Histopathological and Ultrastructural Study of a Canine Langerhans Cell Tumour (Canine Cutaneous Histiocytoma).

Canine cutaneous histiocytoma (CCH) represents a significant proportion of dog skin tumours, often manifesting as the most common neoplastic skin condition in young animals. Predominantly affecting dogs under four, these tumours appear primarily as solitary lesions that may regress spontaneously. This study, conducted over five years at the University of Trás-os-Montes e Alto Douro, involved a detailed histopathological and ultrastructural examination of 93 CCH cases. Histologically, these tumours showed distinct patterns of lymphoid infiltration, which contributed to their classification into four groups based on the inflammatory response and histological architecture. Most tumours displayed signs of epidermal invasion and frequent mitotic figures, with necrosis present in over half of the cases. Ultrastructurally, the neoplastic cells were characterised by pleomorphism, abundant organelles, and adherens-type junctions. This study offers significant insights into the pathophysiology and morphological characteristics of CCH, underscoring the importance of detailed histological and ultrastructural analysis in accurately diagnosing and understanding this common canine tumour.

Effects of Urtica urens in the Feed of Broilers on Performances, Digestibility, Carcass Characteristics and Blood Parameters.

With the aim of improving animal health and productivity, plants or plant extracts that have antimicrobial, antifungal and antioxidant properties are often used in studies with broilers. The aim of this work was to investigate the effect of Urtica urens in broilers. Ninety male Ross chicks were used, randomly placed in 30 pens (three broilers per pen). The broilers were assigned to three treatment groups: group CT (control) received a basal diet; group UU1 received a basal diet with 1% dried Urtica urens; and group UU2 received a basal diet with 2% dried Urtica urens. Each treatment consisted of two feeds, distributed from day 1 to 15 (starter) and from day 15 to 36 (grower). As a result of this study, broilers in the UU1 treatment group had higher weight on day 15 (p = 0.029) and day 36 (p = 0.014) than those in treatments CT and UU2 and a higher daily weight gain between days 1 and 15 (p = 0.028) and days 1 and 36 (p = 0.014). Broilers in the UU1 and UU2 groups had lower serum HDL cholesterol (88.8 and 88.9 mg/dL, respectively) than the CT (96.1 mg/dL). In conclusion, the use of dried Urtica urens at 1% as an additive in broiler diets may affect growth performance and blood HDL cholesterol.

Detection of Abnormal Prion Protein by Immunohistochemistry.

Abnormal prion proteins (PrPSc) are the disease-associated isoform of cellular prion protein and diagnostic markers of transmissible spongiform encephalopathies (TSEs). These neurodegenerative diseases affect humans and several animal species and include scrapie, zoonotic bovine spongiform encephalopathy (BSE), chronic wasting disease of cervids (CWD), and the newly identified camel prion disease (CPD). Diagnosis of TSEs relies on immunodetection of PrPSc by application of both immunohistochemistry (IHC) and western immunoblot methods (WB) on encephalon tissues, namely, the brainstem (obex level). IHC is a widely used method that uses primary antibodies (monoclonal or polyclonal) against antigens of interest in cells of a tissue section. The antibody-antigen binding can be visualized by a color reaction that remains localized in the area of the tissue or cell where the antibody was targeted. As such, in prion diseases, as in other fields of research, the immunohistochemistry techniques are not solely used for diagnostic purposes but also in pathogenesis studies. Such studies involve detecting the PrPSc patterns and types from those previously described to identify the new prion strains. As BSE can infect humans, it is recommended that biosafety laboratory level-3 (BSL-3) facilities and/or practices are used to handle cattle, small ruminants, and cervid samples included in the TSE surveillance. Additionally, containment and prion-dedicated equipment are recommended, whenever possible, to limit contamination. The PrPSc IHC procedure consists of a formic acid epitope-demasking step also acting as a prion inactivation measure, as formalin-fixed and paraffin-embedded tissues used in this technique remain infectious. When interpreting the results, care must be taken to distinguish non-specific immunolabeling from target labeling. For this purpose, it is important to recognize artifacts of immunolabeling obtained in known TSE-negative control animals to differentiate those from specific PrPSc immunolabeling types, which can vary between TSE strains, host species, and prnp genotype, further described herein.

