Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.(AU)

A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.(AU)

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Abstract Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

Resumo A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation

Abstract Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

Resumo A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Carcinoma de células escamosas em gato - tratamento com quimioterapia metronômica / Squamous cell carcinoma in a cat - treatment with metronomic chemotherapy

Background: Squamous cell carcinoma (SCC) is a malignant neoplasm associated with prolonged exposure to ultraviolet light and lack of skin pigment in white-coated animals. The disease diagnosis is based on the animal's history, clinical signs and histopathology of the samples collected. The therapeutic alternatives are diverse. Among them, one of the most used is the metronomic chemotherapy which is based on the use of cytostatics drugs, applied in low and frequent doses for a longer period of time. One of the mechanisms of action of this type of therapy is the decrease in angiogenesis. Non-steroidal anti-inflammatory drugs (NSAIDs) are simultaneously used to maximize the angiogenic effect. The objective of this study is to report the results of a treatment accomplised with metronomic chemotherapy (cyclophosphamide with meloxicam) in a domestic feline which presented a squamous cell tumor. Case: A 7-year-old female cat, mixed breed, not neutered, weighing 3.5 kg, was assisted at the Veterinary Specialties Teaching Clinic (CEEV) of the Technical University of Machala (UTMACH), presenting bilateral ulcerative lesion in the nasal plane for 5 months. Chronic exposure to sunlight is referred to in the patient's anamnesis. Aspiration cytology performed in the patient revealed an inflammatory process with a predominance of neutrophils and macrophages, and showed absence of neoplastic cells. A treatment based on antibiotics and corticosteroids was adopted. As there were no favorable results, an excisional biopsy procedure was chosen to collect the affected tissues. The result of the histopathological analysis was a level 2 squamous cell carcinoma (SCC). The initial treatment consisted of surgical removal of the tumor and later the use of metronomic chemotherapy based on Cyclophosphamide, Meloxicam, DHA (docosahexaenoic acid) and Sucralfate for 5 months was adopted. At the beginning of the second phase of treatment, the patient developed a macula that progressively diminished. Currently, the patient does not have any recurrence of the diagnosed neoplasm. Discussion: The diagnosis of squamous cell carcinoma was based on clinical and histopathological findings. The macroscopic lesions were crusted ulcers that do not heal in the nasal plane and auricular pavilion, as reported in the literature. SCC is characterized by the infiltration of squamous cells in the dermal layer of the skin, forming epithelial islands with keratin pearls. The histopathological findings of this case present the same characteristics. The climatic conditions of Ecuador, high temperatures with greater incidence of sunlight, hypopigmented areas of the animal's body and chronic exposure to solar radiation are risk factors present in this case that foster the development of this malignant neoplasm. There are multiple treatments to be introduced in this sort of pathology. In this case, surgical removal of the tumor was adopted, followed by metronomic chemotherapy based on cyclophosphamide at 10 mg/m2 and meloxicam at a 0.05 mg/kg per day for a period of time of five months, obtaining favorable results and absence of recurrence. Despite metronomic chemotherapy being easy to administer, with fewer side effects, which includes a better acceptance of care takers who are reluctant to administer conventional chemotherapy, further studies are still necessary in what concerns certain variables such as age, carcinoma level, appropriate dosages and drugs according to each neoplasm.

Solid iridociliary carcinoma in a dog

Background: Iridociliary epithelial tumors (ICETs) originate from the iris epithelium or ciliary body. They comprise ciliary body adenoma, carcinoma, pleomorphic adenocarcinomas, medulloepitheliomas, and other primitive neuroectodermal tumors. They are the second most common primary intraocular tumors in dogs and have already been reported in sheep and humans. In dogs, they occur more frequently in middle-aged to elderly animals, and the Labrador and Golden Retriever seem to be more predisposed breeds. This study aimed to describe the clinical and pathological aspects of solid iridociliary carcinoma in a dog. Case: A 3-year-old Poodle bitch was treated for discomfort in the left eyeball region, increased intraocular pressure and moderate buphthalmia. A direct ophthalmological examination was performed without equipment, and a mass was visualized in the posterior chamber, distorting the pupillary cleft. We opted for unilateral enucleation and forwarded the material for histological analysis. Macroscopically, the eyeball measured 3.4 cm (anteroposterior) x 2.6 cm (vertical), with a brownish mass that occupied the entire anterior chamber and part of the posterior chamber. Histologically, there was a neoformation in the ciliary body and iris pigment epithelium, partially well-delimited and densely cellular. The neoplasm was organized into predominantly solid formations interspersed with a discrete amount of blood vessels, rare bundles of fibrous stroma, and amorphous eosinophilic material forming membranes that were positive for PAS. Sections of the neoplasm were subjected to immunohistochemistry using anti-cytokeratin AE1/AE3, anti-S100 protein, anti-vimentin, and anti-Ki-67. Positive cytoplasmic immunostaining for cytokeratin and S-100 was observed. Only 45.6% of cells were positive for Ki-67 (500 cells). No immunostaining was observed for vimentin. Discussion: The diagnosis of solid iridociliary carcinoma was based on the histological features and positive immunostaining for cytokeratin AE1/AE3 and protein S100. Iridociliary carcinomas present positive immunostaining for cytokeratin, whereas adenomas and normal iridociliary epithelium do not present this immunostaining. Moreover, the high rate of cell proliferation was indicative of malignant neoplasia, as observed by the high mitotic count and high positivity for Ki-67. The S100 protein helped in the diagnosis of ICETs, as the iridociliary epithelium showed positive staining for this protein. Some histological features are important to consider in the diagnosis of iridociliary tumors in dogs, such as noninvasive growth in the posterior chamber, pigment epithelium, and thick homogeneous membranes on the cell surface. Furthermore, the presence of positive PAS membranes favors the diagnosis of iridociliary epithelial tumors. ICETs must be differentiated from melanocytomas, anterior uveal melanoma, medulloepitheliomas, and metastatic and pleomorphic carcinomas. The histological characteristics, especially the presence of PAS-positive membranes, associated with the immunohistochemical profile of neoplasm cells, help differentiate the ICETs from these tumors. In general, the prognosis is poor for eyeball and vision maintenance in canine iridociliary tumors, and scleral invasion is associated with a higher recurrence rate.

