A new standardization for the use of chicken embryo: selection of target from the phage display library and infection.

The filamentous bacteriophage M13KO7 (M13) is the most used in phage display (PD) technology and, like other phages, has been applied in several areas of medicine, agriculture, and in the food industry. One of the advantages is that they can modulate the immune response in the presence of pathogenic microorganisms, such as bacteria and viruses. This study evaluated the use of phage M13 in the chicken embryos model. We inoculated 13-day-old chicken embryos with Salmonella Pullorum (SP) and then evaluated survival for the presence of phage M13 or E. coli ER2738 (ECR) infected with M13. We found that the ECR bacterium inhibits SP multiplication in 0.32 (M13-infected ECR) or 0.44 log UFC/mL (M13-uninfected ECR) and that the ECR-free phage M13 from the PD library can be used in chicken embryo models. This work provides the use of the chicken embryo as a model to study systemic infection and can be employed as an analysis tool for various peptides that M13 can express from PD selection. KEY POINTS: • SP-infected chicken embryo can be a helpful model of systemic infection for different tests. • Phage M13 does not lead to embryonic mortality or cause serious injury to embryos. • Phage M13 from the PD library can be used in chicken embryo model tests.

Unlocking the power of Libidibia ferrea extracts: antimicrobial, antioxidant, and protective properties for potential use in poultry production.

Plant extracts are increasingly recognized as potential prophylactic agents in poultry production due to their diverse bioactive properties. This study investigated the phytochemical and biological properties of Libidibia ferrea (L. ferrea), a plant species native to the Caatinga region of northeastern Brazil. The aim of this study was to identify secondary metabolites and to demonstrate the antimicrobial, antioxidant and protective effects of the plant extract. Three extracts were produced: EHMV, a hydroalcoholic extract from the maceration of pods, and EEMC and EEMV ethanolic extracts from the maceration of peels and pods, respectively, from L. ferrea. High-performance liquid chromatography (HPLC-MS/MS) and atomic absorption spectroscopy (AAS) were used to characterize the metabolites and metals. The antimicrobial activity against Salmonella Galinarum (SG), Salmonella pullorum (SP), Salmonella Heidelberg (SH) and Avian pathogenic Escherichia coli (APEC) was evaluated alone and in combination with probiotic bacteria (Bacillus velenzensis) using agar diffusion and the bactericidal minimum concentration (CBM). The antioxidant potential of the extracts was evaluated in 5 in vitro assays and 6 assays in 3t3 cells. The toxicity of EHMV was tested, and its ability to combat SP infection was demonstrated using a chicken embryo model. The results showed that EHMV exhibited significant antimicrobial activity. The combination of EHMV with BV had synergistic effects, increased antimicrobial activity and induced bacterial sporulation. Composition analysis revealed the presence of 8 compounds, including tannins and phenolic compounds. In vitro antioxidant tests demonstrated that total antioxidant capacity(TAC) activity was increased, and the extract had strong reducing power and notable metal chelating effects. Analysis of 3T3 cells confirmed the protective effect of EHMV against oxidative stress. Toxicity assessments in chicken embryos confirmed the safety of EHMV and its protective effect against SP-induced mortality. EHMV from L. ferrea is rich in proteins and contains essential metabolites that contribute to its antimicrobial and antioxidant properties. When associated with probiotic bacteria such as B. velezensis, this extract increases the inhibition of SH, SG, SP, and APE. The nontoxic nature of EHMV and its protective effects on chicken embryos make it a potential supplement for poultry.

Solid epithelioid peritoneal mesothelioma with pulmonary metastasis in feline.