Estimating sequence diversity of prion protein gene (PRNP) in Portuguese populations of two cervid species: red deer and fallow deer.

Among the transmissible spongiform encephalopathies (TSEs), chronic wasting disease (CWD) in cervids is now a rising concern in wildlife within Europe, after the detection of the first case in Norway in 2016, in a wild reindeer and until June 2022 a total of 34 cases were described in Norway, Sweden and Finland. The definite diagnosis is post-mortem, performed in target areas of the brain and lymph nodes. Samples are first screened using a rapid test and, if positive, confirmed by immunohistochemistry and Western immunoblotting. The study of the genetics of the prion protein gene, PRNP, has been proved to be a valuable tool for determining the relative susceptibility to TSEs. In the present study, the exon 3 of PRNP gene of 143 samples from red deer (Cervus elaphus) and fallow deer (Dama dama) of Portugal was analysed. Three single nucleotide polymorphisms (SNPs) were found in red deer - codon A136A, codon T98A, codon Q226E - and no sequence variation was detected in fallow deer. The low genetic diversity found in our samples is compatible with previous studies in Europe. The comparison with results from North America suggests that the free-ranging deer from our study may present susceptibility to CWD, although lack of experimental data and the necessity of continuous survey are necessary to evaluate these populations.

Neoplasms in Domestic Ruminants and Swine: A Systematic Literature Review.

BACKGROUND: Due to the limited information and lack of studies on neoplasms in domestic ruminants, i.e., cattle, sheep, and goats, and domestic swine, the objective of the present study was to systematically review the scientific literature to verify the occurrence, type, organ system, and organs most affected by neoplasms in these animals. METHODS: The recommendations of the PRISMA methodology were followed for the elaboration of this study. The research consisted of a systematic review of neoplasms in domestic cattle, sheep, goats, and swine. RESULTS: The number of neoplasms found was 1873. The most affected organ system was the integumentary system with 35.0%, followed in descending order by the alimentary system with 16.90%, the hematopoietic system with 13.50%, the special senses (i.e., eyes and ears) with 10.51%, the female and male genital systems with 7.31%, the urinary system with 4.38%, the liver and biliary system with 3.152%, the endocrine glands with 3.91%, the respiratory system with 2.67%, the nervous system with 2.35%, bones and joints with 0.43%, muscles and tendons with 0.37%, the cardiovascular system with 0.21%, and the pancreas with 0.16%. Of the animals with neoplasms studied, cattle were affected in 69.80% of cases, goats in 10.52%, sheep in 10.46%, and swine in 9.18%. In all species, the most frequent neoplasms were squamous cell carcinomas in ruminants, while melanoma was the most frequent in swine. Few studies carried out in slaughterhouses were found, and the existing ones referred to cattle and swine. No data were found on economic losses with carcass condemnation. CONCLUSIONS: In view of the above, it is necessary to carry out extensive and detailed studies that provide knowledge about the impact of neoplasms on the production and condemnation of carcasses in domestic cattle, sheep, goats, and swine and the respective risk factors.

Effects of Dietary Incorporation of Grape Stalks Untreated and Fungi-Treated in Growing Rabbits: A Preliminary Study.

This study aimed to evaluate the effect of the incorporation of untreated grape stalks (UGS) and fungi-treated grape stalks (Lentinula edodes, TGS) in rabbits' diets. The control group was fed with a control diet without grape stalks (C), two experimental groups were fed on diets with 5% and 10% incorporation of UGS (5UGS and 10UGS), and two with 5% and 10% incorporation of TGS (5TGS and 10TGS). Rabbits fed with TGS diets showed higher daily weight gain (p = 0.034), feed conversion rate (p = 0.002), carcass weight (p = 0.038), and reference carcass weight (p = 0.03) when compared to the control diet. Moreover, animals fed with TGS diets showed an increase in the caecum (p = 0.015) and small intestine (p = 0.021) lengths and in the total volatile fatty acid content (p = 0.005) compared to animals fed UGS diets. Blood triglyceride levels were lower in animals fed with TGS diets compared to UGS (p = 0.005) and C (p ≤ 0.001) diets (12% and 19% lower, respectively), and a trend to lower cholesterol levels was observed (p = 0.071). Meat from rabbits fed with TGS diets had higher levels of linoleic acid, γ-linolenic, ∑ω-6, ∑PUFA, and ∑PUFA/∑SFA ratio compared to rabbits fed with the C diet. Results indicated that grape stalks (UGS and TGS) could be effectively used as an alternative raw material in rabbits' diets without compromising animal performance.

