Search

Small intestinal bacterial overgrowth (SIBO) is a condition hallmarked by an increase in the concentration of colonic-type bacteria in the small bowel. Watery diarrhea, bloating, abdominal pain and distension are the most common clinical manifestations. Additionally, malnutrition and vitamin (B12, D, A, and E) as well as minerals (iron and calcium) deficiency may be present. SIBO may mask or worsen the history of some diseases (celiac disease, irritable bowel disease), may be more common in some extra-intestinal disorders (scleroderma, obesity), or could even represent a pathogenetic link with some diseases, in which a perturbation of intestinal microbiota may be involved. On these bases, we performed a review to explore the multiple links between SIBO and digestive and extra-intestinal diseases., Competing Interests: The authors declare no conflict of interest.

This article traces the historical co-evolution of microbiology, bacteriology, and virology, framed within industrial and agricultural contexts, as well as their role in colonial and national history between the end of the 19th century and the first decades of the 20th century. The epistemology of germ theory, coupled with the economic interests of European colonies, has shaped the understanding of human-microbial relationships in a reductionist way. We explore a brief history of the medical and biological sciences, focusing on microbes and the difficulty of implementing germ theory outside of biology laboratories. Furthermore, we highlight the work of Lynn Margulis, who conceptualized microbes within their ecological contexts. Such research shows the active role microbes play in handling life-sustaining biological and biochemical processes. We outline how the industrial and technological advancements of the last two centuries not only impacted almost all human societies, but also changed the world on microbial, biological, and geological levels. The narration of these histories is a complex task, and depends on how national, international, and intergovernmental institutions (such as the World Health Organization) conceive of the selective environmental pressures exerted by industry and biotechnological companies., (© 2020 The Authors. Centaurus published by John Wiley & Sons Ltd.)

Puppy neonatal mortality may be related to low birth weight, which has been shown in humans to be linked to placental factors. The relation between the newborn puppy and the placental characteristics has been poorly investigated in dogs. Twenty bitches, 9 toy-sized (i.e. < 5 Kg) and 11 small-sized (i.e. 5 to 10 Kg), were included in this study. During natural delivery or c-section, puppies were identified and their order of birth, sex and weight were recorded. Puppy weights were registered at birth and daily until Day 6. Placentas were weighed after removal of extraplacental adnexa, after which a photo was taken and morphometrically assessed. The Total Placental Area (TPA) and the Transfer Zone Area (TZA) were calculated and their surface expressed in mm 2 . Immunohistochemistry with monoclonal mouse anti CD31 antibody was used to identify fetal and maternal vessels in the placental labyrinth zone. A vascularization index (VI) was determined for each placenta and the Total Vascular Area (TVA) was estimated. Puppies' birth weight correlated positively with placental weight (P < 0.001, r = 0.689). A positive correlation was found between the puppies' birth weight and TPA (P < 0.001, r = 0.786), TZA (P < 0.001, r = 0.772), and TVA (P < 0.001, r = 0.482). A positive correlation was also found between placental weight and TPA (P < 0.001, r = 0.661), TZA (P < 0.001, r = 0.583), and TVA (P < 0.001, r = 0.333). In the small-sized breeds, the placentas of low-weight puppies were lighter and had a smaller TZA and TVA (P < 0.05). The VI was higher in the placentas of the toy-sized compared to small-sized bitches (P < 0.01). No effect of parity, litter size, or sex of the puppy was observed on birth weight or growth rates on Day 6. The growth pattern of low-weight puppies did not differ from that of the other puppies during the first 6 days of life. As in humans, placental weight, the extension of the transfer zone and placental total vascular area correlates closely with the puppies' birth weight in normal pregnancies. Our data could represent reference values for placental weight, TZA, TVA and VI in toy and small-sized dog breeds., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Several studies based on histopathology or molecular investigations suggest a causal relation between Felis catus papillomavirus (FcaPV-2) infection and bowenoid in situ carcinoma (BISC) in cats. Nevertheless, data on distribution of viral DNA for different F. catus papillomavirus types (FcaPV-1, 2, 3, 4, 5) in precancerous skin lesions are lacking. In this study, incisional and excisional skin biopsies from 18 cats with BISC were investigated for the presence of FcaPV DNA by quantitative polymerase chain reaction (qPCR) and chromogenic in situ hybridization (CISH) using specific probes to detect each of the FcaPVs that have been identified so far. By qPCR analysis, 15 of 18 samples were positive for FcaPV-2, 2 were positive for FcaPV-4, and 1 sample was negative for all FcaPVs studied. Two cases were positive for FcaPV-5 by qPCR only. FcaPV-1 and FcaPV-3 were not detected by either method. CISH positivity for FcaPV-2 and FcaPV-4 was 100% concordant with qPCR. FcaPV-2 CISH signal was observed as nuclear dots within grouped neoplastic keratinocytes in 12 BISCs and in the perilesional skin of 9 biopsies. In 3 of these 9 cases, the signal was not observed within the BISC. FcaPV-4 CISH positivity was detected only within BISCs in 2 cases. The overall rate of concordance for FcaPV detection between PCR and CISH was 97.8%. This study suggests that CISH is a reliable method to detect FcaPV-2 and FcaPV-4 infection in cats and provides useful information on the type, rate, and localization of infected cells.

Background: It is commonly accepted that canine dystrophic mineralization of the hair follicle glassy membrane can be seen in hyperadrenocorticism and as a senile change in poodles. Pathology textbooks define this change as deposition of calcium salts in the form of basophilic, amorphous, granular material along collagen fibrils., Hypothesis/objectives: The aim was to evaluate whether the incidence of the lesion is specific to poodles and if it is always associated with calcium deposition., Animals: One hundred and forty-seven dogs divided into three groups: (i) 91 normal poodles; (ii) 40 dogs of other breeds; and (iii) 16 dogs with clinical/histopathological diagnosis of hyperadrenocorticism., Methods and Materials: Retrospective study; with haematoxylin and eosin, and von Kossa staining., Results: Our findings demonstrate that perifollicular changes of the hair follicle glassy membrane of poodles are not always associated with calcium deposition., Conclusions and Clinical Importance: A specific staining is needed to identify true calcium deposition when performing histological examination of the skin of poodles., (© 2018 ESVD and ACVD.)

Background: Several species of nematodes included in the superfamily Metastrongyloidea are recognized agents of parasitic infections in felines. Aelurostrongylus abstrusus is the most prevalent species affecting the respiratory system of domestic cats. The route of infection in cats is supposed to be through ingestion of gastropod intermediate or paratenic hosts. However, because gastropods are not the preferred preys of cats, rodents were suggested to play an important role as paratenic hosts in the biological cycle of A. abstrusus and in the epidemiology of aelurostrongylosis., Results: Two studies were conducted to document histopathological tissue lesions in mice experimentally infected with A. abstrusus third-stage larvae (L3) (Study 1), and to determine larval counts in their organs (Study 2). Additionally, cats were fed with experimentally infected mice to assess their infectivity. Aelurostrongylus abstrusus L3 were recovered from the liver, spleen, brain, skeletal muscle and gastrointestinal tract tissues by artificial digestion, and heart, spleen and brain tested positive for A. abstrusus at molecular diagnosis. Multifocal encephalitis and meningitis and glial nodules were the most common histopathological lesions found in mice inoculated with A. abstrusus. All cats shed first-stage larvae of A. abstrusus after ingestion of mice inoculated with this nematode., Conclusions: In this study, we provide information on the anatomical localization, histopathological alterations and rate of recovery of A. abstrusus L3 in mice, and confirm their infectivity to cats (definitive hosts) after feeding on infected mice (paratenic hosts). Data presented here add knowledge to further understand the biology of A. abstrusus in mice and underline the importance of mice as paratenic hosts of this nematode for the infection of cats.

