Comparing modified Lee and White method against 20-minute whole blood clotting test as bedside coagulation screening test in snake envenomation victims

Abstract Background: Twenty-minute whole blood clotting test (20WBCT) and Modified Lee and White (MLW) method are the most routinely employed bedside tests for detecting coagulopathic snake envenomation. Our study compared the diagnostic utility of MLW and 20WBCT for snakebite victims at a tertiary care hospital in Central Kerala, South India. Methods: This single-center study recruited 267 patients admitted with snake bites. 20WBCT and MLW were performed simultaneously at admission along with the measurement of Prothrombin Time (PT). The diagnostic utility of 20WBCT and MLW was determined by comparing the sensitivity (Sn), specificity (Sp), positive and negative predictive values, likelihood ratios, and accuracy at admission with an INR value > 1.4. Results: Out of 267 patients, 20 (7.5%) patients had VICC. Amongst those who had venom-induced consumption coagulopathy (VICC), MLW was prolonged for 17 patients, (Sn 85% 95% confidence interval [CI]: 61.1-96.0) whereas 20WBCT was abnormal for 11 patients (Sn 55%, 95% CI: 32.04-76.17). MLW and 20WBCT were falsely positive for the same patient (Sp 99.6%, 95% CI: 97.4-99.9%). Conclusion: MLW is more sensitive than 20WBCT to detect coagulopathy at the bedside amongst snakebite victims. However, further studies are necessary for standardizing bedside coagulation tests in snakebite cases.

O1-conotoxin Tx6. 7 cloned from the genomic DNA of Conus textile that inhibits calcium currents

Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the crude venom or gene amplification from the venom duct. Methods: In this study, a novel O1 superfamily conotoxin Tx6.7 was directly cloned from the genomic DNA of Conus textile using primers corresponding to the conserved intronic sequence and 3' UTR elements. The mature peptide of Tx6.7 (DCHERWDW CPASLLGVIYCCEGLICFIAFCI) was synthesized by solid-phase chemical synthesis and confirmed by mass spectrometry. Results: Patch clamp experiments on rat DRG neurons showed that Tx6.7 inhibited peak calcium currents by 59.29 ± 2.34% and peak potassium currents by 22.33 ± 7.81%. In addition, patch clamp on the ion channel subtypes showed that 10 µM Tx6.7 inhibited 56.61 ± 3.20% of the hCaV1.2 currents, 24.67 ± 0.91% of the hCaV2.2 currents and 7.30 ± 3.38% of the hNaV1.8 currents. Tx6.7 had no significant toxicity to ND7/23 cells and increased the pain threshold from 0.5 to 4 hours in the mouse hot plate assay. Conclusion: Our results suggested that direct cloning of conotoxin sequences from the genomic DNA of cone snails would be an alternative approach to obtaining novel conotoxins. Tx6.7 could be used as a probe tool for ion channel research or a therapeutic candidate for novel drug development.(AU)

Dejeto líquido de suínos: efeito no solo e nas perdas de K, Ca e Mg por erosão hídrica / Pig slurry: effect on soil and K, Ca and Mg losses by water erosion

O dejeto de suíno (DLS) é usado como fertilizante cujos nutrientes são perdidos por erosão e contaminam o ambiente se o dejeto é usado inadequadamente. Objetivou-se avaliar o efeito de DLS nosolo e na erosão, num Nitossolo Bruno. Os tratamentos, 0; 50; 100; e 200 m³ ha-1de DLS foram aplicados superficialmente após a semeadura uma vez em cada um dos cinco cultivos, e na palhada do último cultivo, totalizando 0, 250, 500 e 1000 m³ ha-1, na aveia (Avena strigosa), milho (Zea mays), nabo (Raphanus sativus L.) e soja (Glycine max), e nos resíduos de aveia. A chuva simulada (65 mm h-1e 75 minutos) foi aplicada três vezes no milho e quatro vezes no nabo, na soja, e nos resíduos, com um simulador debraços rotativos. Antes e depois da pesquisa, determinou-se o teor de K, Ca e Mg no solo. As chuvas aplicadas em cada momento compuseram um teste (T). O T1 ocorreu após a aplicação do DLS e os demais testes em intervalos que variaram entre 14 e 70 dias, em função do clima. Durante o escoamento, em intervalos de cincominutos coletaram-se amostras de enxurrada para determinar as perdas de água e o teor de K, Ca e Mg na água. O DLS não influenciou o teor dos nutrientes no solo. O teore as perdas totais dos nutrientes foram maiores com 1000 m3ha-1de DLS do que na ausência do dejeto. Com o aumento de escoamento, diminuiu o teor na enxurrada e aumentou a perda total, a qual aumentou também com o aumento de teor dos nutrientes na enxurrada. Recomenda-se aplicar menos do que 500 m3ha-1de DLS total no solo, mesmo em aplicações intervalares, devido à elevada perda de K, Ca e Mg pela erosão hídrica.(AU)