Scrapie at Abattoir: Monitoring, Control, and Differential Diagnosis of Wasting Conditions during Meat Inspection.

Wasting disease in small ruminants is frequently detected at slaughterhouses. The wasting disorder is manifested by the deterioration of the nutritional and physiological state of the animal indicated by thinness, emaciation, and cachexia. Evidence of emaciation and cachexia, alone, are pathological conditions leading to carcass condemnation during an inspection. Several diseases are associated with a wasting condition, including scrapie, pseudotuberculosis, tuberculosis, paratuberculosis, Maedi Visna, and tumor diseases. On the other hand, parasitic diseases, nutrition disorders, exposure or ingestion of toxins, metabolic conditions, inadequate nutrition due to poor teeth, or poor alimentary diet are conditions contributing to poor body condition. Classical and atypical scrapie is naturally occurring transmissible spongiform encephalopathies in small ruminants. The etiological agent for each one is prions. However, each of these scrapie types is epidemiologically, pathologically, and biochemically different. Though atypical scrapie occurs at low incidence, it is consistently prevalent in the small ruminant population. Hence, it is advisable to include differential diagnosis of this disease, from other possibilities, as a cause of wasting conditions detected during meat inspection at the abattoir. This manuscript is a review of the measures in force at the abattoir for scrapie control, focusing on the differential diagnosis of gross lesions related to wasting conditions detected in small ruminants during meat inspection.

Neuropathology of Animal Prion Diseases.

Transmissible Spongiform Encephalopathies (TSEs) or prion diseases are a fatal group of infectious, inherited and spontaneous neurodegenerative diseases affecting human and animals. They are caused by the conversion of cellular prion protein (PrPC) into a misfolded pathological isoform (PrPSc or prion- proteinaceous infectious particle) that self-propagates by conformational conversion of PrPC. Yet by an unknown mechanism, PrPC can fold into different PrPSc conformers that may result in different prion strains that display specific disease phenotype (incubation time, clinical signs and lesion profile). Although the pathways for neurodegeneration as well as the involvement of brain inflammation in these diseases are not well understood, the spongiform changes, neuronal loss, gliosis and accumulation of PrPSc are the characteristic neuropathological lesions. Scrapie affecting small ruminants was the first identified TSE and has been considered the archetype of prion diseases, though atypical and new animal prion diseases continue to emerge highlighting the importance to investigate the lesion profile in naturally affected animals. In this report, we review the neuropathology and the neuroinflammation of animal prion diseases in natural hosts from scrapie, going through the zoonotic bovine spongiform encephalopathy (BSE), the chronic wasting disease (CWD) to the newly identified camel prion disease (CPD).

Uptake and effects of the antimicrobial florfenicol, microplastics and their mixtures on freshwater exotic invasive bivalve Corbicula fluminea.

Microplastics and antimicrobials are widely spread environmental contaminants and more research on their toxicity is needed. The uptake and effects of the antimicrobial florfenicol, microplastics, and their mixtures on Corbicula fluminea were investigated. Bivalves were exposed for 96h to florfenicol (1.8 and 7.1mg/l), microplastics (0.2 and 0.7mg/l), or mixtures of the two substances. After 96h, all bivalves exposed to antimicrobial treatments had florfenicol in their body (e.g. 2±1µg/g). Microplastics were found in the gut, lumen of the digestive gland, connective tissue, hemolymphatic sinuses, and gills surface of animals. Florfenicol caused a significant inhibition of cholinesterase (ChE) activity (~32%). Animals exposed to 0.2mg/l of microplastics showed ChE activity inhibition (31%), and no other significant alterations. Mixtures caused feeding inhibition (57-83%), significant ChE inhibition (44-57%) and of isocitrate dehydrogenase activity, and increased anti-oxidant enzymes activity and lipid peroxidation levels. Overall, the results indicate that C. fluminea take up florfenicol and microplastics from the water and accumulated or at least retained it in their body for some time; both florfenicol (low ppm range) and microplastics (ppb range) were toxic to C. fluminea, with mixtures containing florfenicol and microplastics being more toxic. Thus, the risk of exposure and toxic effects of florfenicol to C. fluminea and other bivalves, and its predators increase in ecosystems contaminated with the antimicrobial and microplastics, as well as to humans consuming contaminated species from these ecosystems.

