Epidemiological, clinical, and histopathological aspects of mammary neoplasms in dogs from Rio Branco, Acre, Brazil / Aspectos epidemiológicos, clínicos e histopatológicos das neoplasias mamárias em cães do município Rio Branco, Acre, Brasil

Mammary neoplasms are very common in the clinical routine of small animals and have an enormous histological diversity and complexity. Most of these neoplasms are classified as malignant and originate from metastases, considered the main cause of death. This study aimed to present and correlate epidemiological data with the malignancy of histological types of mammary neoplasms in dogs in the municipality of Rio Branco, Acre, Brazil. The analyzed samples were collected in collaborating hospitals and veterinary clinics, the data acquired through a questionnaire filled in by them, the clinical staging carried out through the TNM system, and the histological classification according to the Consensus regarding the Diagnosis, Prognosis, and Treatment of Canine and Feline Mammary Tumors. The fixed material was evaluated macroscopically and then subjected to classical histological processing. Histological preparations were observed under optical microscopy. The data were organized statistically using Pearson's chi-square test (p<0.05). Of the 71 samples obtained from dogs, 50 consisted of mammary neoplasms, 18 skin neoplasms, and three non-tumor lesions. Eleven breeds of dogs were identified. Among them, 48 were females and two males with an average age of 10 years and most patients intact (90%) and the minority used progestogens (16%). Of the mammary neoplasms, 94% had a malignant profile, with carcinoma in a mixed tumor being the most frequent (60%). Statistically, no correlation was observed between breed, sex, age, reproductive status, use of progestogens, mammary location, or tumor size with malignancy. In general, dogs can be affected by malignant mammary neoplasms, regardless of the mentioned variables, reinforcing the importance of clinical staging and early diagnosis for the best prognosis within the small animal clinic.

As neoplasias mamárias são muito frequentes na rotina clínica de pequenos animais e possuem uma enorme diversidade e complexidade histológica. A maior parte dessas neoplasias é classificada como malignas e originam às metástases, sendo consideradas a principal causa mortis. Este estudo objetivou apresentar e correlacionar os dados epidemiológicos com a malignidade dos tipos histológicos das neoplasias mamárias em cães dentro do município de Rio Branco, Acre. As amostras analisadas foram coletadas em hospitais e clínicas veterinárias colaboradoras, os dados adquiridos através de questionário preenchido pelos mesmos, o estadiamento clínico realizado através do sistema TNM e a classificação histológica segundo o Consenso de Diagnóstico, Prognóstico e Tratamento de Neoplasias Mamárias Caninas. O material fixado foi avaliado macroscopicamente e, em seguida, submetido ao processamento histológico clássico. As preparações histológicas foram observadas sob microscopia óptica. Os dados obtidos foram organizados na forma estatística utilizando o Teste de Qui-quadrado de Pearson (p<0,05). Das 71 amostras obtidas em cães, 50 foram neoplasias mamárias, 18 foram tumores de pele e três eram lesões não tumorais. Foram identificadas 11 raças caninas, 48 eram fêmeas e dois machos, com idade média de 10 anos, onde a maioria dos pacientes eram inteiros (90%) e a minoria fazia uso de progestágenos (16%). Das neoplasias mamárias, 94% apresentaram o perfil maligno, sendo o carcinoma em tumor misto o mais frequente (60%). Estatisticamente não houve correlação entre raça, sexo, idade, estado reprodutivo, uso de progestágenos, localização das mamas ou tamanho tumoral com a malignidade. A espécie canina de modo geral pode ser acometida de neoplasia mamária maligna, independente das variáveis citadas, reforçando a importância do estadiamento clínico e diagnóstico precoce para o melhor prognóstico dentro da clínica de pequenos animais.

The prognostic significance of immunophenotypes in canine malignant mammary tumors / [Significância prognóstica dos imunofenótipos em tumores mamários malignos caninos]

Canine malignant mammary neoplasms (CMMN) exhibit behavioral variability with the patient survival time depending on several prognostic factors. In the present study, 134 CMMN were selected and different immunophenotypes and their associations with clinical and pathological parameters were identified. The tumors were classified as follows: 46% of luminal B HER2-, 34% of luminal A, 13% of triple-negative, and 7% of luminal B HER2+. Shorter specific survival time were associated with larger tumor sizes (>3.0 cm, HR=1.94; P=0.0209), lymph node metastasis or distant metastasis (HR= 2.82; P <.0001), more aggressive histological types (HR= 7.15, P<0.0001), higher histological grades (HR= 12.97 P=0.011), angiolymphatic invasion (HR=4.68, P<0.0001) and luminal B HER2 - (HR= 3.27, P<0.0001) and luminal B HER2 + (HR= 7.14 P<0.0001) immunophenotypes. In patients with lymph nodal metastasis, shorter survival times were associated with luminal immunophenotype B HER2 + (P=0.003). However, in patients without metastasis, an increased risk of death was associated with the aggressive histological type. In conclusion, the classification in our study allowed us to identify subtypes with different prognoses in canine malignant mammary tumors. Factors such as clinical stage, histological type, luminal B HER2+ subtype, and angiolymphatic invasion were the most important prognostic factors.

Neoplasias mamárias malignas caninas (CMMN) apresentam variável comportamento biológico e o tempo de sobrevida depende de diversos fatores prognósticos. Neste estudo, foram selecionadas 134 CMMN, bem como identificados diferentes imunofenótipos e suas associações com parâmetros clínicos e patológicos. Os tumores foram classificados em: 46% luminal B HER2-, 34% luminal A, 13% triplo negativo e 7% luminal B HER2+. Menores taxas de sobrevida específica foram associadas a tamanhos de tumor maiores (> 3,0cm; HR = 1,94; P = 0,0209), metástases em nodais ou a distância (HR = 2,82; P <0,0001), tipos histológicos mais agressivos (HR = 7,15; P <0,0001), graus histológicos mais elevados (HR = 12,97; P = 0,011), invasão angiolinfática (HR = 4,68; P <0,0001) e aos imunofenótipos luminal B HER2- (HR = 3,27; P <0,0001) e luminal B HER2+ ( HR = 7,14; P <0,0001). Em pacientes com estágio avançado, menor sobrevida específica foi associada ao imunofenótipo luminal B HER2+ (P = 0,003). Entretanto, em estágio inicial, um risco aumentado de óbito foi associado a tipos histológicos agressivos. Em conclusão, a classificação utilizada no presente estudo permitiu identificar subtipos com diferentes prognósticos em CMMN. Estágio clínico, tipo histológico, subtipo luminal B HER2+ e invasão angiolinfática foram os fatores prognósticos mais importantes.

