Evidence and molecular characterization of Bartonella spp. and hemoplasmas in neotropical bats in Brazil.

The order Chiroptera is considered the second largest group of mammals in the world, hosting important zoonotic virus and bacteria. Bartonella and hemotropic mycoplasmas are bacteria that parasite different mammals' species, including humans, causing different clinical manifestations. The present work aimed investigating the occurrence and assessing the phylogenetic positioning of Bartonella spp. and Mycoplasma spp. in neotropical bats sampled from Brazil. Between December 2015 and April 2016, 325 blood and/or tissues samples were collected from 162 bats comprising 19 different species sampled in five states of Brazil. Out of 322 bat samples collected, while 17 (5·28%) were positive to quantitative PCR for Bartonella spp. based on nuoG gene, 45 samples (13·97%) were positive to cPCR assays for hemoplasmas based on 16S rRNA gene. While seven sequences were obtained for Bartonella (nuoG) (n = 3), gltA (n = 2), rpoB (n = 1), ftsZ (n = 1), five 16S rRNA sequences were obtained for hemoplasmas. In the phylogenetic analysis, the Bartonella sequences clustered with Bartonella genotypes detected in bats sampled in Latin America countries. All five hemoplasmas sequences clustered together as a monophyletic group by Maximum Likelihood and Bayesian Inference analyses. The present work showed the first evidence of circulation of Bartonella spp. and hemoplasmas among bats in Brazil.

Detection of Leptospira spp. in wild Phrynops geoffroanus (Geoffroy's side-necked turtle) in urban environment.

Leptospira spp., a zoonotic agent relevant for public health, occurs frequently in tropical regions. The aquatic environment represents a viable survival and transmission pathway. This study aimed to investigate the presence of anti-Leptospira spp. antibodies in Phrynops geoffroanus (Geoffroy's side-necked turtle) serum samples using the microagglutination test (MAT), and Leptospira spp. in gastric and cloacal lavage samples using the polymerase chain reaction (PCR) technique. Antibodies against nine different Leptospira spp. serovars were detected in 45.45% (30/66) of the serum samples. Specific amplification of Leptospira spp. genomic material (331bp) was observed in 16.67% (11/66) of the samples. In conclusion, these freshwater testudines host Leptospira spp. and eliminate them. This situation may represent a risk to public health, especially to people who use urban streams for fishing and recreational activities. Additionally, we described some Leptospira spp. serovars, not yet reported in testudines, detected here in P. geoffroanus.

Humeral fracture repair using a robust fixation in an adult Giant Anteater (Myrmecophaga tridactyla): case report / [Reparo de fratura umeral usando uma fixação robusta em um tamanduá-bandeira adulto (Myrmecophaga tridactyla): relato de caso]

The treatment of fractures from the thoracic limb in giant anteaters is extremely challenging. Unfamiliar and peculiar anatomical characteristics, robust musculature and the imminent need for an early return to limb function highlight such challenges. The objective of this report was to describe the successful use of anatomical osteosynthesis with a robust locking compression plate in a humeral fracture of an adult giant anteater. The patient was rescued on the highway after being run over and presented for treatment at the Veterinary Teaching Hospital. Surgical stabilization was performed using a craniomedial approach to the humerus, using a customized broad 3.5mm locking compression plate. The patient presented early limb support at 24 hours postoperatively. Radiographic monitoring was performed at 30, 60 and 90 days postoperatively, and bone healing was observed without any complications. It is concluded that the treatment of humerus fractures in giant anteaters requires robust fixation. The use of a reinforced locking compression plate system proved to be effective and adequate to the mechanical load that an adult individual of this species needs for early use of the thoracic limb and, at the same time, efficient in controlling interfragmentary movement, which allowed fracture consolidation.(AU)