Metastatic vulvar squamous cell carcinoma in a mare

Background: Squamous cell carcinoma (SCC) is a common cutaneous neoplasm in horses, which mainly affects the external genitalia, oral cavity, and periocular region. The development of SCC metastases is rare in these animals, and the most common occurrence is a marked local infiltration. Exposure to ultraviolet rays and skin depigmentation are the main etiological factors of SCC. Definitive diagnosis of the neoplasm is performed through histological examination of lesions. The present report describes the clinical, anatomopathological, and immunohistochemical (IHC) aspects of a case of metastatic SCC, with the vulva as the primary site in a mare. Case: A 17-year-old mare, mixed breed, was referred to the Veterinary Hospital of the Universidade de Passo Fundo (UPF), with history of areas of depigmented skin (pinto coat), and clinical history of anorexia, frequent episodes of colic, and recumbency. During clinical evaluation, heart rate (68 bpm), rectal temperature (38.4ºC), and respiratory rate (48 mpm) were elevated, and the oral mucosa was pale. The mare also had an ulcerated tumor mass involving the vulva, which extended to the inguinal region and involved the mammary gland. A cytological aspirate of the vulvar tumor was performed, in which no neoplastic cells were found. Next, a biopsy of 2 distinct areas of the vulva was performed. The material was sent for anatomopathological examination, which showed markedly pleomorphic malignant squamous cells, with individual keratinization and high mitotic index, organized in trabeculae with rare keratin pearl-like formations. The exam allowed the diagnosis of SCC Grade II. Due to the poor prognosis and high cost of treatment, the owner consented to euthanasia and necropsy examination. During necropsy, the vulvar tumor mass was grayish, firm to hard, infiltrative, and had friable areas. Tumor foci suggestive of metastasis were also observed in inguinal, mesenteric, mediastinal and renal lymph nodes, adrenal glands, lung, pericardium, medullary canal, intercostal muscles, right 15th rib, and tissue surrounding the azygos vein. Samples from all organs were collected for anatomopathological examination. Diagnosis of metastatic vulvar SCC was confirmed through histological and IHC studies, which evaluated the expression of cytokeratins (AE1/ AE3), as well as the proliferative activity of neoplastic cells through the PCNA marker. Discussion: The diagnosis of metastatic SCC was obtained through the observed clinical, necroscopic, histological, and IHC characteristics. This neoplasm usually appears in depigmented regions exposed to ultraviolet light, and older animals are more likely to be affected. The mare in the present case had areas of depigmented skin (pinto coat). The animal was kept in a paddock outdoor and exposed to constant solar radiation. Although rare in horses, manifestation of anorexia, progressive weight loss, and frequent colic episodes and recumbency may be closely related to the multiple sites of metastasis in the present case. The main histological findings of the neoplasm were the dense proliferation of malignant squamous cells with individual keratinization, arranged in a trabecular pattern and with rare formations of keratin pearls, in line with previous studies. During IHC evaluation, the neoplastic cells showed expression of cytokeratins (AE1/AE3), as well as high proliferative activity evidenced by the PCNA marker. Given this background, the present report describes the clinical, anatomopathological, and IHC aspects of a case of metastatic SCC with a primary site in the vulva of a mare.

