Physiological gait test: an effective method for analyzing balance, locomotion, and neuromuscular disorders in rats and a comparison to the elevated beam test / Teste de marcha fisiológica: um método eficaz para analisar distúrbios de equilíbrio, locomoção e neuromusculares em roedores

The evaluation of animal locomotor activity is a behavioral tool widely used to measure the mechanisms underlying a particular disease, disorder, or injury, as well as the effects of exposure to a xenobiotic. The elevated beam test is one of the most used tests in rodents to assess balance and motor coordination. Despite being inexpensive and utilizing a simple apparatus, the high beam test requires a long period of animal training and habituation. The development and characterization of an alternative test, namely the gait test, has the potential to circumvent the time and effort required for animal training, deeming it an effective, inexpensive, and fast method for the analysis of behaviors that are comparably assessed by the high beam test. Therefore, the present study focused on determining the effectiveness and feasibility of the gait test for assessing rodent locomotion and balance as a replacement for the elevated beam test. For this purpose, male rats were divided into three groups: one control group exposed to a saline solution (NaCl 0.9%) and two experimental groups exposed to a single dose of either 0.2 or 1.0 mg/kg of ivermectin intraperitoneally for induction of locomotor disturbance. The high beam and gait tests were performed 15 min and 24 h after drug administration. Results show that the experimental groups had difficulty performing the tasks of either test at both time points analyzed compared to the control groups. At the high beam, experimental animals had trouble maintaining balance and walking. At the gait test, experimental animals showed alterations in gait, which were quantitated by: (a) shortening of step length, (b) decrease of stride, (c) altered step symmetry, and (d) altered stride area. Such results are indicative of compensatory efforts and were comparable between both tests. Altogether, the data indicate that the gait test meets all requirements for assessing motor coordination in rodents. The gait test is therefore validated as a complement to the elevated beam test for the study and analysis of neurodegenerative impairment and other disorders involving neuromuscular disturbances.(AU)

A avaliação da atividade locomotora animal é uma ferramenta comportamental bastante utilizada para mensurar os mecanismos subjacentes a uma determinada doença, distúrbio ou lesão e efeitos da exposição a um xenobiótico. Um dos testes mais utilizados em roedores para avaliar o equilíbrio e coordenação motora é o teste da trave elevada que, apesar de ser um teste barato e que exige um aparato simples, é necessário um longo período de treino e habituação dos animais. O desenvolvimento e caracterização de um teste alternativo, chamado de teste da marcha, tem o potencial de contornar o tempo e o esforço necessários ao treino dos animais, considerando-o um método eficaz, barato e rápido para a análise de comportamentos avaliados comparativamente pelo alto teste de feixe. Portanto, o presente estudo concentrou-se em determinar a eficácia e viabilidade do teste de marcha para avaliação da locomoção e equilíbrio de roedores em substituição ao teste da trave elevada. Para isso, ratos machos foram divididos em 3 grupos, sendo 1 grupo controle exposto à solução salina (NaCl 0,9%) e 2 grupos experimentais expostos à dose única de 0,2 e 1,0 mg/kg de ivermectina por via intraperitoneal para indução da alteração locomotora. Os testes de trave elevada e marcha foram realizados 15 min e 24 h após a administração da droga. Os resultados mostram que os grupos experimentais tiveram dificuldade em realizar as tarefas de qualquer teste em ambos os momentos analisados em comparação com os grupos de controle. Na trave elevada, os animais experimentais tiveram dificuldade em manter o equilíbrio e andar. No teste de marcha, os animais experimentais apresentaram alterações na marcha, que foram quantificadas por: (a) encurtamento do comprimento da passada, (b) diminuição da passada, (c) alteração da simetria da passada e (d) alteração da área da passada. Tais resultados são indicativos de esforços compensatórios e foram comparáveis entre os dois testes. Em conjunto, os dados indicam que o teste de marcha atende a todos os requisitos para avaliação da coordenação motora em roedores. O teste de marcha é, portanto, validado como um complementar para o teste da trave elevada e para o estudo e análise de comprometimento neurodegenerativo e outros distúrbios envolvendo distúrbios neuromusculares.(AU)

Diagnóstico hematológico de coinfecção por Ehrlichia spp. e Hepatozoon canis em cães no semiárido da Paraíba / Hematogical diagnosis of coinfection by Ehrlichia spp. and Hepatozoon canis in dogs in the semi-arid of Paraiba

Dentre as hemoparasitoses com maior frequência de transmissão por carrapatos pode-se destacar a Erliquiose, a Hepatozoonose, a Anaplasmose e a Babesiose. A coinfecção por esses agentes são frequentes na rotina clínica; no entanto, poucas são as descrições sobre a ocorrência simultânea desses agentes em hemogramas. Sendo assim, o objetivo deste trabalho foi relatar os casos de quatro cães diagnosticados com coinfecção por Ehrlichia spp. e Hepatozoon canis, os quais foram atendidos no Hospital Veterinário Adílio Santos de Azevedo (HV-ASA) do Instituto Federal de Educação, Ciência e Tecnologia da Paraíba (IFPB), campus Sousa. Durante a avaliação clínica, relatou-se que os pacientes apresentavam, de maneira em comum, oligodipsia e hiporexia, além de histórico de presença de carrapatos. Em seguida, foram realizados hemogramas, que revelaram que todos os animais apresentavam anemia leve a moderada. Além disso, notou-se que o animal 1 estava com trombocitopenia e os animais 2 e 3, neutrofilia. Na pesquisa de hemoparasitas em esfregaço sanguíneo, observou-se coinfecção por H. canis e E. spp. nos quatro animais. Desta forma, este relato demonstrou a importância do uso do hemograma e da pesquisa dos agentes causadores das hemoparasitoses, através do esfregaço sanguíneo. Ressalte-se que o esfregaço sanguíneo, embora seja um método inespecífico e de baixa sensibilidade, pode e deve ser adotado como rotina, por ser uma técnica simples, rápida, barata, pouco invasiva e de diagnóstico definitivo.