Performance, nutrient digestibility, and muscular evaluation of female broiler chickens fed different dietary protein levels and slaughtered at 38 or 46 days

This study evaluated the effects of two protein levels and types of formulation on performance, nutrient metabolization, and myopathies of 390 female broilers from 21 to 46 days old distributed in a completely randomized design, with five treatments and six replicates of 13 broilers each. Treatments were two levels of protein (19.0 and 21.0%) in diets using only commercially available ingredients and three experimental formulations designed to keep similar the main nutrients that could influence nutritional performance (starch, fiber, ether extract), using unusual ingredients, with 19.0% (19E) or 21.0% (21E) crude protein and the third with 19.0% crude protein with amino acid levels similar to treatment 21P (19E+Aa). We studied broiler performance, nutrient digestibility, macroscopic muscular evaluation, and histological muscular evaluation. The design for the performance and myopathy assessment was a 5 × 2 factorial scheme, five diets × two ages (38 or 46 days). The worst feed conversion ratio was observed for the 19P treatment. Neither diet nor age had an effect on myopathies, however, older slaughter age increased the intensity of microscopic lesions. The diet 19P should be avoided and increased inclusion of lipids in diets improves nutrient utilization and, consequently, performance.(AU)

Study of the integument that covering back and stinger of the freshwater stingray Potamotrygon rex (Chondricthyes, Potamotrygonidae) / Estudo do tegumento que reveste o dorso e ferrão da arraia de água doce Potamotrygon rex (Chondricthyes, Potamotrygonidae)

The objective of this study was to describe the histology and histochemistry of the integument covering the back and stinger of the freshwater stingray Potamotrygon rex, endemic to the Middle Upper Tocantins River. The species has a dark back and yellowish circular spots that extend to the tail, which has one to two stings located in the median portion of the tail. Through histological study it was observed that the epithelia of the back and stinger are composed of non-keratinized stratified pavement epithelial tissue, and are organized in three layers: basal, intermediate and superficial. The differences between the tissues are related to the cell types that compose them. The back is organized with epithelial cells, mucus cells, granulocyte cells and chromatophores. The mucus cells are distributed in different layers along the animal's back, influencing the thickness of the tissue. The tissue that covers the stinger is composed of epithelial cells, chromatophores and specialized cells in protein synthesis. In the histochemistry, the stinger epithelial cells were stained with Bromophenol Blue, especially those of the intermediate layer, which were called specialized cells. In the back the epithelial cells were stained with Bromophenol Blue, Alcian Blue and PAS, and the mucous cells with PAS. In both tissues the presence of protein reserves was detected, areas so called because they are stained strongly with Bromophenol Blue. The results show that the stinger presents activity directed to the production of proteins, and that the back is organized to produce different components, which constitute the cuticle that covers the animal's body.

O objetivo deste estudo foi descrever a histologia e histoquímica do tegumento que reveste o dorso e o ferrão da arraia de água doce Potamotrygon rex, endêmica do Médio Alto Tocantins. A espécie possui o dorso escuro e manchas circulares amareladas que se estendem até a cauda, que possui de um a dois ferrões localizados na porção mediana da cauda. Através do estudo histológico observou-se que os epitélios do dorso e do ferrão são compostos por tecido epitelial pavimentoso estratificado não queratinizado, e estão organizados em três camadas: basal, intermediária e superficial. As diferenças entre os tecidos estão relacionadas aos tipos de células que os compõem. O dorso é organizado com células epiteliais, células mucosas, células de granulócitos e cromatóforos. As células mucosas estão distribuídas em diferentes camadas ao longo do dorso do animal, influenciando na espessura do tecido. O tecido que reveste o ferrão é composto por células epiteliais, cromatóforos e células proteicas. Na histoquímica, as células epiteliais do ferrão foram coradas com Azul de Bromofenol, principalmente as da camada intermediária, que foram denominadas células proteicas. No dorso as células epiteliais foram coradas com Azul de Bromofenol, Azul Alcian e PAS, e as células mucosas com PAS. Em ambos os tecidos também foi detectada a presença de reservatórios de proteínas, áreas assim denominadas por estarem fortemente coradas com Azul de Bromofenol. Os resultados mostram que o ferrão apresenta atividade direcionada à produção de proteínas, e que o dorso se organiza para produzir diferentes componentes que constituem a cutícula que reveste o corpo do animal.

Morphological Studies on the Testis, Epididymis and Vas Deferens of Al-Ahsa Native Rooster

This study was carried out to investigate the morphological and histological structures of the testis, epididymis and vas deferens of the Al-Ahsa native rooster (ANR). There were two types of ANR; the brown feather one with light yellow shank and the black feather one with grey or dark grey shank. Their body weight was 1840.88± 92.13 g and 1555.66± 82.83g, respectively. The morphology of the testes showed that the black rooster has larger testes than the brown rooster and there was asymmetry in size between the right and left testis in both. They were grey yellowish in color and oval-shaped, situated in the abdomen dorsal to the proventriculus, the liver and the gizzard, cranial to the lungs, caudal to the abdominal sac and ventral to the kidneys. The histology of the testes revealed the capsule, the different cells of the lining epithelium of the seminiferous tubules and the interstitial tissue. The morphology of the epididymis was revealed pseudostratified columnar epithelium, light brown in color with c to L-shaped, located cranial to the testis and extended caudally to continue with vas deferens. The latter has columnar epithelium, light grey in color, run caudally medial to the kidneys and opened in the cloaca.(AU)