Background: Melanomas are rare in cats. The eye is the most commonly involved site, whereas few data are available about feline non-ocular melanomas (NOMs). Ki-67 thresholds with prognostic relevance have been established for canine melanomas, but not in cats. This study was undertaken to investigate the relationship between Ki-67 index, tumour characteristics, and clinical outcome in feline NOMs. Histologic samples were retrospectively reviewed. Amelanotic tumours were admitted upon immunohistochemical positivity for Melan A or S100. Evaluated parameters included morphological diagnosis, histotype, junctional activity, degree of pigmentation, vascular invasion, lymphocytic infiltrate, necrosis, mitotic count (MC) and Ki-67 index. Pigmented tumours were bleached before evaluation. Clinical and follow-up information were retrieved via telephone interviews with the referring veterinarians., Results: Fifty tumours located in skin (n = 33) and mucosae (n = 17) were included. Forty-eight percent and 95% of amelanotic tumours (n = 21) stained positive for Melan A and S100, respectively. Most achromic tumours were mucosal (P < 0.001, Fisher's exact test) and presented a spindle cell morphology (P = 0.002; Fisher's exact test). MC and Ki-67 index were significantly correlated (P < 0.001; R = 0.67; Spearman's rank correlation); median values were 15 (range, 0-153) and 28% (range, 1-78%), respectively. Both were significantly higher in spindle cell melanomas, in tumours lacking junctional activity and in poorly-pigmented tumours. Follow-up information was available for 33 cats (66%). Variables related with a poor clinical outcome included mucosal location, tumour size, spindle, balloon and signet ring cell histotypes, low pigmentation, MC > 5, Ki-67 > 20% and lack of treatment administration. On multivariable analysis, only tumour histotype and treatment retained prognostic significance., Conclusions: Although the majority of feline NOMs behave aggressively, Ki-67 index, together with other parameters, may contribute to prognostic assessment. Prospective studies on homogeneous populations are warranted to identify reliable threshold values for this marker.

Objectives The aim of the study was to investigate, by quantitative PCR (qPCR), the presence of papillomavirus in feline viral plaques (VPs), Bowenoid in situ carcinoma (BISC), squamous cell carcinoma (SCC) and actinic keratosis (AK). Methods Twenty-nine cases with previously established diagnoses of feline VPs, BISC, invasive SCC and AK were selected from a dermatopathological database. A critical re-evaluation of diagnosis was performed by defining clear criteria toward carcinomatous vs non-carcinomatous, in situ vs invasive (if carcinomatous) and viral vs actinic. Cases were evaluated for p16 immunolocalisation. The presence of the target viral genes for Felis catus papillomavirus (FcaPV)-1, FcaPV-2, FcaPV-3 and FcaPV-4 was determined by qPCR. The data generated ΔΔCq values, which represent a normalised measure of DNA viral quantity. Samples with a positive ΔΔCq value were submitted to sequence analysis. Results Four VPs, 19 BISCs, four SCCs and one case of AK were included. By ΔΔCq analysis we found that all VPs were positive for FcaPV-1 or FcaPV-2; eight BISCs were positive for FcaPV-1, FcaPV-2 and FcaPV-4. FcaPV-2 was the most prevalent among the group of VPs and BISCs. Conclusions and relevance Using the ΔΔCq method we report the first evidence of FcaPV-1, FcaPV-2 and FcaPV-4 in Italy. FcaPV-2 was the most frequently detected; to a lesser extent, FcaPV-1 and FcaPV-4 were detected in the examined samples. FcaPV-3 was never associated with viral-induced lesions by ΔΔCq investigation. Compared with conventional PCR the ΔΔCq method has the advantage of establishing a possible role of the virus in the outcome of infection.

Background: Cannabinoid receptors and peroxisome proliferator-activated receptor-alpha (PPAR-α) are gaining recognition as potential therapeutic targets for the treatment of skin disorders., Hypothesis/objectives: The aim of this study was to investigate the distribution of cannabinoid type 1 and 2 receptors (CBR1 and CBR2) and PPAR-α in feline skin and verify whether changes occur in the course of hypersensitivity dermatitis., Animals: Twelve privately owned cats. Skin samples were obtained from five healthy cats with no skin lesions and seven cats clinically diagnosed with hypersensitivity dermatitis., Methods and Materials: Haematoxylin and eosin stained skin sections were investigated for histopathological changes. Indirect immunofluorescence for CBR1, CBR2 and PPAR-α was performed on paraffin-embedded sections, and antibody specificity tested by Western blot analysis., Results: Skin samples from cats with hypersensitivity dermatitis were all histopathologically diagnosed with eosinophilic dermatitis. CB receptors and PPAR-α were distributed throughout the skin in both healthy and allergic cats. In normal feline skin, these receptors were mainly distributed in the epithelial compartment. Receptor expression increased in hypersensitivity compared to healthy skin, with the main distribution changes being suprabasal for CBR1, dermal for CBR2 and marked expression of PPAR-α in hyperplastic epidermis and perivascular infiltrate., Conclusions and Clinical Importance: Increased expression of cannabinoid receptors in the skin of cats with hypersensitivity dermatitis suggests an endogenous protective strategy and may support the use of natural cannabinoid receptor or PPAR-α agonists to treat feline hypersensitivity dermatitis., (© 2018 ESVD and ACVD.)

Millions of people worldwide currently suffer from serious neurological diseases and injuries for which there are few, and often no, effective treatments. The paucity of effective interventions is, no doubt, due in large part to the complexity of the disorders, as well as our currently limited understanding of their pathophysiology. The bleak picture for patients, however, is also attributable to avoidable impediments stemming from quality concerns in preclinical research that often escape detection by research regulation efforts. In our essay, we connect the dots between these concerns about the quality of preclinical research and their potential ethical impact on the patients who volunteer for early trials of interventions informed by it. We do so in hopes that a greater appreciation among preclinical researchers of these serious ethical consequences can lead to a greater commitment within the research community to adopt widely available tools and measures that can help to improve the quality of research., Competing Interests: The authors have declared that no competing interests exist.

Background: Head and neck ulcers in cats can arise from allergic and nonallergic disorders, including feline leishmaniosis (FeL). It is important to rule out this aetiological agent in regions that are endemic for canine leishmaniosis, because the drugs used to treat immune-mediated disorders of cats can be contraindicated in the setting of infection., Hypothesis/objectives: The aim of this study was to evaluate the skin of cats with ulcerative dermatitis of the head or neck for evidence of Leishmania infection using combined immunohistochemistry (IHC) and Polymerase Chain Reaction (PCR). An IHC for tissue histiocytes was also utilized because leishmaniosis may provoke a histiocytic inflammatory response., Animals: Twenty seven cats with head and/or neck ulcers., Methods: Skin biopsy specimens were examined for the presence of Leishmania spp. by routine histopathological evaluation and IHC using a polyclonal anti-Leishmania antibody, and by quantitative PCR (qPCR). The ionized calcium-binding adapter molecule-1 (IBA-1) antibody was used to immunolocalize histiocytes. Selected history and clinical data were recorded., Results: All specimens showed a superficial mid-perivascular mixed inflammatory infiltrate. The presence of histiocytes was confirmed in 23 of 27 cases with the IBA-1 antibody. Immunohistochemistry and qPCR techniques confirmed the absence of Leishmania in all cases., Conclusions and Clinical Importance: Leishmania did not seem to play a role in the pathogenesis of feline ulcerative dermatitis of the head and neck in the subjects studied, despite a lifestyle potentially associated with infection. Histiocytic infiltration of tissue is not a specific marker for Leishmania infection in this population., (© 2018 ESVD and ACVD.)

