent genotype II ASFV strain. From a didactic point of view, this detailed clinical course and macroscopic description may be essential for early postmortem detection of outbreaks in wild boar in the field and contribute to disease surveillance and prevention efforts.

Biochar is a pyrogenous organic material resulting from the pyrolysis of organic residues, which is attracting the interest from researchers and farmers for its potential to sequester carbon and its use as soil ameliorant. Pyrolysis conditions and feedstock determine the properties of the biochars produced. In order to understand the relationship between these variables we analysed in detail the physical, chemical and surface characteristics of biochars produced from three contrasting agronomic residues abundantly generated in South Spain, such as rice husk (RH), olive pit (OP) and pruning remains of olive trees (mainly composed of olive branches and leaves; OB), using a temperature range from 350 to 600 °C and residence times from 0.5 to 4 h. High pyrolysis temperature (600 °C) and time resulted in the greatest pH and C content in the biochars. In general, elemental composition and ash content were dependent on the type of organic waste used as feedstock. 13 C Nuclear Magnetic Resonance Spectroscopy and thermal (TG-DSC) analyses showed that temperatures ≥500 °C are needed to achieve a high degree of aromatization of the chars. Micro-computed tomography and field emission scanning electron microscopy revealed that the structure of RH was preserved during the pyrolysis process, favouring a greater porosity for these biochars. These data are very useful for the production of stable biochars obtained from residual biomass, maximising the value of residual biomass resources. These biochars show physical and chemical properties, such as adequate pH, high water retention capacity or high porosity, of interest for their use as soil amendments., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The olive oil sector is currently adapting its traditional function to also become a supplier of high-value by-products that possess antioxidant, anti-inflammatory and antimicrobial properties. In this study, we evaluated the effect of the fermented defatted alperujo (FDA) on the intestinal health of laying hens. The morphology of the duodenal and cecal mucosa, the composition of the intestinal microbiota and the productivity of a batch of laying hens were evaluated after FDA supplementation. At early life stages, significant differences ( p < 0.001) were observed in duodenal villi height and in crypt depth of both the duodenum and the cecum in the FDA-supplemented group, indicating improved intestinal health in this group. Microbiota composition in the hatchery group supplemented with FDA had a higher abundance of Actinobacteria, Firmicutes and Proteobacteria, and higher bacterial diversity. During the production period, significant differences ( p < 0.05) were observed in the number of broken eggs from the supplemented group. We conclude that FDA supplementation improves the absorption capacity of the intestinal mucosa and modifies the intestinal microbiota to favor a greater immune response, leading to an increase in egg production.

Various different factors have led to the accumulation of biomass in forest soils in the Mediterranean-climate region in the last few decades, thus exacerbating the effects of wildfires. Although prescribed burning is used to decrease the fuel load and reduce the currency of mega-wildfires, the impacts on soil organic matter (SOM) and nutrient cycling, and therefore on forest ecosystem sustainability, are uncertain. The present study was designed to cover a range of conditions and therefore to assess the variability in the responses in similar geographical areas. Three prescribed burning treatments producing different levels of soil burn severity were conducted in two different types of forests (Pinus nigra and Pinus pinaster) and one (previously treated by prescribed burning) shrubland ecosystem (Cytisus oromediterraneus), all characterized by different fuel loads and depths of soil organic layer, in Central Spain. After the treatments, the SOM content, its thermal properties, and the distribution of Phosphorus (P) forms ( 31 P NMR spectroscopy) were measured in the soil organic layer and mineral soils (0-2 cm depth), and the results were related to the temperatures reached. The prescribed burning les to low-moderate perturbations in SOM quality and Carbon (C) and P dynamics. The organic P, which in the unburnt plots represented 70% of the extractable P, was greatly depleted (by 56 and 95% with respect the initials values). This effect was concurrent with decreases in the most thermolabile SOM fractions, suggesting that organic P is readily mineralized, even at relatively low temperatures. Release of large amounts of soluble orthophosphate may occur when the prescribed burning leads to a high level of soil burn severity. The findings show that prescribed burning treatments should be planned carefully in order to prevent long-term perturbation of C and P cycling., (Copyright © 2018. Published by Elsevier Ltd.)

