Management of canine wounds using platelet-rich fibrin (PRF) biomaterial. A case series report.

BACKGROUND: The increasing interest in platelet-based therapies has underwritten the development of novel veterinary regenerative treatments. The haemoderivative platelet-rich fibrin (PRF) comprises abundant concentrations of platelets and leucocytes, above the physiologic baseline, which are considered essential elements for wound regeneration, stimulating local angiogenesis, cellular migration, proliferation and differentiation, considered essential for skin repair. OBJECTIVES: This study aimed to describe the treatment of eight dogs with naturally occurring cutaneous wounds, where autologous PRF therapy was applied, using a protocol developed by our group. METHODS: Eight dogs, aged between 7-month and 9-year old, from different breeds and sexes, were enrolled in this study. Four of these wounds were clinically infected. In three cases, two PRF treatments were performed during the first week of treatment, followed by single weekly treatments from the second week onwards, until exophytic granulation tissues were present. In each case, the treatment was finalized only when complete wound closure was achieved. Wounds did not receive topical antiseptics, antibiotics or topical drugs to promote wound healing during the treatment. RESULTS: PRF-grafting treatments were well tolerated in all treated wounds, inducing significant granulation tissue formation. PRF clots acted as a natural tissue filler, promoting epithelization and wound closure, without the requirement of topical antimicrobial/antiseptics application, or additional surgical debridement. Evident skin contraction was recorded in larger injuries and all the treatments resulted in vestigial aesthetic scars where hair growth was also observed. CONCLUSIONS: PRF-therapy obtained promising results, as an alternative wound treatment, revealing a biological regenerative action, prompting the natural skin healing process.

Updating an Overview of Teratology.

In this chapter, the authors aim to update an overview of the principles of teratology, beginning with the definition of teratology, the critical point at which this process occurs, and some of the most common etiological agents that improve our understanding of teratology.Modern teratology has greatly improved in recent years with advances in new methods in molecular biology, toxicology, animal laboratory science, and genetics, increasing our knowledge of ambient influences. Nevertheless, there is a lot to do to reduce the influence of hazardous intervening agents, whether they target our genetics or not, that can negatively affect pregnancy and induce congenital development disorders, including morphological, biochemical, or behavioral defects.Certain agents might indeed be related to certain defects, but we have not been able to identify the cause of most congenital defects, which highlights the importance of finding and testing out new genetics techniques and conducting laboratory animal science to unravel the etiology and pathogenicity of each congenital defect.

Effect of Immunocastration on Culled Sows-A Preliminary Study on Reproductive Tract, Carcass Traits, and Meat Quality.

The Bísaro pig is a Portuguese autochthonous breed greatly appreciated for its meat quality and is mainly reared outdoors. Immunocastration could be a solution to avoid undesirable pregnancies and boar taint in cull sows. The present study tested three immunocastration protocols (with Improvac®) according to their reproductive cycle. The first inoculation was performed two weeks after farrowing (IM1, n = 5), at the beginning of estrus (IM2, n = 5), and one week after the end of estrus (IM3, n = 5), followed by a second administration four weeks apart. A control group (C, n = 5) was also included in the same housing conditions. The sample collection included the reproductive tract for morphometric evaluation, neck fat for the quantification of boar taint compounds, and a portion of the Longissimus thoracis et lumborum for meat quality trait assessment. The reproductive tracts from intact sows (C) were significantly heavier compared to the immunocastrated groups (p < 0.05) (1.403 kg C to 0.508 kg IM1, 0.590 kg IM2, and 0.599 kg IM3), suggesting the regression of the reproductive tract to nonstimulated conditions due to immunization against GnRH. The IM1 group exhibited significantly smaller reproductive tract measurements compared to group C for most of the evaluated segments (p < 0.05). No marked differences were observed in the meat quality traits. Therefore, immunocastration can be used in culling sows to avoid ovarian activity, and it is not detrimental to pork quality traits.

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.

Seasonal Variations on Testicular Morphology, Boar Taint, and Meat Quality Traits in Traditional Outdoor Pig Farming.

Traditional outdoor pig farming is renowned for its emphasis on animal welfare and the production of highly valued, quality meat. While seasonality is known to impact certain animals, particularly those raised outdoors, there is a lack of research on Bísaro boars, a native Portuguese breed. This research study was conducted on a total of 20 male entire Bísaro pigs, reared in outdoor pens from 4 to 13 months old, and subsequently slaughtered. The animals were divided into two groups: one slaughtered in winter (Wi, n = 9), and the other in summer (Su, n = 11). The objective was to evaluate testicular morphometry, boar taint compounds, and meat quality traits, including sensory analysis and fatty acid profile. Testicles from the Su group exhibited reduced volume, indicating diminished functionality during that season. While no significant differences were observed in the boar taint compound analysis, panelists could discern a more intense aroma and flavor of boar taint in the Su meat. Other meat quality traits showed no significant variations, but the fatty acid profile displayed higher values in the Wi group. This study reveals that Bísaro boars experience reproductive seasonality, leading to variations in boar taint compounds across the seasons. This information is crucial for farm planning.