Among the hemoparasitoses with the highest frequency of transmission by ticks, ehrlichiosis, hepatozoonosis, anaplasmosis, and babesiosis may be highlighted. The coinfection with these agents is frequent in clinical routine; however, there are few descriptions of the simultaneous occurrence of these agents in blood counts. Therefore, this work aimed to report the cases of four dogs diagnosed with Ehrlichia spp. and Hepatozoon canis, which were treated at the Adílio Santos de Azevedo Veterinary Hospital (HV-ASA), of the Federal Institute of Education, Science, and Technology of Paraíba (IFPB), Sousa campus. During the clinical evaluation, it was reported that the patients had oligodipsia and hyporexia in common, in addition to a history of tick presence. Then, blood counts were performed, which revealed that all animals had mild to moderate anemia. In addition, it was noted that animal 1 had thrombocytopenia and animals 2 and 3 had neutrophilia. In the investigation of hemoparasites in blood smears, co-infection with H. canis and E. spp. was observed in the four animals. Thus, this report demonstrated the importance of using the blood count and researching the causative agents of hemoparasitosis, through the blood smear. It should be noted that the blood smear, although it is a non-specific and low-sensitivity method, can and should be adopted as a routine, as it is a simple, fast, inexpensive, minimally invasive, and definitive diagnostic technique.

Qualidade microbiológica de hortaliças comercializadas em diferentes restaurantes do tipo self-service na cidade do Recife-PE / Microbiological quality of vegetables selled in different self service restaurants in the city of Recife-PE

A falta de tempo para o preparo das refeições, devido ao ritmo de vida acelerado, incentiva a procura por serviços de alimentação fora de casa, em busca de alimentos práticos, de rápido consumo e nutritivos. Com o propósito de fazer uma alimentação mais saudável, cresceu a busca por restaurantes self-service, sendo a salada uma das opções usualmente mais consumida nesses estabelecimentos. Neste contexto, este trabalho teve como objetivo avaliar a qualidade microbiológica de 20 amostras de hortaliças cruas servidas em restaurantes self-service da cidade do Recife-PE. As coletas foram realizadas no mês de fevereiro de 2021. As amostras foram submetidas às análises microbiológicas de coliformes totais, termotolerantes, Escherichia coli e Salmonella spp. Para coliformes totais os resultados variaram entre <3 a >1100 NMP/g. A presença de coliformes termotolerantes foi observada em 16 amostras, destas, uma apresentou contagem acima do estabelecido pela RDC 12/2001 se mostrando imprópria para o consumo. Não foi detectada a presença de Escherichia coli e Salmonella spp. nas amostras analisadas, estando estas de acordo com a RDC 331/2019. Estes resultados sugerem que, como houve a presença de coliformes nas amostras, mesmo estando a maioria dos valores em conformidade, é importante que haja maiores cuidados quanto à higienização e manipulação das hortaliças pelos estabelecimentos. Visando diminuir a carga microbiana presente nesses alimentos, portanto, é necessário que as boas práticas de manipulação sejam adotadas pelos restaurantes, para que se evite o aparecimento de doenças transmitidas por alimentos (DTAs).(AU)

The lack of time to prepare meals, due to a fast-paced contemporary lifestyle, encourages people to seek for alternative food services that are fast, practical, and nutritious. With the purpose of maintaining a healthy diet, the search for self-service restaurants has increased, being salad one of the usual options consumed in these restaurants. In this scenario, the purpose of the study was to investigate the microbiological quality of 20 raw vegetables samples served in restaurants in Recife ­ PE. The tests were carried out in February 2021. The samples were submitted to microbiological analysis of Total Coliform, thermotolerant Coliforms, Escherichia coli and Salmonella spp. For Total Coliform the results ranged from <3 to >1100 MPN/g. The presence of thermotolerant coliforms were observed in 16 samples, and 1 of them had a count above the established for the RDC 12/2001, proving to be unsuitable for consumption. The presence of Escherichia coli and Salmonella spp. were detected in none of the analyzed samples, being in accordance with the RDC 331/2019. These results suggest that, because of the presence of coliforms in the samples, even though most of values are in compliance, it is important to be more careful about hygiene and manipulation of vegetables by restaurants. Therefore, aiming to decrease the microbial load present in these foods, it is necessary that the good handling practices should be adopted in restaurants, to avoid the appearance of foodborne diseases.(AU)

Isolation and molecular identification of Mycobacterium bovis in tissue lesions of cattle slaughtered in slaughterhouses located in the State of Ceará / [Isolamento e identificação molecular de Mycobacterium bovis em lesões teciduais de bovinos abatidos em matadouros-frigoríficos localizados no estado do Ceará]

The objective of this research was to identify Mycobacterium bovis in lesions suggestive of tuberculosis in bovine carcasses in the State of Ceará, by means of bacteriological and molecular diagnostic tests. Between August 2017 and January 2019, the State inspection service (SIE) inspected 59,512 cattle, of which 7.4% (44 / 59,512) presented suggestive lesions. Of these animals, 68 samples were sent, of which 4.5% (31/68) located in the lung, 2.9% (20/68) in lymph nodes, 2.0% (14/68) in the liver, and 0.4% in the carcass (3/68). When performing bacteriological isolation, 15.9% (7/44) of bovines showed colony growth in the samples. The smears of the isolates were submitted to Zielh-Neelsen staining and all confirmed acid-fast bacilli. The polymerase chain reaction identified all isolates 100% (7/7) as M. bovis. The association of diagnostic techniques allowed to identify the presence of the agent in the State and the molecular analysis proved to be a beneficial technique in the monitoring of bovine tuberculosis and can be used as an auxiliary method in the bovine tuberculosis control and eradication program in the State of Ceará.

O objetivo do trabalho foi pesquisar Mycobacterium bovis em lesões sugestivas de tuberculose nas carcaças de bovinos no estado do Ceará, por meio dos testes de diagnóstico bacteriológico e molecular. Entre agosto de 2017 e janeiro de 2019, o Serviço de Inspeção Estadual (SIE) inspecionou 59.512 bovinos; destes, 7,4% (44/59.512) apesentaram lesões sugestivas. Desses animais foram enviadas 68 amostras, das quais 4,5% (31/68) estavam localizadas no pulmão, 2,9% (20/68) nos linfonodos, 2,0% (14/68) no fígado e 0,4% (3/68) na carcaça. Ao realizar o isolamento bacteriológico, 15,9% (7/44) dos bovinos evidenciaram crescimento de colônias nas amostras. Os esfregaços dos isolados foram submetidos à coloração de Zielh-Neelsen e todos eles confirmaram bacilo álcool-ácido resistente. A reação em cadeia da polimerase identificou todos os isolados, 100% (7/7), como M. bovis. A associação das técnicas de diagnóstico permitiu identificar a presença do agente no estado, e a análise molecular demonstrou ser uma técnica benéfica no monitoramento da tuberculose bovina, podendo ser utilizada como um método auxiliar no programa de controle e erradicação da tuberculose bovina no estado do Ceará.