The field of epigenetics is leading to new conceptualizations of the role of environmental factors in health and genetic disease. Although more evidence is required, epigenetic mechanisms are being implicated in the link between low socioeconomic status and poor health status. Epigenetic phenomena work in a number of ways: they can be established early in development, transmitted from previous generations and/or responsive to environmental factors. Knowledge about these types of epigenetic traits might therefore allow us to move away from a genetic deterministic perspective, and provide individuals with the opportunity to change their health status. Although this could be equated with patient empowerment, it could also lead to stigmatization and discrimination where individuals are deemed responsible for their health, even if they are not in social situations where they are able to enact change that would alter their health status. In this paper, we will explore the responsibilities of different actors in the healthcare sphere in relation to epigenetics across four different contexts: (1) genetic research, (2) clinical practice, (3) prenatal care and (4) the workplace. Within this exploration of role responsibilities, we will also discuss the potential constraints that might prevent the patient, mother-to-be, research participant or employee, from enacting any necessary steps in order to increase their health status in response to epigenetic information.

Background: Ultramicronized palmitoylethanolamide (PEA-um) has been reported to reduce pruritus and skin lesions in dogs with moderate atopic dermatitis and pruritus., Hypothesis/objectives: A canine ex vivo skin model was used to investigate the ability of PEA-um to counteract changes induced by compound 48/80, a well-known secretagogue that causes mast cell degranulation., Animals: Normal skin was obtained from three donor dogs subjected to surgery for reasons unrelated to the study., Methods: Cultured skin biopsy samples in triplicate were treated with 10 and 100 μg/mL compound 48/80, without or with 30 μM PEA-um. Mast cell (MC) degranulation, histamine release into the culture medium, local microvascular dilatation, epidermal thickness, keratinocyte proliferation and epidermal differentiation markers were evaluated., Results: Exposure of the skin organ culture to PEA-um 24 h before and 72 h concomitantly to compound 48/80 resulted in a significant decrease of degranulating MCs. PEA-um also reduced the histamine content in the culture medium by half, although the effect did not reach statistical significance. PEA-um significantly counteracted vasodilation induced by 100 μg/mL compound 48/80. Finally, PEA-um alone did not induce changes in epidermal thickness, differentiation markers, keratinocyte proliferation, MC density and/or degranulation., Conclusions and Clinical Importance: Collectively, these results support the protective action PEA-um on the skin of dogs undergoing allergic changes., (© 2017 ESVD and ACVD.)

Involvement of the endocannabinoid system in female reproduction has been extensively described in humans with the cognate receptors and ligands being found in the ovaries and genital tract. In human, an imbalance of the endocannabinoid system is linked with both ectopic pregnancy and infertility. In bovine species anandamide levels regulate aspects of sperm-oviduct interaction. Here we report the immunohistochemical distribution of cannabinoid receptor 1 (CB1R) and fatty acid amide hydrolase (FAAH) in cat ovary and oviduct, using paraffin-embedded tissue samples and commercially available antibodies. We found a differential expression of both CB1R and FAAH during different stages of ovarian function and in the oviduct. CB1R was detected only in tertiary follicle granulosa cells while more immature follicles were negative. FAAH was instead found in ovarian pre-antral follicles, the oocyte cytoplasm, and in granulosa cells of primary, secondary and tertiary follicles. Secondary and tertiary follicles were also FAAH immunoreactive. Luteal cells were immunopositive for both CB1R and FAAH. Because CBR1 in oviduct was found only in ciliated cells, it might represent a specific marker at least in cats. In contrast, FAAH immunoreactivity was observed in both ciliated and non-ciliated cells. The present study may thus serve as the starting point for further investigations on the role of the endocannabinoid system in cat reproduction. Additional work will be needed to assess whether the morphological distribution of CB1R and FAAH changes in different conditions such as pre-pubertal age, follicular phase of the sexual cycle and pregnancy., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Background: Innovations in technology have contributed to rapid changes in the way that modern biomedical research is carried out. Researchers are increasingly required to endorse adaptive and flexible approaches to accommodate these innovations and comply with ethical, legal and regulatory requirements. This paper explores how Dynamic Consent may provide solutions to address challenges encountered when researchers invite individuals to participate in research and follow them up over time in a continuously changing environment., Methods: An interdisciplinary workshop jointly organised by the University of Oxford and the COST Action CHIP ME gathered clinicians, researchers, ethicists, lawyers, research participants and patient representatives to discuss experiences of using Dynamic Consent, and how such use may facilitate the conduct of specific research tasks. The data collected during the workshop were analysed using a content analysis approach., Results: Dynamic Consent can provide practical, sustainable and future-proof solutions to challenges related to participant recruitment, the attainment of informed consent, participant retention and consent management, and may bring economic efficiencies., Conclusions: Dynamic Consent offers opportunities for ongoing communication between researchers and research participants that can positively impact research. Dynamic Consent supports inter-sector, cross-border approaches and large scale data-sharing. Whilst it is relatively easy to set up and maintain, its implementation will require that researchers re-consider their relationship with research participants and adopt new procedures.

Equids can be infected by a range of skin-dwelling filarial nematodes, including four species of the genus Onchocerca. Current literature on equine onchocercosis is fragmentary and often limited to isolated case reports. The present study aimed to describe a clinical case of equine onchocercosis caused by Onchocerca boehmi (Supperer, 1953) (syn. Elaeophora boehmi) in an 8-year-old gelding Belgian show jumper from northern Italy. The horse was presented with a firm and painless mass on the proximal third of the right metacarpal region. Ultrasound examination showed a peritendinous enlargement around the palmaro-lateral area of the tendons, characterized by an elongated hypoechoic and well-defined structure, embedding a coiled hyperechoic line. The metacarpal nodule was resected and histologically examined. Fragments of a parasitic nematode were detected, isolated and examined. The morphological analysis allowed identifying the nematode as O. boehmi. In addition, total genomic DNA was extracted from individual fragments using a commercial kit for the nematode identification and a comparative sequence analysis of the nematode cytochrome oxidase subunit 1 (cox1) sequence with data available in the GenBank TM database revealed the closest identity (i.e. 91 %) with that of Onchocerca lupi. Thus far, O. boehmi has only been reported in Austria and Iran, and information about its life-cycle and vectors is lacking. The systematic position of this species within the genus Onchocerca, not in Elaeophora where it was originally described, is in concordance with the morphological and molecular analysis. In this article, we describe the first autochthonous case of equine onchocercosis in Italy caused by O. boehmi and discuss novel parasitological, clinical, and pathological data on these pathogens of horses.

Donation of human tissues for research and ELSI (ethical, legal, and social issues) of biobanking are increasingly debated issues. While several studies have highlighted patients' concerns, little is known about opinions and preferences of healthy potential donors. Further investigations in this respect may allow communication procedures tailored to participants' needs. Based on the Italian Twin Registry, a cross-sectional survey was conducted among twins who had not yet donated biological samples for research. The objective was to assess the importance these potential donors attributed to specific procedures and pieces of communication related to research biobanking. A self-administered questionnaire was constructed and validated. Items were as follows: potential agreement on use of biological samples for research; knowledge of biobanks; and importance given to privacy protection and to communication of general and individual tests results, study objectives, type and amount of participant involvement, location and duration of sample storage, and benefits and potential risks. Multivariate analysis was performed to estimate the association of these items with sociodemographic factors as well as with perceived health status and chronic or long-term diseases. The questionnaire was mailed to 4894 twins aged 18-65 (response rate 34%). One-third of subjects already knew about biobanks, 52% had some knowledge, and 20% were uninformed. A majority expressed unconditional agreement to sample use for research. Only 6% of respondents considered privacy protection not important in research biobanking. Knowledge of biobanks predicted attention to most of the issues. Higher education was associated with more frequent concern about type and amount of involvement, but less frequent concern about place and time of storage, and presence of benefits. Women were more attentive to research biobanking. This study supports the need of procedures tailored on different donors' concerns and highlights the social value of population biobanks. Furthermore, the results call for greater efforts in the promotion of research biobanking.