Primary Tracheal Squamous Cells Carcinoma in a Dog

Primary tracheal neoplasms are uncommon and poorly documented in veterinary medicine, being most frequently reported in domestic cats. Squamous cell carcinoma (SCC) is a malignant neoplasm that originates from the stratified squamous epithelium, considered one of the most common skin neoplasms in dogs and cats. This paper aims to report the anatomopathological and immunohistochemical findings of a clinical case of primary SCC in the trachea of ​​a female Schnauzer canine, attended at the Veterinary Hospital Luiz Quintiliano de Oliveira of the Faculty of Veterinary Medicine of Araçatuba (FMVA ­ UNESP), complaining of choking, coughing and dyspnea for 1 month. The tracheal portion affected by the neoplasm was sent to the Veterinary Pathology department of the FMVA after surgical excision. Fragments of the neoplasm were collected and fixed in 10% formaldehyde for further histopathological and immunohistochemical analysis. Microscopically, the neoplasm was well differentiated, being characterized by the presence of keratin pearls, low degree of pleomorphism and rare mitotic figures. In the immunohistochemical analysis, there was immunoexpression of anti-cytokeratin antibodies AE1AE3, 34BE12, CK14 and CK5/6, confirming the diagnosis of squamous cell carcinoma. In about 30% of the cells there was immunostaining for Ki67 antibodies, justifying the low mitotic index of tumor cells and the few images of mitosis seen. Due to the rare occurrence of primary tracheal SCC in dogs, the use of diversified diagnostic techniques is important in order to better understand the biological behavior of this neoplasm in unusual anatomical locations.(AU)

Carcinoma de células escamosas em pálpebra com metástase ocular em um gato doméstico / Squamous cell carcinoma of the eyelid with ocular metastases in a domestic cat

Background: Eye neoplasms are infrequent in felines and usually affect the eyelids. Squamous cell carcinoma (SCC) is a neoplasm that occurs owing to recurrent lesions caused by solar radiation and affects the margins of the ears, eyelids, nose and lips of animals with light-colored coat. The clinical signs consist of ulcerated, crusty lesions that may form masses. The diagnosis is established by means of histopathological analysis, and the prognosis depends on the area affected and tumor infiltration. This work aimed to describe a case of SCC in the eyelid with invasion of the eye and its adnexa in a cat, highlighting clinical aspects and the importance of the anatomopathological examination. Case: A female mixed breed senile cat that had been rescued from the streets was brought in for examination. Dehydration and a low body condition score were observed at the general clinical examination. A physical inspection revealed the presence of an ulcerated periocular lesion on the eyelid; the right eye exhibited increased size and loss of definition of its internal structures. The presence of malodorous cerumen in the right ear canal, a tilted head, and walking in circles to the right were also noted. A dysplastic epithelium along with neutrophilic inflammation was observed at the cytopathological analysis. The treatment instituted consisted of administration of antibacterial and anti-inflammatory drugs, and cleaning of the ear. On the follow-up appointment, 2 days later, the patient's condition had worsened and in spite of the supportive therapy administered, the animal died. An anatomopathological study of the body identified, in addition to the alterations in the eyelid and the eye mentioned above, a traumatic lesion with perforation of the mandibular bone, and presence of a brownish content with caseous consistency in the right ear canal. After visualization of the microscopic alterations, the diagnosis established was squamous cell carcinoma of the eyelid with metastases in the optical nerve and choroid blood vessels. Discussion: Squamous cell carcinoma is one of the most common epithelial neoplasms in felines, and can occur in any region of the face; however, the eyelids and the eyes are less frequently affected, which confirms that the case reported here is uncommon. SCC has a higher incidence in female cats older than 10 years of age with light-colored coat and highly exposed to ultraviolet radiation. The case reported here exhibited these characteristics, as the patient had been rescued from the streets where exposition to sunlight is intense. The clinical presentation was consistent with that described in other works, with ulcerated crusty lesions on the eyelid and the nose. Vestibular syndrome, as inferred from the tilted head and circular gait (walking in circles to the right), could be explained by internal otitis, which was observed at necropsy. The cytological analysis of the eye lesion demonstrated the presence of a neutrophilic inflammatory process along with a dysplastic or immature squamous epithelium, which prompted for therapy using antimicrobial and anti-inflammatory drugs. The definitive diagnosis was established with basis on the histopathological analysis, which revealed a pattern of neoplastic cells arranged in clusters or strands connected to the epidermal surface, and the presence of "keratin pearls" consistent with SCC affecting not only the eyelid and the eye, but also invading the optical nerve and the choroid blood vessels. The prognosis was unfavorable owing to the advanced stage of the tumor and concomitant diseases. Even though the development of SCC in the eyelid is relatively common, the occurrence of metastasis in the eye adnexa is rare in felines. An anatomopathological study was required to establish the diagnosis of SCC as the patient's clinical history data were scarce.