Pigg slurry (PS) is used as a fertilizer whose nutrients are lost through erosion and contaminate the environment if the manure is used inappropriately. The objective was to evaluate the effect of PS on soil and erosion, in a Alfisol soil. The treatments, 0; 50; 100; and 200 m³ ha-1of PS were applied superficially after sowing once in each of the five crops, and in the straw of the last crop, totaling 0, 250, 500 and 1000 m³ ha-1, in oat (Avena strigosa), corn (Zea mays), turnip (Raphanus sativusL.) and soybean (Glycine max), and in oat residues. The simulated rain (65 mm h-1and 75 minutes) was applied three times to corn and four times to turnip, soybeans, and residues, with a rotating arms simulator. Before and after the research, the content of K, Ca and Mg in thesoil was determined. The rainfall applied at each moment made up a test (T). T1 occurred after the application of the PS and the other tests at intervals that varied between 14 and 70 days, depending on the weather. During the runoff, at five-minute intervals, runoff samples were collected to determine water losses and the content of K, Ca and Mg in the water. The PS did not influence the nutrient content in the soil. The content and total losses of nutrients were higher with 1000 m3ha-1of PS than in theabsence of manure. With the increase in runoff, the content inthe runoff decreased and the total loss increased, which also increased with the increase in the nutrient content in the runoff. It is recommended to apply less than 500 m3ha-1of total PS inthe soil, even in interval applications, due to the high loss of K, Ca and Mg by water erosion.(AU)

Treatment of chronic immunomediated thrombocytopenia with mesenquimal stem cells in dog: case report / Tratamento de trombocitopenia imunomediada crônica com células-tronco mesenquimais em cão: relato de caso

Immunomediated thrombocytopenia is a systemic metabolic disorder in which the platelet count falls below reference values, as the patient's immune system destroys them. The main clinical signs in thrombocytopenia are petechial, hemorrhages, ecchymoses and suffusions. Hematomas can also occur in coagulation disorders. The diagnosis is based on clinical findings and hematological examinations. The treatment consists of the use of corticosteroids and immunosuppressants, delaying cell destruction, and may last for months, not always obtaining a cure for the disease. The present work reports the use of therapy with allogeneic mesenchymal stem cells, derived from the adipose tissue of dogs, for the treatment of chronic immunomediated thrombocytopenia, with an evolution of more than one year, in a Pinscher dog. The alternative treatment showed a good evolution, keeping platelets within the reference values during the treatment, giving the patient quality of life and removing the need for continuous medication for homeostasis after treatment.

A trombocitopenia imunomediada é uma desordem metabólica sistêmica, na qual a contagem plaquetária fica abaixo dos valores de referência, pois o sistema imunológico do paciente a destrói. O principal sinal clínico na trombocitopenia são hemorragias, petequiais, equimoses e sufusões. Hematomas podem ocorrer também em alterações da coagulação. O diagnóstico baseia-se nos achados clínicos e nos exames hematológicos. O tratamento consiste na utilização de corticosteroides e imunossupressores, o que retarda a destruição celular, mas pode se prolongar por meses, nem sempre obtendo cura da doença. O presente trabalho relata a utilização da terapia com células-tronco mesenquimais alogênicas, oriundas do tecido adiposo de cães, para tratamento de trombocitopenia imunomediada crônica, com evolução de mais de um ano, em um cão da raça Pinscher. O tratamento alternativo revelou boa evolução, pois manteve as plaquetas dentro dos valores de referência durante o tratamento, o que proporcionou qualidade de vida ao paciente e tornou desnecessárias medicações de uso contínuo para a homeostase após o tratamento.

Effect of calcium on Pseudomonas aeruginasa and Bacillus cereus metabolites / Efeito do cálcio nos metabólitos de Pseudomonas aeruginasa e Bacillus cereus

The effects of Calcium (Ca+2) on virulence and some parameters should be analyzed in this study. Pseudomonas aeruginosa Gram (-) and Bacillus cereus Gram (+) were used. Both bacteria are soil bacteria. In this study; the effect of Ca+2 on protease, amylase, LasB elastolytic assay, H2O2, pyorubin and biofilm on metabolites of these bacteria were investigated during 24 hour time. In this study, the effect of Ca+2 on the production of some secondary metabolites on P. aeruginosa and B. cereus was investigated and presented for the first time by us.

Os efeitos do cálcio (Ca+2) na virulência e alguns parâmetros devem ser analisados neste estudo. Pseudomonas aeruginosa Gram (-) e Bacillus cereus Gram (+) foram usados. Ambas as bactérias são bactérias do solo. Neste estudo, o efeito do Ca+2 sobre a protease, amilase, ensaio elastolítico LasB, H2O2, piorubina e biofilme nos metabólitos dessas bactérias foram investigados durante 24 horas. Neste estudo, o efeito do Ca+2 na produção de alguns metabólitos secundários em P. aeruginosa e B. cereus foi investigado e apresentado pela primeira vez por nós.