The Western Honeybee is a key pollinator for natural as well as agricultural ecosystems. In the last decade massive honeybee colony losses have been observed worldwide, the result of a complex syndrome triggered by multiple stress factors, with the RNA virus Deformed Wing Virus (DWV) and the mite Varroa destructor playing crucial roles. The mite supports replication of DWV to high titers, which exert an immunosuppressive action and correlate with the onset of the disease. The aim of this study was to investigate the effect of 1,3-1,6 β-glucan, a natural innate immune system modulator, on honeybee response to low-titer natural and high-titer experimental DWV infection. As the effects exerted by ß-glucans can be remarkably different, depending on the target organism and the dose administered, two parallel experiments were performed, where 1,3-1,6 ß-glucan at a concentration of 0.5% and 2% respectively, was added to the diet of three cohorts of newly emerged honeybees, which were sampled from a Varroa-free apiary and harboured a low endogenous DWV viral titer. Each cohort was subjected to one of the following experimental treatments: no injection, injection of a high-copy number DWV suspension into the haemocel (experimental DWV infection) or injection of PBS into the haemocoel (physical injury). Control bees fed a ß-glucan-free diet were subjected to the same treatments. Viral load, survival rate, haemocyte populations and phenoloxidase activity of each experimental group were measured and compared. The results indicated that oral administration of 0.5% ß-glucan to naturally infected honeybees was associated with a significantly decrease of the number of infected bees and viral load they carried, and with a significant increase of the survival rate, suggesting that this natural immune modulator molecule might contribute to increase honeybee resistance to viral infection., Competing Interests: The authors have declared that no competing interests exist.

Cardiopulmonary infections by Angiostrongylus chabaudi affect domestic and wild felids but, due to limited information on the biology of this nematode, its pathogenicity remains unclear. This article describes the histopathological alterations associated with Angiostrongylus infection in a wildcat from Bulgaria, and reviews current literature on this feline angiostrongylid. Nematodes were isolated from lung lavage and faecal samples of a road killed wildcat in Southern Bulgaria. The morphological identification of parasite larvae as A. chabaudi was confirmed by molecular analysis of part of the 18S ribosomal RNA gene. Upon histopathological examination, severe granulomatous pneumonia, ranging from multifocal to coalescing, and pulmonary vascular lesions were observed. Extensive alveolar collapse, alveolar emphysematous changes, parenchymal haemorrhages and small artery wall hyperplasia were observed in the parenchyma adjacent to the granulomas. Histopathological examination revealed the presence of cross-sections of adult female parasites within the lumen of the pulmonary artery branches, the intima altered markedly by subendothelial proliferation and oedematous changes. This study compliments current knowledge of the pathogenesis of feline angiostrongylosis by A. chabaudi in wildcats, as well as of the distribution of this little-known parasite., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Although canine skin models are already available as either monocellular or organotypic cultures, they only partly recapitulate normal skin morphological features and function. The objective of this study was to establish a canine serum-free skin organ culture model and verify whether dexamethasone could rescue epidermal growth factor-induced changes. The study of morphological changes as a response to pharmacological substances may indeed help to investigate skin physiology and pathology. Normal skin was obtained from five client-owned dogs subjected to surgical procedures unrelated to dermatological conditions. Two experimental designs were performed: (i) two-week viability of the skin culture; (ii) dexamethasone (DMS) inhibition of epidermal growth factor (EGF)-induced effects. Serum-free submerged organ cultures were established in Williams' E medium supplemented with penicillin-streptomycin, insulin, hydrocortisone and l-glutamine. General morphological features of skin anatomical structures were well maintained up to day 14, scattered pyknotic nuclei were visible in the epidermis from day 7. Normal keratinocyte differentiation was confirmed by cytokeratin (K) 10, K14 and loricrin immunostaining. Epidermal thickness did not decrease throughout the study. A decrease in keratinocyte proliferation was observed at day 7 and 14. Treatment with EGF induced both keratinocyte proliferation and thickening of the epidermis; both responses were counteracted by DMS. Treatment with EGF increased the length of epithelial tongues at the edge of the skin explants; this effect was further enhanced by DMS supplementation. Our findings demonstrate the potential use of a full-thickness canine skin organ culture model for the study of skin physiology and pharmacological response to exogenous compounds, especially in the field of re-epithelialisation and keratinization disorders., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Feline oral squamous cell carcinoma (FOSCC) is an aggressive neoplasm in cats. Little is known about the possible molecular mechanisms that may be involved in the initiation, maintenance and progression of FOSCC. Wnt signalling is critical in development and disease, including many mammalian cancers. In this study, we have investigated the expression of Wnt signalling related proteins using quantitative immunohistochemical techniques on tissue arrays. We constructed tissue arrays with 58 individual replicate tissue samples. We tested for the expression of four key Wnt/ß-catenin transcription targets, namely Cyclin D1 (CCND1 or CD1), FRA1, c-Myc and MMP7. All antibodies showed cross reactivity in feline tissue except MMP7. Quantitative immunohistochemical analysis of single proteins (expressed as area fraction / amount of tissue for normal vs tumor, mean ± SE) showed that the expression of CD1 (3.9 ± 0.5 vs 12.2 ± 0.9), FRA1 (5.5 ± 0.6 vs 16.8 ± 1.1) and c-Myc (5.4 ± 0.5 vs 12.5 ± 0.9) was increased in FOSCC tissue by 2.3 to 3 fold compared to normal controls (p<0.0001). By using a multilabel, quantitative fluorophore technique we further investigated if the co-localization of these proteins (all transcription factors) with each other and in the nucleus (stained with 4',6-diamidino-2-phenylindole, DAPI) was altered in FOSCC compared to normal tissue. The global intersection coefficients, a measure of the proximity of two fluorophore labeled entities, showed that there was a significant change (p < 0.01) in the co-localization for all permutations (e.g. CD1/FRA1 etc), except for the nuclear localization of CD1. Our results show that putative targets of Wnt signalling transcription are up-regulated in FOSCC with alterations in the co-localization of these proteins and could serve as a useful marker for the disease., Competing Interests: Competing Interests: The authors of this manuscript have the following competing interests: AA (principal inventor) and CT and JRM (co-inventors) have filed for a patent through UCL Business for targeting Wnt signaling pathway for ß-catenin related disorders (New British Patent Application no. 1318659.8). AA is an associate editor of PLOS ONE. This do not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials. One co-author (Jenny McKay) works for a commercial organization called IDEXX; another co-author (Malcolm Weetman) works for a commercial organization Independent Vetcare. The corresponding author declares, categorically, that none of the issues stated in these statements alter the authors' adherence to all policies of PLOS ONE.