Mesothelioma is a rare malignant neoplasm that affects the mesothelial cells lining the thoracic and abdominal cavities, such as the pleura, peritoneum, and pericardium. It is most prevalent in dogs and cattle, but the causes of this disease in animals are uncertain. In felines, it mainly affects the pleura, with an unfavorable prognosis. This paper explores a rare case of metastatic peritoneal mesothelioma in a 2-year-old female mixed breed cat, emphasizing its uniqueness due to the feline's age. The patient, previously treated at a private clinic, presented moderate abdominal distension as the only clinical sign. Abdominal ultrasound and peritoneal fluid cytology led to the provisional diagnosis of mesothelioma/carcinomatosis. One day after exploratory laparotomy, the animal died and was subsequently sent for necropsy. During macroscopic analysis, nodules were observed in the peritoneum, diaphragm, omentum, stomach serosa, and large intestine, and the diagnosis of solid epithelioid peritoneal mesothelioma with lung metastasis was confirmed after microscopic analysis. The diagnosis of mesothelioma is challenging, and the importance of immunohistochemical panels with specific markers such as cytokeratin AE1/AE3 and calretinin is highlighted. Considering that mesothelioma is a pathology with a poor prognosis, it is essential to include this disease in the list of differential diagnoses within veterinary oncology.

O mesotelioma é uma neoplasia maligna rara que afeta as células mesoteliais que revestem as cavidades torácica e abdominal, como a pleura, o peritônio e o pericárdio. É mais prevalente em cães e bovinos, mas as causas desta doença em animais são incertas. Nos felinos acomete principalmente a pleura, com prognóstico desfavorável. Este artigo explora um caso raro de mesotelioma peritoneal metastático em uma gata sem raça definida de 2 anos de idade, enfatizando sua singularidade devido à idade do felino. O paciente, previamente atendido em clínica particular, apresentava distensão abdominal moderado como único sinal clínico. A ultrassonografia abdominal e a citologia do líquido peritoneal levaram ao diagnóstico provisório de mesotelioma/carcinomatose. Um dia após a laparotomia exploratória, o animal veio a óbito e posteriormente encaminhado para necropsia. Durante a análise macroscópica, foram observados nódulos no peritônio, diafragma, omento, serosa estomacal e intestino grosso e o diagnóstico de mesotelioma peritoneal epitelioide sólido com metástase pulmonar foi confirmado após análise microscópica. O diagnóstico do mesotelioma é desafiador, sendo destacada a importância de painéis imunohistoquímicos com marcadores específicos como citoqueratina AE1/AE3 e calretinina. Considerando que o mesotelioma é uma patologia de prognóstico ruim, é fundamental incluir esta doença na lista de diagnósticos diferenciais dentro da oncologia veterinária.

Chicken embryos are a valuable model for the selection of Bacillus subtilis for probiotic purposes.

Bacillus subtilis (BS) has been used as an excellent probiotic; however, some BS strains seem to be opportunist pathogens or do not present inhibitory effects in the pathogenic bacteria, so the characterization of BS strains for use in animals is mandatory. This study aimed to select nonpathogenic strains of BS, which can inhibit Salmonella spp., avian pathogenic Escherichia coli (APEC), and Campylobacter jejuni (CJ) using a chicken embryo as a model. We tested nine (9) strains of BS isolated from several sources (named A to I) in in vitro by tests of mucin degradation activity, haemolytic activity, apoptosis, and necrosis in fibroblasts from chickens. After the in vitro test, we tested the remaining seven (7) strains (strains A to G) in a chicken embryo (CE) as an in vivo model and target animal. We inoculated 3 log CFU/CE of each strain via allantoic fluid at the 10th day postincubation (DPI). Each treatment group consisted of eight CEs. At the 17th DPI we checked CE mortality, gross lesions, CE weight, and whether BS strains were still viable. To perform the cytokine, total protein, albumin, and reactive C protein analysis, we collected the CE blood from the allantoic vessel and intestine fragments in the duodenum portion for histomorphometric analysis. After the results in CEs, we tested the inhibition capacity of the selected BS strains for diverse strains of Salmonella  Heidelberg (SH), S. Typhimurium (ST), S. Enteritidis (SE), S. Minnesota (SM), S. Infantis (SI), Salmonella var. monophasic (SVM), APEC and C. jejuni. After the in vitro trial (mucin degradation activity, haemolytic activity, apoptosis, and necrosis), we removed two (2) strains (H and I) that showed ß-haemolysis, mucin degradation, and/or high apoptosis and necrosis effects. Although all strains of BS were viable in CEs at the 17th DPI, we removed four (4) strains (A, B, D, F) once they led to the highest mortality in CEs or a high albumin/protein ratio. C. jejuni inoculated with strain G had greater weight than the commercial strain, which could be further used for egg inoculation with benefits to the CE. From the tests in CEs, we selected the strains C, E, and G for their ability to inhibit pathogenic strains of relevant foodborne pathogens. We found that the inhibition effect was strain dependent. In general, strains E and/or G presented better or similar results than commercial control strains in the inhibition of SH, ST, SI, APEC, and two (2) strains of CJ. In this study, we selected BS strains C, E and G due to their in vitro and in vivo safety and beneficial effects. In addition, we emphasize the value of CE as an in vivo experimental model for assessing BS's safety and possible benefits for poultry and other animals.