Pituitary carcinoma in a bitch: clinical, tomographic, histopathological and immunohistochemistry findings

Background: Primary pituitary carcinoma is rarely reported in dogs and only few reports describe its malignancy. In veterinary literature, primary pituitary carcinomas correspond up to 2.4% to 3.4% of intracranial neoplasms found in dogs andinformation regarding its biological behavior is quite limited. In humans, primary pituitary carcinomas represent less than1.0% of all tumors found in the pituitary gland. The proposed classification for pituitary carcinoma in humans and dogsdetermines that the tumor must have its origin in adenohypophyseal region and disseminated metastasis by cerebrospinalfluid or systemically to other organs must be observed. In dogs, a few reports have described primary pituitary carcinoma.The goal of this report was to describe clinical, tomographic, histopathological and immunohistochemistry features of abitch with primary pituitary carcinoma with adjacent invasion.Case: A 7-year-old female spayed Golden Retriever dog was assessed by general practice due progressive weight loss,muscular atrophy, lethargy, blindness, head pressing, and hyporexia for 21 days. Computed tomography (CT) showeda cerebral parenchyma with expansive extra-axial base formation, originating from sella turcica topography, measuringabout 2.0 cm dorsally, displacing the third ventricle, suggesting the diagnosis of pituitary neoplasia. The hormones thyroidstimulating hormone (TSH) and total thyroxine (T4) as well as stimulation ACTH test were unremarkable. After 7 days,neurological clinical signs progressed and unfortunately the patient died ten days later after hospitalization. A necropsyexam revealed pituitary gland with increased dimensions (2.5 x 2.0 cm). Histopathological fi ndings revealed tumor proliferation in pituitary gland. The neoplasm showed invasion to the nervous parenchyma and metastatic foci between the brainlobes. Immunohistochemistry was positive for keratin and neuron-specifi c enolase...

The effect of androgen on wool follicles and keratin production in Hetian sheep / Efeito do androgênio na produção de folículos e queratina em ovinos Hetianos

To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.(AU)

Investigar a concentração ideal de andrógenos em cultura de folículos pilosos de carneiro Hetiano e detectar os efeitos da concentração de andrógenos na proliferação e apoptose de células foliculares, por meio de imunofluorescência e de determinação quantitativa, em tempo real, da fluorescência dos níveis de expressão gênica de proteína associada à queratina. Folículos pilosos foram isolados por microdissecção, e folículos de lã e pedaços de pele foram cultivados em várias concentrações de di-hidrotestosterona (DHT) em meio de cultura. Em seguida, medições diárias de crescimento longitudinal dos folículos capilares foram obtidas usando um micrômetro microscópico. Folículos de lã cultivados de Hetianos foram corados pelo método SACPIC para revelar a estrutura do folículo piloso, enquanto fatias de pele de carneiro foram usadas para observar a proliferação celular por imunocoloração e apoptose celular por meio do método TUNEL. Em âmbito da biologia molecular, a expressão gênica da proteína associada à queratina (Kap) foi estudada usando folículos capilares cultivados por vários dias, in vitro. Os efeitos das concentrações de andrógenos no crescimento e desenvolvimento dos folículos de lã de Hetianos foram estudados experimentalmente. Ensaios de proliferação de EdU revelaram que o andrógeno promoveu a proliferação celular dentro das papilas dérmicas do folículo piloso. A detecção de apoptose por TUNEL demonstrou que o tratamento com andrógeno poderia atrasar a apoptose celular. Os resultados da reação em cadeia da polimerase transcrição reversa quantitativa (qPCR) demonstraram que os padrões de expressão gênica da proteína de enxofre Kap1.1, KIF1.2, Kap2.12 e Kap4.2 foram significativamente maiores no grupo de ovinos Hetianos de montanha. Uma concentração de androgênio de 100 nM pode promover o crescimento de células foliculares de lã de Hetianos in vitro, resultando na superexpressão de alguns genes da família Kap.(AU)

Intracranial epidermoid cyst in a cat / Cisto epidermoide intracraniano em um gato

Epidermoid cysts, also known as epidermal and keratin cysts, or congenital cholesteatomas are benign congenital non-neoplastic tumors, rarely observed in the brain of humans and some animal species (dogs, horses, mice, and rats). Histologically, they are composed of laminated, concentrically arranged keratin surrounded by a thin layer of stratified squamous epithelium. We describe a case of intracranial epidermoid cyst in a 6-year-old mixed-breed male cat in southern Brazil. The patient presented central vestibular syndrome. Given the poor prognosis and the fact that it belonged to a shelter with many dogs and cats, the owner requested euthanasia, and a thorough post-mortem examination was conducted immediately after death. The definitive diagnosis was based on histopathological findings. To the best of our knowledge, this is the first report of an intracranial epidermoid cyst in a cat.(AU)