Canine mammary neoplasms - evaluation of tumor microenvironment

Background: The tumor microenvironment is an important target of studies in different types of neoplasms. Understanding the role of general components such as immune, vascular and fibroblastic cells has the objective of contributing to prognosis and treatment. The aim of this study was to evaluate the relationship between mast cells and angiogenesis in benign and malignant mammary neoplasms by investigating the role of degranulation and microlocation of mast cells and neoformed vessels in canine mammary neoplasms. Materials, Methods & Results: Mammary glands (n = 122) from 50 female dogs submitted to mastectomy without chemotherapy were evaluated and categorized into 3 groups: control group (n = 46); malignant group (n = 57) and benign group (n = 19). Lymph nodes without changes (n = 59) and with metastases (n = 6) were also evaluated. To evaluate the MCD (mast cell density) and angiogenesis, Toluidine Blue (0.1%) and Gomori’s Trichrome techniques were performed and adapted from previous studies. Photomicrographs of 10 hotspot areas on a 40x objective lens of the mammary glands and lymph nodes were captured to assess MCD and angiogenesis. In the absence of these areas, random fields were captured. For the mammary glands of the malignant and benign groups, 20 fields were analyzed, as the analysis considered the microlocation (peritumoral and intratumoral). Counting was performed manually using ImageJ software version 1.42q by 2 observers. The statistical analysis were performed using SPSS software version 19.0. The most frequent histological type in the malignant group was carcinoma in mixed tumor (68.42%; 39/57) and in the benign group was benign mixed tumor (57.89%; 11/19). Female dogs without breed pattern were more frequently affected represented 70% of the animals and the mean age was 9 years and 8 months ± 3 years and 1 month...

Canine mammary neoplasms - evaluation of tumor microenvironment

Background: The tumor microenvironment is an important target of studies in different types of neoplasms. Understanding the role of general components such as immune, vascular and fibroblastic cells has the objective of contributing to prognosis and treatment. The aim of this study was to evaluate the relationship between mast cells and angiogenesis in benign and malignant mammary neoplasms by investigating the role of degranulation and microlocation of mast cells and neoformed vessels in canine mammary neoplasms. Materials, Methods & Results: Mammary glands (n = 122) from 50 female dogs submitted to mastectomy without chemotherapy were evaluated and categorized into 3 groups: control group (n = 46); malignant group (n = 57) and benign group (n = 19). Lymph nodes without changes (n = 59) and with metastases (n = 6) were also evaluated. To evaluate the MCD (mast cell density) and angiogenesis, Toluidine Blue (0.1%) and Gomoris Trichrome techniques were performed and adapted from previous studies. Photomicrographs of 10 hotspot areas on a 40x objective lens of the mammary glands and lymph nodes were captured to assess MCD and angiogenesis. In the absence of these areas, random fields were captured. For the mammary glands of the malignant and benign groups, 20 fields were analyzed, as the analysis considered the microlocation (peritumoral and intratumoral). Counting was performed manually using ImageJ software version 1.42q by 2 observers. The statistical analysis were performed using SPSS software version 19.0. The most frequent histological type in the malignant group was carcinoma in mixed tumor (68.42%; 39/57) and in the benign group was benign mixed tumor (57.89%; 11/19). Female dogs without breed pattern were more frequently affected represented 70% of the animals and the mean age was 9 years and 8 months ± 3 years and 1 month...(AU)

O efeito de probióticos fúngicos adicionados a uma dieta rica em grãos no trato gastrointestinal de ovinos / The effect of fungal probiotics added to a high-grain diet on the gastrointestinal tract of sheep

This study aimed to evaluate the microbiological and physicochemical characteristics of the ruminal fluid and histological characteristics of the gastrointestinal tract (GIT) of sheep on a high-grain diet containing the probiotic fungi Aspergillus terreus and Rhizomucor spp. The study included four treatment groups (without probiotic, with Rhizomucor spp., with Aspergillus terreus, and with a mixture of both fungi), and two types of corn (ground/whole), in a Completely Randomized Design (CRD) arranged in 4 x 2 factorial design. Santa Inês x Dorper lambs were housed in eight pens with five lambs each for 75 days. Rumen fluid was collected to study the rumen microbiological profile, macroscopic characteristics, ammonia nitrogen concentration, and microbiological activity. In addition, GIT samples were taken for histological analysis. Fluid analyses showed that the animals presented a low acidosis index. The samples presented a predominantly aromatic odor and blackish-brown color, indicating a neutral pH and high microbial activity. The rumen pH differed (P < 0.05) according to the level of processed corn consumed, being higher for ground grain corn (GGC). There was no difference for any of the microbiological communities analyzed (P > 0.05) (Lac+ and Lac- bacteria, fungi, yeasts, and protozoa). Six genera of facultative anaerobic fungi were identified in 15 observations. Cladosporium spp. was the most prevalent genus (46.66%), followed by Aspergillus spp. (26,66%). The width of the base of rumen papillae showed significant correlation being greater for GCG (P < 0.05) with Rhizomucor and for the control (P < 0.05). The rumen fluid of sheep on a high-grain diet with added Aspergillus terreus and Rhizomucor spp. showed no microbiological and physicochemical changes.

Objetivou-se avaliar as características microbiológicas e físico-químicas do fluido ruminal e histológicas do trato gástrico intestinal (TGI) de ovinos sob dieta de alto grão com probiótico fungos Aspergillus terreus e/ou Rhizomucor spp. Analisou-se quatro probióticos (sem inóculos, com Rhizomucor spp., com Aspergillus terreus e com mistura dos dois fungos) e dois processamentos de milho (moído/inteiro), em fatorial 4x2 em em Delineamento Inteiramente Casualizados (DIC). Borregos Santa Inês/Dorper foram alojados em oito baias com cinco borregos em cada, durante 75 dias. Coletou-se fluido ruminal para o estudo do perfil microbiológico do rúmen, da característica macroscópica, da concentração de nitrogênio amoniacal e da atividade microbiológica, além dos fragmentos do TGI para análises histológicas. Pelas análises dos fluidos, os animais apresentaram baixo índice de acidose. O odor aromático e a cor castanho-enegrecido predominaram, o que caracteriza ambiente com pH neutro. As amostras do fluido apresentaram alta atividade microbiana. O pH ruminal diferenciou-se (P<0,05) quanto ao tipo de processamento, sendo maior para milho grão moído (MGM). Não houve diferença para nenhuma das comunidades microbiológicas analisadas (P>0,05) (bactérias Lac+ e Lac-, fungos, leveduras e protozoários). Seis gêneros de fungos anaeróbicos facultativos foram identificados num total de 15 observações. O Cladosporium spp. foi o gênero mais prevalente (46,66%), seguido do Aspergillus spp. (26,66%). A largura da base das papilas ruminais apresentou interação significativa, sendo maior para MGM (P<0,05) com Rhizomucor e o controle (P<0,05). O fluido ruminal de ovinos sob dieta de alto concentrado de grão com adição dos fungos Aspergillus terreus e Rhizomucor spp. não tiveram afetadas as características microbiológicas e físico-químicas.