Water uptake dynamics in soybean seeds: influence in seeds performance and DNA integrity / Dinâmica de absorção de água em sementes de soja: influência no desempenho de sementes e integridade do DNA

Relationships among water uptake rate and temperatures were investigated in five soybean seeds lots cv. M 6410 IPRO. Germination, field emergence and electrical conductivity tests were applied to determine seed performance after fast (complete immersion in water), control (moist substrate) and slow (moist atmosphere) imbibitions both at 20 °C and 30 °C. The fresh weight increment over time in soybean seeds during imbibition was recorded. In parallel, analysis of the DNA integrity of seedlings obtained from such seeds was performed in gel electrophoresis. Absorption pattern of soybean seeds were similar among temperatures, but absorption characteristics vary with time progress according to the way of water entrance into the seeds. Upon fast imbibition seeds germinated lower than non-fast-imbibed seeds, besides loss of seed performance. In contrast, slow-imbibed seeds showed high germination, low abnormal seedlings and maintenance of seeds performance. DNA isolation from fast imbibed seeds was highly degraded; although, some considerably degraded samples were reported in controlled imbibed seeds and the best preserved DNA was found in slow-imbibed seeds. The pattern of DNA degradation is typically passive or non programmed cell death. Our results showed it is important to consider the water uptake rate during germination test, since DNA integrity plays a critical role during seed imbibition, preserving soybean seed performance.(AU)

As relações entre a taxa de embebição em água e temperaturas foram estudadas em cinco lotes de sementes de soja cv. M 6410 IPRO. Testes de germinação, emergência a campo e condutividade elétrica foram aplicados para avaliar o desempenho das sementes após rápida embebição (imersão direta em água), controlada (substrato úmido) e lenta (atmosfera úmida), ambos a 20 °C e 30 °C. O incremento no peso fresco das sementes durante a embebição foi registrado. Paralelamente, a integridade do DNA de plântulas oriundas das sementes foi avaliado em gel por eletroforese. O padrão de embebição das sementes de soja foi similar entre as temperaturas, mas houve variações com o tempo de acordo com o modo de entrada de água nas sementes. Após a embebição rápida, as sementes apresentam menor germinação e desempenho que as sementes submetidas aos demais métodos de embebição. O DNA isolado a partir de sementes que embeberam rapidamente encontrava-se degradado, embora em sementes que foram hidratadas de forma controlada também fosse encontrada degradação do DNA. O DNA mais preservado foi encontrado em sementes que embebem lentamente. O padrão de degradação do DNA é típico de morte passiva ou não programada de células. Os resultados indicam que é importante considerar a taxa de embebição em água durante o teste de germinação, pois a integridade do DNA desempenha uma função importante durante a embebição, preservando o desempenho das sementes de soja.(AU)

Free-living capybaras (Hydrochoerus hydrochaeris) in an urban area in Brazil: biochemical and hematological parameters

Capybaras have found favorable conditions for survival and reproduction in green urban environments. In recent years, the population of these large rodents has been increasingly abundant in several brazilian cities such as Uberlândia, a municipality of the southeastern region with a Cerrado biome. Capybaras are important in the Brazilian Spotted Fever epidemiological chain, by amplifying infection rates of the vector population. However, knowledge of this host's physiology is scarce. Thus, the aim of this work was to describe hematological and biochemical parameters of free-living capybaras groups in urbanized areas in the city of Uberlândia, Minas Gerais State, Brazil. Capybaras were captured in 4 different locations of Uberlândia city, Minas Gerais state, including 1 Condominium (P1), 1 Private Market Garden (P2), 1 Private Club (P3) and 1 Municipal Park (P4). The animals were baited into an octagonal iron corral and chemically contained with anesthetic darts. After sedated, blood was collected from the femoral vein in tubes with and without EDTA. Biochemical evaluation, hematological analysis with differential leukocyte counts and search for Dirofilaria sp. were done. The blood count and biochemistry values obtained from animals of different ages, sex and sectors (P1, P2, P3 and P4) were submitted to the Shapiro-Wilk normality test, considering 95% significance. Values that had a normal distribution were subjected to ANOVA tests followed by Student's t-test. Values that did not follow normality were submitted to the Kruskal-Wallis test, to obtain a P-value, with a significance level of 95%. A total of 19 capybaras were captured: 4 in P1, 6 in P2, 4 in P3 and 5 in P4. From the 19 animals, 13 were females (68.42%) and 6 were males (31.57%), 12 adults (63.15%) and 7 juveniles (36.84%). Apart from occasional skin scars and moderate to intense Ambyomma spp. tick infestations, all captured animals were healthy on a broad examination. From 5 animals captured in P4, despite the use of anticoagulant, blood from 4 animals clotted fast. No microfilariae were found in the thick drop test in any of the 19 animals sampled, and in 2 adult female capybaras captured in P1, Kurloff cells were observed. Hematological and biochemical values presented no major differences when comparing sex and age. Nevertheless, differences in liver and kidney profile were observed between the capybara groups, including ALT, alkaline phosphorus, BUN and creatinine. Blood from 4 animals clotted fast, despite the use of EDTA tubes. Blood clotting of samples with anticoagulant in this work could be associated with some physiological features inherent to capybaras. Many attempts were required to obtain enough blood from each individual due to the rapid hemostasis, what come in accordance with reports in literature. Kurloff cells were observed in 2 adult female capybaras captured in P1, which can be found in peripheral blood of female rodents during follicular phase of estrous cycle. Hematological and biochemical values differences in liver enzymes such as ALT and alkaline phosphorus, and kidney profile enzymes including BUN and creatinine could be associated to capture stress or dietetic variation between groups. Despite statistical relevant, the values were still in accordance with other works, although comparisons should be done with caution since various environments exert a diverse array of stimulus upon the animals such as parasitic, infective, stress, nutritional, social and undoubtedly blood parameters mirror them. In conclusion, this work contributes to the standardization of free-living capybaras' physiological parameters in urban areas.(AU)

Detection of Mycobacterium sp. by multiplex PCR directly from suspicious granulomas from cold chambers in the state of Bahia, Brazil