Alternative use of phage display: phage M13 can remain viable in the intestines of poultry without causing damage.

Phage display (PD) is a tool for developing new molecules to control pathogens. Peptides selected by PD are commonly synthesised and tested, but the use of phage M13 displaying the selected peptides as a direct biding in the intestinal tract has not yet been tested. This study evaluated whether phage M13 can remain viable in the chicken gastrointestinal tract and whether it causes injury or humoral immune response. We inoculated phage M13 or E. coli ER2738 (ECR) infected with M13 into birds at different ages. We found the virus in faeces at 5 or 13 days after inoculation, just when it infected the ECR. The presence of phage M13 or ECR did not result in gut injuries and had no impacts on weight gain and bird health. Furthermore, the levels of IgY were similar in all treatments, which indicates that the virus can be used in chicken until 42 days without being recognised by the immune system. This work provides a scientific basis for the use of PD as a tool in numerous applications to control different pathogens.

Physiological Changes in Chicken Embryos Inoculated with Drugs and Viruses Highlight the Need for More Standardization of this Animal Model.

Several studies have been developed using the Gallus gallus embryo as an experimental model to study the toxicity of drugs and infections. Studies that seek to standardize the evaluated parameters are needed to better understand and identify the viability of CEs as an experimental model. Therefore, we sought to verify whether macroscopic, histopathological, blood count, metabolites and/or enzymes changes and oxidative stress in CE of different ages are specific to the model. To achieve this goal, in ovo assays were performed by injecting a virus (Gammacoronavirus) and two drugs (filgrastim and dexamethasone) that cause known changes in adult animals. Although congestion and inflammatory infiltrate were visible in the case of viral infections, the white blood cell count and inflammation biomarkers did not change. Filgrastim (FG) testing did not increase granulocytes as we expected. On the other hand, CE weight and red blood cell count were lower with dexamethasone (DX), whereas white blood cell count and biomarkers varied depended on the stage of CE development. Our work reinforces the importance of standardization and correct use of the model so that the results of infection, toxicity and pharmacokinetics are reproducible.

Leptospirosis in slaughtered cows in the Triangulo Mineiro, Minas Gerais: prevalence, serological profile and renal lesions / Leptospirose em vacas abatidas no Triângulo Mineiro, Minas Gerais: prevalência, perfil sorológico e lesões renais

Leptospirosis is a zoonosis that affects several species of domestic and wild animals and is an important cause of economic losses in cattle in Brazil. In this study, we determined the prevalence of bovine leptospirosis in the Triângulo Mineiro region, Minas Gerais, Brazil, identified the most frequent serovars of Leptospira interrogans, and examine the renal pathological changes associated with the disease. Samples of blood serum and kidney fragments from 100 bovine females were collected in cattle abattoir. In the serological investigation 48% of the cows were positive. The serovars for which there were more reactive animals were Wolffi (24%), Hardjo (21%) and Hebdomadis (18%). Among the positive samples, 14/48 showedantibody titers greater than 1:100, and 70.83% of the seropositive animals responded to more than one Leptospira interrogans serovar. Only one farm did not have seropositive cows and in nine farms studied, six (66.66%) presented seropositive animals to the Hebdomadis serovar. At the histopathological examination, the most frequent microscopic lesions in positive animals were hyalinization (81.25%), congestion (81.25%) and hydropic degeneration (70.83%). However, these histopathological alterations were also found in kidneys of animals negative to serology, such as hyalinization (80.77%), congestion (48.07%) and hydropic degeneration (55.77%) and these findings are unrelated to positivity. Histopathological examination of the kidneys is not indicated to replace the serological diagnosis of leptospirosis, and may be used only as a complementary examination. Despite the low frequency of seropositive animals in the Triângulo Mineiro region, the disease is present in a large number of farms. Noteworthy is the high frequency of serovar Hebdomadis and it can be considered an emerging serovar in the region. The evaluation of the frequency of this serovar in other regions becomes important, and once verified should result in the recommendation of the inclusion of this serovar in the leptospirosis control.