Presença da mutação do gene BRCA1 em cadelas com tumores mamários malignos / Presence of BRCA1 gene mutation in bitches with malignant mammary tumors

Mammary tumors (MTs) in bitches are similar to breast cancers in women. Thus, they can be used as a model for human breast cancer and findings can be extrapolated for use in human medicine. BRCA1 is a tumor suppressor gene. When the gene has a mutation, it cannot repair damaged DNA, which causes genetic instability and tumorigenesis. Therefore, we aimed to study the frequency of single nucleotide polymorphisms (SNPs) in the BRCA1 gene that are associated with distinct histological types of malignant MT in bitches. The study population consisted of 91 bitches, including a control group of 6 animals with healthy mammary glands and 85 animals with MTs. All animals underwent a presurgery evaluation consisting of a questionnaire administered to the person responsible for the animal, a physical examination, collection of peripheral blood for hematological and serum biochemistry evaluations, an electrocardiogram, and a preanesthesia evaluation. In addition, distant metastasis was studied via chest radiography and abdominal ultrasound. After evaluations were complete, the animals that could undergo surgery were administered general anesthesia and underwent a mastectomy or mammary gland sample collection. Histopathological examination and molecular analysis were performed to identify mutations in the BRCA1 gene. Histopathological examinations found 10 different types of malignant tumors in 36 sick animals. Tumor samples plus samples from the 6 control animals were subjected to DNA extraction, polymerase chain reaction (PCR) analysis, and genetic sequencing. The tumor with the highest incidence (33.33%) was a complex carcinoma, followed by carcinoma in mixed tumor (13.88), tubular carcinoma (13.88) and carcinosarcoma (13.88). Molecular analysis revealed 3 different SNP points in 5 samples (4006G>A, 3619A>G, and 3761C>T). The allelic variant 4006G>A (1/36) resulted in the alteration of the amino acid valine by isoleucine (V1336 I). The mutation 3619A>G (2/36) inserted the amino acid alanine instead of threonine (T1207 A). The mutation 3761C>T (2/36) led to the alteration of the amino acid serine by phenylalanine (S1254 F), a mutation for which there are no published reports. The histological types that showed BRCA1 mutations were complex carcinoma (1/5), carcinoma in mixed tumor (1/5), papillary carcinoma (1/5) and tubular carcinoma (2/5). Software analysis identified the new SNP (nucleotide 3761) in BRCA1 and 2 point mutations in nucleotides 4006 and 3619 and responsible for genetic instability. The development of breast cancer is caused by many endogenous and exogenous factors. The results of our study show that these factors have a greater presence in female, mixed breed, uncastrated, and older dogs, confirming the data in the veterinary literature. In the present study, we found different histological types of malignant breast tumors with mutations in the BRCA1 gene, as other authors have reported. However, we also found the mutation 3761C>T, which, to the best of our knowledge, has not been reported in the literature. This shows the need for studies in veterinary medicine that assess mutations in the BRCA1 gene and the most common histological types. In conclusion, SNPs in the BRCA1 gene cause genetic instability, resulting in additional mutations that lead to the development of breast tumors. They are point mutations that affect transcription, resulting in truncated proteins. These proteins may have a loss of function, leading to carcinogenesis.(AU)

Histological evaluation of capsules formed by texturized silicone implants with and without polyester mesh coverage (Parietex®). A study on female rats

ABSTRACT Purpose To evaluate capsules formed by microtextured silicone implants with and without Parietex® mesh coverage histologically. Methods Sixty Wistar rats were divided in two groups (meshed and unmeshed). Each group was, then, divided into two subgroups for evaluation at 30 and 90 days. Capsules were analyzed based on hematoxylin and eosin (HE) and picrosirius staining. Results The number of fibroblasts, neutrophils and macrophages was similar among all subgroups. There was a higher lymphocyte reaction in the 30-day meshed group (p = 0.003). Giant cell reaction, granulation tissue and neoangiogenesis were similar among the subgroups. Synovial metaplasia was milder at 90-day in the unmeshed (p = 0.002) and meshed group (p < 0.001). Capsular thickness was significantly greater in the meshed samples (30-day p < 0.001 and 90-day p < 0.001). There was a similar amount of collagen types I and III in both groups. Conclusions The mesh-covered implants produced capsules similar to the microtextured ones when analyzing inflammatory variables. Synovial metaplasia was milder at 90 than at 30 days, and the capsular thickness was significantly greater in the meshed group. A similar amount of collagen types I and III was observed. Due to these characteristics, the mesh coverage did not seem to significantly affect the local inflammatory activity.