Background: Litter decomposition is a key process in the functioning of forest ecosystems, because it strongly controls nutrient recycling and soil fertility maintenance. The interaction between the litter chemical composition and the metabolism of the soil microbial community has been described as the main factor of the decomposition process based on three hypotheses: substrate-matrix interaction (SMI), functional breadth (FB) and home-field advantage (HFA). The objective of the present study was to evaluate the effect of leaf litter quality (as a direct plant effect, SMI hypothesis), the metabolic capacity of the microbial community (as a legacy effect, FB hypothesis), and the coupling between the litter quality and microbial activity (HFA hypothesis) on the litter decomposition of two contiguous deciduous oak species at a local scale., Methods: To accomplish this objective, we performed a litterbag experiment in the field for 270 days to evaluate mass loss, leaf litter quality and microbial activity in a complete factorial design for litter quality and species site., Results: The litter of Quercus deserticola had higher rate of decomposition independently of the site, while the site of Quercus castanea promoted a higher rate of decomposition independently of the litter quality, explained by the specialization of the soil microbial community in the use of recalcitrant organic compounds. The Home-Field Advantage Index was reduced with the decomposition date (22% and 4% for 30 and 270 days, respectively)., Discussion: We observed that the importance of the coupling of litter quality and microbial activity depends on decomposition stage. At the early decomposition stage, the home-advantage hypothesis explained the mass loss of litter; however, in the advanced decomposition stage, the litter quality and the metabolic capacity of the microbial community can be the key drivers., Competing Interests: The authors declare there are no competing interests.

Simple, rapid and reliable methods of assessing soil burn severity (SBS) are required in order to prioritize post-fire emergency stabilization actions. SBS proxies based on visual identification and changes in soil organic matter (SOM) content and quality can be related to other soil properties in order to determine the extent to which soil is perturbed following fire. This task is addressed in the present study by an approach involving the use of differential scanning calorimetry-thermogravimetric analysis (DSC-TGA) to determine changes in SOM generated in soils subjected to different levels of SBS. Intact topsoil monoliths comprising the organic horizons and the surface mineral soil (alumic-humic umbrisols) were collected from a representative P. pinaster stand in NW Spain. The monoliths were experimentally burned in a combustion wind tunnel to simulate different fire conditions (fuel bed comprising forest pine litter and wood; air flow, 0.6 m s -1 ). Changes in OM properties in the soil organic layer and mineral soils samples (0-2 cm) at the different temperatures and SBS levels were identified. For both duff and mineral soil, the data revealed a temperature-induced increase in aromatic compounds and a concomitant decrease of carbohydrates and alkyl products. However, for a given temperature, the degree of carbonization/aromatization was lower in the mineral soil than in the duff, possibly due to the different composition of the OM and to the different combustion conditions. The low degree of aromatization of the organic matter suggests that this soil component could undergo subsequent biological degradation. SOM content and thermal recalcitrance (measured as T50) discriminated the SBS levels. Use of visual identification of SBS levels in combination with DSC-TGA enables rapid evaluation of the spatial variability of the effects of fire on SOM properties. This information is useful to predict soil degradation process and implement emergency soil stabilization techniques., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The effect of precipitation regime on the C cycle of tropical forests is poorly understood, despite the existence of models that suggest a drier climate may substantially alter the source-sink function of these ecosystems. Along a precipitation regime gradient containing 12 mature seasonally dry tropical forests growing under otherwise similar conditions (similar annual temperature, rainfall seasonality, and geological substrate), we analyzed the influence of variation in annual precipitation (1240 to 642 mm) and duration of seasonal drought on soil C. We investigated litterfall, decomposition in the forest floor, and C storage in the mineral soil, and analyzed the dependence of these processes and pools on precipitation. Litterfall decreased slightly - about 10% - from stands with 1240 mm yr(-1) to those with 642 mm yr(-1), while the decomposition decreased by 56%. Reduced precipitation strongly affected C storage and basal respiration in the mineral soil. Higher soil C storage at the drier sites was also related to the higher chemical recalcitrance of litter (fine roots and forest floor) and the presence of charcoal across sites, suggesting an important indirect influence of climate on C sequestration. Basal respiration was controlled by the amount of recalcitrant organic matter in the mineral soil. We conclude that in these forest ecosystems, the long-term consequences of decreased precipitation would be an increase in organic layer and mineral soil C storage, mainly due to lower decomposition and higher chemical recalcitrance of organic matter, resulting from changes in litter composition and, likely also, wildfire patterns. This could turn these seasonally dry tropical forests into significant soil C sinks under the predicted longer drought periods if primary productivity is maintained., (© 2016 John Wiley & Sons Ltd.)

