Diagnosis of transmissible venereal tumors in bitches - platelet indices are a remarkable marker?

Background: Hematological analyses are seen as more preferred laboratory analyses in canine transmissible venereal tumor studies. There is no information about the availability of platelets and their indices in routine practice in canine transmissible venereal tumor cases. Taking this as a starting point, this study analyzed the usefulness of platelet indices in dogs with transmissible venereal tumor in clinical laboratory diagnosis as well as examined the relationship between white blood cells, red blood cells, platelets (PLT), main platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT), and the ratio of main platelet volume to platelets (MPV/PLT). Materials, Methods & Results: In the study, a total of 42 bitches of various breeds were used. Nineteen healthy bitches were used as a control group, and the others 23 with cTVT as a study group. Metastasis was not observed in any of the bitches involved in the study. History, clinical findings, and cytological examinations were evaluated for the diagnosis of cTVT. In animals with hemorrhagic discharge and neoplastic lesions, a vaginal cytological examination was performed. Typical TVT cells with large nuclei and intracytoplasmic vacuoles were observed in the vaginal cytological examinations, and the diagnosis of TVT was made. Healthy bitches (19) and those with TVT (23) were 39.16 5.37 months and 47.61 5.14 months old, respectively. From all animals, 2 mL blood samples were collected from V. cephalica to evaluate PIs in the complete blood count (CBC). Collected blood samples were analyzed using an automated hematology analyzer. As a result of the analysis, WBC, RBC, HGB, HCT, MCV, MCHC, RDW, PLT, MPV, PDW, PCT, and MPV/PLT data were obtained. Mild leukocytosis, an increase in PLT, and a decrease in MCV and MPV/PLT were determined in the study group compared to the control group. Cut-off values in CBC of bitches with TVT were determined as WBC: 13.35 (sensitivity: 78%; specificity: 90%); MCV: 67 (sensitivity: 57%; specificity: 95%); PLT: 315.50 (sensitivity: 65%; specificity: 74%); and MPV/PLT: 0.028 (sensitivity: 78%; specificity: 58%). In CBC analyses, a strong negative correlation between PLT and MPV/PLT was detected in both groups. Discussion: Canine transmissible venereal tumors are common in both stray and pet dogs. It is naturally transferred from animal to animal during mating by live tumor cells. This tumor can commonly affect the external genitalia and internal organs in some cases. It generally has the look of cauliflower, and its surface is ulcerated, inflammatory, hemorrhagic, and infectious. More preferred laboratory analyses are complete blood count and blood chemistry analysis in cTVT for to evaluate the success of treatments. Platelet indices have been investigated in many diseases such as endotoxemia, chronic enteropathy, mammary tumor, parvoviral enteritis, septic peritonitis, lymphoma, pyometra, visceral leishmaniasis, and babesiosis in dogs. There is no information available for either diagnostic or prognostic use of the PIs in canine TVT cases. Ultimately, in light of the presented study's results, platelet indices, especially PLT and the MPV/PLT ratio, seem to be notable laboratory markers in terms of easy accessibility and low-cost assessment techniques in canine transmissible venereal tumor cases. New data, however, should be established by a thorough follow-up study using a larger sample size and addressing its usefulness as a diagnostic or prognostic marker in canine transmissible venereal tumors.

Ozonioterapia, PRP, fitoterapia, misoprostol e imunomodulação: Novas abordagens terapêuticas para a solução de velhos problemas / Ozone therapy, PRP, phytotherapy, misoprostol and immunomodulation: New therapeutic approaches to solving old problems

A resistência aos antimicrobianos representa uma ameaça sanitária global crescente, que deve ser abordada com urgência na saúde pública, na produção animal, agrícola e no meio ambiente. Essa circunstância põe em risco a eficiência do tratamento e diminui a eficácia contra infecções causadas pelos mais diversos agentes etiológicos, resultando em enfermidades mais prolongadas afetando diretamente os índices de fertilidade. Dessa forma, o objetivo dessa revisão foi abordar diferentes formas terapêuticas para sub/infertilidade em equinos, com o intuito de minimizar os problemas acarretados pelo uso indiscriminado de fármacos, relatando novas terapias alternativas, tais como ozonioterapia, plasma rico em plaquetas, uso de prostaglandina E1 e imunomoduladores na tentativa de solucionar velhos problemas.(AU)

Antimicrobial resistance represents a growing global health threat that must be urgently addressed in public health, animal production, agriculture and the environment. This circumstance jeopardizes the efficiency of the treatment and decreases the effectiveness against infections caused by the most diverse etiological agents, resulting in more prolonged illnesses, directly affecting fertility rates. Thus, the objective of this review was to address different therapeutic forms for sub/infertility in horses, with the aim of minimizing the problems caused by the indiscriminate use of drugs, reporting new alternative therapies, such as ozone therapy, platelet-rich plasma, use of prostaglandin E1 and immunomodulators in an attempt to solve old problems.(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.