Evaluation of platelet indices and complete blood count in canine mammary tumors

Malignant mammary tumors in humans and bitches cause hematological disorders such as anemia, erythrocytosis, thrombocytosis, hyperproteinemia, and leucopenia. Novel studies have been conducted on the predictive and prognostic values of platelet (PLT) indices in human breast cancer (HBC). However, there is little information about the alterations in hematological parameters in canine mammary tumors (CMTs). The aims of this study were to evaluate the platelet indices and complete blood count (CBC) parameters in bitches with and without mammary tumor and to assess the above mentioned parameters with regard to histological tumor types and grades. A total of 71 bitches were enrolled in this study. The bitches in the study group were divided into 2 groups which consisted of malignant epithelial mammary tumors (group EMT; n = 43) and malignant mixed mammary tumors (group MMT; n = 12). Control group (group C) consisted of clinically and gynaecologically healthy 16 bitches. Blood samples were obtained to perform the CBC and PLT indices analysis. Histopathological examinations were carried out under a light microscope. Histological tumor types and malignancy grades were classified. The bitches with mammary tumor showed significantly increased PLT values and decreased hematocrit (HCT), hemoglobin (HGB) and mean corpuscular hemoglobin (MCH) values versus the healthy ones, regardless of the tumor type. However, in comparisons with the group C, mean platelet volume (MPV) and mean corpuscular hemoglobin concentration (MCHC) values were different only in the group MMT, while plateletcrit (PCT) and mean corpuscular volume (MCV) values were different only in the group EMT. Also white blood cell (WBC), PLT and PCT values were higher than the referenced laboratory ranges in grade 3 tumors. In the presented study, MPV was considerably correlated with PLT, platelet distribution width (PDW) and PCT. Also, PCT and PLT had high sensitivity and specificity to distinct EMT and MMT from the healthy bitches. Microcytic and hypochromic anemia occurs due to the decrease in the amount of HGB. Levels of MCV, MCH, and MCHC in the HBC group were reported to be significantly lower than in humans without breast cancer. Although anemia did not occur in EMT and MMT groups, obtained significances in the HCT, HGB, MCV, MCH, and MCHC levels between the bitches with and without mammary tumor were in line with the previous reports. In this study, WBC levels in grade 3 tumors were significantly higher than grade1 tumors (P < 0.05). Whereas levels of WBC in grade 1 and grade 2 tumors were in referenced laboratory ranges, it was higher in grade 3. Increased level of WBC in grade 3 was supposed to be due to the rise in malignancy as previously reported. Thrombocytosis was detected in 48.83% and 41.66% of the bitches in EMT and MMT groups, respectively. The higher percentage of CMTs with thrombocytosis in this study might be due to the difference in referenced upper limit of PLT in previous studies. The elapsed time between tumor formation and clinical presentation could be another influencing factor. Although PLT and PCT values were not significant according to the histological grading in this study, both parameters were found to be higher in grade 3 than the normal reference values. Further studies conducted with higher populations may lead the differences in these parameters to significance. With the support of further studies, alterations in the above mentioned parameters in bitches may contribute in the diagnosis process, management of treatment and may constitute an easy way to have an idea about the prognosis of mammary tumors.(AU)

Evidence of cytogenetic and histological damage in specimens of Astyanax lacustris (Pisces, Characidae) exposed to the hydrogen cyanide-based herbicide Dormex®

The herbicide Dormex®, a solution of hydrogen cyanamide, is a growth regulator capable of breaking the dormancy of fruit plants, and is commonly applied in agriculture. However, the biological effects of this product on non-target organisms are unknown. The present study investigated the biological response of Astyanax lacustris (Lütken, 1875) specimens exposed to Dormex® using a chromosome aberration test, the mitotic index, and the histological analysis of the gills. Forty specimens of Astyanax lacustris were obtained from a local breeding facility and divided into 10 groups (nine experimental and one control) with four fish in each aquarium (group). The control group was maintained for 24 hours in dechlorinated water while the experimental groups were allocated to one of nine different treatments, with three concentrations of Dormex®, 0.05, 0.1 and 0.5 mL L-1, and exposure for 24, 48 and 72 hours. The fish exposed to Dormex® presented chromosomal aberrations of a number of types, including chromosomal breaks, acentric fragments, decondensation, and gaps at the three Dormex® concentrations, at all exposure times. The mitotic index decreased significantly in comparison with the control group. The histological preparations of the gills revealed alterations such as hyperplasia, and lamellar fusion and edema, whereas in the control group the structure of the gills was preserved. The cytogenetic analysis revealed the genotoxic potential of the herbicide Dormex® and the morphological alterations of the gills demonstrated the sensitivity of the fish, which responded rapidly to the stressor. These findings reinforce the need for special care and restrictions on the use of these herbicides in agricultural areas located near aquatic environments.

Evidence of cytogenetic and histological damage in specimens of Astyanax lacustris (Pisces, Characidae) exposed to the hydrogen cyanide-based herbicide Dormex®

The herbicide Dormex®, a solution of hydrogen cyanamide, is a growth regulator capable of breaking the dormancy of fruit plants, and is commonly applied in agriculture. However, the biological effects of this product on non-target organisms are unknown. The present study investigated the biological response of Astyanax lacustris (Lütken, 1875) specimens exposed to Dormex® using a chromosome aberration test, the mitotic index, and the histological analysis of the gills. Forty specimens of Astyanax lacustris were obtained from a local breeding facility and divided into 10 groups (nine experimental and one control) with four fish in each aquarium (group). The control group was maintained for 24 hours in dechlorinated water while the experimental groups were allocated to one of nine different treatments, with three concentrations of Dormex®, 0.05, 0.1 and 0.5 mL L-1, and exposure for 24, 48 and 72 hours. The fish exposed to Dormex® presented chromosomal aberrations of a number of types, including chromosomal breaks, acentric fragments, decondensation, and gaps at the three Dormex® concentrations, at all exposure times. The mitotic index decreased significantly in comparison with the control group. The histological preparations of the gills revealed alterations such as hyperplasia, and lamellar fusion and edema, whereas in the control group the structure of the gills was preserved. The cytogenetic analysis revealed the genotoxic potential of the herbicide Dormex® and the morphological alterations of the gills demonstrated the sensitivity of the fish, which responded rapidly to the stressor. These findings reinforce the need for special care and restrictions on the use of these herbicides in agricultural areas located near aquatic environments.(AU)

Comparison of two different biomaterials in the bone regeneration (15, 30 and 60 days) of critical defects in rats