Background: There is growing consensus that individual genetic research results that are scientifically robust, analytically valid, and clinically actionable should be offered to research participants. However, the general practice in European research projects is that results are usually not provided to research participants for many reasons. This article reports on the views of European experts and scholars who are members of the European COST Action CHIP ME IS1303 (Citizen's Health through public-private Initiatives: Public health, Market and Ethical perspectives) regarding challenges to the feedback of individual genetic results to research participants in Europe and potential strategies to address these challenges., Materials and Methods: A consultation of the COST Action members was conducted through an email survey and a workshop. The results from the consultation were analyzed following a conventional content analysis approach., Results: Legal frameworks, professional guidelines, and financial, organizational, and human resources to support the feedback of results are largely missing in Europe. Necessary steps to facilitate the feedback process include clarifying legal requirements to the feedback of results, developing harmonized European best practices, promoting interdisciplinary and cross-institutional collaboration, designing educational programs and cost-efficient IT-based platforms, involving research ethics committees, and documenting the health benefits and risks of the feedback process., Conclusions: Coordinated efforts at pan-European level are needed to enable equitable, scientifically sound, and socially robust feedback of results to research participants.

Personalized medicine is a new field based on molecular biology and genomics in which targeted tumor therapies are administered to patients. Psycho-oncology is a complementary approach that considers social and psychological aspects of patients as part of the treatments for cancer patients. The aim of this mini-review is to weigh clinical benefits for breast cancer patients of both treatments and possibly enhance benefits by modulating the use of both interventions. We have compared and evaluated on the one hand the use of anti Vascular Endothelial Growth Factor and, on the other hand, psycho-oncological interventions in metastatic and non-metastatic breast cancer patients.Both treatments did not increase survival of metastatic breast cancer patients, while in a selected study psycho-oncological interventions extended lifespan of non-metastatic breast cancer patients and ameliorate psychological and social factors of metastatic breast cancer patients. Because the two approaches address completely different aspects of cancer patients, if the comparison is limited to the extension of survival, the value of these two treatments cannot be assessed and compared.It is likely that by comparing patients reported outcomes, possibly by using standardized Quality of Life questionnaires, both patients and health care providers can weigh the benefits of the two treatments. It is therefore important to evaluate the use of cancer patients' quality of life measures as a mean to improve their experiences about life and treatment, and possibly to extend their survival.

Biobanks, which contain human biological samples and/or data, provide a crucial contribution to the progress of biomedical research. However, the effective and efficient use of biobank resources depends on their accessibility. In fact, making bio-resources promptly accessible to everybody may increase the benefits for society. Furthermore, optimizing their use and ensuring their quality will promote scientific creativity and, in general, contribute to the progress of bio-medical research. Although this has become a rather common belief, several laboratories are still secretive and continue to withhold samples and data. In this study, we conducted a questionnaire-based survey in order to investigate sample and data accessibility in research biobanks operating all over the world. The survey involved a total of 46 biobanks. Most of them gave permission to access their samples (95.7%) and data (85.4%), but free and unconditioned accessibility seemed not to be common practice. The analysis of the guidelines regarding the accessibility to resources of the biobanks that responded to the survey highlights three issues: (i) the request for applicants to explain what they would like to do with the resources requested; (ii) the role of funding, public or private, in the establishment of fruitful collaborations between biobanks and research labs; (iii) the request of co-authorship in order to give access to their data. These results suggest that economic and academic aspects are involved in determining the extent of sample and data sharing stored in biobanks. As a second step of this study, we investigated the reasons behind the high diversity of requirements to access biobank resources. The analysis of informative answers suggested that the different modalities of resource accessibility seem to be largely influenced by both social context and legislation of the countries where the biobanks operate.

Background: Skin lesions in canine leishmaniosis (CanL) are diverse, including exfoliative, ulcerative, nodular and papular dermatitides. An uncommon pustular form has also been reported., Hypotheses/objectives: We hypothesized that CanL infection can produce a pustular reaction pattern in the skin of dogs. The aim of this retrospective study was to describe the clinicopathological features of dogs with CanL infection and pustular dermatitis, and correlate them with response to therapy., Animals: Twenty two affected dogs., Methods: Retrospective review of medical records and examination of archived biopsy materials or previously processed glass slides was performed. Cytological examinations had been recorded for all cases. Specimens were available for histopathological examination in 17 of 22 cases and for immunohistochemical detection of Leishmania amastigotes in 13 of 22 cases., Results: All dogs presented with multifocal to diffuse pustular dermatitis. CanL was diagnosed by IFAT serology (20 cases), bone marrow cytology (one case) or bone marrow PCR (one case). Cytological and/or histopathological examinations revealed acantholytic keratinocytes within pustules in 18 of 22 cases. Bacterial and fungal cultures were not performed. Leishmania amastigotes were identified by histopathology within the dermis in three cases; immunohistochemistry was positive in four cases. All dogs underwent concurrent anti-leishmanial and immunosuppressive therapy to control the pustular dermatitis, with favourable outcome in 11 of 22 cases., Conclusions and Clinical Importance: Due to the retrospective nature of this study it is not possible to either accept or reject the hypothesis that CanL is the direct cause of pustular dermatitis., (© 2015 ESVD and ACVD.)

With the rise of -omics disciplines and biobank research, personal data and biosamples crossing national borders pose new ethical questions. In this article, informed consent, as originally conceived, is shown as not being sufficient to address aims of research and interests of patients any more. Therefore the author has, after having scrutinised issues in biobanking, sketched a model of dynamic consent and a manner of scrutinising ethical issues through empirical data., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.)

Background: Appropriate information and consent has been one of the most intensely discussed topics within the context of biobank research. In parallel to the normative debate, many socio-empirical studies have been conducted to gather experiences, preferences and views of patients, healthy research participants and further stakeholders. However, there is scarcity of literature which connects the normative debate about justifications for different consent models with findings gained in empirical research. In this paper we discuss findings of a limited review of socio-empirical research on patients' and healthy research participants' experiences and views regarding consent to biobank research in light of ethical principles for appropriate information and consent., Methods: Review question: Which empirical data are available on research participants' perceptions and views regarding information and elicitation of consent for biobank research? Search of articles published till March 1st 2014 in Pubmed. Review of abstracts and potentially relevant full text articles by two authors independently. As categories for content analysis we defined (i) understanding or recall of information, (ii) preferences regarding information or consent, and (iii) research participants' concerns., Results: The search in Pubmed yielded 337 abstracts of which 10 articles were included in this study. Approaches to information and consent varied considerably across the selected studies. The majority of research participants opted for some version of limited consent when being informed about such possibility. Among the factors influencing the type of preferred consent were information about sponsoring of biobank research by pharmaceutical industry and participants' trade-off between privacy and perceived utility. Studies investigating research participants' understanding and recall regarding the consent procedure indicated considerable lack of both aspects. Research participants' perceptions of benefits and harms differ across those studies., Conclusion: The knowledge, perceptions and views of research participants who have undergone a consent procedure within the context of biobank research raise several questions on the issue of how to inform and elicit consent in an ethically acceptable way. In our empirical-ethical analysis we develop suggestions on how the practice of eliciting consent in the biobank context should be improved.

The masked palm civet (Paguma larvata) is a small Asian mammal (order Carnivora, family Viverridae) uncommon in Italy. Limited information is available about management and sanitary maintenance in captivity. A 4-mo-old masked palm civet presented with pruritus, itch, scratching, and disorexia. On physical examination, alopecia and crusts were detected on the ventral and lateral trunk, tail, legs, and lips. Skin scrapings and cytology revealed Notoedres spp. and bacterial infection. On histopathology, parasitic dermatitis was observed with the presence of a Sarcoptidae mite and Demodex spp. Selamectin spot-on (15 mg/kg every 2 wk, three applications) and marbofloxacin per os (2.5 mg/kg once daily for 2 wk) were administered, and the animal recovered in 1 mo. With the good response to this therapy, a notoedric mange was thought to be the main problem. This is the first report about the use of selamectin to treat a mite infection in masked palm civet.