Nasal mucosal mineralization, ulceration and epistaxis in a dog with uremia caused by chronic renal failure / Mineralização, ulceração e epistaxe em um cão com uremia causada por insuficiência renal crônica

ABSTRACT: Soft tissue mineralization and epithelial ulceration are common findings in dogs with uremia, being commonly reported in the gastrointestinal tract, lungs and pleura. This report described a case of nasal mucosal mineralization and ulceration contributing to recurrent epistaxis in a dog with chronic renal failure and uremia. A dog with recurrent epistaxis accompanied by elevated urea and creatinine was hospitalized. Platelet count and coagulation tests were within normal limits. Chronic renal failure was diagnosed, and the dog was euthanized. On necropsy, the kidneys were small, with an irregular capsular surface. The nasal conchae were slightly reddish. Histopathology revealed chronic glomerulonephritis, with gastric mineralization and bilateral parathyroid hyperplasia. Vascular and basal lamina mineralization, epithelial ulceration and hemorrhage were seen in the nasal conchae. The observed findings indicated that nasal mineralization and ulceration were caused by uremia. The severity of histopathological findings suggested that nasal mineralization/ulceration may have caused or at least contributed to epistaxis in this dog. We hope to stimulate further investigations into possible association between uremia, nasal mucosa mineralization/ulceration and epistaxis in dogs.

RESUMO: Mineralização dos tecidos moles e ulceração epitelial são achados comuns em cães com uremia, sendo geralmente observados no trato gastrointestinal, pulmões e pleura. O objetivo desse relato é reportar um caso de mineralização e ulceração da mucosa nasal contribuindo para epistaxe recorrente em um cão com insuficiência renal crônica e uremia. Um cão com epistaxe recorrente e aumento da ureia e creatinina foi hospitalizado. A contagem plaquetária e os testes de coagulação não tinham alterações. Foi diagnosticado insuficiência renal crônica, e o cão foi submetido a eutanásia. Na necropsia, o cão tinha os rins diminuídos, com superfície irregular. As conchas nasais estavam levemente avermelhadas. Histologicamente, foi diagnosticada uma glomerulonefrite crônica com mineralização gástrica e hiperplasia das paratireoides. As conchas nasais tinham mineralização da parede de vasos e membrana basal, úlceras e hemorragia. Os achados histopatológicos indicam que a mineralização e ulceração nasal foram causadas pela uremia. A severidade das lesões histológicas sugere que a mineralização/ulceração nasal pode ter causado, ou pelo menos contribuído, para a epistaxe deste cão. Espera-se, com esse relato, estimular futuros estudos que investiguem uma possível associação entre uremia, mineralização/ulceração nasal e epistaxe em cães.

Feline leishmaniosis: hematological and biochemical analysis / Leishmaniose felina: análises de hematologia e bioquímica

One hundred and sixty-six cats from two animal shelters were subjected to enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence antibody test (IFAT), conventional polymerase chain reaction (cPCR), quantitative PCR (qPCR) and parasitological tests (PA) for the diagnosis of Leishmania spp. Among them, 15% (25/166), 53.6% (89/166), 3.6% (06/166) and 1.8% (03/166) were positive by ELISA, IFAT, both PCRs and PA, respectively. The sequencing of ITS-1 PCR amplicons revealed a 100% match with Leishmania infantum. After the Leishmania spp. survey, 12 cats were selected and divided into two groups for clinical, hematological, and biochemical analysis: six L. infantum positive cats (G1) and six Leishmania spp. negative cats (G2). All the cats were negative for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV). A statistical analysis indicated significantly low platelet counts and significant hyperproteinemia associated with hypoalbuminemia in positive cats (p<0.05). Our results suggest that in endemic areas, cats with clinical signs of feline leishmaniosis (such as skin lesions, weight loss and/or enlarged lymph nodes) and that exhibit hematological and biochemical changes, such as low platelet counts and hyperproteinemia with hypoalbuminemia, should be tested for Leishmania spp. infection.(AU)

Cento e sessenta e seis gatos de dois abrigos foram submetidos ao diagnóstico de Leishmania spp. por ensaio imunoenzimático (ELISA), imunofluorescência indireta (RIFI), reação em cadeia pela polimerase convencional (cPCR) e quantitativa (qPCR) e métodos parasitológicos (PA). Destes, 15% (25/166), 53,6% (89/166), 3,6% (06/166) e 1,8% (03/166) foram positivos por ELISA, RIFI, as duas PCRs e PA, respectivamente. O sequenciamento dos produtos amplificados da PCR ITS-1 foi 100% idêntico à Leishmania infantum. Após o inquérito, 12 gatos foram selecionados para compor dois grupos para análises de hematologia e bioquímica: 6 gatos positivos para L. infantum (G1) e 6 gatos Leishmania spp. negativos (G2). Todos os gatos foram negativos para o vírus da imunodeficiência felina (FIV) e o da leucemia felina (FeLV). Foi observada uma diminuição na contagem de plaquetas e uma hiperproteinemia e hipoalbuminemia significativas em gatos positivos (p<0,05). Esses resultados sugerem que, em áreas endêmicas, os gatos com sinais clínicos de leishmaniose felina (tais como lesões dermatológicas, perda de peso e/ou linfonodos aumentados), associados a alterações hematológicas e bioquímicas, como contagem reduzida de plaquetas e hiperproteinemia com hipoalbuminemia, devem ser testados para leishmaniose felina.(AU)

Síndrome da feminização em cão com sertolioma / Feminization syndrome in a dog with Sertoli cell tumor