Coxiella burnetii in Dogs and Cats from Portugal: Serological and Molecular Analysis.

Dogs and cats are potential sources of infection for some zoonotic diseases such as Q fever, caused by Coxiella burnetii, a multiple host pathogen. Q fever outbreaks in dogs and cats have been related with parturition and abortion events, and ticks have a potential role in the transmission of this pathogen. This study aimed to screen for C. burnetii in dogs and cats, and in ticks collected from infested animals. An observational descriptive study was conducted in Portugal at two time points nine years apart, 2012 and 2021. Sera obtained from dogs and cats (total n = 294) were tested for C. burnetii antibodies using a commercial ELISA adapted for multi-species detection. C. burnetii DNA was screened by qPCR assay targeting IS1111 in uterine samples and in ticks. A decrease in the exposure to C. burnetii was observed in cats from 17.2% (95% CI: 5.8−35.8%) in 2012 to 0.0% in 2021, and in dogs from 12.6% (95% CI: 7.7−19.0%) in 2012 to 1.7% (95% CI: 0.3−9.1%) in 2021 (p < 0.05). Overall, and despite differences in the samples, rural habitat seems to favour the exposure to C. burnetii. The DNA of C. burnetii was not detected in ticks. The low seropositivity observed in 2021 and the absence of C. burnetii DNA in the tested samples, suggest that dogs and cats from Portugal are not often exposed to the pathogen. Nevertheless, the monitoring of C. burnetii infection in companion animals is an important tool to prevent human outbreaks, considering the zoonotic potential for owners and veterinarians contacting infected animals, mainly dogs and cats from rural areas which often come into contact with livestock.

Feline Oral Squamous Cell Carcinoma: A Critical Review of Etiologic Factors.

Feline oral squamous cell carcinoma (FOSCC) is the most common oral neoplasia in cats. This malignant tumor is locally invasive, has a high mortality rate, and its etiology is not yet known. In humans, head and neck squamous cell carcinoma is associated with tobacco smoke, alcohol consumption, and human papillomavirus infection. Herein, a critical review about the potential etiologic factors of FOSCC was performed, considering publications between 2000 and 2022, aiming to synthesize all available scientific evidence regarding this issue. Recommendations of the PRISMA statement and the Cochrane Collaboration were followed and the PubMed database searched by using the MeSH terms MeSH terms "oral", "mouth", "lingual", "labial", "gingiva", "carcinoma", "squamous", and "feline". The selection process for eligible studies was based on specific inclusion and exclusion criteria and the quality of the studies assessed. The initial search resulted in 553 publications, with only 26 of these being included in the review. Sixteen studies were related to viral etiology and nine related to environmental factors such as exposure to tobacco smoke, ectoparasitic products, and the presence of oral comorbidities. When evaluated, feline papillomavirus was detected in 16.2% of samples of FOSCC. In the three studies focused on exposure to tobacco smoke, 35.2% (30/85) of cats with FOSCC had a history of this exposure. The consumption of canned food and the use of deworming collars were associated, in only one publication, with a risk of neoplasia increased by 4.7 and 5.3 times, respectively. Among 485 cats with FOSCC, 6.4% had dental and oral pathology (i.e., periodontal disease or feline chronic gingivostomatitis). The present study demonstrates that the available evidence on the etiology of FOSCC is still limited, however, there has been an increasing interest on this topic. To better understand the role of the possible etiological factors of this aggressive disease, and model for its human counterpart, large, prospective multi-institutional studies are needed.

Pathological Findings in African Pygmy Hedgehogs Admitted into a Portuguese Rehabilitation Center.

Most of the pathologies that affect hedgehogs are diagnosed postmortem; thus, it is essential to share knowledge between clinicians and pathologists in order to recognize predispositions to diseases and to establish adequate diagnostic and therapeutic plans. This study aimed to describe the most relevant postmortem pathological conditions in a group of six rescued African pygmy hedgehogs, performed over a period of four months. Hedgehogs were submitted to necropsy examinations and subsequent histopathological analyses. Microscopically, all the studied hedgehogs revealed alterations in one or more organ systems. Although a significant and diverse number of pathological conditions were obtained, this study focused on less common or more relevant pathologies found in African pygmy hedgehogs-namely, wobbly hedgehog syndrome, squamous cell carcinoma and mast cell tumors. Furthermore, this study constitutes the first report of Mycobacterium spp. in hedgehogs in Portugal, the second report of follicular thyroid carcinoma in an African pygmy hedgehog, the description of a lipoid pneumonia for the first time in this species and a lung adenocarcinoma-a pathology rarely reported in African pygmy hedgehogs.