O tratamento das fraturas do membro torácico dos tamanduás-bandeira (Myrmecophaga tridactyla) é extremamente desafiador. Características anatômicas pouco familiares e peculiares, musculatura muito desenvolvida e necessidade iminente de retorno precoce à função do membro destacam tais desafios. Objetivou-se, neste relato, descrever a utilização com sucesso da osteossíntese anatômica com placa bloqueada robusta em fratura umeral de um tamanduá-bandeira. O paciente foi resgatado em rodovia após atropelamento e atendido no Hospital Veterinário Universitário. Após sedação e avaliação física completa, foi realizado exame radiográfico, o qual revelou fratura completa oblíqua curta em diáfise média de úmero esquerdo. A estabilização cirúrgica foi realizada por abordagem craniomedial ao úmero, utilizando-se placa bloqueada (LCP) do sistema 3,5mm customizada. O paciente apresentou apoio precoce do membro com 24 horas de pós-operatório. Realizou-se acompanhamento radiográfico aos 30, 60 e 90 dias, sendo observada união clínica sem quaisquer complicações. Conclui-se que o tratamento das fraturas do úmero em tamanduás-bandeira exige robustez na fixação. A utilização de sistema reforçado de placa bloqueada mostrou-se efetiva e adequada à carga mecânica de que um indivíduo adulto dessa espécie necessita para uso precoce do membro torácico e, ao mesmo tempo, eficiente no controle da movimentação interfragmentária, o qual permitiu consolidação da fratura.(AU)

Ultrasound evaluation of female reproductive system in free ranging Giant Anteater (Myrmecophaga tridactyla): case report / Avaliação ultrassonográfica do sistema reprodutor feminino em tamanduá-bandeira de vida livre (Myrmecophaga tridactyla): relato de caso

An adult, female, 31kg body weight, free range Myrmecophaga tridactyla was referred for medical consultation due to apathy, dehydration, intense flatulence and fetid stools. The animal was submitted to chemical restraint and physical examination, blood count, fecal analysis, and vaginal cytology and abdominal ultrasonography were performed. Good nutritional status and clinical variables within the normal range were observed at physical examination. At vaginal cytology leukocytes, spermatozoa and a large proportion of cornified cells (superficial) were observed, indicating estrus and recent copula. At ultrasound examination it was possible to locate, identify, evaluate and measure the ovaries and the uterine structures, cervix, body, lumen, myometrium and endometrium, a fact never reported in the literature for this species. These data can be used as reference for clinical evaluation of the reproductive tract in Myrmecophaga tridactyla females considered vulnerable species, for the diagnosis of reproductive pathologies, biotechnologies application or estrous cycle evaluation.(AU)

Uma fêmea, com 31kg de peso corporal, de Myrmecophaga tridactyla, de vida livre, foi apresentada para consulta médica em razão de apatia, desidratação, flatulência intensa e fezes fétidas. O animal foi submetido à contenção química e foram realizados exame físico, hemograma, análise fecal, citologia vaginal e ultrassonografia abdominal. Ao exame físico, foi determinado bom estado nutricional e variáveis clínicas dentro da faixa de normalidade. Na citologia vaginal, foram observados leucócitos, espermatozoides e uma grande proporção de células cornificadas (superficiais), indicando estro e cópula recente. No exame ultrassonográfico, foi possível localizar, identificar, avaliar e mensurar os ovários e as estruturas uterinas, o colo do útero, o corpo do útero, o lúmen, o miométrio e o endométrio, fato nunca relatado na literatura para essa espécie. Esses dados podem ser utilizados como referência para avaliação clínica do trato reprodutivo de fêmeas de Myrmecophaga tridactyla considerada espécie vulnerável, para o diagnóstico de patologias reprodutivas, aplicação de biotecnologias ou avaliação do ciclo estral.(AU)

The use of the CELLection kit in the isolation of carcinoma cells from mononuclear cell suspensions.