Influence of Phytase with Or without Organic Acid (Sodium Di-Formate) Supplementation on Growth Performance, Carcass Response, Protein and Mineral Digestibility in Starter Phase of Broilers

This experiment was planned to evaluate the effects of phytase supplementation with or without organic acid (OA) on performance, protein, calcium (Ca), phosphorus (P) and sodium (Na) digestibility and carcass parameters in broiler chickens fed low Ca and low P diets in comparison to a high Ca and high P diet with standard specifications. For this purpose, two iso-nitrogenous and iso-caloric diets were prepared in such a way that one diet had high Ca (9.80) and high P (4.50) and second diet had low Ca (8.50) and low P (2.40), respectively. Low Ca and low P diet was further supplemented with enzyme phytase @ 500 FTU/kg, and phytase @ 500 FTU/kg + organic acid (1 kg/ton). Rations were offered to seven replicates of ten birds each, from day 1 to 21. Results revealed that the experimental treatments had no effect on feed intake and growth performance of birds (p>0.05). Carcass parameter results showed highest thigh meat yield % and liver weight % in the birds fed diet with low Ca & P and supplemented with enzymes phytase (p<0.05). Fecal P and Na digestiblities were high (p<0.05) in experimental broilers fed low Ca low P with enzyme phytase supplementation as compared to the diet without phytase supplementation. It is concluded that phytase and OA supplementation in the starter diet with low Ca and low P level did not influence intake, growth and feed conversion of the broilers. However, thigh meat yield percent and mineral digestiblities (P and Na) increased in the broilers that received the diet with low Ca and low P level and supplemented with enzyme phytase.(AU)

Bothrops envenomation in dogs: local and systemic manifestations

Background: Snakebite envenoming is a condition that affects humans and domestic animals worldwide. Identification of the snake species involved in the envenomation is infrequent. Bothrops envenomation presents typical clinicopathological features. This report describes epidemiological, clinical, and pathological data of 2 cases of Bothrops envenomation in dogs, including the first case of Bothrops moojeni snake striking a domestic animal in Brazil. Cases: Case 1. A dog was witnessed to have a Bothrops moojeni snakebite on a farm. In the first 24 h, acute lameness, pain, diffuse swelling, focal bleeding at the left forelimb, and increased whole-blood clotting time were observed in the envenomed dog. Polyvalent antivenom was administered in addition to fluid therapy, analgesics, corticosteroids, and antibiotics. On the 5th day, the animal presented spontaneous bleeding at the wound site, thrombocytopenia, and increased whole-blood clotting time. An additional dose of polyvalent antivenom was administered, and local treatment at the snakebite site was initiated. After 13 days, the dog showed no clinical or laboratory changes and recovered entirely. Case 2. A mongrel dog was taken for a necropsy to determine the cause of death. Grossly, major findings included swelling in the nasal plane that extended to the neck and dissecting hemorrhage in the subcutaneous tissue and adjacent musculature. Hemorrhages were observed in the heart, parietal pleura, left forelimb, lumbar region, and perirenal tissue. Marked necrosis and disruption of small blood vessels and lymphatics within the deep dermis and subcutaneous tissue were the main microscopic findings close to the snakebite site. Additionally, degeneration and necrosis of muscle fibers and dissecting hemorrhage were observed in the head and neck tissues surrounding the snakebite site. Kidneys showed marked interstitial hemorrhage and acute tubular nephrosis. Discussion: Bothrops envenoming is characterized by local (hemorrhage, dermonecrosis, and myonecrosis) and systemic (coagulative disorders, systemic hemorrhage, and acute kidney injury) changes due to the effect of the main venom components such as phospholipase A2 and metalloproteinases. These changes are hallmarks for the bothropic envenomation, supporting the diagnosis in cases 1 and 2. In case 1, the dog developed a Bothrops moojeni snakebite envenomation, but the immediate treatment with antivenom allowed a favorable outcome. In case 2, gross and microscopic findings supported the presumptive diagnosis of fatal bothropic envenomation. A marked local reaction such as swelling, pain, bleeding, bruising, and tissue necrosis was observed in case 1. In case 2, the most significant local changes were swelling and edema at the head and neck, hemorrhage in the subcutaneous tissue, and adjacent musculature. Systemic effects were observed clinically as spontaneous bleeding, thrombocytopenia, increased whole-blood clotting time (Case 1), systemic hemorrhages, and acute tubular nephrosis (Case 2). A proper treatment probably prevented the development of acute renal failure in Case 1. Herein, we show the first case of accidental snakebite envenomation by B. moojeni in a dog in Brazil. Information is scarce on the identification of venomous snake species striking domestic animals. Fast detection of well-determined clinical and pathological findings of Bothrops envenomation is essential for a correct diagnosis, therapeutics, and a good prognosis, even in cases with an unknown history.

Different Calcium Levels and Two Limestone Granulometries in the Diet of Laying Hens: Performance and Bone Characteristics

The effect of different calcium levels and two limestone granulometries on performance, egg quality, biometry of digestive organs and bone characteristics of light laying hens were evaluated. A total of 270 laying hens were used during 112 days, distributed in a completely randomized design in a 3 x 2 factorial scheme, with 3 levels of calcium (3.8, 4.0 and 4.2%) and 2 limestone granulometries (0.222 and 1.922 mm), totaling 6 treatments with 5 repetitions each. The parameters evaluated were: feed intake, egg production, egg weight and mass, feed conversion per mass and per dozen eggs, albumen, yolk and shell percentage, specific gravity of eggs and shell thickness. In addition, the relative weight of the heart, proventricle, gizzard, liver, pancreas, intestine weight and length, and bone characteristics were also evaluated. A significant interaction was observed for tibia mineral matter. Egg production was influenced by the limestone granulometry, presenting greater value in the treatments which contained fine granulometry limestone. Higher calcium levels of 4.0 and 4.2% provide greater specific density of eggs. Biometric variables and bone characteristics were not influenced by the treatments. In conclusion, it is recommended to use fine-grained limestone (0.222 mm) and a 4% calcium level in diets for light commercial hens, as they improve performance and quality of eggs, without interfering in biometrics of digestive organs and bone characteristics.(AU)