A leptospirose é uma zoonose que afeta várias espécies de animais domésticos e silvestres e importante causa de prejuízos à bovinocultura nacional. Objetivou-se determinar a prevalência da leptospirose e identificar os sorovares de Leptospira interrogans mais freqüentes na região do Triângulo Mineiro, além de avaliar microscopicamente as lesões renais e correlacionar estas lesões com sorovares específicos. Foram coletadas amostras de soro sanguíneo e fragmentos de rim de 100 fêmeas bovinas em abatedouro e, destas 100 amostras, 48% foram positivas. Os sorovares para os quais houve mais animais reagentes foram Wolffi (24%), hardjo (21%) e Hebdomadis (18%). Dentre as amostras positivas, 14/48 apresentaram títulos de anticorpos aglutinantes superiores a 1:100 e notou-se que 70,83% dos animais soropositivos reagiram a mais de um sorovar de Leptospira interrogans. Somente uma propriedade não possuía vacas sororreagentes para leptospirose e das nove propriedades estudadas, seis (66,66%) apresentaram animais soropositivos ao sorovar Hebdomadis. Ao exame histopatológico, as alterações microscópicas mais encontradas em animais positivos foram hialinização (81,25%), congestão (81,25%) e degeneração hidrópica (70,83%). Porém, essas alterações histopatológicas também foram encontradas em rins de animais negativos à sorologia, como hialinização (80,77%), congestão (48,07%) e degeneração hidrópica (55,77%) e estes achados não apresentaram correlação com positividade. O exame histopatológico dos rins não é indicado para substituir o diagnóstico sorológico da leptospirose, podendo ser utilizado somente como um exame complementar. Apesar da frequência de animais sororreagentes ser baixa na região do Triângulo Mineiro, a doença está presente em um grande número depropriedades. Chama a atenção a alta frequência do sorovar Hebdomadis, podendo ser considerado um sorovar emergente na região. A avaliação da frequência deste sorovar em outras regiões se torna importante, pois uma vez verificada, deve resultar na recomendação da inclusão deste sorovar no controle da leptospirose.

Imunomarcação da transição epitélio-mesenquima na neoplasia mamária primária de cadelas e sua metástase em linfonodo / Epithelial to mesenchymal transition immunostaining in canine primary mammary tumor and lymph node

Os carcinomas mamários em cães apresentam alta capacidade metastática o que confere menor sobrevida para os pacientes com este tipo de neoplasia. O fenótipo transição epitélio-mesênquima, caracterizado pela troca dos filamentos intermediários de citoqueratina por vimentina, além da perda da proteína de adesão entre células (E-caderina) está relacionado com a maior ocorrência de metástase. Diante disto, objetivou-se avaliar, por meio de imunomarcações, a expressão de vimentina, citoqueratina e E-caderina nos tumores mamários caninos e suas metástases em linfonodo, a fim de avaliar o comportamento celular frente a esta neoplasia. Foram analisados cinco casos de neoplasias mamárias primárias caninas e suas respectivas metástases em linfonodos. Foram comparadas as médias de imunomarcações do grupo de neoplasias primárias com as médias do grupo metástase. Não houve diferença estatística nas imunomarcações da citoqueratina (p=0,1407) e E-caderina (p= 0,312) entre os grupos, apesar da média de expressão da E-caderina ter sido maior no grupo de metástases. A expressão da vimentina foi maior nos sítios das metástases (p=0,0462). Conclui-se que a expressão de vimentina aumenta no foco da metástase em relação aos seus respectivos tumores primários mamários caninos, caracterizando alteração estrutural celular, conferindo um fenótipo transição epitélio-mesênquima. Além da E-caderina apresentar fortes indícios de aumento no foco da metástase caracterizando maior adesão.(AU)