Background: Gastropod-borne parasites may cause debilitating clinical conditions in animals and humans following the consumption of infected intermediate or paratenic hosts. However, the ingestion of fresh vegetables contaminated by snail mucus and/or water has also been proposed as a source of the infection for some zoonotic metastrongyloids (e.g., Angiostrongylus cantonensis). In the meantime, the feline lungworms Aelurostrongylus abstrusus and Troglostrongylus brevior are increasingly spreading among cat populations, along with their gastropod intermediate hosts. The aim of this study was to assess the potential of alternative transmission pathways for A. abstrusus and T. brevior L3 via the mucus of infected Helix aspersa snails and the water where gastropods died. In addition, the histological examination of snail specimens provided information on the larval localization and inflammatory reactions in the intermediate host., Methodology/principal Findings: Twenty-four specimens of H. aspersa received ~500 L1 of A. abstrusus and T. brevior, and were assigned to six study groups. Snails were subjected to different mechanical and chemical stimuli throughout 20 days in order to elicit the production of mucus. At the end of the study, gastropods were submerged in tap water and the sediment was observed for lungworm larvae for three consecutive days. Finally, snails were artificially digested and recovered larvae were counted and morphologically and molecularly identified. The anatomical localization of A. abstrusus and T. brevior larvae within snail tissues was investigated by histology. L3 were detected in the snail mucus (i.e., 37 A. abstrusus and 19 T. brevior) and in the sediment of submerged specimens (172 A. abstrusus and 39 T. brevior). Following the artificial digestion of H. aspersa snails, a mean number of 127.8 A. abstrusus and 60.3 T. brevior larvae were recovered. The number of snail sections positive for A. abstrusus was higher than those for T. brevior., Conclusions: Results of this study indicate that A. abstrusus and T. brevior infective L3 are shed in the mucus of H. aspersa or in water where infected gastropods had died submerged. Both elimination pathways may represent alternative route(s) of environmental contamination and source of the infection for these nematodes under field conditions and may significantly affect the epidemiology of feline lungworms. Considering that snails may act as intermediate hosts for other metastrongyloid species, the environmental contamination by mucus-released larvae is discussed in a broader context.

Background: Canine noninflammatory alopecia (CNA) is a heterogeneous group of skin diseases with different underlying pathogenesis. The therapeutic approach is challenging, and new options for treatment are desirable., Hypothesis/objectives: To test the clinical efficacy of low-level laser therapy (LLLT) on hair regrowth in CNA., Animals: Seven dogs of different ages, breeds and genders with a clinical and histopathological diagnosis of noninflammatory alopecia., Methods: Each dog was treated twice weekly for a maximum of 2 months with a therapeutic laser producing the following three different wavelengths emerging simultaneously from 21 foci: 13 × 16 mW, 470 nm; 4 × 50 mW, 685 nm; and 4 × 200 mW, 830 nm. The fluence given was 3 J/cm(2) , frequency 5 Hz, amplitude of the irradiated area was 25 cm(2) and application time was 1.34 min. A predetermined alopecic area was left untreated and served as a control area. From one dog, post-treatment biopsies of treated and untreated sites were obtained for histological evaluation of hair density and the percentage of haired and nonhaired follicles., Results: At the end of the study, coat regrowth was greatly improved in six of seven animals and improved in one of seven. By morphometry, the area occupied by hair follicles was 18% in the treated sample and 11% in the untreated one (11%); haired follicles were (per area) 93% in the treated sample and only 9% in the control sample., Conclusions and Clinical Importance: Our clinical and histological data document promising effects of LLLT on hair regrowth in CNA. Further studies investigating the biological mechanism underlying the effect of LLLT on hair follicle cycling are warranted., (© 2014 ESVD and ACVD.)

Background: Hair cycle arrest (alopecia X) refers to a canine alopecic condition of unknown pathogenesis, characterized by symmetrical, nonpruritic and noninflammatory alopecia that spares the head and distal extremities., Hypothesis/objectives: The objective of this study was prospectively to evaluate the efficacy of a 4.7 mg deslorelin implant in the treatment of intact male and neutered female dogs affected by hair cycle arrest., Animals: Ten Pomeranian dogs (eight intact males, two neutered females), four Italian spitz (three intact males, one neutered female), three miniature poodles (two intact males, one neutered female), two Siberian huskies (both intact males) and one intact male chow chow dog with confirmed hair cycle arrest were included in the study., Methods: Each dog was treated with a subcutaneous sterile implant containing 4.7 mg deslorelin. Responder dogs were re-implanted 6 months after the first implant in order to obtain a 1 year pharmacological exposure and 1 year of follow-up., Results: Hair regrowth was visible within 3 months in 12 of 16 intact male dogs (75%); no hair regrowth was noted in any neutered female dogs. The overall response to therapy was 60%. No adverse effects were noted., Conclusions and Clinical Importance: These findings suggest that deslorelin may be a treatment option for intact male dogs with idiopathic hair cycle arrest. Deslorelin is an alternative to current therapies and castration., (© 2014 ESVD and ACVD.)

To investigate larval development of Acanthocheilonema reconditum in the cat flea Ctenocephalides felis, fleas were fed through an artificial feeding system with dog blood containing different concentrations of microfilariae (i.e. low, group L = 250; medium, group M = 500; high, group H = 1500 microfilariae per mL) or no microfilariae (group C). Fleas were sampled at 12 different time-points throughout the study period (D1-D28) and A. reconditum was detected by dissection, PCR and histology. Of 2105 fleas fed with infected dog blood, 891 (38·7%) died during the study before being sampled whilst the remaining (n = 1214) were examined for A. reconditum. Upon dissection, first-stage larvae (L1) were identified after 2 days post infection (D2), second-stage (L2) at D13 and infective third-stage larvae (L3) at D15. Eighteen (30%) of 60 pools of fleas molecularly examined tested positive. Histologically, L2 were detected at D13 in the sub-cuticle region embedded in the back muscle of one female flea. This study provides original data on larval development of A. reconditum in C. felis and reports on the usefulness of the artificial feeding system.

A two or one pot synthesis has been used for the reaction of hyaluronic acid (HA) with octadecylamine (C 18 ) and hydrazine (Hy). In both cases, the chemical derivatization involved primary hydroxyl groups of hyaluronic acid and not its carboxyl groups, whose presence is important for receptor interaction. In this way, Hy-HA-C 18 derivatives have been obtained with appropriate hydrophobic and hydrophilic character. Their ability to form homogeneous physical hydrogels has been evaluated as well as the possibility to obtain porous sponges through salt leaching technology. Sponges showing the highest porosity, potentially compatible with cell entrapment, have been characterized with regard to their physicochemical and biological properties. Swelling ability under simulated physiological conditions and stability in the absence or in the presence of hyaluronidase have been investigated. Bovine chondrocytes were viable in Hy-HA-C 18 sponges as determined with MTS assay and were able to produce collagen and glycosaminoglycans, as assessed by using Masson's trichrome and Alcian blue, respectively. Finally, in vivo degradation of Hy-HA-C 18 sponges has been confirmed after subcutaneous implantation in mice until 6 weeks.

Background: Canine dermatomyositis is a hereditary disease described in collies and Shetland sheep dogs and their cross-breeds. A similar disease, called dermatomyositis-like disease, has been described occasionally in other breeds but never in the Rottweiler., Hypothesis/objectives: We report on the clinicopathological findings associated with dermatomyositis-like disease in a Rottweiler., Animal: A 7-month-old female Rottweiler was referred for dermatological abnormalities, regurgitation and weakness. Cutaneous abnormalities included alopecia, crusting and scaling on the inner surface of the pinnae, the tip of the tail, periorbital and perilabial skin. The dog also presented onychogryphosis and onychalgia., Methods: Complete blood count, serum biochemistry panel, thoracic radiographs, electromyography, nerve-conduction studies and skin and muscle biopsies were performed., Results: Megaoesophagus, pneumonia, ischaemic dermatopathy and generalized myositis were documented. The final diagnosis was dermatomyositis-like disease., Conclusions and Clinical Importance: This is the first report of dermatomyositis-like disease in a Rottweiler., (© 2014 ESVD and ACVD.)