Correlation and path analysis of assessment methodologies of bone quality from brown egg layers at final of the production cycle

This study aimed to investigate direct and indirect correlations of methodologies of bone quality analysis from brown egg layers, at final of the production cycle. Twelve femurs ofDekalb Brown laying hens, euthanized at 85-week-old, were assessed to evaluate breaking strength (BS), Seedor index (SI), mineral matter (MM), calcium (Ca) and phosphorus (P) contents, besides cortical (CorD), medullar (MedD)and epiphysis (EpiD) diameters. Correlations and path analysis were obtained with the aid of SAS®University (p≤0.05). The BS directly represented the bonequality and was compared toother methodologies. Greater linear correlations occurred betweenBS and MM (r = 0.82), MM and Ca (r= 0.72), and BS and Ca (r =0.70). The MM content displayedthe greatest direct effect on the BS (r = 0.53). The Ca content showeda reduced direct effect on the BS (r = 0.18), with indirect effects throughMM content (r = 0.44) and EpiD (r = 0.15), however, presented a great total correlation (r = 0.78). Determination of mineral matter content is the main methodology associated with femur breaking strength from brown egg layers at final of the productive cycle. Because of that, this methodologyis more reliable to determine bone quality.(AU)

Hemangiossarcoma pouco diferenciado grau II em seio nasal esquerdo de cão: relato de caso / Grade II poorly differentiated hemangiosarcoma in the left nasal sinus of dog: case report

Hemangiossarcoma é um câncer com alto poder de malignidade e metástase, tendo origem em células endoteliais de vasos sanguíneos, é frequentemente encontrado em baço, pele e raramente em cavidade nasal de cães. Foi atendido um cão, SRD, de 13 anos, com secreção nasal hemorrágica em narina esquerda e aumento de volume dorsal ao nariz que se estendia até seio frontal, com exoftalmia de globo ocular esquerdo, tendo se instalado de forma aguda. Pela radiografia de crânio nas projeções latero-lateral obliqua e dorso ventral, observou-se lise óssea dos seios nasal, frontal e orbital. Colhido material para Citopatologia, o diagnóstico presuntivo foi osteossarcoma. A fim de se planejar o ato cirúrgico, realizou se a tomografia computadorizada de crânio, que evidenciou formação na cavidade nasal esquerda, com expansão até o osso orbital, sem acometimento de linfonodos regionais. A fim de se avaliar a hemostasia e possibilidades de tromboembolismo no pós-cirúrgico, realizou-se o tromboelastograma e o paciente tinha possibilidades de realizar trombo. Ele foi operado com sucesso, removida a maior parte da massa tumoral e utilizada a eletroquimioterapia no leito cirúrgico. O diagnóstico histopatológico foi hemangiossarcoma pouco diferenciado grau 2. O paciente recebeu tratamento com anti-coagulante nos dias seguintes, teve uma recuperação satisfatória e iniciou-se a quimioterapia adjuvante com doxorubicina após 1 mês da cirurgia, entretanto, ele apresentou quadro de sangramento difuso compatível com CID e veio a óbito. Os hemangiossarcomas são raros na cavidade nasal de cães e parece adotar um pior comportamento nesta região, quando comparado ao baço e pele, inclusive com possibilidade de síndrome para-neoplásica. O trombolestograma tem se mostrado um exame importante para avaliação do equilíbrio hemostático no controle as doenças neoplásicas.

Hemangiosarcoma is a cancer with high power of malignancy and metastasis, originating in endothelial cells of blood vessels, it is often found in the spleen, skin and rarely in the nasal cavity of dogs. A dog, SRD, 13 years old, was treated with hemorrhagic nasal discharge in the left nostril and increase in volume dorsal to the nose that extended to the frontal sinus, with exophthalmos of the left eyeball, which had been installed acutely. On the skull X-ray in the lateral oblique and dorsal-ventral projections, bone lysis of the nasal, frontal and orbital sinuses was observed. After collecting material for cytopathology, the presumptive diagnosis was osteosarcoma. In order to plan the surgical act, a computed tomography scan of the skull was performed, which showed formation in the left nasal cavity, with expansion up to the orbital bone, without involvement of regional lymph nodes. In order to assess hemostasis and possibilities of thromboembolism in the postoperative period, a thromboelastogram was performed and the patient had possibilities of having a thrombus. He was successfully operated on, removing most of the tumor mass and using electrochemotherapy in the surgical bed. The histopathological diagnosis was grade 2 poorly differentiated hemangiosarcoma. The patient received treatment with anticoagulants in the following days, had a satisfactory recovery and adjuvant chemotherapy with doxorubicin was started 1 month after surgery, however, he presented compatible diffuse bleeding. with ICD and died. Hemangiosarcomas are rare in the nasal cavity of dogs and seem to have a worse behavior in this region when compared to the spleen and skin, including the possibility of paraneoplastic syndrome. The thrombolestogram has been shown to be an important test for evaluating the hemostatic balance in the control of neoplastic diseases.