The diagnosis of bovine tuberculosis (TB) by molecular techniques has been broadly studied. These methods allow accelerating the diagnosis, in addition to presenting high specificity and sensitivity in the identification of the pathogen, critical characteristic for public health, especially when it comes to the direct diagnosis of the biologic samples, which has been little explored. This paper has evaluated a multiplex polymerase chain reaction (mPCR) as a tool to diagnose TB, which was performed directly on the granulomatous material of suspicious lesions collected in a cold chamber under state inspection in the state of Bahia, Brazil. Of the 74 samples evaluated, 14.86% were positive, with 10.81% positive for mPCR and culture, 4.05% negative for cultivation and positive for mPCR. The correlation between the cultivation and the mPCR presented agreeance higher than 61.54% of the cases. The results have indicated that the protocol proved itself effective, fast and very promising in the surveillance in slaughterhouses for the diagnosis of tuberculosis directly from the granuloma.(AU)

Anthelmintic resistance of Haemonchus contortus from sheep flocks in Brazil: concordance of in vivo and in vitro (RESISTA-Test©) methods / Resistência anti-helmíntica de Haemonchus contortus em rebanhos ovinos noBrasil: concordância de métodos in vivo e in vitro (RESISTA-Test©)

This study evaluated the resistance status of Haemonchus contortus from sheep flocks in the state of São Paulo, Brazil, through comparison between the fecal egg count reduction test (FECRT) and the larval development test (LDT). For the FECRT, 35 sheep were selected in each of five flocks and divided into groups treated with: benzimidazole, levamisole, ivermectin, monepantel and control. Feces were collected for EPG and fecal cultures. The LDT was performed using thiabendazole (TBZ), levamisole (LEV), ivermectin aglycone (IVM-A) and Zolvix (ZLV). Resistance to all drugs was detected using FECRT in 100% of the flocks, except in relation to ZLV (40% resistant and 20% suspected of resistance). LDT indicated resistance to TBZ and IVM-A in all flocks, to LEV in 80% of flocks and to ZLV in 10%. Total agreement was obtained between the two tests for TBZ and IVM (k = 1.0), while for LEV (k = 0.8) and ZLV (k = 0.9), substantial and almost perfect agreement were obtained, respectively. The concordance between the tests was significant, thus showing that it is possible to use the outcome of the LDT to predict the FECRT, and hence validating the former as a fast diagnostic test for use by sheep farmers in Brazil.(AU)

Este estudo avaliou o status de resistência de Haemonchus contortus em rebanhos ovinos do estado de São Paulo, Brasil, através de um estudo comparativo entre o teste de redução da contagem de ovos nas fezes (TRCOF) e o teste de desenvolvimento larvar (TDL). Para o TRCOF, 35 ovinos foram selecionados em cada um dos cinco rebanhos e divididos em grupos tratados com: benzimidazol, levamisol, ivermectina, monepantel e controle. Fezes foram coletadas para OPG e coproculturas. O TDL foi realizado com tiabendazol-TBZ, levamisol-LEV, ivermectina aglicona-IVM-A e Zolvix-ZLV. Resistência a todos os fármacos foi detectada por meio do TRCOF em 100% dos rebanhos, exceto para ZLV (40% resistentes e 20% suspeitos de resistência). O LDT indicou resistência ao TBZ e IVM-A em todos os rebanhos; em 80% dos rebanhos ao LEV e 10% ao ZLV. Foi obtida concordância total entre os dois testes para TBZ e IVM (k = 1,0), enquanto para LEV (k = 0,8) e ZLV (k = 0,9) foram obtidas concordância substancial e quase perfeita, respectivamente. A concordância entre os testes foi significante, sendo possível usar o resultado do LDT para prever o do TRCOF, validando-se o primeiro como um teste diagnóstico rápido para criadores de ovinos no Brasil.(AU)

Central nervous system infiltration in acute lymphoblastic leukemia in a dog

Background: Acute lymphoblastic leukemia (ALL) is a malignant neoplasia in which there is proliferation of lymphoid progenitor cells in the bone marrow, blood, and extramedullary sites. This disorder has a fast and progressive development; in dogs, cases of infiltration of ALL cells in the central nervous system (CNS) are uncommon and rare. Diagnosis can be achieved with the help of the clinical history and physical, radiographic, hematological, myelographic, and cerebrospinal fluid (CSF) tests in patients with or without neurological clinical signs. The present report aims to describe a case of ALL and the presence of lymphoblasts in the CSF of a dog with neurological clinical signs. Case: An 8-year-old Lhasa Apso dog was examined at the Veterinary Hospital of Universidade Federal do Paraná, Curitiba campus. At the physical examination, the animal exhibited apathy and paralysis of pelvic limbs, which progressed to tetraplegia. Abdominal palpation revealed presence of hepatosplenomegaly and absence of lymphadenomegaly. No alterations were observed in radiographs of the cervical, thoracic or lumbar spine. A complete blood count revealed presence of non-regenerative anemia (hematocrit = 22%), extreme lymphocytosis (185,229 cells/µL), lymphoblasts at a level of 72% (133,364 cells/µL), and thrombocytopenia (66,000 platelets/µL). The biochemical tests revealed increased alkaline phosphatase (859 IU/L). The levels of alanine aminotransferase, creatinine, urea, total protein, albumin, and globulin were normal. The diagnosis of ALL was achieved with the help of a myelogram. The myelogram findings included 39% of mature lymphocytes and 59% of lymphoblasts exhibiting large size, spherical shape, poorly delimited borders, with a high nucleus/cytoplasm ratio, marked cytoplasmic basophilia, and 2 to 3 evident nucleoli; metarubricytes (1%) and promyelocytes (0.6%) were also observed. The CSF contained an increased number of nucleated cells (27 cells/µL) comprising lymphocytes (43%), macrophages (33%), and segmented neutrophils (24%). Of the 11.6 lymphocytes per µL of CSF, 8.1 were lymphoblasts, which indicates infiltration of ALL cells in the CNS. The animal died one day after collection of bone barrow and CSF. Discussion: Relevant alterations observed in this case included the neurological signs caused by the infiltration of neoplastic cells in the CNS, severe leukocytosis and lymphocytosis, with large amounts of lymphoblasts in the blood and predominance of lymphoblasts in the bone marrow, which are alterations typically found in ALL. The animal also exhibited non-regenerative anemia and thrombocytopenia, which were secondary to infiltration of leukemic cells in the bone marrow. The CSF exhibited pleocytosis (27 cells/ µL), and 30% of the cells observed were lymphoblasts. Lymphoblast infiltration in the CNS of leukemic dogs is rare, and other studies have reported absence of neurological signs or neurological signs different from those observed in the present study. CSF analysis in indicated in cases of leukemia to assess leukemic cell infiltration in the CNS. In the case reported here, the plasma level of alkaline phosphatase was increased (859 IU/L) as a consequence of hepatomegaly and hepatic cholestasis. ALL is a very aggressive, proliferative neoplasia, and the resulting lymphoblasts infiltrated the CNS of the animal. In cases of ALL, performing complete blood count, myelogram, and CSF analysis is indicated whether the patients exhibit neurological signs or not.(AU)