Mammary carcinomas in dogs have a high metastatic capacity which gives a shorter survival rate for patients with this type of tumor. The epithelial-mesenchymal transition phenotype, characterized by the trade of intermediary filaments of cytokeratin by vimentin, also by the loss of the adhesion protein between cells (E-cadherin) is associated with metastasis. Due to this fact, it was aimed to evaluate, by immunostaining, the expression of vimentin, cytokeratin and E-cadherin in canine mammary tumors and the metastasis in lymph node, in order to assess the cell behavior when facing this cancer. Five cases of canine mammary tumors and metastasis in lymph node were evaluated. The averages of immunostainings of the group of primary neoplasms were compared with the averages of the lymph node group. The results showed that immunostaining for cytokeratins (p=0,1407) and E-caderina (p=0,312) were not significant between the groups, despite the expression mean of cadherin was higher in the metastase group. The expression of vimentin (p=0,04) was greater at sites of metastases. It is concluded that the expression of vimentin increases in the focus of the metastase in relation to their respective primary canine mammary tumors, characterizing cellular structural alteration, conferring a transient epithelial-mesenchymal phenotype. And cadherin present strong evidence of increased focus on metastasis characterizing increased adhesion.(AU)

Patologias pancreáticas em cães: Revisão de literatura / Pancreatic diseases in dogs: Literature review / Patologías pancreáticas en perros: Revisión de literatura

O pâncreas é conhecido como uma glândula mista, uma vez que atua tanto na secreção de enzimas digestivas através de sua porção exócrina, quanto na secreção de insulina e glucagon pela porção endócrina. Envolvendo a porção exócrina, nos cães a principal alteração evidenciada é a pancreatite, enquanto que no pâncreas endócrino a principal moléstia é o Diabetes mellitus. Não obstante, o pâncreas pode ser acometido por diversas patologias, dentre elas neoplasias, processos isquêmicos, traumas ou ainda por doenças idiopáticas. Dessa forma, o diagnóstico das disfunções deste órgão torna-se um desafio na Medicina Veterinária, tornando relevante a realização de maiores estudos acerca deste tema. Tendo em vista que abordagens sobre as patologias pancreáticas em cães são escassas, propõe-se assim a realização de uma revisão de literatura mensurando as principais enfermidades pancreáticas diagnosticadas em cães.(AU)

The pancreas is known as a mixed gland, since it acts both in the secretion of digestive enzymes through its exocrine portion, and in the secretion of insulin and glucagon through its endocrine portion. Pancreatitis is main disease developed in the pancreas exocrine portion in dogs, while Diabetes mellitus is the main disease in the endocrine portion. Nevertheless, the pancreas may be affected by several pathologies, among them neoplasms, ischemic processes, injuries or idiopathic diseases. The diagnosis of disorders in this organ becomes a challenge in veterinary medicine, thus the relevance of holding further studies on this subject. Considering the scarcity of approaches on pancreatic diseases in dogs, this study proposes to hold a literature review, addressing the major pancreatic diseases diagnosed in dogs.(AU)

El páncreas es conocido como una glándula mixta, una vez que actúa en la secreción de enzimas digestivas a través de su porción exocrina, así como en la secreción de insulina y glucagón por la porción endocrina. Involucrando la porción exocrina, en perros, la principal alteración observada es la pancreatitis, mientras que en el páncreas endócrino la molestia más común es el Diabetes mellitus. Sin embargo, el páncreas puede ser afectado por diversas patologías, tales como neoplasias, procesos isquémicos, traumas o por trastornos idiopáticos. De este modo, el diagnóstico de las disfunciones de este órgano se convierte en un reto para la Medicina Veterinaria, haciendo relevante la realización de nuevos estudios sobre este tema. Teniendo en cuenta que los enfoques sobre enfermedades pancreáticas en perros son escasos, se propone la realización de una revisión de literatura abordando las principales enfermedades pancreáticas diagnosticadas en perros.(AU)