Calcium Requirement and Vitamin D Supplementation in Meat-Type Quail at First Stage of Growth

Currently, feed formulations for quail are based on foreign data or data from other species; this undermines productivity with the use of excessive or deficient levels of some nutrients. Therefore, this study aimed to determine the nutritional requirement levels of calcium (Ca) and supplementation vitamin D in meat-type quail from 1 to 14 d of age. The experiment was a 4 × 4 factorial arrangement of treatments (levels of Ca: 0.42, 0.58, 0.74 and 0.90% and levels of vitamin D: 1,000, 2,000, 3,000 and 4,000 IU) with 3 replicate pens and 43 quails per pen. Weight gain increased quadratically with increased levels of Ca and vitamin D; this variable yielded estimated levels of 0.73% Ca and 2,883 IU vitamin D. There was no Ca x vitamin D interaction effect on bone variables. Bone Ca, bone phosphorus, bone ash, femoral bone strength, tibial bone strength and the femoral Seedor index increased quadratically with an increase in Ca levels. These variables yielded estimated values of 0.78, 0.71, 0.78, 0.77, 0.83, and 0.71% Ca, respectively. It is concluded that the requirements of Ca and vitamin D for the maximum weight gain of meat-type quails, in the period from one to 14 days of age, were 0.73% Ca (p=0.001) and 2,883 IU vitamin D (p=0.02), respectively.(AU)

Calcium Requirement and Vitamin D Supplementation in Meat-Type Quail at First Stage of Growth

Currently, feed formulations for quail are based on foreign data or data from other species; this undermines productivity with the use of excessive or deficient levels of some nutrients. Therefore, this study aimed to determine the nutritional requirement levels of calcium (Ca) and supplementation vitamin D in meat-type quail from 1 to 14 d of age. The experiment was a 4 × 4 factorial arrangement of treatments (levels of Ca: 0.42, 0.58, 0.74 and 0.90% and levels of vitamin D: 1,000, 2,000, 3,000 and 4,000 IU) with 3 replicate pens and 43 quails per pen. Weight gain increased quadratically with increased levels of Ca and vitamin D; this variable yielded estimated levels of 0.73% Ca and 2,883 IU vitamin D. There was no Ca x vitamin D interaction effect on bone variables. Bone Ca, bone phosphorus, bone ash, femoral bone strength, tibial bone strength and the femoral Seedor index increased quadratically with an increase in Ca levels. These variables yielded estimated values of 0.78, 0.71, 0.78, 0.77, 0.83, and 0.71% Ca, respectively. It is concluded that the requirements of Ca and vitamin D for the maximum weight gain of meat-type quails, in the period from one to 14 days of age, were 0.73% Ca (p=0.001) and 2,883 IU vitamin D (p=0.02), respectively.

Effect of calcium on Pseudomonas aeruginasa and Bacillus cereus metabolites / Efeito do cálcio nos metabólitos de Pseudomonas aeruginasa e Bacillus cereus

The effects of Calcium (Ca+²) on virulence and some parameters should be analyzed in this study. Pseudomonas aeruginosa Gram (-) and Bacillus cereus Gram (+) were used. Both bacteria are soil bacteria. In this study; the effect of Ca+² on protease, amylase, LasB elastolytic assay, H2O2, pyorubin and biofilm on metabolites of these bacteria were investigated during 24 hour time. In this study, the effect of Ca+² on the production of some secondary metabolites on P. aeruginosa and B. cereus was investigated and presented for the first time by us.

Os efeitos do cálcio (Ca+²) na virulência e alguns parâmetros devem ser analisados neste estudo. Pseudomonas aeruginosa Gram (-) e Bacillus cereus Gram (+) foram usados. Ambas as bactérias são bactérias do solo. Neste estudo, o efeito do Ca+² sobre a protease, amilase, ensaio elastolítico LasB, H2O2, piorubina e biofilme nos metabólitos dessas bactérias foram investigados durante 24 horas. Neste estudo, o efeito do Ca+² na produção de alguns metabólitos secundários em P. aeruginosa e B. cereus foi investigado e apresentado pela primeira vez por nós.

Effect of calcium on Pseudomonas aeruginasa and Bacillus cereus metabolites / Efeito do cálcio nos metabólitos de Pseudomonas aeruginasa e Bacillus cereus

The effects of Calcium (Ca+²) on virulence and some parameters should be analyzed in this study. Pseudomonas aeruginosa Gram (-) and Bacillus cereus Gram (+) were used. Both bacteria are soil bacteria. In this study; the effect of Ca+² on protease, amylase, LasB elastolytic assay, H2O2, pyorubin and biofilm on metabolites of these bacteria were investigated during 24 hour time. In this study, the effect of Ca+² on the production of some secondary metabolites on P. aeruginosa and B. cereus was investigated and presented for the first time by us.(AU)