A study was performed to evaluate in vitro the sensitivity, specificity and variability of a new immunomagnetic microbead isolation technique which provides subsequent immunological staining of captured carcinoma cells. In a mixture of peripheral blood mononuclear cells (PBMCs) and human carcinoma cells the epithelial cancer cells were isolated with the Dynal((R)) RAM IgG1 CELLection Kit using Dynabeads M-280 coated with a rat monoclonal antibody (Mab) against mouse IgG1. The rat Mab was biotinylated and attached to Dynabeads via streptavidin and a DNA linker. The anti-epithelial monoclonal mouse antibody Ber-EP4 was used as the primary capture antibody. In order to permit phenotyping of the isolated carcinoma cells the magnetic beads were removed from the carcinoma cells by DN'ase digestion of the DNA linker between the magnetic bead and the secondary antibody. In an ex vivo model system an average recovery of approximately 60% of a human colon carcinoma cell line HCC-2998 seeded in 5.10(6) PBMCs was obtained, and the recovered cells could subsequently be immunologically stained for the surface antigen CD87 (urokinase plasminogen activator receptor). No positive stained cells were found in control experiments with PBMCs without carcinoma cells. Despite a relatively low recovery, the described method will be valuable for the detection of carcinoma cells in cytospin preparations with subsequent phenotyping of the cells for expression of surface antigens. Depending on the chosen antibodies, the method may be useful for the isolation and characterisation of other cell types in various cell suspensions.

Immunoglobulin and enzyme-conjugated dextran polymers enhance u-PAR staining intensity of carcinoma cells in peripheral blood smears.

The presence of disseminated carcinoma cells in bone marrow and peripheral blood has prognostic importance in patients with carcinomas. Much evidence indicates that dissemination of tumor cells may depend on activation of a variety of degradative enzymes. A strong positive correlation has been shown between the expression of tumor cell proteases and tumor invasion. Therefore, phenotypic characterization of disseminated carcinoma cells for expression of protease activators might define the invasive potential of the cells. We present an immunocytochemically enhanced staining method that allows phenotyping of disseminated carcinoma cells in bone marrow and peripheral blood smears. In the first step, the cells were incubated with antibodies against urokinase plasminogen activator receptor (u-PAR) and subsequently with secondary antibodies conjugated to peroxidase-labeled dextran polymers. A brown color reaction was developed with diaminobenzidine as chromogen. In the second step, the cells were incubated with alkaline phosphatase-conjugated murine monoclonal antibodies against a common cytokeratin epitope and a red color reaction was developed with new fuchsin as substrate. This method allows simultaneous and unambiguous immunolabeling of intracellular cytokeratin and of u-PAR intracellularly and on the surface of carcinoma cells. This novel approach can be used for detection and phenotyping of carcinoma cells in blood smears for u-PAR or, presumably, for any other heterogeneously expressed antigen on the surface of the detected cells.

Soluble vascular endothelial growth factor levels in patients with primary colorectal carcinoma. The Danish RANX05 Colorectal Cancer Study Group.

INTRODUCTION: Angiogenesis is decisive in tumour progression and metastasis. Vascular endothelial growth factor (VEGF) is a potent angiogenic factor, and increased VEGF levels in patients with carcinomas may facilitate growth of both primary and secondary tumours. METHODS: Soluble (s) VEGF levels were determined in serum from 91 volunteer healthy blood donors and from 614 patients scheduled to undergo resection for primary colorectal cancer. None of the patients received pre- and/or post-operative chemo- and/or radiotherapy. The results of sVEGF were analysed with respect to Dukes>> stage, gender, age and topographical tumour localization. RESULTS: Patients with colorectal cancer had significantly (P<0.0001) higher levels of sVEGF, compared to healthy blood donors. Patients with Dukes>> stage D disease had significantly (P=0.01) higher values than patients with Dukes>> stage A, B and C disease, who had comparable values. Patients with the primary tumour localized in the colon had significantly (P<0.0001) higher levels of sVEGF than patients with the primary tumour localized in the rectum. By classifying the patients into two groups, based on the upper limit of the 95(th)percentile of sVEGF of healthy individuals (sVEGF=465 pg/ml), we found a significantly (P<0.0001) reduced overall survival in patients with sVEGF >465 pg/ml compared to patients with sVEGF values below this level. Moreover, in the subgroup of patients with the carcinoma localized in the colon and sVEGF levels above 465 pg/ml, we found a significantly (P<0.0001) reduced overall survival compared to colon cancer patients with lower sVEGF values. In conclusion, data from the present study suggest a biological significance of VEGF in patients with colorectal cancer, and indicate that a high pre-operative sVEGF value is associated with poor outcome, but further research is needed to validate sVEGF as a cancer marker.