Background: The uncontrolled multiplication of Sertoli cells causes Sertoli cell tumor or Sertolioma. Because of this, the level of estrogen in the bloodstream increases rapidly and approximately 25% of dogs with this tumor develop feminization syndrome. Testicular neoplasms are more common in dogs than cats, and are often found in elderly patients. This work aims to describe the clinical signs of the feminization syndrome and the treatment instituted in a canine diagnosed with sertolioma. Case: A 18-year-old male canine, 19.5 kg of body mass, with an increase in testicular volume for about 2 years, was treated at the University Veterinary Hospital. On clinical examination, a matte and brittle coat, alopecia on the hind limbs and gynecomastia were observed. Also noted, non-harmonious aspect of the scrotum, pendular foreskin, atrophied right testicle and hyperplastic left, scrotal hyperthermia and absence of pain. In addition, as a result of the hyperestrogenism resulting from the neoplasm, the paraneoplastic syndrome of feminization, the patient also presented galactorrhea, pendular foreskin, atrophy of the penis and the contralateral testicle, dermatopathies, such as bilateral symmetrical alopecia of the flanks, easily removable hair and variable hyperpigmentation. Rectal body temperature of 38.6°C, clear lung auscultation and muffled cardiac auscultation. The results of laboratory tests showed changes such as thrombocytopenia, platelet counts below the reference levels, platelet count of 163,000/uL. There were no alterations that represented metastases in the imaging exams, such as in the chest X-ray in three incidences and in the abdominal ultrasonography. Then, we opted for the surgical procedure of orchiectomy, with the traditional technique of three clamps, associated with total ablation of the scrotum. Samples were sent to the histopathology laboratory and the diagnosis of sertolioma was confirmed. At 10, 30 and 90 days after the operation, the patient was reassessed for possible recurrences or alterations, but there were no complications or recurrence after the procedure. Discussion: Neoplasms of the male reproductive system are common in dogs. Sertolioma is considered one of the most frequent neoplasms in elderly dogs and that results in systemic clinical signs. This is in line with the 18-year-old dog described in the present report. In addition, it may result in clinical signs resulting from hyperestrogenism resulting from the neoplasm that is called paraneoplastic feminization syndrome. The characteristics of this syndrome are: gynecomastia, galactorrhea, pendular foreskin, atrophy of the penis and contralateral testicle, associated with dermatopathies, such as symmetrical bilateral alopecia. All these clinical signs were present. The diagnosis is made through complete anamnesis, complete clinical examination and complementary examination such as ultrasound help in the presumptive diagnosis, but only with histopathology can it be confirmed. In the clinical approach, histopathology was performed to close the diagnosis. Treatment is behind orchiectomy and total ablation of the scrotum, which was performed in the reported case. The treatment of choice was easy to apply, in addition to improving the patient's quality of life, promoting rapid post-surgical healing and an early return to normal life. However, for the effectiveness of the technique, the early diagnosis and collaboration of tutors is fundamental.

Nasal mucosal mineralization, ulceration and epistaxis in a dog with uremia caused by chronic renal failure / Mineralização, ulceração e epistaxe em um cão com uremia causada por insuficiência renal crônica

Soft tissue mineralization and epithelial ulceration are common findings in dogs with uremia, being commonly reported in the gastrointestinal tract, lungs and pleura. This report described a case of nasal mucosal mineralization and ulceration contributing to recurrent epistaxis in a dog with chronic renal failure and uremia. A dog with recurrent epistaxis accompanied by elevated urea and creatinine was hospitalized. Platelet count and coagulation tests were within normal limits. Chronic renal failure was diagnosed, and the dog was euthanized. On necropsy, the kidneys were small, with an irregular capsular surface. The nasal conchae were slightly reddish. Histopathology revealed chronic glomerulonephritis, with gastric mineralization and bilateral parathyroid hyperplasia. Vascular and basal lamina mineralization, epithelial ulceration and hemorrhage were seen in the nasal conchae. The observed findings indicated that nasal mineralization and ulceration were caused by uremia. The severity of histopathological findings suggested that nasal mineralization/ulceration may have caused or at least contributed to epistaxis in this dog. We hope to stimulate further investigations into possible association between uremia, nasal mucosa mineralization/ulceration and epistaxis in dogs.

Mineralização dos tecidos moles e ulceração epitelial são achados comuns em cães com uremia, sendo geralmente observados no trato gastrointestinal, pulmões e pleura. O objetivo desse relato é reportar um caso de mineralização e ulceração da mucosa nasal contribuindo para epistaxe recorrente em um cão com insuficiência renal crônica e uremia. Um cão com epistaxe recorrente e aumento da ureia e creatinina foi hospitalizado. A contagem plaquetária e os testes de coagulação não tinham alterações. Foi diagnosticado insuficiência renal crônica, e o cão foi submetido a eutanásia. Na necropsia, o cão tinha os rins diminuídos, com superfície irregular. As conchas nasais estavam levemente avermelhadas. Histologicamente, foi diagnosticada uma glomerulonefrite crônica com mineralização gástrica e hiperplasia das paratireoides. As conchas nasais tinham mineralização da parede de vasos e membrana basal, úlceras e hemorragia. Os achados histopatológicos indicam que a mineralização e ulceração nasal foram causadas pela uremia. A severidade das lesões histológicas sugere que a mineralização/ulceração nasal pode ter causado, ou pelo menos contribuído, para a epistaxe deste cão. Espera-se, com esse relato, estimular futuros estudos que investiguem uma possível associação entre uremia, mineralização/ulceração nasal e epistaxe em cães.