Cisto epidermoide, também denominado cisto epidermal, cisto de queratina ou colesteatoma congênito é um tumor não neoplásico, benigno e congênito raramente encontrado no encéfalo de humanos e de algumas poucas espécies animais (cães, equinos, camundongos e ratos). Histologicamente, esse tumor é composto por queratina laminada concentricamente arranjada e circundada por uma fina parede de epitélio escamoso estratificado. Descreve-se um caso de cisto epidermoide intracraniano em um gato, macho, sem raça definida, de seis anos de idade, no sul do Brasil. O paciente foi encaminhado para atendimento veterinário por apresentar sinais de síndrome vestibular central. Devido ao mau prognóstico e por pertencer a um abrigo com muitos cães e gatos, o proprietário optou pela eutanásia e o paciente foi encaminhado para a realização de necropsia. O diagnóstico definitivo foi baseado nos achados histopatológicos. Pelo conhecimento dos autores, este é o primeiro relato de um cisto epidermoide intracraniano em um gato.(AU)

Epidermal renewal after treatment of primary canine hypothyroidism using levothyroxine

Background: Primary canine hypothyroidism, an endocrine disorder that causes imbalances in the hypothalamus-pituitarythyroid axis, is a common cause of endocrine dermatoses, which frequently presents with opaque dry brittle hair. Tissue changes are also visible, such as myxedema, hyperkeratosis, epidermal atrophy, alopecia, and others. This paper describes the skin changes caused by primary hypothyroidism in a female dog before and after treatment with levothyroxine. Case: This case study involved a 7-year-old Dalmatian bitch with a history of weight gain and changes such as rough dry brittle hair. For about a year, the dog also had also presented symmetrical erythematous and alopecia skin lesions in the regions of the hind limbs, lower back and tail, progressive lethargy and fatigue after exercise. A hemogram showed mild normocytic and normochromic regenerative anemia, as well as mild leukopenia and neutropenia. Biochemical tests revealed increased levels of creatinine, total cholesterol, and triglycerides. Skin biopsy revealed the presence of comedones in the epidermis and acanthosis and trichilemmal keratinization. Hormonal tests revealed high TSH and low free T4 and total T4 levels. A cervical ultrasound scan showed changes in the thyroid glands, with hypoechoic parenchyma, in addition to increased size of the right lobe, suggesting thyroiditis. Based on these exams, the patient was diagnosed with primary hypothyroidism, and treatment with levothyroxine was instituted. Discussion: Around 90% of dogs with hypothyroidism show a decrease in total T4 and about 65-75% of them show an increase in TSH levels. In endocrinopathy, there is a decrease in the expression of T3 receptors in the keratinocytes and genes responsible for epidermal renewal. Also, molecules that affect epidermal differentiation bind to intracellular receptors belonging to the steroid/thyroid hormone superfamily. In this study, however, the application of the minimum levothyroxine dose restored the normal epidermal pattern for the species and the remaining parameters returned to normal. The hormone T3 acts in the differentiation of keratinocytes, a fact that, although not yet proven, is believed to be indirect and mediated by the epidermal growth factor or by the expression of the genes responsible for the renewal of the epidermis. Besides, some effects on the epidermis caused by deficiency of thyroid hormones may be due to secondary vitamin A deficiency, which is necessary for epithelial differentiation and binding epidermal growth factor (EGF) to the cell. Still, there is a theory that sex hormones also affect the differentiation of the epidermis, as studies show that deficiency causes effects similar to those caused by hypothyroidism. Some studies indicate the investigation of the relationship between secondary vitamin D deficiency and the increased cellularity of the epidermis of the spayed hypothyroid female rats. Additionally, the mechanism of the formation of hyperkeratosis in hypothyroidism is not yet elucidated. T3 is known to regulate keratin gene expression and perhaps also epidermal maturation, epithelial cycle, and normal keratin synthesis. The single treatment with levothyroxine in the female dog was effective in promoting the restoration of gene expression to T3 in keratinocytes.