Os efeitos do cálcio (Ca+²) na virulência e alguns parâmetros devem ser analisados neste estudo. Pseudomonas aeruginosa Gram (-) e Bacillus cereus Gram (+) foram usados. Ambas as bactérias são bactérias do solo. Neste estudo, o efeito do Ca+² sobre a protease, amilase, ensaio elastolítico LasB, H2O2, piorubina e biofilme nos metabólitos dessas bactérias foram investigados durante 24 horas. Neste estudo, o efeito do Ca+² na produção de alguns metabólitos secundários em P. aeruginosa e B. cereus foi investigado e apresentado pela primeira vez por nós.(AU)

Anticoagulants and their effects on the hematological and biochemical parameters of yellow-spotted amazon river turtle / Anticoagulantes e seus efeitos em parâmetros hematológicos e bioquímicos de tracajá

Knowledge of suitable methods and reagents for assessing the health condition of specimens of a given species is essential. The present study evaluated the efficacy of the heparin anticoagulants 5,000 I.U. mL-1, Na2EDTA, and K3EDTA on the blood parameters of yellow-spotted amazon river turtle Podocnemis unifilis, employing different solutions for red blood cells count. The use of the various anticoagulants evaluated after 10 hours of storage efficiently inhibited coagulation in blood samples from P. unifilis. An increased number of erythrocytes was observed with the use of K3EDTA 5% when compared with heparin. Statistically significant changes in the erythrocyte number were observed with the use of the different solutions. Solutions which featured sodium citrate and formaldehyde in their composition, allowed erythrocytes counting up to 120 hours after blood collection, without a change in values. The use of the heparin anticoagulants 5,000 I.U. mL-1, Na2EDTA 3%, Na2EDTA 5%, K3EDTA 3% was recommended in the hematological analysis of P. unifilis. Also recommended was the use of the formaldehyde-citrate solution containing 1.9 g of sodium citrate and 1.0 mL of formaldehyde (in 50 mL of distilled water) to perform red blood cells counts in yellow-spotted amazon river turtle.

O conhecimento sobre métodos e reagentes apropriados para as avaliações da condição de saúde de exemplares de determinada espécie é fundamental. Este estudo avaliou a eficácia dos anticoagulantes heparina 5.000 U.I. mL-1, Na2EDTA (3% e 5%), e K3EDTA (3 e 5%) em parâmetros sanguíneos de tracajá (Podocnemis unifilis), bem como diferentes soluções para contagem de eritrócitos totais. A coagulação foi eficientemente inibida nas amostras de sangue de P. unifilis com o uso dos diferentes anticoagulantes avaliados após 10 horas de armazenamento. Maior número de eritrócitos com o uso de K3EDTA 5% foi observado quando comparado com a coleta de sangue realizada com heparina. Diferenças, estatisticamente significativas, entre as contagens de eritrócitos com o uso das diferentes soluções de reagentes avaliadas foram verificadas. As soluções contendo citrato de sódio e formol na composição possibilitaram contagens de eritrócitos até 120 horas após a coleta, sem alteração em seus valores. Recomenda-se o uso dos anticoagulantes heparina 5000 U.I. mL-¹, Na2EDTA 3%, Na2EDTA 5%, K3EDTA 3% nas análises hematológicas de tracajá. Assim como o uso da solução de formol-citrato contendo 1,9 g de citrato de sódio e 1,0 mL de formol (em 50 mL de água destilada) para realização das contagens de eritrócitos totais em tracajá.

Chronic expanding hematoma secondary to excision of recurrent myxomas in a bitch

Background: Expanding chronic hematoma is a relatively common postoperative complication in humans, not in dogs, nevertheless, is important understand this condition because it may affect the animal's health. A hematoma could be formed within hours and usually stabilizes spontaneously with the reabsorption of its content. However, occasionally the hematoma slowly expands, becomes encapsulated and forms a mass, causing a chronic expanding hematoma. Possibly skin and adipose tissue are displaced secondary to trauma, causing the formation of blood-filled cysts surrounded by a fibrous capsule. The aim of this document is to present the first report of a postsurgical chronic expanding hematoma in dog. Case: A case of chronic expanding hematoma secondary to excision of recurrent myxoma in a bitch was described. Physical examination showed a hard, firm, non-mobile mass enclosed within the deep muscular layers. This mass was between the sternum and the cranial abdomen, similar to previous lesions excised by another veterinarian. Fine needle aspiration indicated the presence of blood. Radiology demonstrated that the bone-cartilage tissue was not involved. Due to the size of the mass, a skin stretching pre-suturing technique using self-adhesive tape was applied around the mass 48 h before surgery. The mass was removed with wide excision margins and portions of the affected muscles, but a large wound with great tension was created; consequently, a parallel to the wound incision and the thoraco-lumbar vertebrae was made. Six months after surgery, the animal presented recurrence of the same lesion, which was also removed. The bleeding times were within the normal range, but the animal was diagnosed with hyperadrenocorticism. After both surgeries histopathological exam revealed a hematoma, that had thrombosis and granulation tissue with fibrosis and severe panniculitis, in addition of multifocal, chronic, moderate lymphohistiocytic myositis; all these changes were compatible with a chronic expanding hematoma. Discussion: Chronic expanding hematoma is a mass that grows slowly secondary to hemorrhage and does not exhibit elements of malignancy. However, it can occasionally be confused for a soft-tissue neoplasm. It has been reported frequently in humans but not in small animals. The first case was reported in 2002: 5 puppies with a cervical hematoma classified as a chronic expanding hematoma was described. In addition, 3 cases were reported: 1 cat with perirenal chronic expanding hematoma; 1 bitch with intra-abdominal presentation, and 1 cat with this pathology on the right pelvic limb after an intramuscular injection. The etiology of chronic expansive hematoma is unidentified in dogs, nevertheless, it is stipulated that is not an inflammatory process as previously assumed, but rather a neoplastic process. As in other cases, it was difficult to determine the origin of the chronic expanding hematoma. The coagulation times and platelets count were within ranges. Similarly, it is very complicated to relate hyperadrenocorticism with chronic expanding hematoma because dogs with Cushing's disease generally present hypercoagulopathies. Myxomas are associated with endocrinopathies in humans and dogs, therefore, hyperadrenocorticism is possibly related with this neoplasia. Myxomas have an expansive growth pattern, contrasting with myxosarcomas, which are infiltrating; Although in the present case the skin mass appeared to be invasive, both macroscopical and histopathological appearance of it corresponded to a well-shaped capsule compatible with chronic expanding hematoma. In summary, to the best of our knowledge, this is the first report of a post-surgical chronic expanding hematoma in a dog.