Carvedilol exacerbate gentamicin-induced kidney mitochondrial alterations in adult rat.

Gentamicin is an aminoglycoside antibiotic widely used to treat many types of bacterial infections. Although its properties, his clinical use is limited due to the occurrence of nephrotoxicity, which has been related to mitochondrial dysfunction. Carvedilol, an antihypertensive drug with strong antioxidant properties, has been tested in order to prevent gentamicin nephrotoxicity. This study aimed to test this hypothesis using a rat model of gentamicin-induced nephrotoxicity. Animals were treated subcutaneously with DMSO (control) (0.4%/kg/24h bw) for 11days; with carvedilol (2mg/kg/24h bw) for 11days; with gentamicin (60mg/kg/24h bw) for the last 8days and with carvedilol (2mg/kg/24h bw) for 11days and with gentamicin (60mg/kg/24h bw) for the last 8days. Estimations of urine creatinine, urine carboxylic acids, blood urea, serum creatinine and glomerular filtration rate were carried out after the last administered dose of gentamicin. Mitochondria functionality was analyzed by monitoring its bioenergetics function and cardiolipin oxidized products were analyzed by ESI-MS. The kidneys were also examined for morphological changes. Gentamicin caused marked nephrotoxicity and mitochondrial dysfunction as evidenced by several mitochondrial parameters. Carvedilol did not induce significant changes while the co-treatment exacerbated the negative effect of gentamicin although maintaining ATP levels and membrane potential. Kidneys from gentamicin treated rats, with and without carvedilol, showed necrosis of tubular cells in renal cortex. Higher values on relative abundance of cardiolipin oxidation products identified as [M-2H]2- ions, at m/z 771 were observed in the groups treated with gentamicin. The observed effects were associated to a possible interaction of carvedilol with F1F0-ATP synthase that merit further investigation. In conclusion, carvedilol may contribute to the exacerbation of renal dysfunction induced by gentamicin, at least in some physiological and biochemical parameters. From a clinical perspective, and until further conclusions, cautious use of both drugs in combination is advised with particular emphasis in patients presenting mitochondrial disorders.

Workplace ergonomics in lean production environments: A literature review.

BACKGROUND: Lean Production Systems (LPS) have become very popular among manufacturing industries, services and large commercial areas. A LPS must develop and consider a set of work features to bring compatibility with workplace ergonomics, namely at a muscular, cognitive and emotional demands level. OBJECTIVE: Identify the most relevant impacts of the adoption of LPS from the ergonomics point of view and summarizes some possible drawbacks for workplace ergonomics due to a flawed application of the LPS. The impacts identified are focused in four dimensions: work pace, intensity and load; worker motivation, satisfaction and stress; autonomy and participation; and health outcome. This paper also discusses the influence that the work organization model has on workplace ergonomics and on the waste elimination previewed by LPS. METHODS: Literature review focused LPS and its impact on occupational ergonomics conditions, as well as on the Health and Safety of workers. The main focus of this research is on LPS implementations in industrial environments and mainly in manufacturing industry workplaces. This is followed by a discussion including the authors' experience (and previous research). RESULTS: From the reviewed literature it seems that there is no consensus on how Lean principles affect the workplace ergonomics since most authors found positive (advantages) and negative (disadvantages) impacts. CONCLUSIONS: The negative impacts or disadvantages of LPS implementations reviewed may result from the misunderstanding of the Lean principles. Possibly, they also happen due to partial Lean implementations (when only one or two tools were implemented) that may be effective in a specific work context but not suitable to all possible situations as the principles of LPS should not lead, by definition, to any of the reported drawbacks in terms of workplace ergonomics.