ABSTRACT Purpose To evaluate and compare two types of different scaffolds in critical bone defects in rats. Methods Seventy male Wistar rats (280 ± 20 grams) divided into three groups: control group (CG), untreated animals; biomaterial group 1 (BG1), animals that received the scaffold implanted hydroxyapatite (HA)/poly(lactic-co-glycolic) acid (PLGA); and biomaterial group 2 (BG2), animals that received the scaffolds HA/PLGA/Bleed. The critical bone defect was induced in the medial region of the skull calotte with the aid of an 8-mm-diameter trephine drill. The biomaterial was implanted in the form of 1.5 mm thick scaffolds, and samples were collected after 15, 30 and 60 days. Non-parametric Mann-Whitney test was used, with the significance level of 5% (p ≤ 0.05). Results Histology revealed morphological and structural differences of the neoformed tissue between the experimental groups. Collagen-1 (Col-1) findings are consistent with the histological ones, in which BG2 presented the highest amount of fibers in its tissue matrix in all evaluated periods. In contrast, the results of receptor activator of nuclear factor kappa-Β ligand (Rank-L) immunoexpression were higher in BG2 in the periods of 30 and 60 days, indicating an increase of the degradation of the biomaterial and the remodeling activity of the bone. Conclusions The properties of the HA/PLGA/Bleed scaffold were superior when compared to the scaffold composed only by HA/PLGA.

Aspergilose sistêmica em potro (Equus caballus) / Systemic aspergillosis in a foals - Equus caballus

O gênero Aspergillus compreende espécies de fungos saprotróficos, globalmente distribuídos, sendo os Aspergillus seção Fumigati conhecidos por causarem doença em humanos e animais imunocomprometidos. Os sinais clínicos dependem do sistema acometido, podendo apresentar febre branda, cólicas intensas, pneumonias, micose de bolsa gutural, e, ocasionalmente, lesões no sistema nervoso central. O presente trabalho objetiva relatar os sinais clínicos, os achados de necropsia e a histologia de um caso de aspergilose sistêmica em um potro de oito meses de idade. O animal apresentava refluxo enterogástrico, peristaltismo aumentado, fezes pastosas, dor abdominal e desidratação intensa. O quadro clínico evoluiu para óbito e o cadáver foi encaminhado para a necropsia. Macroscopicamente, havia sufusões e equimoses na serosa do trato gastrointestinal e ulcerações na mucosa do intestino delgado. No pulmão, havia nódulos multifocais, esbranquiçados e firmes que, ao corte, apresentavam-se císticos, com revestimento interno vinhoso e friável. O coração exibia áreas esbranquiçadas multifocais no miocárdio e os rins continham nódulos avermelhados nas regiões cortical e medular. No encéfalo, havia áreas multifocais amareladas e hemorrágicas com bordos avermelhados. Microscopicamente, observaram-se áreas multifocais de necrose com infiltrado inflamatório granulomatoso no intestino, pulmão, encéfalo, miocárdio e nos rins. Hifas fúngicas intralesionais e intravasculares foram observadas no encéfalo e no pulmão. Estruturas de fenótipo compatível com Aspergillus fumigatus foram observadas na cultura fúngica. Amostras encaminhadas para virologia apresentaram-se positivas para Herpesvírus Equino (HVE) tipo 1 e 4. O diagnóstico de aspergilose sistêmica foi determinado pelo histórico clínico, imunossupressão, achados macroscópicos, histológicos, isolamento do fungo e à infecção concomitante por HVE.

The Aspergillus genus comprises globally distributed species of saprotrophic fungi, with the Aspergillus section Fumigatus known to cause diseases in humans and immunocompromised animals. Clinical signs depend on the affected system, exhibiting mild fever, severe colic, pneumonia, guttural bag mycosis and occasionally lesions in the central nervous system. The present work aims to report the clinical signs, necropsy findings and histology of a case of systemic aspergillosis in an eight-months-old foal. The animal had enterogastric reflux, increased peristalsis, pasty feces, abdominal pain and severe dehydration. The condition evolved to death and the cadaver was sent for necropsy. Macroscopically, there were suffusions and ecchymosis in the serosa of the gastrointestinal tract and ulcerations in the mucosa of the small intestines. In the lung, there were multifocal, whitish and firm nodules that were cystic when cut with a friable winey internal lining. The heart had whitish multifocal areas in the myocardium and the kidneys contained reddish nodules in the cortical and medullary regions. In the brain there were yellowish and hemorrhagic multifocal areas with reddish borders. Microscopically, multifocal areas of necrosis with granulomatous inflammatory infiltrate were observed in the intestine, lung, brain, myocardium and kidneys. Intralesional and intravascular fungal were observed in the brain and lung. Structures with phenotype compatible with Aspergillus fumigatus were observed in the fungal culture. In virology, samples were positive for equine herpesvirus (HVE) types 1 and 4. The diagnosis of systemic aspergillosis was determined due to the clinical history, immunosuppression, macroscopic and histological findings, fungal isolation and concomitant HVE infection.

Aspergilose sistêmica em potro (Equus caballus) / Systemic aspergillosis in a foals - Equus caballus

O gênero Aspergillus compreende espécies de fungos saprotróficos, globalmente distribuídos, sendo os Aspergillus seção Fumigati conhecidos por causarem doença em humanos e animais imunocomprometidos. Os sinais clínicos dependem do sistema acometido, podendo apresentar febre branda, cólicas intensas, pneumonias, micose de bolsa gutural, e, ocasionalmente, lesões no sistema nervoso central. O presente trabalho objetiva relatar os sinais clínicos, os achados de necropsia e a histologia de um caso de aspergilose sistêmica em um potro de oito meses de idade. O animal apresentava refluxo enterogástrico, peristaltismo aumentado, fezes pastosas, dor abdominal e desidratação intensa. O quadro clínico evoluiu para óbito e o cadáver foi encaminhado para a necropsia. Macroscopicamente, havia sufusões e equimoses na serosa do trato gastrointestinal e ulcerações na mucosa do intestino delgado. No pulmão, havia nódulos multifocais, esbranquiçados e firmes que, ao corte, apresentavam-se císticos, com revestimento interno vinhoso e friável. O coração exibia áreas esbranquiçadas multifocais no miocárdio e os rins continham nódulos avermelhados nas regiões cortical e medular. No encéfalo, havia áreas multifocais amareladas e hemorrágicas com bordos avermelhados. Microscopicamente, observaram-se áreas multifocais de necrose com infiltrado inflamatório granulomatoso no intestino, pulmão, encéfalo, miocárdio e nos rins. Hifas fúngicas intralesionais e intravasculares foram observadas no encéfalo e no pulmão. Estruturas de fenótipo compatível com Aspergillus fumigatus foram observadas na cultura fúngica. Amostras encaminhadas para virologia apresentaram-se positivas para Herpesvírus Equino (HVE) tipo 1 e 4. O diagnóstico de aspergilose sistêmica foi determinado pelo histórico clínico, imunossupressão, achados macroscópicos, histológicos, isolamento do fungo e à infecção concomitante por HVE.(AU)