Purpose: Wood ash, characterized by high content of certain nutrients and charcoal, can be applied to soils as a means of managing this waste product improving the soil quality. The associated environmental risk must be assessed. The objective of this study was to characterize the bottom and fly ash collected from 15 biomass power plants in Spain by determining the benzene, toluene, ethylbenzene, xylene and styrene (BTEX+S), PAHs and aliphatic hydrocarbon contents of both types of ash. Biochar was also used for comparison purposes., Methods: Gas chromatography-mass spectrometric methods were used for the identification and determination of both BTEX+S and aliphatic hydrocarbon contents in bottom and fly ashes, as well as biochar. High performance liquid chromatography with fluorescence detection was used for PAHs measurements. Multivariate correlation analysis was used to determine the relationship between sample characteristics and pollutants identified by partial least squares regression analysis., Results and Discussion: In general, the degree to which organic matter in the sample is burned increases with T50 or the "50% burn off" temperature (possibly due to the addition of fuel), and the BTEX+S also tended to increase. However, as the Q/MO (the heat of combustion divided by organic matter mass) increased, the combustion decreased or proceeded with less oxygen, which appears to be related to an increased presence of PAHs. The results confirm that the amounts of organic pollutants (PAHs and BTEX+S, together with total aliphatic hydrocarbons) in the wood ash do not exceed limits established for different soil or industrial uses., Conclusions: Both types of ash, together with biochar, may therefore be suitable for application to soil as a fertilizer and an organic amendment, taking into account the target organic pollutants., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Amendment of forest soils with dewatered dairy factory sludge (DDFS), characterized by low heavy metal contents and high amounts of degradable C, can prevent the depletion of soil nutrients that results from intensive harvesting in forest plantations. However, this practice involves environmental risks when N supplies exceed the demand of plants or when the strong acidity of the soil favors the mobility of trace metals. These aspects were assessed in a young radiata pine plantation growing in a sandy, acidic, and organic N-rich soil for the 7 yr after application of a DDFS. The supply of limiting nutrients (mainly P, Mg, and Ca) provided by application of the DDFS, along with control of the ground vegetation, improved the nutritional status of the stand and led to increases in timber volume of more than 60 to 100%. Increases in soil inorganic N were observed during the first months after amendment. Data from soil incubation experiments revealed that some of the additional N was immobilized and, to a lesser extent, denitrified due to the readily available organic C content of the DDFS. Leaching and increased plant uptake of N were prevented by a combination of the latter processes and the low rate of nitrification. The strong acidity of the soil enhanced the availability of Mn and Zn to plants, although the maximum concentrations did not reach levels harmful to organisms. We conclude that although application of DDFS has positive effects on tree nutrition and growth and the environmental risks are low, repeated application may favor mobility of N and availability of heavy metals.

Wood ash, a by-product generated in power plants, can be used to fertilize forest plantations to replenish nutrients lost during harvesting. Although wood ash generally contains low levels of trace metals, release of some of these may occur soon after ash application in acid soils. The risk of heavy metal contamination associated with application of mixed wood ash was assessed in six Pinus radiata D. Don plantations, on two types of mineral soil differing in texture, drainage and CECe. Four of the stands received a single application of 4500 kg ha(-1) (March 2003), and in the other two stands the same treatment was applied over three consecutive years (2003-2005). Trace metal (Cd, Cr, Cu, Mn, Ni, Pb, Zn) concentrations were monitored throughout the 3 years in different components of the forest ecosystem--soil solid fraction, soil solution, tree needles, ground vegetation and different mushroom species. Repeated applications of wood ash led to moderate increases in soil extractable Mn and Zn, and Mn in all mushrooms species. However, the maximum concentrations did not reach levels potentially harmful to organisms. Concentrations of Zn, Cu and Cd decreased in some mushroom species, probably because of increased soil pH caused by the treatment. Heavy metal concentrations in tree needles and ground vegetation were not altered. Although the risk of heavy metal contamination appears to be low, the long-term effects of wood ash application must be assessed.