Tratamento de tromboembolismo e preparo para implantação de prótese osteointegrada com oxigenioterapia hiperbárica em uma gata / Treatment of tromboembolism and preparation for an osseointegrated prothesis fixation with hyperbaric oxygen therapy in a kitten

Background: Ischemic neuromyopathy is the most common reason for amputation in cats. In veterinary medicine, the use of prosthetic limbs is not widespread; therefore, in most cases total limb amputation is indicated. However, hyperbaric oxygen therapy (HBOT) is an alternative with several benefits for the treatment of vascular disorders with reperfusion, ischemia, and infection. Therefore, this study aimed to report the positive effects of HBOT on the treatment of ischemic neuromyopathy secondary to arterial thromboembolism on the patient's clinical improvement, and on the preparation of the patient for insertion of an osseointegrated prosthesis. Case: A 6-month-old mixed-breed kitten returned for treatment after undergoing surgery seven days earlier for reduction of traumatic diaphragmatic hernia, during which it suffered a cardiorespiratory arrest. The patient presented with acute pelvic limb paralysis with 24-h evolution, absent femoral pulse, plantar cushions and dorsal part of the limbs cold and pale. After supportive therapy and diagnosis of aortic thromboembolism by arterial Doppler, the patient started adjunctive treatment with HBOT from the first day of hospitalization. Sessions took place in an exclusive hyperbaric chamber for animals and lasted 60 min at a pressure of 2.5 absolute atmospheres and 100% oxygen, initially every 12 h. However, during the first 5 days of hospitalization, the distal region of both pelvic limbs began to show tissue devitalization and edema, and hematologic parameters showed changes on the 7th day. The right pelvic limb (RPL) showed more involvement of superficial tissues, extending to the tarsometatarsal joint region. After 8 days of hospitalization, the devitalized tissue was debrided. The RPL had an extensive devitalized area with exposed bone in the phalanges and necrosis in the pads. The left pelvic limb (LPL) suffered minor complications, with involvement of the phalangeal region. After 12 days, with HBOT every 48 h, exuberant granulation tissue was observed. After 17 days, the patient was discharged, and HBOT sessions were performed weekly. Gangrene of the midfoot and lack of proprioception were observed in RPL, while LPL showed bone divulsion of the 1st, 3rd, and 4th phalanges. Because of the poor prognosis for limb viability, the RPL was partially amputated, and a self-threaded intraosseous prosthesis was inserted. Discussion: The cardiorespiratory arrest that occurred during the surgical procedure to reduce the diaphragmatic hernia without thromboprophylaxis may have contributed to the peripheral ischemia. HBOT was proposed for the adjuvant treatment of ischemic injury because it is especially indicated for cases of ischemia-reperfusion injury. The main hematological parameters were evaluated at an average interval of 7 days. While the platelet count and hematocrit increased, the leukocytosis decreased. This demonstrates the benefit of oxygen therapy in the reported patient. The use of HBOT in orthopedic injuries is known to result mainly in stimulation of osteoblasts, promoting osseointegration of the prosthesis. We conclude that the adjuvant treatment with HBOT helped to preserve a large segment of both pelvic limbs, prevent the progression of necrosis, and provide a healthy bed for fixation of an osseointegrated prosthesis in the RPL, resulting in clinical improvement of the patient.

Dr Association of bone morphogenetic protein 15 and growth differentiation factor 9 with litter size in livestock: a review study

Litter size is one of the crucial factors in livestock production and is of high economic value, which is affected by ovulation rate, hormones, and growth factors.Growth factors play a multifaceted role in reproductive physiology. This review aims to investigate the association of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9)with litter size in livestock.The transforming growth factor ß(TGF-ß) superfamily includes more than 34 members; GDF9 and BMP15 are among the most significantfactors for regulating fertility and litter size in most livestock species. Ovarian follicles release BMP15 and GDF9 that are involved in the maturation of primary follicles into the basal form, proliferation of granulosa and theca cells, steroidogenesis, ovulation, and formation of the corpus luteum. Besides, these factors are highly expressed in oocytes and are necessary for female fertility and multiple ovulation in several livestock species. Animals with two inactive copies of these factors are sterile, while those with one inactive copy are fertile. Thus, the present reviewprovides valuable information on the association of BMP15 and GDF9with litter size in livestock that can be used as biological markers of multiple ovulation or for improving fertility in livestock.(AU)

Effects of breed, age and gender on equine platelet rich plasma and correlation of platelet count with its physical aspect / [Efeitos de raça, idade e gênero sobre o plasma rico em plaquetas em equinos e correlação da contagem plaquetária com seu aspecto físico]

Platelet rich plasma samples from 50 healthy horses of five different breeds (Thoroughbreds - TB, Brazilian Criollo Horses - BCH, Brazilian Sport Horses - BSH, Miniature Horses - MH and Crossbred Horses - CB), were investigated as to breed, age, and gender effect for platelet concentration. Moreover, a score for physical analysis was established to correlate PRP physical aspect with platelet count. Platelet count was performed by an automatic hematology analyzer and by manual count. PRP physical analysis was based on color, aspect, and capacity to separate blood components. MH showed significant higher platelet concentration than BSH (p<0.05), while the other breed comparisons showed no significant difference. There was no significant difference for gender but there was a weak correlation of age with PRP platelet concentrations (rs = -0.24). Most of the PRP presented yellow color, the separation of blood components showed no correlation, but the aspect showed a moderate correlation (rs = 0.30) with platelet count. Results suggest that PRP platelet concentration can be influenced by intrinsic factors such as breed. Additionally, the analysis of PRP aspect can help to evaluate the quality of the product when there is no access to platelet counts.