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).

Co-existing monophasic teratoma and uterine adenocarcinoma in a female dog.

Ovarian teratomas are occasionally reported in dogs; the rarest type is the monophasic teratoma, composed of tissues originating from only one germ layer. Canine endometrial adenocarcinomas are also rare in dogs and mainly affect geriatric females. This report describes the case of co-existing ovarian teratoma and uterine adenocarcinoma in a 10-year-old nulliparous female Boxer presented with lethargy, anorexia and purulent vaginal discharge. Abdominal ultrasonography evidenced pyometra and a mass in the left ovary. This was composed of a uniform whitish tissue with multiple cystic structures. The histology revealed an atrophy of the ovarian parenchyma, compressed by a proliferation of well-differentiated nervous tissue staining positively to vimentin, S100 and neuronal specific enolase (NSE), and negatively to keratin and inhibin. The left uterine horn, whose diameter was markedly increased, showed foci of endometrial cellular atypia, evident nucleoli and mitoses, at light microscopy. To our best knowledge, this is the first report of a co-existing ovarian monophasic teratoma and endometrial adenocarcinoma, two rare reproductive neoplasia in dogs.

The eyeworm Thelazia callipaeda in Portugal: Current status of infection in pets and wild mammals and case report in a beech marten (Martes foina).

Ocular thelaziosis is caused by nematodes of the genus Thelazia (Spirurida, Thelaziidae), which inhabit the surface of the eyes and associated tissues. Thelazia callipaeda affects a range of mammal species, including humans, and in the last two decades has been reported in multiple European countries, being classified as an emergent vector-borne pathogen. In Portugal T. callipaeda is endemic in north-eastern areas, where it has been reported in domestic dogs, cats, red foxes (Vulpes vulpes) and wild European rabbits (Oryctolagus cuniculus). We report, for the first time, T. callipaeda in a beech marten (Martes foina) from Portugal and highlight the presence of haplotype 1 as the only one found in Europe, irrespective of the host species and geographical area of provenience.

Distribution of superoxide dismutase 1 and glutathione peroxidase 1 in the cyclic canine endometrium.

Superoxide dismutase (SOD) and glutathione peroxidase (GPx) are two important antioxidant enzymes involved in tissue homeostasis by protecting cells and tissues from an accumulation of reactive oxygen species. Information concerning antioxidant enzymes in the canine uterus is almost inexistent. This work intends to establish the pattern of distribution of SOD1 and GPx1 immunoreaction in canine endometrium throughout the estrous cycle, using 46 endometrium samples of healthy dogs representing different cycle stages (anestrus-10, proestrus-10, estrus-10, early diestrus-7, and diestrus-9). SOD1 distribution in canine endometrium showed cyclic variations (P ≤ 0.001), with higher immunoscores in the progesterone-associated stages. Changing immunoreaction also concerned the different epithelial structures considered (surface epithelium, superficial glandular epithelium, and deep glandular epithelium) (P ≤ 0.001), but it was always higher than in the stroma (P ≤ 0.001). Deep glandular epithelial cells usually showed higher scores of immunoreaction compared with the other epithelial cells. Interestingly, in epithelial cells, distinct subcellular patterns for SOD1 were seen: the nuclear labeling was observed in estrus and early diestrus (P ≤ 0.001), whereas an apical reinforcement was observed in estrus (P = 0.011) in the glandular epithelia but not in the surface epithelia. In general, GPx1 distribution in canine endometrium remained relatively unchanged throughout the estrous cycle (P = 0.169) despite the slight decrease observed from proestrus to early diestrus. The highest scores were found in anestrus and diestrus (P < 0.05), varying with of the structure considered. An apical reinforcement pattern was also found for this molecule, which peaked in proestrus and estrus (P < 0.005). In summary, the present study showed that SOD1 and GPx1 are consistently distributed in the canine endometrium. The cyclic changes registered for both molecules suggest that they may play important roles in endometrial physiology, probably in apoptosis and proliferation.

An immunohistochemical study on the expression of sex steroid receptors, Ki-67 and cytokeratins 7 and 20 in feline endometrial adenocarcinomas.