Trypanosoma evansi experimental infection in the South American coati (Nasua nasua): clinical, parasitological and humoral immune response.

The course of Trypanosoma evansi infection in coatis (Carnivora, Procionidae) was followed for 262 days. Parasites were detected in all infected animals from day 2 post infection until the end of the study. No correlation between temperature and parasitemia was observed. Animals of the infected group demonstrated depression, weakness, lethargy and pale mucous membranes. Indirect fluorescent antibody tests detected anti-T. evansi antibodies within 7 to 14 days post infection and showed high levels until the end of the experimental period. The persistent parasitemia in coati and their relative tolerance to clinical signs suggested that this species develops a chronic disease and plays an important role in the epidemiology of trypanosomosis due to T. evansi in enzootic regions.

Arthroscopic shoulder surgery with epinephrine saline irrigation.

PURPOSE: To determine whether dilute epinephrine saline irrigation (0.33 mg/L) delivered by a pressure-controlled pump would significantly reduce intraoperative bleeding during routine arthroscopic shoulder surgery and, in addition, to investigate potential adverse cardiovascular reactions of adding epinephrine to the irrigation fluid. TYPE OF STUDY: Prospective, randomized, double-blinded, placebo-controlled study. METHODS: Fifty-four patients requiring arthroscopic shoulder surgery were randomly assigned to either an epinephrine group that received dilute epinephrine irrigation by a pressure-controlled pump or to a placebo group that received plain saline irrigation by a pressure-controlled pump. Intraoperative, intra-articular bleeding was estimated by multiplying the total volume of the irrigation fluid used with the hemoglobin concentration in the irrigation fluid. Postoperatively, the surgeon rated the clarity of the visual field during the arthroscopy by a visual analogy scale. RESULTS: Intraoperative bleeding was significantly reduced (P =.008) and the clarity of the visual field was significantly better (P =.0007) in the group of patients receiving dilute epinephrine irrigation compared with the group of patients without epinephrine added to the irrigation fluid. CONCLUSIONS: The addition of epinephrine to irrigation fluid seems to reduce intra-articular bleeding during routine arthroscopic shoulder surgery and may improve visualization. Furthermore, no cardiovascular adverse reactions were observed resulting from the intra-articular epinephrine administration.

Cat flea (Ctenocephalides felis) infestation in quarantined marsh deer (Blastocerus dichotomus) populations.

Marsh deer (Blastocerus dichotomus) were captured for a research program in Brazil and maintained in quarantine stations. After 60 days, fleas were detected on animals and identified as Ctenocephalides felis felis. Elimination of the infestation was difficult. Animal treatment with a fipronil-based compound was effective, and subsequently captured animals were treated prophylactically. Some animals remained infested, and some died from the infestation.

[Recurrent infantile digital fibromatosis]. / Recidiverende infantil digital fibromatose.

In reflection of a current case, the latest literature concerning recurring infantile digital fibromatosis (Reye's tumour) are summed up. It is a non-malignant, rarely encountered tumour observed in children, appearing on the hands and feet. The tumour is almost exclusively diagnosed before the age of three. The etiology is unknown. Approximately 100 cases have been reported in the literature. Eosinophilic inclusion bodies near the nucleus of the tumourcells are pathognomonic. According to the latest literature, the content of the inclusions is mostly actin filaments between which the formed elements of the cell are entrapped. Previous treatment was repeated excision of the recurring tumours, until spontaneous cessation of disease activity. According to the latest literature, a more conservative treatment is recommended, because the fibromas often regress spontaneously over few years. Therefore surgical treatment is only needed in rare cases with aggressive or uncontrollable growth of the fibromas.

[Osteochondritis dissecans of the acetabulum]. / Osteochondritis dissecans acetabuli.

A case of osteochondritis dissecans (OD) of the acetabulum in a 14-year-old girl is presented. The initial clinical manifestation was hip pain and restricted movements of the joint. Magnetic Resonance Imaging confirmed the diagnosis. The patient was treated conservatively, using crutches. One year after the first examination, no clinical improvements were found. X-rays of the hip showed increased sclerosis around the osteochondritis-segment, indicating incipient healing. The patient is still conservatively treated, with regular clinical examinations. The diagnosis should be remembered in the case of patients with unexplained hip-pain, and may require magnetic resonance imaging for confirmation.