Cutaneous squamous cell carcinoma with mucinous metaplasia in dogs

Background: Squamous cell carcinoma (SCC) is one of the most common malignant skin tumors in domestic animals. Histologically, they are characterized by a proliferation of neoplastic keratinocytes with varied keratin production. Some SCCs have peculiar histological characteristics that permit them to be classified into uncommon to rare histological subtypes, reported in animals and humans. However, according to the authors' knowledge, the mucin-producing subtype described in humans has not yet been reported in animals. In this study, we report the occurrence of two mucin-producing SCCs in dogs, a histological presentation similar to that seen in cutaneous SCCs with mucinous metaplasia in humans. Cases: Two dogs, a 5-year-old Yorkshire female and a 17-year-old Dachshund male, had a skin nodule near the tail and on the right eyelid. The nodules varied from 1 to 5 cm in diameter, were firm and covered with skin and hair. The cut surface was firm and white. Histological findings were compatible with squamous cell carcinoma, characterized by a neoplastic proliferation of keratinocytes originating in the epidermis and infiltrating the dermis. The keratinocytes were arranged in islands and occasional anastomosed cords, supported by a fibrous stroma. The formation of pearls varied from moderate to sparse. The nuclear and cellular pleomorphism was accentuated in case two and moderate in case one. Mitosis figures ranged from two to five in a high magnification field. Within the neoplasm, there were large vacuolated neoplastic cells with slightly fibrillar intracytoplasmic basophilic content. This content has been rarely observed in an extracellular medium. The presence of mucin was confirmed by positive Alcian Blue (AB) staining. In immunohistochemistry (IHC), tumor cells showed strong immunostaining for pancitokeratin, and in areas with marked mucin deposition, immunostaining was predominantly moderate to weak. No tumor cells were immunostained for CD34 and Bcl-2 antibodies. Compared to AB and Harris' hematoxylin, it was possible to demonstrate the presence of mucin in the cytoplasm of neoplastic keratinocytes using IHC. No vascular or lymphatic invasion by neoplastic cells was observed. The average cell proliferation index assessed by counting the nucleolar argyrophilic organizing regions (AgNOR) was 3.4 in case 1 and 4.5 in case 2. Discussion: Although the SCC routinely does not present a diagnostic challenge in veterinary practice, the histological presentation of the reported cases does not fit the current classification available in veterinary medicine. The histological presentation observed in these two dogs is similar to that described for cutaneous SCCs with mucinous metaplasia in humans, and so far not described in animals. The observation of intracytoplasmic mucin in humans is an essential finding for the diagnosis of SCC with mucin metaplasia. In the present cases, we observed a slightly basophilic amorphous substance in the cytoplasm of proliferated neoplastic keratinocytes, which stained strongly in blue when applied the Alcian Blue (AB) histochemical technique. This observation became more evident when using IHC counterstained with AB and Harris hematoxylin. In the histological analysis, the absence of an adenoid growth pattern or glandular formation amid neoplastic proliferation ruled out the possibility of a mucinous adenocarcinoma or a mucoepidermoid carcinoma. In addition, we could rule out a follicular neoplasia, including keratinizing infundibular acanthoma due to the absence of a central pore and the absence of immunostaining for CD34 and Bcl-2. These immunohistochemical findings, together with histological findings, reinforce the diagnosis of SCC with mucinous metaplasia in our dogs.(AU)

Prótese de rinoteca em carcará (Caracara plancus) / Rhinotheca prosthesis in a carcará (Caracara plancus)

Background: The bird's beak is a structure in constant growth, covered by keratinized epidermal sheaths called rhamphotheca. When subjected to certain degrees of injury, birds can suffer from lesions and fractures in different parts of the body, including the beak. One can treat simple ranch lesions by antisepsis and covering it with resin while the keratin is replaced; yet in complete fractures, with segment avulsion, they need complex prostheses to restore the functions of the nozzle. The main goal of this study was to report a case of a synthetic rhinotheca prosthesis placement in a wild carcará (Caracara plancus). Case: The carcará was referred to the Veterinary Medical Teaching Hospital of the UNIUBE, at the end of May 2018, by Uberaba's Environmental Police, with a history of having been run over. On the physical examination, the animal showed aggressive behavior, low body weight, increased heart and respiratory rates, mild dehydration and complete fracture of the rhinotheca, with avulsion of the mid-distal portion and presence of necrosis in the remaining proximal remnant, the last one seen after debridement and complete cleansing of the lesion. In the following days, complementary exams were performed aiming a complete evaluation of the animal, these revealed the following: oral cavity swab, positive for Candida sp.; radiography of thoracic and pelvic limbs, without any changes; research of hemoparasites, with a negative result; and complete blood count showing marked leukocytosis. The animal was kept in the hospital's wild animal ward, with a daily handling of 400 g of chicken neck and heart, processed on a blender and was also supplemented with a variety of minerals. After the wild animal was managed for eleven months, evolving to the ideal weight set for the species, the process for making the prosthesis started. At first, the prosthesis was made manually with epoxy resin and polyamide, being molded on the animal's rhinotheca, before that, the carcará was sedated with intranasal midazolam, and then modeled in the ideal shape for the beak. Then, the definitive prosthesis was made with acrylic resin, using the previous mold as a base. The surgical procedure was performed in April 2019, with an anesthetic protocol composed of midazolam, dexmedetomidine, ketamine and morphine for sedation, and maintenance in sevoflurane. Prosthesis and rhinotheca were both fixated by bilateral perforation with a 1.0 mm drill, and later on 2 titanium screws were inserted for dental use, these measuring 1.5 mm. After being correctly threated, the screws received a thin layer of acrylic resin on top, for better fixation. Immediately after the operation, the animal was able to use the prosthesis to feed itself. After the surgical procedure, the bird was destined for a sanctuary, and months later the prosthesis eventually fell. Since then, the bird has remained without it. Discussion: Because the beak did not grow due to trauma and consequent necrosis, and the animal did not adapt well with the remnant still present, the manufacture of the prosthesis was the solution so that it could return to its natural habits and behaviors. For greater adherence and fixation, we opted for the use of 2 titanium screws for dental use with the deposition of a thin layer of acrylic resin on top of both. Prostheses can be successful in fixing, but there are no studies indicating how long they will remain viable. Therefore, even with the success of the procedure, allowing the bird to return to its natural habits and behaviors, until after the fall of the prosthesis, it is an animal that needs to be kept in captivity suitable for daily observation.