Nelore females along a new Brazilian agricultural frontier: hematological and clinical-biochemical approaches / [Abordagens hematológicas e bioquímicas clínicas de fêmeas Nelore na nova fronteira agrícola brasileira]

The Nelore breed is the second largest bovine breed in the world and has actively participated in the expansion of new Brazilian agricultural frontiers. In this context, the purpose of this study was to determine the hematological and biochemical reference intervals of healthy Nelore matrices raised under an extensive regime without supplementation along southwest of Piauí state. Blood samples were collected from fifty-five multiparous female of the Nelore breed. Biochemical and hematological parameters were analyzed using a parametric statistical method with 95% CI of reference limits. The average values of red blood cells, hemoglobin as well as hematimetric indices showed reference ranges similar to reference standards. The hematocrit as well as granulocytes and agranulocytes presented alterations typical of animals raised in environments with higher temperatures. Mineral, enzymatic, protein and metabolic profiles were similar to other bovine breeds but with a narrower range of values. However, lower mean values were observed for levels of ionized calcium, total protein and urea. Nelore females present slightly different biochemical and hematological profiles from other breeds, which might result from the environmental and nutritional management applied and the natural deficiency of nitrogen, phosphorus and calcium in the region's pastures.(AU)

Nelore é a segunda maior raça bovina do mundo e tem participado ativamente da expansão das novas fronteiras agrícolas brasileiras. Nesse contexto, o objetivo do presente estudo foi determinar os intervalos de referência hematológicos e bioquímicos de matrizes Nelore criadas em regime extensivo sem suplementação, ao longo do sudoeste do estado do Piauí. Amostras de sangue foram coletadas de 55 fêmeas multíparas da raça Nelore. Os parâmetros bioquímicos e hematológicos foram analisados por método estatístico paramétrico com IC 95% para os limites de referência. Os valores médios de hemácias, hemoglobina e índices hematimétricos apresentaram intervalos de referência semelhantes aos padrões de referência. Tanto o hematócrito quanto os granulócitos e os agranulócitos apresentaram alterações típicas de animais criados em ambientes com temperaturas mais elevadas. Os perfis mineral, enzimático, proteico e metabólico foram semelhantes aos de outras raças bovinas, mas com uma faixa de valores mais estreita. No entanto, valores médios mais baixos foram observados para os níveis de cálcio ionizado, proteína total e ureia. Fêmeas Nelore apresentam perfis bioquímicos e hematológicos ligeiramente diferentes de outras raças, o que pode resultar dos manejos ambiental e nutricional aplicados e da deficiência natural de nitrogênio, fósforo e cálcio nas pastagens da região.(AU)

A novel proline-rich M-superfamily conotoxin that can simultaneously affect sodium, potassium and calcium currents

Background Conotoxins have become a research hotspot in the neuropharmacology field for their high activity and specificity in targeting ion channels and neurotransmitter receptors. There have been reports of a conotoxin acting on two ion channels, but rare reports of a conotoxin acting on three ion channels. Methods Vr3a, a proline-rich M-superfamily conotoxin from a worm-hunting Conus varius, was obtained by solid-phase synthesis and identified by mass spectrometry. The effects of synthesized Vr3a on sodium, potassium and calcium currents were tested on rat DRG cells by patch clamp experiments. The further effects of Vr3a on human Cav1.2 and Cav2.2 currents were tested on HEK293 cells. Results About 10 μM Vr3a has no effects on the peak sodium currents, but can induce a ~10 mV shift in a polarizing direction in the current-voltage relationship. In addition, 10 μM Vr3a can increase 19.61 ± 5.12% of the peak potassium currents and do not induce a shift in the current-voltage relationship. An amount of 10 μM Vr3a can inhibit 31.26% ± 4.53% of the peak calcium currents and do not induce a shift in the current-voltage relationship. The IC50 value of Vr3a on calcium channel currents in rat DRG neurons is 19.28 ± 4.32 μM. Moreover, 10 μM Vr3a can inhibit 15.32% ± 5.41% of the human Cav1.2 currents and 12.86% ± 4.93% of the human Cav2.2 currents. Conclusions Vr3a can simultaneously affect sodium, potassium and calcium currents. This novel triple-target conotoxin Vr3a expands understanding of conotoxin functions.(AU)