Potenciais usos do modelo animal Zebrafish Danio rerio em pesquisas na Medicina Veterinária / Potential uses of the animal model Zebrafish Danio rerio in Veterinary Medicine researches

Zebrafish (Danio rerio) é um proeminente organismo utilizado como modelo para a avaliação da segurança e eficácia de novos compostos na saúde animal de maneira rápida e econômica. Considerando que, anualmente, mais de mil novas substâncias são introduzidas no mercado para o desenvolvimento de produtos como vacinas, medicamentos, aditivos alimentares e agroquímicos, o objetivo deste artigo é apresentar uma revisão do emprego do Zebrafish como modelo animal destinado a avaliação da eficácia e da biotoxidade de compostos em pesquisas em Medicina Veterinária. Diversos estudos confirmam que os perfis entre Zebrafish e mamíferos são surpreendentemente semelhantes, sendo que sua transparência, fertilização externa, pequeno porte e curto ciclo de vida permitem a avaliação direta in vivo e em tempo real dos (i) efeitos de compostos químicos no desenvolvimento animal, (ii) eficácia de novas drogas no tratamento de doenças específicas, (iii) eficácia e segurança no desenvolvimento de vacinas contra doenças infecciosas, (iv) direcionamento de tratamento do câncer animal de forma direcionada e específica. Desta forma, a praticidade e eficiência deste modelo animal nas pesquisas pode acelerar o processo de desenvolvimento de novos compostos veterinários quando comparado com pesquisas realizadas em outros modelos animais, e com maior valor preditivo e informativo que pesquisas conduzidas com ensaios in vitro.

Zebrafish (Danio rerio) is an excellent model body to evaluate the safety and efficacy of new chemicals in animal health in a fast and economical way. Considering that more than 1,000 new chemicals are marketed annually for the development of products such as vaccines, drugs, food additives and agrochemicals, the purpose of this review is to present the Zebrafish as a model for assessing the efficacy and biotoxicity of compounds in the veterinary scope. Numerous studies have confirmed that profiles between Zebrafish and mammals are surprisingly similar, and their transparency, external fertilization, small size and short life cycle allow direct in vivo and real-time evaluation of (i) effects of chemical compounds in the development of animals, (ii) efficacy of new drugs in the treatment of specific diseases, (iii) efficacy and safety in the development of vaccines against infectious diseases, (iv) directing the treatment of animal cancer in a more targeted and specific way. Therefore, thepracticality and efficiency of its use in research can accelerate the process of developing new veterinaryand agricultural compounds with less expenses when compared to tests carried out in other animalmodels and with higher predictive and informative value than in vitro tests.

Potenciais usos do modelo animal Zebrafish Danio rerio em pesquisas na Medicina Veterinária / Potential uses of the animal model Zebrafish Danio rerio in Veterinary Medicine researches

Zebrafish (Danio rerio) é um proeminente organismo utilizado como modelo para a avaliação da segurança e eficácia de novos compostos na saúde animal de maneira rápida e econômica. Considerando que, anualmente, mais de mil novas substâncias são introduzidas no mercado para o desenvolvimento de produtos como vacinas, medicamentos, aditivos alimentares e agroquímicos, o objetivo deste artigo é apresentar uma revisão do emprego do Zebrafish como modelo animal destinado a avaliação da eficácia e da biotoxidade de compostos em pesquisas em Medicina Veterinária. Diversos estudos confirmam que os perfis entre Zebrafish e mamíferos são surpreendentemente semelhantes, sendo que sua transparência, fertilização externa, pequeno porte e curto ciclo de vida permitem a avaliação direta in vivo e em tempo real dos (i) efeitos de compostos químicos no desenvolvimento animal, (ii) eficácia de novas drogas no tratamento de doenças específicas, (iii) eficácia e segurança no desenvolvimento de vacinas contra doenças infecciosas, (iv) direcionamento de tratamento do câncer animal de forma direcionada e específica. Desta forma, a praticidade e eficiência deste modelo animal nas pesquisas pode acelerar o processo de desenvolvimento de novos compostos veterinários quando comparado com pesquisas realizadas em outros modelos animais, e com maior valor preditivo e informativo que pesquisas conduzidas com ensaios in vitro.(AU)

Zebrafish (Danio rerio) is an excellent model body to evaluate the safety and efficacy of new chemicals in animal health in a fast and economical way. Considering that more than 1,000 new chemicals are marketed annually for the development of products such as vaccines, drugs, food additives and agrochemicals, the purpose of this review is to present the Zebrafish as a model for assessing the efficacy and biotoxicity of compounds in the veterinary scope. Numerous studies have confirmed that profiles between Zebrafish and mammals are surprisingly similar, and their transparency, external fertilization, small size and short life cycle allow direct in vivo and real-time evaluation of (i) effects of chemical compounds in the development of animals, (ii) efficacy of new drugs in the treatment of specific diseases, (iii) efficacy and safety in the development of vaccines against infectious diseases, (iv) directing the treatment of animal cancer in a more targeted and specific way. Therefore, thepracticality and efficiency of its use in research can accelerate the process of developing new veterinaryand agricultural compounds with less expenses when compared to tests carried out in other animalmodels and with higher predictive and informative value than in vitro tests.(AU)

Clomipramina no tratamento da perseguição compulsiva da cauda em Bull Terrier - relato de caso / Clomipramine in the treatment of compulsive tail pursuit in bull terrier - case report