Causes of death of domestic cats in a veterinary hospital of Uberlândia, Minas Gerais state: a retrospective study / Causas de óbito em gatos domésticos em um hospital veterinário de Uberlândia, Minas Gerais: estudo retrospectivo

Cats play an important role as pets in the present society. Thus, the knowledge about their main diseases and causes of death is fundamental to prevent various pathologies. However, when dealing with domestic cats there are very few researches in Veterinary Medicine. The aim of this study is to identify the causes of death of cats through a retrospective study of necropsies in the Animal Pathology Laboratory in the Federal University of Uberlândia (Brazil), in a period of 36 years, in which 350 cat necropsies were conducted. The most frequent causes of death of these animals were digestive system diseases (48 cats ­ 13.71%), and among them, feline hepatic lipidosis was the most frequent etiology (18.75%). The second most affected system was the respiratory (10.86%) and acute pneumonia, pulmonary edema, and diaphragmatic hernia were the main diseases to cause death of cats with some kind of disorder in this system. Other decease causes were rabies, squamous cell carcinoma, fibrous osteodystrophy, fractures and traumatism, hypovolemic shock, and feline lower urinary tract disease. This type of analysis can provide basic information about the most common diseases in felines, contributing with veterinarians as they adopt control and prevention actions against diseases.(AU)

Os gatos apresentam papel relevante como animal de estimação na sociedade atual. Neste sentido, o conhecimento das principais enfermidades e causas de morte desses animais é fundamental para prevenção de várias patologias. No entanto, quanto ao gato doméstico, essas pesquisas ainda são escassas na medicina veterinária. Assim, este trabalho realizou um estudo retrospectivo das causas de morte de gatos registradas das necropsias efetuadas pelo Laboratório de Patologia Animal da Universidade Federal de Uberlândia, em um período de 36 anos, no qual 350 gatos foram necropsiados. As doenças do sistema digestivo foram as mais frequentes (48 gatos ­ 13,71%) e destas, a lipidose hepática foi a etiologia de maior ocorrência (18,75%). O sistema respiratório foi o segundo mais afetado (10,86%), sendo pneumonia aguda, edema pulmonar e hérnia diafragmática as principais doenças deste sistema responsáveis pela morte de gatos. Outras causas de óbito importantes foram raiva, carcinoma de células escamosas, osteodistrofia fibrosa, fraturas e traumatismos, choque hipovolêmico e síndrome urológica felina. Este tipo de análise pode fornecer informações básicas sobre as enfermidades que mais acometem os felinos de uma determinada região, contribuindo para que os profissionais médicos veterinários passem a adotar as respectivas ações de prevenção e controle.(AU)

Case report and immunohistochemical study of a malignant peripheral nerve sheath tumor in a dog / Relato de caso e estudo imuno-histoquímico de tumor maligno da bainha do nervo periférico em um cão

Malignant peripheral nerve sheath tumor is rare and rarely reported in dogs. The term neurofibroma/sarcoma is classically used when the tumor is composed of Schawann and perineural cells. This work describes the clinical, histopathological, and immunohistochemical case of a subcutaneous malignant peripheral nerve sheath tumor, most likely a malignant neurofibroma located in the pelvic member of a Rottweiler dog. Histopathological features and immunohistochemical analysis corroborated the diagnosis, showing positivity for S-100 protein, vimentin, and CD57, and was useful to distinguish this type of neoplasm from other malignancies of similar morphologies.(AU)

O tumor maligno da bainha do nervo periférico é raro e pouco descrito em cães. Classicamente, o termo "neurofibroma/sarcoma" é empregado quando o tumor é composto por células de Schwann e células perineurais. Neste relato são descritos os achados clínicos, histopatológicos e imuno-histoquímicos de um caso de tumor maligno da bainha do nervo periférico, provavelmente neurofibrossarcoma, localizado no subcutâneo do membro pélvico de um cão da raça Rottweiler. Os achados histopatológicos associados às observações imuno-histoquímicas contribuíram para o diagnóstico, sendo observada positividade para proteína S-100, vimentina e CD57, permitindo a diferenciação da neoplasia em questão de outros tumores malignos com características morfológicas similares.(AU)