Amostras de plasma rico em plaquetas de 50 cavalos saudáveis, de cinco raças diferentes (Puro Sangue de Corrida, Crioula, Brasileiro de Hipismo, Pônei Brasileiro e sem raça definida), foram investigadas quanto à concentração plaquetária em relação à raça, à idade e ao sexo. Além disso, um escore foi estabelecido para correlacionar o aspecto físico do PRP com a contagem plaquetária. A contagem celular foi realizada por um analisador hematológico automático e pelo método manual. A análise física baseou-se na cor, no aspecto e na capacidade de separar hemocomponentes. Pôneis tiveram concentração plaquetária significativamente maior que os Brasileiros de Hipismo (P<0,05), mas as demais comparações entre raças não apresentaram diferença quanto à contagem plaquetária. Não houve diferença para o gênero, mas a idade demonstrou correlação fraca com as concentrações plaquetárias do PRP (rs = -0,24). A maioria dos PRPs apresentaram coloração amarela, e a separação dos hemocomponentes não mostrou correlação com a contagem plaquetária, mas o aspecto apresentou uma correlação moderada (rs = 0,30). Os resultados deste estudo sugerem que a concentração plaquetária do PRP pode ser influenciada por fatores intrínsecos, como a raça. Além disso, a análise do aspecto do PRP pode ajudar na avaliação da qualidade do produto quando não há acesso à contagem plaquetária.

Cellular components and TGF-beta1 content of a closed Tube system for Platelet Rich Plasma acquisition in horse] / [Componentes celulares e TGF-beta1 do plasma rico em plaquetas obtido por sistema fechado em equinos]

Platelet-rich plasma (PRP) has been proposed as an agent to accelerate the healing process and stimulate the regenerative capacity of tissues due to its abundance of growth factors. A large variety of kits and protocols are available to obtain PRP by different cell-separation systems. However, the lack of standardization may lead to inconsistent results. The aim of this study was to characterize cellular composition, platelet parameters using the ADVIA 120 flow cytometer, and TGF-ß1 concentration from the PRP product obtained through a closed system, using simple centrifugation. Six clinically healthy horses were used in this study. The protocol in the closed system resulted in approximately 1.6-fold higher platelet and approximately 2.0-fold lower white blood cell concentrations in comparison with whole blood values. The evaluated system was efficient in concentrating platelets and in retrieving a small number of leukocytes, using a protocol of single centrifugation at low speed.

O plasma rico em plaquetas (PRP) tem sido proposto como um agente para acelerar o processo de cicatrização e estimular a capacidade regenerativa dos tecidos, devido à sua abundância de fatores de crescimento. Uma grande variedade de kits e de protocolos está disponível para obtenção de PRP, utilizando-se diferentes sistemas de separação de células. No entanto, a falta de padronização pode levar a resultados inconsistentes. O objetivo deste estudo foi caracterizar a composição celular, os parâmetros plaquetários pelo citômetro de fluxo ADVIA 120 e a concentração de TGF-ß1 do PRP obtido em sistema fechado, por centrifugação simples. Seis cavalos clinicamente saudáveis ​​foram usados ​​neste estudo. O protocolo no sistema fechado resultou em concentrações de plaquetas aproximadamente 1,6 vez maior e concentrações de leucócitos aproximadamente 2,0 vezes menores em comparação com os valores do sangue total. O sistema avaliado foi eficiente na concentração de plaquetas e na recuperação de um pequeno número de leucócitos, utilizando um protocolo de centrifugação única em baixa velocidade.

Biochemical energy and hematological profile of sheep supplemented with soy molasses

and proteins. Despite the great variety of by-products and being potentially beneficial, it is not yet fully understood how the interaction between the nutrients in this product and the physiological changes in sheep occurs. Thus, the objective was to evaluate the effects of supplementation levels with soy molasses under the energy metabolites and hematological parameters of sheep. The treatments consisted of 0, 3, 6, 9, and 12% of inclusion of soy molasses in the dry matter for a basal diet of corn silage, which met the nutritional needs of the sheep in maintenance, distributed in a 5 × Latin square design. 5. The energy metabolites were evaluated: cholesterol, triglycerides, very-low-density lipoproteins (VLDL), and fructosamine. The hematological parameters were red blood cells, hemoglobin, hematocrits, and platelets, in addition to leukocytes, rods, neutrophils, eosinophils, monocytes, and lymphocytes. Supplementation with soy molasses did not significantly alter (P> 0.05) the serum concentrations of cholesterol, triglycerides, VLDL, and fructosamine. There was no significant difference in the blood cell, hemoglobin, hematocrit, and platelet concentrations of sheep with soy molasses supplementation (P> 0.05), which may indicate that the level of inclusion was sufficient to meet the nutritional needs of the animals, without damage. Leukocyte, rod, neutrophil, e

Soy molasses is a by-product of the soy industry, with low commercial cost and rich in carbohydrates and proteins. Despite the great variety of by-products and being potentially beneÞ cial, it is not fully understood how the interaction between the nutrients in this product and the physiological changes in sheep occurs. Thus, the objective was to evaluate the effects of supplementation levels with soy molasses on the energy metabolites and hematological parameters of sheep. The treatments consisted of 0, 3, 6, 9, and 12% of inclusion of soy molasses in dry matter basis for a basal diet of corn silage, which met the nutritional needs of the sheep in maintenance, distributed in a 5 × 5 Latin square design. The energy metabolites evaluated were: cholesterol, triglycerides, very-low-density lipoproteins (VLDL), and fructosamine. The hematological parameters were: red blood cells, hemoglobin, hematocrits, and platelets, in addition to leukocytes, rods, neutrophils, eosinophils, monocytes, and lymphocytes. Supplementation with soy molasses did not signiÞ cantly alter (P> 0.05) the serum concentrations of cholesterol, triglycerides, VLDL, and fructosamine. There was no signiÞ cant difference in the blood cell, hemoglobin, hematocrit, and platelet concentrations in sheep receiving soy molasses supplementation (P> 0.05), which may indicate that the level of inclusion was