Background: Despite the precise pathogenesis of atopic dermatitis (AD) is unknown, an immune dysregulation that causes Th2-predominant inflammation and an intrinsic defect in skin barrier function are currently the two major hypotheses, according to the so-called outside-inside-outside model. Mast cells (MCs) are involved in AD both by releasing Th2 polarizing cytokines and generating pruritus symptoms through release of histamine and tryptase. A link between MCs and skin barrier defects was recently uncovered, with histamine being found to profoundly contribute to the skin barrier defects.Palmitoylethanolamide and related lipid mediators are endogenous bioactive compounds, considered to play a protective homeostatic role in many tissues: evidence collected so far shows that the anti-inflammatory effect of palmitoylethanolamide depends on the down-modulation of MC degranulation.Based on this background, the purpose of the present study was twofold: (a) to determine if the endogenous levels of palmitoylethanolamide and other bioactive lipid mediators are changed in the skin of AD dogs compared to healthy animals; (b) to examine if MC number is increased in the skin of AD dogs and, if so, whether it depends on MC in-situ proliferation., Results: The amount of lipid extract expressed as percent of biopsy tissue weight was significantly reduced in AD skin while the levels of all analyzed bioactive lipid mediators were significantly elevated, with palmitoylethanolamide showing the highest increase.In dogs with AD, the number of MCs was significantly increased in both the subepidermal and the perifollicular compartments and their granule content was significantly decreased in the latter. Also, in situ proliferation of MCs was documented., Conclusions: The levels of palmitoylethanolamide and other bioactive lipid mediators were shown to increase in AD skin compared to healthy samples, leading to the hypothesis that they may be part of the body's innate mechanisms to maintain cellular homeostasis when faced with AD-related inflammation. In particular, the increase may be considered a temptative response to down-regulating the observed elevation in the number, functionality and proliferative state of MCs in the skin of AD dogs. Further studies are warranted to confirm the hypothesis.

Macrophage aggregates (MA) occur in various organs of fish as discrete aggregations of pigmented macrophages. The study presented herein investigates the quantitative modifications from normal anatomical condition, of interrenal gland (IG) and kidney MA in six treatment groups of adult rainbow trout submitted to either specific or aspecific immune stimulation and subsequently challenged with Yersinia ruckeri. Routinely stained tissue sections from both IG and kidney were analysed. The percentage of tissues occupied by MA and the MA density (number/mm(2)) were calculated on at least 10 randomly selected nonoverlapping fields taken from each tissue section. MA morphometric findings from challenged fish were compared to those from a control group. Results showed that fish from control group displayed a statistically significant (P < 0.05) higher percentage of tissue occupied by MA and MA density. Among different treatment groups, anti-Yersinia ruckeri immunized fish, which did not show clinical signs of disease after bacterial challenge, displayed higher values of morphometric parameters compared with symptomatic fish from other groups. Our study demonstrates that the quantification of the area occupied by MA might be an efficient parameter to evaluate the general condition of a salmonid population since it positively correlates with the health status and negatively with stress factor such as the acute bacterial infection.

Background: Among the arthropod-borne nematodes infesting dogs, Onchocerca lupi (Spirurida: Onchocercidae) is of increasing zoonotic concern, with new human cases of infection diagnosed in Turkey, Tunisia, Iran and the USA. Knowledge of the biology of this nematode is meagre. This study aimed at assessing the distribution and periodicity of O. lupi microfilariae from different body regions in naturally infested dogs., Methodology/principal Findings: Skin samples were collected from six dogs infested with O. lupi but without apparent clinical signs. Two skin samples were collected from 18 anatomical regions of dog 1 at necropsy. In addition, single skin biopsies were performed from the forehead, inter-scapular and lumbar regions of dogs 2-6, in the morning, afternoon, and at night. Two aliquots of the sediment of each sample were microscopically observed, microfilariae counted and morphologically and molecularly identified. Most of the 1,667 microfilariae retrieved from dog 1 were in the right ear (59.6%), nose (26.5%), left ear (6.7%), forehead (3.0%), and inter-scapular (2.9%) regions. In dogs 2-6, the overall mean number of microfilariae was larger on the head (n = 122.8), followed by the inter-scapular (n = 119.0) and lumbar (n = 12.8) regions. The overall mean number of microfilariae was larger in the afternoon (153.4), followed by night (75.4) and morning (25.8)., Conclusions: Onchocerca lupi microfilariae were more common in the head (i.e., ears and nose) than in the remaining part of the dog's body, indicating they tend to aggregate in specific body regions, which are the best sites to collect skin samples for diagnostic purposes. The periodicity pattern of microfilariae of O. lupi and their concentration in specific body regions is most likely a result of the co-evolution with their as-yet-unknown vector. The detection of skin microfilariae in asymptomatic animals, suggests the potential role of these animals as carriers and reservoirs of O. lupi.

Background: The conversion of arachidonic acid into prostaglandin is catalysed by the cyclo-oxygenases (COX-1/COX-2). Several studies indicate that COX-2 is overexpressed in actinic keratosis in humans and dogs. Firocoxib is a COX-2-selective inhibitor that blocks the biochemical activity of COX-2., Hypothesis/objectives: To evaluate the efficacy of firocoxib (5 mg/kg orally once daily) for the treatment of dogs with solar dermatitis/actinic keratosis., Methods: Firocoxib 5 mg/kg was given orally once daily for 180 days to five dogs with clinical signs and histopathological lesions consistent with solar dermatitis/actinic keratosis. On days 0, 50 and 180, the severity of erythema, skin shine, induration and the number of comedones were evaluated by a clinical scoring system. On the same days, samples were collected for histopathology from 'target lesions' and COX-2 expression was evaluated by immunohistochemistry., Results: The clinical follow-up showed that four of five dogs improved with the treatment; improvement in terms of histological findings was correlated with the regularization of the epidermal proliferation rather than the recovery of dermal changes., Conclusions and Clinical Importance: A role for COX-2 might thus be hypothesized in the pathogenesis of canine solar dermatitis., (© 2013 ESVD and ACVD.)

Background: Onchocerca lupi is a dog parasite of increasing zoonotic concern, with new human cases diagnosed in Turkey, Tunisia, Iran, and the United States. Information about the morphology of this nematode is scant and a detailed re-description of this species is overdue. In addition, histopathological data of potential usefulness for the identification of O. lupi infections are provided., Methods: Male and female nematodes, collected from the connective tissue of a dog, were examined using light microscopy and scanning electron microscopy (SEM), and an histological evaluation was performed on biopsy samples from periocular tissues., Results: The morphological identification was confirmed by molecular amplification and partial sequencing of cytochrome oxidase subunit 1 gene. This study provides the first comprehensive morphological and morphometric description of O. lupi from a dog based on light microscopy, SEM, molecular characterization, and histological observations., Conclusions: Data herein presented contribute to a better understanding of this little known parasitic zoonosis, whose impact on human and animal health is still underestimated. The presence of granulomatous reactions only around the female adult suggests that the release of microfilariae from the uterus might be the cause of the inflammatory reaction observed.