Regression of canine cutaneous histiocytoma: reduced proliferation or increased apoptosis?

BACKGROUND/AIM: Canine cutaneous histiocytoma (CCH) is a tumour that undergoes spontaneous regression. The aim of this study was to establish a possible relationship between regression of CCH and tumoural cell proliferation and apoptosis. MATERIALS AND METHODS: Immunostaining with Ki-67 antigen and the terminal deoxytransferase (TdT) deoxyuridine-5'-triphosphated (dUTP) nick-end labelling (TUNEL) method were performed on 93 specimens of CCH, grouped into four histological groups. RESULTS: The proliferative index evaluated with Ki-67 antigen expression was on average 23.56 ± 7.91%. The apoptotic index determined by the TUNEL method was on average 39.37 ± 5.87%. Neither the proliferative nor apoptotic index differed between histological groups. Moreover, the proliferative and apoptotic indices did not correlate significantly. However, apoptotic activity was higher than proliferative activity in almost all tumours. CONCLUSION: A reduction of proliferation or an increase of apoptosis does not appear to justify regression of CCH. However, our results suggest that an imbalance between cell proliferation and apoptotic cell death plays a significant role in spontaneous regression of CCH.

Immunohistochemical and immunoelectron study of major histocompatibility complex class-II antigen in canine cutaneous histiocytoma: its relation to tumor regression.

In order to investigate the immune mechanisms involved in regression of canine cutaneous histicytoma (CCH), major histocompatibility complex (MHC) class-II immuno-expression and the number of T- and B-lymphocytes and macrophages were analyzed in 93 cases of CCH. MHC class-II was also studied in 16 cases of CCH by immunoelectron microscopy. All tumors expressed MHC class-II, and two major staining patterns were identified: focal juxtanuclear cytoplasmic staining and rim-like staining along the cell periphery. The MHC class-II labelling pattern and T- and B-lymphocyte infiltrates were associated with tumor regression. In regressing lesions, MHC class-II molecules shift to the cell surface and an increase of both T- and B-lymphocytes were noted. The increasing expression of MHC class-II molecules on the cell surface could be a significant factor for the onset and progression of tumour regression.

Processing of degradable ulvan 3D porous structures for biomedical applications.

The interest in ulvan within a biomedical framework increases as the knowledge of this polysaccharide evolves. Ulvan has been recently proposed as a potential biomaterial, and structures based on this polysaccharide are now being studied for different biomedical applications. In this work, a novel porous structure based on cross-linked ulvan was designed and characterized. Its mechanical performance, water-uptake ability and weight loss were assessed, morphology analyzed through scanning electron microscopy, and morphometric parameters quantified by microcomputed tomography. Cell viability and cell proliferation were evaluated in order to estimate the cytotoxicity of these structures and respective degradation products. Produced ulvan structures revealed remarkable ability to uptake water (up to ∼ 2000% of its initial dry weight) and are characterized by a highly porous and interconnected structure. Furthermore, these ulvan structures underwent nontoxic degradation, and cells remained viable through the time of culture. These results position ulvan structures as prospective blocks that can be further functionalized in order to acquire the desired stability and needed biological interactivity to be used as tissue-engineered structures. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013.

In vitro cytotoxicity assessment of ulvan, a polysaccharide extracted from green algae.

Sustainable exploitation and valorization of natural marine resources represents a highly interesting platform for the development of novel biomaterials, with both economic and environmental benefits. In this context, toxicity data is regarded as a crucial and fundamental knowledge prior to any advances in the application development of natural derived polymers. In the present work, cytotoxicity of ulvan extracted from green algae Ulva lactuca was assessed by means of standard in vitro cytotoxicity assays. Fibroblast-like cells were incubated in the presence of this green algae's polysaccharide, and cell viability was assayed through 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium test. In addition, double stranded DNA and total protein were quantified in order to assess cell number. In order to establish ulvan's non-cytotoxic behaviour, the effect of this polysaccharide on cellular metabolic activity and cell number was directly compared to hyaluronic acid (HA), used as a non-cytotoxic control material. In this study, ulvan demonstrated promising results in terms of cytotoxicity, comparable to the currently used HA, which suggests that ulvan can be considered as non-toxic in the range of concentrations studied.