Characterization of macrophage polarization in lesions of dogs inoculated with Leishmania (Leishmania) infantum (BH401) strain

The aim of this work was to describe the anatomical pathology of dogs experimentally infected with Leishmania (Leishmania) infantum strain MCAN / BR / 2002 / BH401, a Brazilian form of L. infantum isolated from a symptomatic dog from an endemic area. For this, five beagles (three months old and both sexes) composed the experimental group. Markers of macrophage subpopulations M1 and M2 (related to resistance and susceptibility to visceral leishmaniasis) and the tissue cytokine transforming growth factor beta 1 (TGF-ß1) (one of the main cytokines related to the fibrosis process and anti-inflammatory action) were evaluated in livers, lungs and kidneys. The BH 401 L. infantum strain induced classical lesions of the visceral disease where all evaluated organs showed a chronic inflammatory reaction and tissue parasitism associated with a higher expression of CD163 and TGF-ß1 markers, might be related to the progression of the disease. In this work it was possible to conclude that the BH 401 strain reproduces canine visceral leishmaniasis that occurs naturally.(AU)

Expression of TGF-ß/Smads in Cecum and Spleen of Chicken Infected with E. Tenella

Chicken coccidiosis is a common and severe parasitic disease caused by infection from Eimeria spp., which affects the integrity of the intestinal mucosa. TGF-ß has been shown to play an important role in the healing of intestinal mucosas, immunity, and the maintenance of bowel mucosa integrity. Very little is known about the presence of the components of TGF-ß/Smads signaling pathway of chicken at different times following coccidian infection. In the present study, we measured the levels of TGF-ß2, 3, 4, receptor TßRI, II, down-stream Smad 2, 3, 7 in cecum and spleen of chicken at different times after inoculation with Eimeria tenella using quantitative real-time PCR. The results showed that the TGF-ß/Smads signaling pathway was not activated in cecum in the early stage of infection. However, on the 8th day, the expression of TGF-ß2, 4, down-stream protein Smad 2, 7 were significant up-regulated in the spleen, which indicated that the TGF-ß/Smads signaling was changed in the E. tenella infection and was differentially expressed in various tissues in the early stages of infection.(AU)

Biochemical energy and hematological profile of sheep supplemented with soy molasses / Perfil bioquímico energético e hematológico de ovinos suplementados com melaço de soja

Soy molasses is a by-product of the soy industry, with low commercial cost and rich in carbohydrates and proteins. Despite the great variety of by-products and being potentially beneÞ cial, it is not fully understood how the interaction between the nutrients in this product and the physiological changes in sheep occurs. Thus, the objective was to evaluate the effects of supplementation levels with soy molasses on the energy metabolites and hematological parameters of sheep. The treatments consisted of 0, 3, 6, 9, and 12% of inclusion of soy molasses in dry matter basis for a basal diet of corn silage, which met the nutritional needs of the sheep in maintenance, distributed in a 5 × 5 Latin square design. The energy metabolites evaluated were: cholesterol, triglycerides, very-low-density lipoproteins (VLDL), and fructosamine. The hematological parameters were: red blood cells, hemoglobin, hematocrits, and platelets, in addition to leukocytes, rods, neutrophils, eosinophils, monocytes, and lymphocytes. Supplementation with soy molasses did not signiÞ cantly alter (P> 0.05) the serum concentrations of cholesterol, triglycerides, VLDL, and fructosamine. There was no signiÞ cant difference in the blood cell, hemoglobin, hematocrit, and platelet concentrations in sheep receiving soy molasses supplementation (P> 0.05), which may indicate that the level of inclusion was sufÞ cient to meet the nutritional needs of the animals, without damage. Leukocyte, rod, neutrophil, eosinophil, monocyte, and lymphocyte concentrations did not change (P> 0.05) with the inclusion of soy molasses in the sheep's diet, indicating the absence of intoxication, diseases, and infections. It can be concluded that supplementation with soy molasses does not alter the energy metabolites and hematological parameters of sheep in maintenance.(AU)