The Aspergillus genus comprises globally distributed species of saprotrophic fungi, with the Aspergillus section Fumigatus known to cause diseases in humans and immunocompromised animals. Clinical signs depend on the affected system, exhibiting mild fever, severe colic, pneumonia, guttural bag mycosis and occasionally lesions in the central nervous system. The present work aims to report the clinical signs, necropsy findings and histology of a case of systemic aspergillosis in an eight-months-old foal. The animal had enterogastric reflux, increased peristalsis, pasty feces, abdominal pain and severe dehydration. The condition evolved to death and the cadaver was sent for necropsy. Macroscopically, there were suffusions and ecchymosis in the serosa of the gastrointestinal tract and ulcerations in the mucosa of the small intestines. In the lung, there were multifocal, whitish and firm nodules that were cystic when cut with a friable winey internal lining. The heart had whitish multifocal areas in the myocardium and the kidneys contained reddish nodules in the cortical and medullary regions. In the brain there were yellowish and hemorrhagic multifocal areas with reddish borders. Microscopically, multifocal areas of necrosis with granulomatous inflammatory infiltrate were observed in the intestine, lung, brain, myocardium and kidneys. Intralesional and intravascular fungal were observed in the brain and lung. Structures with phenotype compatible with Aspergillus fumigatus were observed in the fungal culture. In virology, samples were positive for equine herpesvirus (HVE) types 1 and 4. The diagnosis of systemic aspergillosis was determined due to the clinical history, immunosuppression, macroscopic and histological findings, fungal isolation and concomitant HVE infection.(AU)

Relationship of obesity with clinical and laboratory changes in female dogs with mammary neoplasia

ABSTRACT: Obesity may be associated with the onset of mammary tumors in women. In companion animals, these data are still scarce, mainly associating the clinic of the patient with laboratory and histological findings. This study aimed to evaluate the clinical and laboratory aspects of female dogs with mammary neoplasia, investigating whether they are related to obesity. Four groups of spayed or non-spayed female dogs were studied, divided into (G1) female dogs without mammary tumor and normal body condition score (BCS), (G2) female dogs without mammary tumor and obese, (G3) female dogs with mammary tumor and normal BCS and (G4) female dogs with mammary tumor and obese. BCS, canine body mass index (CBMI), blood count, glucose, total cholesterol, triglycerides and cytopathological and histopathological examinations were evaluated. Mixed-breed animals were the most prevalent in this study (67.5%). There was a limitation regarding the use of CBMI related to body proportionality. Considering the 28 tumors evaluated, carcinoma in mixed tumor was the most prevalent malignant histological type, while adenomyoepithelioma was the most prevalent benign histological type. It was possible to conclude that obesity was not related to clinical and laboratory changes in the female dogs affected with mammary neoplasias. In addition, no relationship was noted between BCS and CBMI with the histological type or malignancy of mammary neoplasia, since the malignant histological types were the most prevalent tumors in both the ideal score group and the overweight to obese group.

RESUMO: A obesidade pode estar associada ao aparecimento de tumores mamários em mulheres. Em animais de companhia, esse dado ainda é escasso, principalmente associando a clínica da paciente aos achados laboratoriais e histológicos. Este estudo objetivou avaliar os aspectos clínicos e laboratoriais de cadelas com neoplasia mamária, investigando se estão relacionadas à obesidade. Foram estudados quatro grupos de cadelas castradas ou não castradas, divididas em (G1) cadelas sem tumor de mama e Escore de Condição Corporal normal (BCS), (G2) cadelas sem tumor de mama e obesa, (G3) cadelas com tumor de mama e ECC normal e (G4) cadelas com tumor de mama e obesa. Foram avaliados o Escore de Condição Corporal (ECC), Índice de Massa Corporal Canina (IMC), hemograma, glicose, colesterol total, triglicerídeos e exames citopatológicos e histopatológicos. Os animais de raça mista foram os mais prevalentes neste estudo (67,5%). Houve uma limitação quanto ao uso do IMC relacionado à proporcionalidade corporal. Considerando os 28 tumores avaliados, o carcinoma em tumor misto foi o tipo histológico maligno mais prevalente, enquanto o adenomioepitelioma foi o tipo histológico benigno mais prevalente. Foi possível concluir que a obesidade não esteve relacionada a alterações clínicas e laboratoriais nas cadelas acometidas por neoplasias mamárias. Além disso, não foi observada relação entre BCS e CBMI com o tipo histológico ou malignidade das neoplasias mamárias, uma vez que os tipos histológicos malignos foram os tumores mais prevalentes no grupo escore ideal e no grupo com sobrepeso e obesidade.