A perseguição compulsiva da cauda consiste em um distúrbio caracterizado por movimentos giratórios em círculos, lentos a rápidos com ou sem foco na cauda. É uma desordem comportamental comumente observada em cães da raça Bull Terrier, sendo mais frequente em machos, com início entre três e seis meses de idade. O diagnóstico é realizado com base no histórico do paciente junto com exames físico, neurológico e laboratoriais. O tratamento pode ser feito por meio do manejo ambiental e da retirada de fatores estressantes aliados a medicamentos antidepressivos. O objetivo deste artigo é relatar o caso de um Bull Terrier que apresentava perseguição compulsiva da cauda e teve evidente melhora clínica 75 dias após início do tratamento com clomipramina. A melhora foi ainda maior após a orquiectomia. A clomipramina consiste em um antidepressivo tricíclico promissor no tratamento de perseguição compulsiva da cauda em cães, devendo-se associar alteração no manejo ambiental e evitar gatilhos que induzam frustração, ansiedade e conflito.(AU)

The compulsive pursuit of the tail is characterized by a rotating movement in circles, slow to fast with or without focus on the tail. It is a disorder commonly observed in bull terrier dogs, being more frequent in males, beginning between 3 and 6 months of age. Diagnosis can be made based on patient history along with physical, neurological, and laboratory tests. Treatment can be done through environmental management and withdrawal of stressors allied with antidepressant drugs. The objective of this article is to report the case of a Bull Terrier that had compulsive pursuit of the tail and had evident clinical improvement 75 days after starting treatment with clomipramine. The improvement was even greater after orchiectomy. Clomipramine is the promising tricyclic antidepressant in the treatment of compulsive tail chasing in dogs. It should be associated with changes in environmental management and avoid triggers that induce frustration, anxiety and conflict.(AU)

Clomipramina no tratamento da perseguição compulsiva da cauda em Bull Terrier - relato de caso / Clomipramine in the treatment of compulsive tail pursuit in bull terrier - case report

A perseguição compulsiva da cauda consiste em um distúrbio caracterizado por movimentos giratórios em círculos, lentos a rápidos com ou sem foco na cauda. É uma desordem comportamental comumente observada em cães da raça Bull Terrier, sendo mais frequente em machos, com início entre três e seis meses de idade. O diagnóstico é realizado com base no histórico do paciente junto com exames físico, neurológico e laboratoriais. O tratamento pode ser feito por meio do manejo ambiental e da retirada de fatores estressantes aliados a medicamentos antidepressivos. O objetivo deste artigo é relatar o caso de um Bull Terrier que apresentava perseguição compulsiva da cauda e teve evidente melhora clínica 75 dias após início do tratamento com clomipramina. A melhora foi ainda maior após a orquiectomia. A clomipramina consiste em um antidepressivo tricíclico promissor no tratamento de perseguição compulsiva da cauda em cães, devendo-se associar alteração no manejo ambiental e evitar gatilhos que induzam frustração, ansiedade e conflito.(AU)

The compulsive pursuit of the tail is characterized by a rotating movement in circles, slow to fast with or without focus on the tail. It is a disorder commonly observed in bull terrier dogs, being more frequent in males, beginning between 3 and 6 months of age. Diagnosis can be made based on patient history along with physical, neurological, and laboratory tests. Treatment can be done through environmental management and withdrawal of stressors allied with antidepressant drugs. The objective of this article is to report the case of a Bull Terrier that had compulsive pursuit of the tail and had evident clinical improvement 75 days after starting treatment with clomipramine. The improvement was even greater after orchiectomy. Clomipramine is the promising tricyclic antidepressant in the treatment of compulsive tail chasing in dogs. It should be associated with changes in environmental management and avoid triggers that induce frustration, anxiety and conflict.(AU)

A review of simulation analyses of economics and genetics for the use of in-vitro produced embryos and artificial insemination in dairy herds

The use of in-vitro produced (IVP) embryo transfer (ET) in dairy herds is growing fast. Much of this growth is on dairy farms where the focus is on milk production and not on selling breeding stock. The value of implementing IVP-ET in a dairy herd arises from a higher genetic merit of the IVP-embryo, but the cost to produce a pregnancy with an IVP embryo is greater than the cost of artificial insemination (AI). The first objective of this study was to review estimates of the net benefit of using IVP-ET over AI in dairy herds using existing literature. Another objective was to show how much IVP-ET use in a herd is optimal. Most of the literature is based on simulation modeling, including our own work that focuses on the dairy industry in the USA. We found that the most profitable use of AI and IVP-ET is often a combination of the two. More IVP-ET should be used when the value of surplus calves is greater and the cost of IVP-ET is lower, among many other factors. In the future, use of IVP-ET will be further improved by more accurately identifying superior donors and recipients, reducing the generation interval, and achieving greater efficiency in embryo production.(AU)

A review of simulation analyses of economics and genetics for the use of in-vitro produced embryos and artificial insemination in dairy herds

The use of in-vitro produced (IVP) embryo transfer (ET) in dairy herds is growing fast. Much of this growth is on dairy farms where the focus is on milk production and not on selling breeding stock. The value of implementing IVP-ET in a dairy herd arises from a higher genetic merit of the IVP-embryo, but the cost to produce a pregnancy with an IVP embryo is greater than the cost of artificial insemination (AI). The first objective of this study was to review estimates of the net benefit of using IVP-ET over AI in dairy herds using existing literature. Another objective was to show how much IVP-ET use in a herd is optimal. Most of the literature is based on simulation modeling, including our own work that focuses on the dairy industry in the USA. We found that the most profitable use of AI and IVP-ET is often a combination of the two. More IVP-ET should be used when the value of surplus calves is greater and the cost of IVP-ET is lower, among many other factors. In the future, use of IVP-ET will be further improved by more accurately identifying superior donors and recipients, reducing the generation interval, and achieving greater efficiency in embryo production.