Ocorrência simultânea de teratoma ovariano e hiperplasia endometrial cística com piometra em cadela Labrador Retriever / Simultaneous occurrence of ovarian teratoma and endometrial cystic hyperplasia with pyometra in a Labrador Retriever bitch

Background: Ovarian teratoma is a rare tumor that occurs in dogs. Its origin comes from embryonic cells of the notochord and it is a unipotent tumor. Pyometra is the accumulation of purulent content in the uterine lumen. Cystic endometrial hyperplasia is one of the factors predisposing a dog to the development of pyometra. The safest and most efficient treatment for pyometra is ovariohysterectomy. The purpose of this report was to describe a case of ovarian teratoma and cystic endometrial hyperplasia with concomitant pyometra in a Labrador Retriever Bitch. Case: A 10-year-old bitch Labrador Retriever, weighing 42 kg, was evaluated at Animal Care Barueri Veterinary Clinic, in Barueri, São Paulo. The patient was referred from another veterinary service to our clinic for an ovariohysterectomy and removal of an intestinal foreign body as previous ultrasonography (US) had indicated the presence of pyometra and a foreign body in the descending colon (she had ingested cloth according to the owner's report). Laboratory tests, complete blood count, alanine aminotransferase, alkaline phosphatase, urea, and creatinine levels did not show any significant changes. Upon examination, the clinical signs were mucopurulent vaginal discharge, 5% dehydration, rectal temperature 39.1°C, mild abdominal pain on palpation, normophagy, normodipsia, and normal defecation and urination. Laboratory tests and a new US were requested on which the uterus was observed with the presence of luminal anechoic content and increased uterine diameter (uterine horns 3.97 cm), an acoustic intestinal shadow supposedly from a foreign body, and on the leukogram, leukocytosis (23,600/mm3) due to neutrophilia with a right shift (20,532/mm3) was observed. Therapeutic ovariohysterectomy and exploratory celiotomy were chosen as therapeutic options. Celiotomy with caudoumbilical mid ventral access was performed. When the left uterine horn was identified, an attempt was made to pull it in order to expose the left ovary, but this maneuver was not successful. For this reason, the abdominal incision was enlarged cranially. At that point, a huge mass was observed in the ovary (which was supposedly the acoustic shadow of the foreign body in the descending colon/left ovary). The intestines appeared normal. Macroscopically, the mass had an irregular surface, round shape, firm consistency, and was 15 cm long x 10 cm wide. Once removed, the mass was incised in the transversal direction, purulent content, hair, and mineralized areas inside it were observed. Histopathological examination showed neoplastic proliferation, consisting of cells with elongated epithelioid shape that were arranged in long irregular bundles among well-differentiated adipose tissue. An exuberant eosinophilic matrix with extensive cystic areas filled with lamellar keratin and hairy stems covered by squamous epithelium was also observed. Extensive areas exhibiting nervous tissue with minimal atypia are shown. The morphological picture was compatible with teratoma. Discussion: The present report describes the concomitant presence of ovarian teratoma and pyometra in a dog. The diagnosis of pyometra requires complementary tests (laboratory and imaging). The intestinal acoustic shadow observed on the two US images obtained by two different professionals was suggestive of intestinal foreign body but was also a wrong diagnosis, which was confirmed after an exploratory celiotomy when the mass in the left ovary was identified. On the histopathological examination of this present case, epithelial, nervous, and cartilaginous tissues were observed. This differentiation of tissues corroborated the teratoma diagnosis. Thus, the importance of an accurate diagnosis contributes to the resolution of a surgical conditions that may have had a poor prognosis when the procedure took a longer time to perform.(AU)

Structural description of sea turtle fibropapilloma

Fibropapillomatosis is a neoplastic disease that affects sea turtles. It is characterized by multiple papillomas, fibropapillomas and cutaneous and/or visceral fibromas. Although its etiology has not been fully elucidated, it is known that there is a strong involvement of an alpha herpesvirus, but the influence of other factors such as parasites, genetics, chemical carcinogens, contaminants, immunosuppression and ultraviolet radiation may be important in the disease, being pointed out as one of the main causes of a reduction in the green turtle population. Thus, the objective of this article was to describe the morphology of cutaneous fibropapillomas found in specimens of the green turtle (Chelonia mydas), using light and scanning electron microscopy in order to contribute to the mechanism of tumor formation. Microscopically, it presented hyperplastic stromal proliferation and epidermal proliferation with hyperkeratosis. The bulky mass was coated with keratin, with some keratinocyte invaginations, that allowed the keratin to infiltrate from the epidermis into the dermis, forming large keratinized circular spirals. Another fact that we observed was the influence of the inflammation of the tumors caused by ectoparasites.(AU)