O melaço de soja é um subproduto da indústria da soja, com baixo custo comercial e rico em carboidratos e proteínas. Apesar da grande variedade de subprodutos e de serem potencialmente benéÞ cos, ainda não está totalmente elucidado como ocorre a interação entre os nutrientes deste produto e as mudanças Þ siológicas em ovinos. Dessa forma, objetivou-se avaliar os efeitos de níveis de suplementação com melaço de soja sob os metabólitos energéticos e parâmetros hematológicos de ovinos. Os tratamentos consistiram em 0, 3, 6, 9 e 12% de inclusão de melaço de soja na base da matéria seca para uma dieta basal de silagem de milho, que atendia as necessidades nutricionais das ovelhas em mantença, distribuídas em delineamento em quadrado latino 5 × 5. Foram avaliados os metabolitos energéticos: colesterol, triglicérides, lipoproteínas de muito baixa densidade (VLDL) e frutosamina. Já os parâmetros hematológicos foram: hemácias, hemoglobinas, hematócritos e plaquetas, além de leucócitos, bastonetes, neutróÞ los, eosinóÞ los, monócitos e linfócitos. A suplementação com melaço de soja não alterou signiÞ cativamente (P>0,05) as concentrações séricas de colesterol, triglicérides, VLDL e frutosamina. Não houve diferença signiÞ cativa nas concentrações de hemácias, hemoglobinas, hematócritos e plaquetas das ovelhas com suplementação de melaço de soja (P>0,05), podendo ser indício que o nível de inclusão foi suÞ ciente para atender as necessidades nutricionais dos animais, sem causar balanço energético positivo. As concentrações de leucócitos, bastonetes, neutróÞ los, eosinóÞ los, monócitos e linfócitos não alteraram (P>0,05) com a inclusão de melaço de soja na dieta das ovelhas, não indicando a presença de intoxicação, doenças e infecções. Conclui-se que a suplementação com melaço de soja não altera os metabólitos energéticos e parâmetros hematológicos de ovinos em mantença.(AU)

Acute erythroid leukemia in a cat infected with feline leukemia virus

Background: Erythroid leukemia is a myeloproliferative hematopoietic disorder considered acute when there is a predominance of blasts in the bone marrow. It is frequently reported in cats infected with feline leukemia virus, but it is unclear whether this virus is involved in the oncogenesis. The clinical signs in cats are anorexia, apathy, weight loss, with evolution from 2 weeks to 2 months, pale mucous membranes, hemorrhages, ascites, salivation, and dyspnea due to pleural effusion. This affection responds little to chemotherapy with an unfavorable prognosis. The aim of this study is to report a case of a feline leukemia virus infected cat with the onset of severe hemolytic anemia. Case: A 8-year-old male mixed breed cat was attended with a history of anorexia, oligodipsia, apathy, progressive weight loss, and yellowish color of urine for 7 days. Laboratorial exams showed anemia (with metarubricytes, acanthocytes and ghost cells), leukocytosis and FeLV reagent test. The cat underwent treatment with methylprednisolone acetate and supportive care. One day later, the animal returned with icteric mucous membranes, and emesis. A blood count was performed that found worsening anemia, increased leukocytosis, and lymphocytosis. Abdominal ultrasound showed cholangiohepatitis and lymphadenomegaly in mesenteric lymph nodes. Treatment was started with ondansetron, metronidazole, and amoxicilin with potassium clavulanate. The cat returned after 3 days and laboratorial exams revealed worsening of blood parameters, so blood transfusion was performed. After 2 days, the patient started with dyspnea and hypothermia, that evolved to cardiorespiratory arrest. The body was sent to necropsy and histopathology, where blast cells and rubricytes were found in blood vessels of various organs. The bone marrow was markedly cellular with complete disappearance of adipose tissue. Most of the cells were blasts with abundant and eosinophilic cytoplasm, central nucleus with finely dotted chromatin and a large nucleolus. There were rubricytes, which made possible to confirm acute erythroid leukemia as a morphological diagnosis. Discussion: The clinical signs observed in acute erythroid leukemia are lethargy, inappetence, fever, splenomegaly, mild lymphadenomegaly, associated with leukocytosis, severe anemia, and thrombocytopenia. The reported animal presented signs similar to those described in the literature except that there was no change in platelet counts. The diagnosis of leukemia was reached after histopathology, and it is made when is observed more than 30% of myeloblasts and monoblasts together or when the blast cells count including rubriblasts is greater than 30%. Although chemotherapy, the prognosis is usually poor. It is essential to perform the myelogram for the diagnosis of myeloid leukemias in vivo. In this report, we only achieve final diagnosis after the cat's death, due to the aggressive behavior of the disease. Clinicians must be aware of the likely development of acute erythroid leukemia whenever a feline leukemia virus infected cat presents hemolytic anemia to get an early diagnosis, since this is an extremely aggressive disease, to propose prompt chemotherapy and give the patient a longer survival period.

Hormona anti-Mülleriana en felinos domésticos / Anti-Müllerian hormone in domestic felids

La hormona anti-Mülleriana (HAM) es una glicoproteína dimérica de 140 kDa que pertenece a la superfamilia del factor de crecimiento transformante ß. La HAM es producida por las células de Sertoli de los testículos fetales siendo responsable de la regresión de los conductos müllerianos durante la diferenciación sexual masculina. En la hembra la HAM circula a niveles casi indetectable al nacer, con un ligero aumento en los primeros años de vida hasta la pubertad cuando se expresa en las células de la granulosa de folículos preantrales y antrales pequeños del ovario. En hembras adultas, la HAM actúa como un regulador autocrino y paracrino inhibiendo el reclutamiento de folículos primordiales y reduciendo la respuesta a la hormona folículo estimulante. En el gato doméstico el número de estudios de HAM es muy escaso quedando aún abundantes aspectos por definir en la especie. También la validación de un único inmunoensayo que permita la comparación de valores séricos entre los trabajos es necesaria. El objetivo de esta revisión fue explicar aspectos básicos de la fisiología de la HAM y resumir las principales publicaciones en el gato doméstico. Finalmente, se proponen aplicaciones prácticas para esta hormona en la especie.(AU)