BACKGROUND: Endometrial adenocarcinomas are a rare type of tumour in cats. Though different morphologies have been reported, the most frequent histological type of feline endometrial adenocarcinoma (FEA) is the papillary serous. Characterization of molecular markers expression in FEA may contribute to clarify the pathogenesis of these tumours and to assess the differences between normal endometrium and FEA regarding the expression pattern of several proteins. Therefore, this study aimed to evaluate the immunohistochemical profile of a wide panel of antibodies (specific for ER-α, PR, Ki-67, CK7 and CK20) in twenty-four cases of FEA. Comparisons were made between FEA and feline normal cyclic endometrium in follicular (n = 13) and luteal (n = 10) stages. Except for Ki-67, all other molecular markers were assessed independently for the intensity of immunolabeling and for the percentage of cells expressing the protein. RESULTS: This study showed that in FEA a loss of expression occurs for ER-α (P ≤ 0.0001) and less markedly also for PR. The lost in sex steroid receptors concerns a decrease in both the proportion of labelled cells and the intensity of immunolabelling (P = 0.002 and P = 0.024, respectively). Proliferative activity, estimated via Ki-67 immunoreaction, significantly increased in FEA as compared to normal endometrium (P ≤ 0.0001). Feline endometrial adenocarcinomas maintained the CK7+/CK20+ status of normal endometrium. However, FEA showed decreased CK7 intensity of labelling compared to normal endometria (P ≤ 0.0001) and loss of CK20 expression, both in intensity (P ≤ 0.0001) and in percentage of positive cells (P = 0.01), compared to normal tissues. CONCLUSIONS: Data gathered in this study suggest that proliferation in FEA accompanies ER-α down-regulation, possibly following activation of pathways mediated by local growth factors. Moreover, FEA retains combined expression of CK7 and CK20, as evidenced in normal endometrial epithelia, although a decrease in CK7 expression was observed.

Lipid digestion, absorption and uptake in Solea senegalensis.

Dietary lipids are the major energy source for metabolic purposes in most fish species, and improve dietary protein utilization for growth. In a previous study we have reported a low tolerance of Senegalese sole juveniles to dietary lipid levels and suggested a maximal dietary inclusion level of 8% lipids for both optimal growth and nutrient utilization. The mechanisms behind this apparent poor utilization of the dietary lipids are still to be elucidated. The primary aim of the present study was to investigate the overall process of digestion and lipid absorption in relation to dietary lipid levels. Triplicate groups of twenty fish (mean initial mass 29g) were fed two isonitrogenous diets (54% of protein dry matter basis) with different lipid levels (L4 and L17, 4 and 17% lipids dry matter basis), for 88days. Protein and lipid apparent digestibility coefficients as well as lipase activity were similar in both groups suggesting that Solea senegalensis has the ability to digest equally well a low fat or a high fat diet. Plasma triglyceride concentrations were significantly higher 5 and 16h after feeding in fish fed the L17 compared to those fed L4, following dietary lipid supply, demonstrating effective lipid absorption. Expression of proteins related to lipid transport (microsomal triglyceride transfer protein), trafficking (Fatty acid binding protein 11) and fatty acid uptake (VLDL-r) was significantly higher in liver of fish fed the high fat diet 16h after the meal, but remained unchanged in muscle. In conclusion, it seems that high fat diets do not impair lipid digestion and absorption.

A case of pulmonary cryptococcosis in a free-living toad (Bufo bufo).

Pulmonary cryptococcosis was observed in a free-living adult female common toad (Bufo bufo) that was killed by a vehicle. Both lungs had various eosinophilic, monomorphic, and spherical to elliptical organisms identified as Cryptoccocus spp. The yeasts were demonstrated by Grocott's silver method and the periodic acid-Schiff reaction and the capsule was positive for mucin with a mucicarmine stain. The agent was confirmed by immunohistochemistry, using the monoclonal antibody anti-Cryptococcus neoformans, and by a polymerase chain reaction-based method using a C. neoformans-specific primer. This report, to the best of our knowledge, represents the first case of cryptococcosis in a common toad.

Prenatal administration of vitamin A alters pulmonary and plasma levels of vascular endothelial growth factor in the developing mouse.

Vitamin A and the retinoids play a unique role in mammalian embryonic and foetal development and are essential for both cellular differentiation and the establishment of normal morphogenesis. Vascular endothelial growth factor (VEGF) is a known potent mitogenic factor that plays a key role in lung development and function maintenance. In order to contribute to a better knowledge of the modulating effects of vitamin A in lung development, we investigated the effects of the antenatal administration of vitamin A on VEGF expression in lungs and plasma from foetuses and neonates. Pregnant mice were subjected to subcutaneous administration of vitamin A on the 12th gestational day. The lungs and plasma from foetuses and neonates were collected daily from the 15th gestational day till the day of birth. Our results show that vitamin A modulates VEGF concentrations both in lungs and plasma. Statistically significant differences were observed at gestational days 15 (P = 0.004 for lungs; P < 0.0001 for plasma), 16 (P < 0.0001 for lungs and plasma) and 18 (P < 0.0001 for lungs; P < 0.05 for plasma). Vitamin A tends to increase the expression of this factor in the lung, particularly during the critical period of perinatal adaptation to postnatal life. These effects seem to be spatial and temporally regulated, and point out to the important role of vitamin A during lung development.