Structural description of sea turtle fibropapilloma

Fibropapillomatosis is a neoplastic disease that affects sea turtles. It is characterized by multiple papillomas, fibropapillomas and cutaneous and/or visceral fibromas. Although its etiology has not been fully elucidated, it is known that there is a strong involvement of an alpha – herpesvirus, but the influence of other factors such as parasites, genetics, chemical carcinogens, contaminants, immunosuppression and ultraviolet radiation may be important in the disease, being pointed out as one of the main causes of a reduction in the green turtle population. Thus, the objective of this article was to describe the morphology of cutaneous fibropapillomas found in specimens of the green turtle (Chelonia mydas), using light and scanning electron microscopy in order to contribute to the mechanism of tumor formation. Microscopically, it presented hyperplastic stromal proliferation and epidermal proliferation with hyperkeratosis. The bulky mass was coated with keratin, with some keratinocyte invaginations, that allowed the keratin to infiltrate from the epidermis into the dermis, forming large keratinized circular spirals. Another fact that we observed was the influence of the inflammation of the tumors caused by ectoparasites.

Mucocutaneous horn in dog

Background: Cutaneous horn is a circumscribed exophytic lesion composed of dense, compact keratin with hyperplasticepidermis, which is primarily orthokeratotic and may include foci of parakeratosis. The hyperkeratotic protuberance resembles a horn but lack bone. In humans, it is well-documented with a wide range of primary epidermal lesions identified.In dogs, the reports are rare and brief. The diagnosis is based on its appearance and excisional biopsy reveals the triggeringlesion. The objective of the present work is to describe the clinical presentation, predictive benignancy finds, treatmentand follow-up of a case of cutaneous horns that arose from anal mucocutaneous boundary in a dog.Case: A 6-month-old male Pug presented two tumors in the anus noted in the early months of life. Complete blood count,serum chemistry profile, surgery, histopathological analysis and postoperative outcome were performed. Possible relapseswere follow-up until 11 months after surgery. On physical examination, the dog was otherwise healthy. The two horn-liketumors were protruding from the mucocutaneous junction of the external anal sphincter in a sun-protected area. Both werehigher than wide in base or, in other words, they had a high height-to-base ratio. During surgery, the masses were excisedusing an electronic scalpel and sent to histopathological analysis. The post-operative care consisted of cephalexin andmeloxicam prescriptions. Also, it was recommended 0.9% NaCl solution wound flushing followed by topical chlorhexidinedigluconate solution (1%) whenever dirt or defecation were noticed. The surgical wound healed after two weeks by secondintention. Histopathology found well-delimited masses that covers the epidermis and dermis and displaces the cutaneousappendages towards the underlying musculature with an aggregate of lymphoplasmohistiocytic inflammatory infiltrate inthe adjacent...

Mucocutaneous horn in dog

Background: Cutaneous horn is a circumscribed exophytic lesion composed of dense, compact keratin with hyperplasticepidermis, which is primarily orthokeratotic and may include foci of parakeratosis. The hyperkeratotic protuberance resembles a horn but lack bone. In humans, it is well-documented with a wide range of primary epidermal lesions identified.In dogs, the reports are rare and brief. The diagnosis is based on its appearance and excisional biopsy reveals the triggeringlesion. The objective of the present work is to describe the clinical presentation, predictive benignancy finds, treatmentand follow-up of a case of cutaneous horns that arose from anal mucocutaneous boundary in a dog.Case: A 6-month-old male Pug presented two tumors in the anus noted in the early months of life. Complete blood count,serum chemistry profile, surgery, histopathological analysis and postoperative outcome were performed. Possible relapseswere follow-up until 11 months after surgery. On physical examination, the dog was otherwise healthy. The two horn-liketumors were protruding from the mucocutaneous junction of the external anal sphincter in a sun-protected area. Both werehigher than wide in base or, in other words, they had a high height-to-base ratio. During surgery, the masses were excisedusing an electronic scalpel and sent to histopathological analysis. The post-operative care consisted of cephalexin andmeloxicam prescriptions. Also, it was recommended 0.9% NaCl solution wound flushing followed by topical chlorhexidinedigluconate solution (1%) whenever dirt or defecation were noticed. The surgical wound healed after two weeks by secondintention. Histopathology found well-delimited masses that covers the epidermis and dermis and displaces the cutaneousappendages towards the underlying musculature with an aggregate of lymphoplasmohistiocytic inflammatory infiltrate inthe adjacent...(AU)