A novel proline-rich M-superfamily conotoxin that can simultaneously affect sodium, potassium and calcium currents

Conotoxins have become a research hotspot in the neuropharmacology field for their high activity and specificity in targeting ion channels and neurotransmitter receptors. There have been reports of a conotoxin acting on two ion channels, but rare reports of a conotoxin acting on three ion channels. Methods Vr3a, a proline-rich M-superfamily conotoxin from a worm-hunting Conus varius, was obtained by solid-phase synthesis and identified by mass spectrometry. The effects of synthesized Vr3a on sodium, potassium and calcium currents were tested on rat DRG cells by patch clamp experiments. The further effects of Vr3a on human Cav1.2 and Cav2.2 currents were tested on HEK293 cells. Results About 10 μM Vr3a has no effects on the peak sodium currents, but can induce a ~10 mV shift in a polarizing direction in the current-voltage relationship. In addition, 10 μM Vr3a can increase 19.61 ± 5.12% of the peak potassium currents and do not induce a shift in the current-voltage relationship. An amount of 10 μM Vr3a can inhibit 31.26% ± 4.53% of the peak calcium currents and do not induce a shift in the current-voltage relationship. The IC50 value of Vr3a on calcium channel currents in rat DRG neurons is 19.28 ± 4.32 μM. Moreover, 10 μM Vr3a can inhibit 15.32% ± 5.41% of the human Cav1.2 currents and 12.86% ± 4.93% of the human Cav2.2 currents. Conclusions Vr3a can simultaneously affect sodium, potassium and calcium currents. This novel triple-target conotoxin Vr3a expands understanding of conotoxin functions.(AU)

Blood parameters of crioulo breed horse / Parâmetros sanguíneos de potros da raça Crioula

Plasma levels of hematocrit, total plasma protein, fibrinogen, creatine phosphokinase, aspartate transferase, and lactate were analyzed in blood samples of 85 Crioula breed foals, from birth to two years of age. The animals were divided into age groups: G1 (up to 15 days of age; n=70), G2 (from 16 days to one month of age; n=67), G3 (between one and three months of age; n=75), G4 (between three and six months of age; n=64), G5 (between six and nine months of age; n=59), G6 (between nine and 18 months of age; n=39), and G7 (between 18 months and two years of age; n=17). These groups were statistically analyzed by one-way variance analysis (ANOVA) and Tukey's test. Male and female means were compared by Student's t-test. Hematocrit levels were significantly higher up to 90 days of age and in G7 females. Total plasma proteins increased significantly in groups 3, 4, 6, and 7. The highest fibrinogen levels were found in G1. Yet for creatine phosphokinase, the highest concentrations were detected in G5, whereas those of aspartate aminotransferase in G7. The levels of this enzyme remained similar from 30 days to two years of age. Lactate concentrations were higher in G3. We concluded that the sex of the animal had no significant effect on laboratory test interpretations. By contrast, the age of the animal should be considered since relevant variations were observed with time. Nevertheless, specific tables for each analysis should be consulted for interpretation of results.

Com o propósito de estabelecer valores de hematócrito, proteínas plasmáticas totais, fibrinogênio, creatina quinase , aspartato transferase e lactato em potros da raça Crioula, do nascimento até os dois anos, utilizaram-se amostras sanguíneas de 85 animais, divididos pela estratificação etária: Grupo 1 (G1) Até 15 dias de vida (n=70); grupo 2 (G2), entre 16 dias até um mês (n=67); grupo 3 (G3), entre 1 e 3 meses (n=75); grupo 4 (G4), entre 3 e 6 meses (n=64); grupo 5 (G5), entre 6 e 9 meses (n=59); grupo 6 (G6), entre 9 e 18 meses (n=39); e grupo 7 (G7), entre 18 meses até 2 anos (n=17). Foi realizado estudo estatístico entre os grupos pela análise de variância unidirecional (one-wayANOVA), complementada pelo teste de Tukey. Para comparação das médias entre os sexos utilizou-se o teste t de Student. O hematócrito foi significativamente mais elevado até os 90 dias e nas fêmeas do G7. Para proteínas plasmáticas totais, notou-se aumento significativo nos grupos 3, 4, 6 e 7. Os valores de fibrinogênio foram maiores no G1. A CK apresentou maior concentração no G5 e a AST no G7. A AST assumiu valores semelhantes dos 30 dias até os 2 anos. A concentração de lactato foi mais elevada no G3. Conclui-se que na interpretação dos exames laboratoriais de potros da raça crioula, o gênero não interfere significativamente nos resultados, porém a idade deve ser considerada devido à ocorrência de variações relevantes. Recomenda-se que para interpretação sejam consultadas tabelas específicas para cada análise.