Bovine tuberculosis: diagnosis in dairy cattle through the association of analyzes / Tuberculose bovina: diagnóstico em bovinos leiteiros através da associação de análises

Tuberculosis is a chronic anthropozoonosis of worldwide occurrence, caused by the bacterium Mycobacterium tuberculosis and its variants. In Brazil, the National Program for the Control and Eradication of Brucellosis and Tuberculosis in cattle, is responsible for diagnosing and the correctly allocate positive animals, but there is still a lack of definitive diagnosis of the disease. This study described the use of five diagnostic tools that can be used, preferably together, for the confirmation of suspected cases. These tools included the clinical examination comparative cervical tuberculin test, macroscopic findings during the slaughtering and histopathology of the damaged tissues followed by histochemistry. We evaluated a total of 211 dairy cattle, where 15.1% (32/211) had classic clinical signs of bovine tuberculosis, 74 (35%) showed reactivity in the comparative cervical tuberculin test. Of the total number of animals, 141 (66.8%) were referred for sanitary slaughter due to legal and control issues in the outbreaks of the disease. In the follow-up of slaughtering and inspection of viscera and carcasses, 74 (52.5%) had macroscopic lesions compatible with bovine tuberculosis, while 67 (47.5%) showed no visible changes. During the inspection, fragments of lymph nodes and liver and lung parenchyma were collected from five cattle with macroscopic lesions and five with no lesions. The histopathological analysis showed numerous areas of caseous necrosis with or without central calcification and granulomatous inflammatory infiltrate. In the special staining of Ziehl-Neelsen, numerous acid-fast bacilli were evidenced in all cases.(AU)

A tuberculose é uma antropozoonose crônica de ocorrência mundial, causada pela bactéria Mycobacterium tuberculosis e suas variantes. No Brasil existe o Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose em bovinos que viabiliza o diagnóstico e a destinação correta dos animais positivos, porém ainda há carência quanto ao diagnóstico da doença. Assim, este trabalho descreve a utilização de cinco ferramentas diagnósticas para a confirmação de casos suspeitos de tuberculose. As ferramentas utilizadas compreenderam o exame clínico, teste tuberculínico cervical comparativo, os achados macroscópicos durante o abate sanitário e a histopatologia dos tecidos lesados seguido de histoquímica. O estudo avaliou um total de 211 bovinos leiteiros, dos quais 15,1% (32/211) apresentaram sinais clínicos clássicos de tuberculose bovina, 35,1% (74/211) apresentaram reatividade no teste tuberculínico cervical comparativo, e 143 animais (67,8%) foram encaminhados para abate sanitário devido a questões legais e de controle nos focos da doença. No acompanhamento do abate e inspeção sanitária de vísceras e carcaças verificou-se que 51,8% (74/143) dos bovinos abatidos apresentavam lesões macroscópicas compatíveis com tuberculose bovina, enquanto 48,2% (69/143) não apresentavam alterações visíveis. Durante a inspeção foram coletados fragmentos de linfonodos e parênquima de fígado e pulmão de cinco bovinos com lesões macroscópicas e de cinco sem lesões, que na análise histopatológica apresentaram numerosas áreas de necrose caseosa com ou sem calcificação central e infiltrado inflamatório granulomatoso. Na coloração de Ziehl-Neelsen foram evidenciados numerosos bacilos álcool-ácido resistentes em todos os casos. Assim, diante dos resultados obtidos verifica-se que as análises empregadas no presente estudo foram de extrema importância para o diagnóstico acurado de tuberculose em bovinos.(AU)

Bovine tuberculosis: diagnosis in dairy cattle through the association of analyzes / Tuberculose bovina: diagnóstico em bovinos leiteiros através da associação de análises

Tuberculosis is a chronic anthropozoonosis of worldwide occurrence, caused by the bacterium Mycobacterium tuberculosis and its variants. In Brazil, the National Program for the Control and Eradication of Brucellosis and Tuberculosis in cattle, is responsible for diagnosing and the correctly allocate positive animals, but there is still a lack of definitive diagnosis of the disease. This study described the use of five diagnostic tools that can be used, preferably together, for the confirmation of suspected cases. These tools included the clinical examination comparative cervical tuberculin test, macroscopic findings during the slaughtering and histopathology of the damaged tissues followed by histochemistry. We evaluated a total of 211 dairy cattle, where 15.1% (32/211) had classic clinical signs of bovine tuberculosis, 74 (35%) showed reactivity in the comparative cervical tuberculin test. Of the total number of animals, 141 (66.8%) were referred for sanitary slaughter due to legal and control issues in the outbreaks of the disease. In the follow-up of slaughtering and inspection of viscera and carcasses, 74 (52.5%) had macroscopic lesions compatible with bovine tuberculosis, while 67 (47.5%) showed no visible changes. During the inspection, fragments of lymph nodes and liver and lung parenchyma were collected from five cattle with macroscopic lesions and five with no lesions. The histopathological analysis showed numerous areas of caseous necrosis with or without central calcification and granulomatous inflammatory infiltrate. In the special staining of Ziehl-Neelsen, numerous acid-fast bacilli were evidenced in all cases.(AU)

A tuberculose é uma antropozoonose crônica de ocorrência mundial, causada pela bactéria Mycobacterium tuberculosis e suas variantes. No Brasil existe o Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose em bovinos que viabiliza o diagnóstico e a destinação correta dos animais positivos, porém ainda há carência quanto ao diagnóstico da doença. Assim, este trabalho descreve a utilização de cinco ferramentas diagnósticas para a confirmação de casos suspeitos de tuberculose. As ferramentas utilizadas compreenderam o exame clínico, teste tuberculínico cervical comparativo, os achados macroscópicos durante o abate sanitário e a histopatologia dos tecidos lesados seguido de histoquímica. O estudo avaliou um total de 211 bovinos leiteiros, dos quais 15,1% (32/211) apresentaram sinais clínicos clássicos de tuberculose bovina, 35,1% (74/211) apresentaram reatividade no teste tuberculínico cervical comparativo, e 143 animais (67,8%) foram encaminhados para abate sanitário devido a questões legais e de controle nos focos da doença. No acompanhamento do abate e inspeção sanitária de vísceras e carcaças verificou-se que 51,8% (74/143) dos bovinos abatidos apresentavam lesões macroscópicas compatíveis com tuberculose bovina, enquanto 48,2% (69/143) não apresentavam alterações visíveis. Durante a inspeção foram coletados fragmentos de linfonodos e parênquima de fígado e pulmão de cinco bovinos com lesões macroscópicas e de cinco sem lesões, que na análise histopatológica apresentaram numerosas áreas de necrose caseosa com ou sem calcificação central e infiltrado inflamatório granulomatoso. Na coloração de Ziehl-Neelsen foram evidenciados numerosos bacilos álcool-ácido resistentes em todos os casos. Assim, diante dos resultados obtidos verifica-se que as análises empregadas no presente estudo foram de extrema importância para o diagnóstico acurado de tuberculose em bovinos.(AU)