O hormônio anti-Mülleriano (HAM) é uma glicoproteína dimérica de 140 kDa que pertence a superfamília do fator de crescimento transformador ß. O HAM é produzido pelas células de Sertoli dos testículos fetais, sendo responsável pela regressão dos ductos de Müller durante a diferenciação sexual masculina. Na fêmea, o HAM circula a níveis quase indetectáveis ao nascimento, com um ligeiro aumento nos primeiros anos de vida até a puberdade, quando se expressa nas células da granulosa de folículos préantrais e antrais pequenos do ovário. Nas fêmeas adultas, o HAM atua como um regulador autócrino e parácrino inibindo o recrutamento de folículos primordiais e reduzindo a resposta ao hormônio folículoestimulante. No gato doméstico o número de estudos do HAM é muito escasso, faltando vários aspectos a serem definidos na espécie. Também é necessária a validação de um único imunoensaio que permita a comparação de valores séricos entre trabalhos. O objetivo desta revisão foi explicar aspectos básicos da fisiologia do HAM e resumir as principais publicações no gato doméstico. Finalmente, se propõe aplicações práticas para este hormônio na espécie.(AU)

Influencing factors for preparation of platelet-rich plasma in horses / Fatores que influenciam o preparo do plasma rico em plaquetas na espécie equina

There is no consensus in the literature as to the best platelet-rich plasma (PRP) acquisition method or the ideal platelet concentration to stimulate tissue repair. Besides that, most studies do not provide a clear and replicable description of the methods used, which makes standardization and result comparison difficult. Thus, this study aimed to accurately describe the method and evaluate factors influencing PRP preparation in equines. In this regard, two protocols were used, P1 and P2, based on two centrifugation methods, which differed in blood volume, speed and time, and platelet pipetting and resuspension. In conclusion, factors such as centrifugation strength and time are essential to obtain PRP with adequate platelet count. However, other factors such as tube type and plasma pipetting and resuspension methods directly interfere with the replicability of the technique, and hence influencing PRP applicability.(AU)

Não há consenso na literatura quanto ao melhor método de obtenção do plasma rico em plaquetas (PRP) ou à concentração plaquetária ideal com o objetivo de estimular o reparo tecidual. Além disso, grande parte dos trabalhos não faz descrição clara e reprodutível dos métodos utilizados, o que dificulta a padronização e a comparação dos resultados. Dessa forma, este trabalho teve por objetivo descrever detalhadamente a metodologia e avaliar fatores que influenciam o preparo do PRP em equinos. Para isso, foram utilizados dois protocolos, P1 e P2, baseados em duas centrifugações, que diferiram quanto ao volume de sangue colhido, velocidade e tempo das centrifugações e método de pipetagem e ressuspensão das plaquetas. Conclui-se que fatores como força e tempo de centrifugação são essenciais à obtenção de um PRP com concentração plaquetária adequada. Contudo, fatores adicionais como tipo de tubo e métodos de pipetagem e ressuspensão do plasma interferem diretamente na repetibilidade da técnica, influenciando a aplicabilidade prática do PRP.(AU)

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

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

Allogenic platelet-rich plasma in induced ulcers in rat's cornea / [Plasma rico em plaquetas alogênico em úlceras de córnea induzidas em ratos]

The objective in this study was to evaluate the clinic effect of applying allogenic platelet-rich plasma (PRP) heated or not, for treating cornea ulcers, including the dosage of PDGF-BB in the cornea. The ulcers were induced, standardizing the left eye from 81 rats (Ratus norvegicus, albinus variety), assigned randomly into three groups (N=27): control group (CG) which did not receive any topic treatment; heated PRP group (GA) and PRP group (GP), which received topical treatment every eight hours for five days. Each group underwent evaluation at 24 hours (M1), three days (M3) and five days (M5). The clinical exam evaluated the opacity, vascularization and corneal repair. The corneal PDGF-BB was dosed through the ELISA method. The corneal opacity was decreased in PRP-treated animals (GA and GP) and corneal repair time reduced when compared to CG at M1 and M5. Furthermore, GP showed greater vascularization at M3 compared to M1. Applied allogenic PRP eye drops, heated or not, speed up corneal healing, and reduce corneal repair time. However, the corneal PDGF concentration was not altered in any of the treatments.(AU)

Objetivou-se avaliar o efeito clínico da aplicação de plasma rico em plaquetas alogênico (PRP) aquecido ou não, no tratamento de úlceras de córnea, como a dosagem de PDGF-BB na córnea. As úlceras foram induzidas, padronizando-se o olho esquerdo de 81 ratos (Rattus norvegicus, variedade albinus), aleatoriamente, nos três grupos (N = 27): grupo controle (CG), que não recebeu nenhum tratamento tópico; grupo PRP aquecido (GA) e grupo PRP (GP), que receberam tratamento tópico a cada oito horas, durante cinco dias. Cada grupo foi subdividido em 24 horas (M1), três dias (M3) e cinco dias (M5). O exame clínico avaliou a opacidade, a vascularização e o reparo corneano. O PDGF-BB corneano foi dosado pelo método Elisa. Houve diminuição da opacidade da córnea nos animais tratados com PRP (GA e GP) e diminuição do tempo de reparo da córnea em comparação com CG, M1 e M5. Além disso, foi observada maior vascularização no GP no momento M3 em relação ao M1. A aplicação de colírios de PRP alogênico, aquecidos ou não, acelera a cicatrização da córnea, além de reduzir o tempo de reparo da córnea. No entanto, a concentração de PDGF na córnea não se alterou em nenhum dos tratamentos.(AU)