Tissues from 34 naturally feline immunodeficiency virus (FIV)-infected cats, 13 asymptomatic cats and 21 cats with signs of feline acquired immunodeficiency syndrome (F-AIDS), and 35 FIV-seronegative subjects were examined to determine the presence of amyloid deposits. Twenty experimentally FIV-infected cats and five specific pathogen-free (SPF) control cats were also included in the study. Paraffin-embedded sections from kidney and other organs were submitted to histological and histochemical analysis. Amyloid deposits were identified by a modified Congo red stain and confirmed by electron microscopy to demonstrate the presence of amyloid fibrils in amyloid positive glomeruli. In all positive cases, secondary amyloidosis was identified with potassium permanganate pretreatment and amyloid type was further characterised by immunohistochemistry using primary antibodies against human AA and feline AL amyloids. Amyloid deposits were present in different tissues of 12/34 (35%) naturally FIV-infected cats (seven presenting F-AIDS and five in asymptomatic phase) and in 1/30 FIV-seronegative cats. All the experimentally FIV-infected and SPF subjects showed no amyloid deposits. Amyloidosis has been reported in human lentiviral infections, and the data reported here demonstrate the need, in naturally FIV-infected cats, to consider the presence of amyloidosis in differential diagnosis of hepatic and renal disorders to better assess the prognosis of the disease.

Background: Pili torti is a congenital or acquired human disease characterized by flattening and twisting of the hair shaft by 180 degrees. A similar condition has been previously reported in a litter of kittens with generalized hair loss, associated with systemic signs and followed by death at a young age., Animal: A 1-year-old castrated male domestic short-haired cat was presented for noninflammatory and nonpruritic symmetrical multifocal alopecia involving the head, pinnae, forelimbs, tail and chest. The cat was otherwise healthy., Methods and Results: Microscopic examination of hair shafts revealed pili torti and a typical combination of anagen and telogen roots, occasionally spiral in shape, without evidence of mites or dermatophytes. Histopathological examination showed the same hair shaft abnormality, with normal epidermis, dermis, hair follicles and other adnexa. Scanning electron microscopy confirmed a hair shaft dysplasia characterized by pili torti, similar to that described in humans., Conclusions and Clinical Importance: To the best of the authors' knowledge, this is the first case report of pili torti in a healthy young adult cat., (© 2013 The Authors. Veterinary Dermatology © 2013 ESVD and ACVD.)

Proliferative and necrotising otitis externa is a rare and recently described disease affecting the ear canals and concave pinnae of kittens. This article describes a case of proliferative and necrotising otits externa in a young adult cat. In this case, the lesions did not affected the pinnae, but both ear canals were severely involved. Video-otoscopy revealed a digitally proliferative lesion, growing at 360° all around the ear canals for their entire length, without involvement of the middle ear. Histopathological examination confirmed the diagnosis, and the cat responded completely to a once-daily application of 0.1% tacrolimus ointment diluted in mineral oil in the ear canals. Video-otoscopy findings, not described previously, were very peculiar and may help clinicians to diagnose this rare disease.

An eleven year-old mongrel dog was referred with a history of left forelimb lameness and an ulcerated mass on the neck. Histologically, the cutaneous neoplasm revealed cystic lobules composed of basaloid cells with abrupt transition to central keratotic material, containing pycnotic and shadow cells. Approximately 3 months after primary diagnosis, a lesion of the cortical bone on the left humerus was observed using X-ray. Samples obtained from the humerus were processed for histopathological examination and the neoplastic tissue was observed to be similar to the type identified in the neck. Based on these findings, the tumor was diagnosed as a malignant pilomatricoma (MP) with bone metastasis. MP is a rare skin tumor that originates from hair matrix cells. To date, only nine reports have been presented in dogs. In the present study, we discuss the cytological and histological patterns of MP, confirmed by immunohistochemistry using β catenin antibody.

Background: Canine subcutaneous dirofilariosis caused by Dirofilaria repens is an important cause of human dirofilariosis in the Old World., Objectives: To describe the clinical and pathological features of infestation by D. repens in dogs with cutaneous nodules and to discuss key diagnostic procedures., Animals: Sixteen privately owned dogs of different breeds, ages and genders with subcutaneous nodules were positive for D. repens on at least one diagnostic test., Methods: Diagnostic tests included cytological examination of fine-needle aspiration biopsies, detection of circulating larvae by Knott's test, morphological and molecular identification of adult parasites and microfilariae, and histopathological examination of excised nodules., Results: Twelve animals were positive for D. repens microfilariae, using cytology and at least one other diagnostic test. Four of twelve dogs screened by Knott's test had circulating D. repens microfilariae, and three of them were co-infected with Dirofilaria immitis. In eight dogs, adult nematodes were morphologically identified as D. repens. Nodular panniculitis was observed in all cases, with nodules containing either one or several cross-sections of nematodes and microfilariae. The nematodes extracted were identified as D. repens by an RT-PCR on the second internal transcribed spacer (ITS-2)., Conclusions and Clinical Importance: Adults and larvae of D. repens can induce skin nodules in dogs, and these lesions should be carefully evaluated in endemic areas. Clinicians should be aware that correct tissue sampling (including the storage of part of a sample in ethanol instead of formalin) is pivotal to achievement of a specific diagnosis. Cytology of fine-needle tissue aspirates was shown to be useful., (© 2013 The Authors. Veterinary Dermatology © 2013 ESVD and ACVD.)

The aim of this study was to investigate the anatomical distribution of microfilariae of a recently described Cercopithifilaria sp. and the relationship with the preferred attachment sites of its vector, the brown dog tick Rhipicephalus sanguineus. Skin samples from 20 dogs were collected from eight anatomical sites and soaked twice in saline solution at 37°C. All samples were also molecularly processed for the specific amplification of partial cytochrome c oxidase 1 gene. Microfilariae were unevenly distributed on the body, with higher frequencies on interscapular region (n=13; 68.4%) and on the head (n=9; 47.4%). Larval abundance in dogs was positively correlated with the number of positive anatomical sites with a higher percentage of skin samples scoring positive at the microscopic examination of the first sediment (n=47; 30.9%) than of the second (n=8; 5.3%). Histological examination revealed that microfilariae were scattered in the dermis, in association with inflammatory cells. Molecular analysis of skin samples showed a lower frequency of positive sites (n=37; 24.3%) when compared to the microscopic examination (n=53; 34.9%) with five haplotypes sequenced. The results here presented suggested a close co-evolution of this filarioid with its vector., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Objective: To determine the distribution of cannabinoid receptor type 1 (CB1) and cannabinoid receptor type 2 (CB2) in skin (including hair follicles and sweat and sebaceous glands) of clinically normal dogs and dogs with atopic dermatitis (AD) and to compare results with those for positive control samples for CB1 (hippocampus) and CB2 (lymph nodes)., Sample: Skin samples from 5 healthy dogs and 5 dogs with AD and popliteal lymph node and hippocampus samples from 5 cadavers of dogs., Procedures: CB1 and CB2 were immunohistochemically localized in formalin-fixed, paraffin-embedded sections of tissue samples., Results: In skin samples of healthy dogs, CB1 and CB2 immunoreactivity was detected in various types of cells in the epidermis and in cells in the dermis, including perivascular cells with mast cell morphology, fibroblasts, and endothelial cells. In skin samples of dogs with AD, CB1 and CB2 immunoreactivity was stronger than it was in skin samples of healthy dogs. In positive control tissue samples, CB1 immunoreactivity was detected in all areas of the hippocampus, and CB2 immunoreactivity was detected in B-cell zones of lymphoid follicles., Conclusions and Clinical Relevance: The endocannabinoid system and cannabimimetic compounds protect against effects of allergic inflammatory disorders in various species of mammals. Results of the present study contributed to knowledge of the endocannabinoid system and indicated this system may be a target for treatment of immune-mediated and inflammatory disorders such as allergic skin diseases in dogs.