Seed vigor of Piptadenia moniliformis Benth. subjected to thermal stress

The objective of this work is to evaluate the influence of accelerated aging test at 41ºC on the germination and seed vigor of Piptadenia moniliformisBenth. Two experiments were conducted. Experiment 1: seeds were submitted to dormancy overcoming treatments. They were immersed into sulfuric acid for 0 (intact seeds), 5, 10, 15, 20 and 25 minutes. Experiment 2: two methods were evaluated for the accelerated aging test at 41ºC, that is, the traditional method and the saturated NaCl solution method with exposure periods of 0, 24, 48, 72 and 96 hours. After each treatment, the seeds were subjected to electrical conductivity and germination tests. Seeds overcame their dormancy when immersed into concentrated sulfuric acid for 5, 10, 15, 20 or 25 minutes, but thegermination was faster when immersed for 20 minutes. The different methods for the accelerated aging test, decreased the germination potential and seed vigor of P. moniliformisafter 24 hours of exposure. Regarding the method used, the saturated solution provided, in general, the best results since the seed water content was lower, thus reducing the proliferation of fungi. The electrical conductivity test showed that, by increasing the aging period regardless of the method, there is an increase in seed deterioration, corroborating with germination results, which showed that the correlation was moderate and negative and indicated that the higher the conductivity, the lower the percentage and the germination speed index. The immersion of seeds of P. moniliformisinto concentrated sulfuric acid for 20 minutes provides a fast and increased germination. The methods for accelerated agingat 41ºC, traditional and saturated solution, can be used as vigor tests to evaluate the physiological quality of seeds.

Imagens digitalizadas na avaliação do teste de tetrazólio em sementes de trigo / Digitized images in the evaluation of the wheat seeds tetrazolium test

O teste de tetrazólio com avaliação através de imagens digitalizadas pode ser uma alternativa para obtenção de resultados rápidos do potencial fisiológico de sementes. Nesse contexto, o objetivo do presente estudo foi avaliar a eficiência do teste de tetrazólio através da avaliação por análise de imagens digitalizadas na determinação da qualidade fisiológica de sementes de trigo. O experimenteo foi realizado com 22 lotes de sementes de trigo, submetidos à determinação do potencial fisiológico, incluindo o teste de tetrazólio avaliado por meio de análise de imagens digitalizadas. Para digitalização, após bissecção longitudinal, e coloração em solução de 0,075% de tetrazólio, as sementes foram agrupadas sobre vidro de scanner de mesa, armazenadas e classificadas em quatro classes de vigor. Os dados foram submetidos à análise de variância, com médias agrupadas pelo teste Scott-Knott, a 5% de significância; também foi realizada análise dos coeficientes de correlação simples entre os resultados dos testes de tetrazólio e demais testes. A avaliação do teste de tetrazólio através da análise de imagens digitalizadas agrupou os lotes em três níveis de vigor. O método é eficiente na determinação de vigor e viabilidade de sementes de trigo, devido a possibilidade de ampliação das imagens que permite análise precisa das estruturas do eixo embrionário.

The tetrazolium test can be an alternative to obtain fast results of the physiological potential of wheat seeds. In this context, the objective of the present study was to evaluate the efficiency of the tetrazolium test through the evaluation by analysis of digitized images in the determination of the physiological quality of wheat seeds. The experiment was carried out with 22 lots of wheat seeds, submitted to the determination of the physiological potential, including the tetrazolium test evaluated by means of digitized image analysis. For digitization, after longitudinal bisection, and staining in 0.075% tetrazolium solution, the seeds were grouped on tabletop scanner glass, stored and classified into four vigor classes. Data were submitted to analysis of variance, with means grouped by the Scott-Knott test, at 5% of significance; we also performed an analysis of the simple correlation coefficients between tetrazolium test results and other tests. The evaluation of the tetrazolium test through the analysis of digitized images grouped the lots in three levels of vigor. The method is effective in determining the vigor and viability of wheat seeds, because the possibility of expanding the images that allows precise analysis of the embryonic axis structures.

Imagens digitalizadas na avaliação do teste de tetrazólio em sementes de trigo / Digitized images in the evaluation of the wheat seeds tetrazolium test

O teste de tetrazólio com avaliação através de imagens digitalizadas pode ser uma alternativa para obtenção de resultados rápidos do potencial fisiológico de sementes. Nesse contexto, o objetivo do presente estudo foi avaliar a eficiência do teste de tetrazólio através da avaliação por análise de imagens digitalizadas na determinação da qualidade fisiológica de sementes de trigo. O experimenteo foi realizado com 22 lotes de sementes de trigo, submetidos à determinação do potencial fisiológico, incluindo o teste de tetrazólio avaliado por meio de análise de imagens digitalizadas. Para digitalização, após bissecção longitudinal, e coloração em solução de 0,075% de tetrazólio, as sementes foram agrupadas sobre vidro de scanner de mesa, armazenadas e classificadas em quatro classes de vigor. Os dados foram submetidos à análise de variância, com médias agrupadas pelo teste Scott-Knott, a 5% de significância; também foi realizada análise dos coeficientes de correlação simples entre os resultados dos testes de tetrazólio e demais testes. A avaliação do teste de tetrazólio através da análise de imagens digitalizadas agrupou os lotes em três níveis de vigor. O método é eficiente na determinação de vigor e viabilidade de sementes de trigo, devido a possibilidade de ampliação das imagens que permite análise precisa das estruturas do eixo embrionário.(AU)

The tetrazolium test can be an alternative to obtain fast results of the physiological potential of wheat seeds. In this context, the objective of the present study was to evaluate the efficiency of the tetrazolium test through the evaluation by analysis of digitized images in the determination of the physiological quality of wheat seeds. The experiment was carried out with 22 lots of wheat seeds, submitted to the determination of the physiological potential, including the tetrazolium test evaluated by means of digitized image analysis. For digitization, after longitudinal bisection, and staining in 0.075% tetrazolium solution, the seeds were grouped on tabletop scanner glass, stored and classified into four vigor classes. Data were submitted to analysis of variance, with means grouped by the Scott-Knott test, at 5% of significance; we also performed an analysis of the simple correlation coefficients between tetrazolium test results and other tests. The evaluation of the tetrazolium test through the analysis of digitized images grouped the lots in three levels of vigor. The method is effective in determining the vigor and viability of wheat seeds, because the possibility of expanding the images that allows precise analysis of the embryonic axis structures.(AU)