Age-dependent histomorphometric evolution of the corpus spongiosum

Purpose: To quantify the age-dependent changes in the relative volume of elastic fibers, collagen fibers and the proportion of collagen types I/III in the corpus spongiosum of rats. Methods: Forty-eight rats, raised under similar conditions, were divided into four groups (G1 to G4) and underwent penectomy at the ages of 6, 9, 12 and 24 months, respectively. Histological sections from the middle segment of the penis were stained with Weigerts resorcin-fuchsin, Massons trichrome and Picrosirius red, the volumetric density of elastic fibers, collagen fibers and the proportion of collagen types I and III in the corpus spongiosum were determined by stereological analysis. Results: A reduction in the proportion of collagen I/III between the groups G3 and G4 (p < 0.048) was observed. In the volumetric analysis of elastic fiber, we observed a significant rise between the groups G2 and G3 (p < 0.03) and a reduction of the volume between the groups G3 and G4 (p < 0.01). However, there was no difference in the quantity of total collagen between the groups (p > 0.54). Conclusions: Aging in rats did not change the quantity of total collagen but reduced the proportion of collagen types I/III and the volume of elastic fibers.(AU)

Testicular tumors in 190 dogs: clinical, macroscopic and histopathological aspects

ABSTRACT: This study aimed to characterize the prevalence and clinical, macroscopic and histopathological aspects of dogs affected by testicular tumors based on biopsy specimens from the Laboratório de Patologia Veterinária of the Universidade Federal de Santa Maria (LPV-UFSM) over 19 years. Parameters regarding the age, size, and breed of the affected dogs were also established. Of all dogs with some type of neoplasm submitted to histopathological analysis at the LPV over these 19 years (n=1,900), 213 (11.2%) had at least one testicular neoplasm. The tissues of 190 dogs (with 220 neoplasms) were available for histological reassessment. The dogs in this study had different types of testicular tumors with relatively similar frequencies. In descending order, the most frequent testicular neoplasms were seminomas (88/220), Leydig (interstitial) cell tumor (LCT; 64/220), Sertoli cell tumor (SCT; 61/220), and mixed germ cell-sex cord stromal tumor (MGSCT) (07/220). Among the dogs of defined breed (119 cases), large breeds had the largest number of cases (50/119), followed by small (47/119) and medium-sized (22/119) breeds. The ages of dogs affected by testicular tumors ranged from 10 months to 18 years. Increased testicular volume was the most common clinical manifestation. Eleven dogs presented information about clinical signs suggestive of hyperestrogenism syndrome (feminization). In seminomas, the diffuse pattern predominated over the intratubular pattern. Two sites (luminal and basal compartments) suggestive of the onset of neoplastic transformations in germ cells were observed in intratubular seminomas. They corroborate the hypothesis that canine seminomas possibly have pathogenesis similar to that observed in human spermatocytic seminomas. The SCTs and LCTs presented high cell morphology variation. SCTs had neoplastic cells organized in five different histological arrangements. As for LCT, solid-diffuse and cystic-vascular histological patterns were the most commonly observed. Through this study, it was possible to establish some of the leading clinical, macroscopic, and histopathological aspects of testicular neoplasms diagnosed over 19 years in the area covered by the LPV-UFSM.

RESUMO: Este estudo teve por objetivo caracterizar a prevalência, aspectos clínicos, macroscópicos e histopatológicos dos cães acometidos por neoplasmas testiculares, a partir dos espécimes de biópsias do Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV-UFSM) em 19 anos. Parâmetros quanto à idade, porte, raça dos cães acometidos também foram estabelecidos. De todos os cães com algum tipo de neoplasma submetido à análise histopatológica no LPV nesses 19 anos (n=1.900), 213 (11,2%) tinham ao menos um neoplasma testicular. Os tecidos de 190 cães (com 220 neoplasmas) estavam disponíveis para reavaliação histológica. Os cães deste estudo apresentaram diferentes tipos de neoplasmas testiculares com frequências relativamente semelhantes. Em ordem decrescente, os neoplasmas testiculares mais frequentes foram: seminomas (88/220), leydigomas (64/220), sertoliomas (61/220) e o tumor misto de células germinativas e do estroma do cordão sexual (MGSCT; 07/220). Dentre os cães com raça definida (119 casos), as raças de grande porte tiveram o maior número de casos (50/119), seguido das raças de pequeno (47/119) e médio porte (22/119). As idades dos cães acometidos por neoplasmas testiculares variaram de 10 meses a 18 anos. Aumento de volume testicular foi a manifestação clínica mais comum. Onze cães tinham informações sobre sinais clínicos sugestivos da síndrome da feminilização. Nos seminomas, houve o predomínio do padrão difuso sobre o intratubular. Dois locais (compartimentos luminal e basal) sugestivos de início das transformações neoplásicas nas células germinativas foram observados nos seminomas intratubulares, corroborando com a hipótese de que os seminomas caninos possivelmente tem patogênese semelhante à observada nos seminomas espermatocíticos humanos. Sertoliomas e leydigomas foram neoplasmas com alta variação na morfologia celular. Os sertoliomas tinham células neoplásicas dispostas em cinco arranjos histológicos distintos. Quanto aos leydigomas, os padrões histológicos sólido-difuso e cístico-vascular foram os mais comumente observados. Através deste estudo foi possível estabelecer alguns dos principais aspectos clínicos, macroscópicos e histopatológicos dos neoplasmas testiculares diagnosticados em 19 anos na área de abrangência do LPV-UFSM.

Metalloproteinases and colorectal cancer. Correlation of gene expression and clinical-pathological parameters

Purpose: To analyze gene and protein expression of metalloproteinases 1, 2, 9, 11 and 16 and their correlation with clinicopathological variables in colorectal adenocarcinoma. Methods: A retrospective study of 114 patients with colorectal adenocarcinoma treated surgically in the period 2006 to 2008 in Hospital de Câncer de Barretos - Fundação Pio XII. The evaluation of gene expression was performed by RT-PCR, and protein by immunohistochemistry. The analysis of gene expression was classified as overexpressed genes and poorly expressed (fold change of approximately 2, p 0.05). The positivity of the markers in the immunohistochemical study was performed by semi-quantitative analysis. The tissue of TMA (Tissue Microarray) was done by two independent pathologists. Results: The gene expression validated by immuno - histochemical was MMP-1(p= 0.00 and 1.57 fold change) and MMP 2 (p= 0.01 and 1.84 to fold change) when correlated with the histological types mucinous and adenocarcinoma NOS, MMP9 (p=0.01 and fold change of 1.13) and MMP-16 (p=0.03 and 1.61 fold change) when compared with the histological types villous and adenocarcinoma NOS, MMP - 11 statistically significant in relation to male (p = 0.04 and 1.65 fold change). Conclusions: The MMPs 1, 2, 9, 11 and 16 gene and protein expression with statistical significance in at least one of the clinicopathological variables studied. Thus, we conclude that these MMPs have potential as a prognostic factor in colorectal adenocarcinoma.(AU)