[Non-traumatic subcutaneous emphysema of the femur. A rare complication of colorectal cancer]. / Ikketraumatisk subkutant emfysem på femur. En sjaelden komplikation til kolorektal cancer.

A case of subcutaneous emphysema of the left thigh as the initial symptoms of a perforated cancer of the descending colon is presented The literature on the subject is briefly reviewed. The resemblance to nontraumatic gas gangrene, and the importance of early therapy is discussed.

[Can detection of cancer cells in bone marrow and peripheral blood be used in diagnosis and patient monitoring?]. / Kan detektion af karcinomceller i knoglemarv og perifert blod benyttes til diagnostik og som recidivmarkør?

The prognosis of patients with carcinomas is determined by whether there is dissemination of the primary tumour at the time of diagnosis. A substantial number of patients develop recurrent carcinoma, even though there were no clinical signs of dissemination at the time of primary surgery. Standard methods of tumor staging fail to identify those patients, who are at risk of recurrent disease. Recently several studies have demonstrated that the presence of occult metastatic carcinoma cells in the bone marrow increases the risk of recurrence. This risk is independent of traditional tumour staging, although the findings do not necessarily indicate later development of solid metastases. This review outlines the range of current applications of immunological and molecular methods for detecting disseminated carcinoma cells in blood and bone marrow aspirations, and discusses the applicability and significance of the findings.

Genetic characterization of Toxoplasma gondii from Brazilian wildlife revealed abundant new genotypes.

This study aimed to isolate and genotype T. gondii from Brazilian wildlife. For this purpose, 226 samples were submitted to mice bioassay and screened by PCR based on 18S rRNA sequences. A total of 15 T. gondii isolates were obtained, including samples from four armadillos (three Dasypus novemcinctus, one Euphractus sexcinctus), three collared anteaters (Tamandua tetradactyla), three whited-lipped peccaries (Tayassu pecari), one spotted paca (Cuniculus paca), one oncilla (Leopardus tigrinus), one hoary fox (Pseudalopex vetulus), one lineated woodpecker (Dryocopus lineatus) and one maned wolf (Chrysocyon brachyurus). DNA from the isolates, originated from mice bioassay, and from the tissues of the wild animal, designated as "primary samples", were genotyped by PCR-restriction fragment length polymorphism (PCR/RFLP), using 12 genetic markers (SAG1, SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L258, PK1, CS3 and Apico). A total of 17 genotypes were identified, with 13 identified for the first time and four already reported in published literature. Results herein obtained corroborate previous studies in Brazil, confirming high diversity and revealing unique genotypes in this region. Given most of genotypes here identified are different from previous studies in domestic animals, future studies on T. gondii from wildlife is of interest to understand population genetics and structure of this parasite.

Experimental infection of crested caracara (Caracara plancus) with Toxoplasma gondii simulating natural conditions.

Toxoplasma gondii affects mainly warm-blooded animals, including birds. Even though previous experimental data indicate that raptors are resistant to clinical infection, there is no information regarding the susceptibility of Brazilian birds of prey to T. gondii. The present study aimed to observe how the crested caracara, a common raptor in Brazil, interacts with T. gondii using an experimental model. Seven crested caracaras, seronegative for T. gondii, were separated into infected (n=5) and control groups (n=2). Birds from the infected group were fed T. gondii-infected Calomys callosus, a rodent present in Brazilian savanna and described as highly susceptible to infection by the parasite, for three consecutive days, while control animals were fed non-infected rodents. All infected birds produced T. gondii-specific IgG antibodies that were firstly detected at day 7 post-infection, with peak production detected between 15 and 30dpi. No significant alterations in clinical and hematological parameters were observed throughout the experimental period, and parasites were sparsely found in muscular tissues after the birds were euthanized. In conclusion, our results demonstrated that crested caracaras are resistant to oral infection with T. gondii, suggesting that the host-parasite relationship between both species has reached a remarkable equilibrium.