Processing ulvan into 2D structures: cross-linked ulvan membranes as new biomaterials for drug delivery applications.

The polysaccharide ulvan, composed of sulphated rhamnose, glucoronic and iduronic acids was used to produce polymeric membranes by solvent casting. As ulvan is soluble in water, a cross-linking step was necessary to render the membrane insoluble in water and stable at physiological conditions. Cross-linked ulvan membranes were characterized by FTIR, SEM, swelling behaviour was investigated and the mechanical performance assessed by quasi-static tensile testing. Furthermore, the ability and mechanism of sustained release of a model drug from ulvan membranes was investigated. Produced membranes revealed remarkable ability to uptake water (up to ∼1800% of its initial dry weight) and increased mechanical performance (1.76 MPa) related with cross-linking. On the other hand, medicated ulvan dressings demonstrate the potential as drug delivery devices. Using a model drug we have observed an initial steady release of the drug - of nearly 49% - followed by slower and sustained release up to 14 days. The properties of ulvan membranes herein revealed suggest a great potential of this natural sulphated polysaccharide as a wound dressing.

Marine algae sulfated polysaccharides for tissue engineering and drug delivery approaches.

Biomedical field is constantly requesting for new biomaterials, with innovative properties. Natural polymers appear as materials of election for this goal due to their biocompatibility and biodegradability. In particular, materials found in marine environment are of great interest since the chemical and biological diversity found in this environment is almost uncountable and continuously growing with the research in deeper waters. Moreover, there is also a slower risk of these materials to pose illnesses to humans.   In particular, sulfated polysaccharides can be found in marine environment, in different algae species. These polysaccharides don't have equivalent in the terrestrial plants and resembles the chemical and biological properties of mammalian glycosaminoglycans. In this perspective, are receiving growing interest for application on health-related fields. On this review, we will focus on the biomedical applications of marine algae sulfated polymers, in particular on the development of innovative systems for tissue engineering and drug delivery approaches.

Extraction and physico-chemical characterization of a versatile biodegradable polysaccharide obtained from green algae.

During the last years, considerable attention has been given to different marine organisms, like algae, as potential sources of valuable materials. The continuous demand for novel materials and technologies is high and research on the underexploited marine green algae, including its polysaccharidic part-ulvan, has increased accordingly. In this research work, a novel method for extraction of ulvan from green algae is proposed and demonstrated successfully. Different characterization techniques were employed to characterize the isolated algal polysaccharide, namely, on what concerns its thermal trace and crystallinity. Upon heating, ulvan behaves as a non-meltable polysaccharide that is thermally stable before degradation at 220°C. Ulvan is semi-crystalline in nature and possesses high hygroscopic features, as revealed in this research work. Due to its properties, ulvan can be considered, pure or modified, as a versatile biodegradable polymer for different applications, including tissue engineering and regenerative medicine.

Comparative study about the effects of pollution on glass and yellow eels (Anguilla anguilla) from the estuaries of Minho, Lima and Douro Rivers (NW Portugal).

The health status of eels (Anguilla anguilla) developing in three estuaries of the NW Portuguese coast with different types and levels of pollution was compared in relation to morphometric parameters, Fulton condition index (F index) and several biomarkers. Relatively to the reference population, glass eels from the Lima estuary had lower weight and length, cholinesterase (ChE) and lactate dehydrogenase (LDH) inhibition, and lower levels of some anti-oxidant parameters, while yellow eels also showed a decreased F index, and increased Na(+)/K(+)-ATPase and lipid peroxidation (LPO) levels. Relatively to the reference population, glass eels from the Douro estuary had increased Na(+)/K(+)-ATPase and glutathione-S-transferase activities and LDH inhibition, while yellow eels also had ChE inhibition and increased LPO. Overall, these results indicate that eels from polluted estuaries showed a poor health status than those from a reference estuary, and that adverse effects become more pronounced after spending several years in polluted estuaries.