Comparative study of bullfrog (Rana catebeiana) skin preservation for use as a biomaterial / Estudo comparativo de meios de conservação de pele de rã-touro (Rana catesbeiana) para utilização como biomaterial

The use of biological membranes in wound dressings has increasingly become a reality. Accordingly, an ideal means of preservation is sought that can provide tissue maintenance for long periods without interfering with its quality or clinical applicability. Therefore, the objective of the present study was to evaluate and histologically and microbiologically compare frog skins subjected to two different preservation methods. Sixteen frog skins were evaluated and, depending on the preservation method, subdivided into two groups with eight skins each, namely, the Freezing Group, in which the skins were frozen at -4º in a 20% glycerin solution; and the Glycerin Group, whose skins were kept in 98% glycerin at room temperature (average of 28 ºC and average humidity of 78%). The skins were analyzed fresh (T0) and at 30 (T1), 60 (T2), 90 (T3), and 120 (T4) days of preservation. Data were analyzed comparatively. There was no bacterial or fungal growth, and the skin structure and collagen arrangement remained intact at all time points in both treatments. In conclusion, both preservation methods are efficient and capable of maintaining the tissue morphological structure and preventing the growth and proliferation of contaminants for up to 120 days.

A utilização de membranas biológicas em curativos tem se tornado cada vez mais uma realidade. Concomitante, busca-se um meio de conservação ideal que possa proporcionar a manutenção do tecido por longos períodos de tempo sem interferir em sua qualidade e aplicabilidade clínica. Desta forma, o objetivo deste trabalho foi o de avaliar e comparar histologicamente e microbiologicamente peles de rã submetidas a dois diferentes métodos de conservação. Foram avaliadas 16 peles de rã-touro as quais foram, em função do método de conservação, subdivididas em dois grupos com 08 peles cada: O Grupo Congelamento (GC) no qual as peles foram submetidas ao congelamento a -4º, em solução de glicerina a 20%; e o grupo Glicerina (GG), no qual as peles foram conservadas em temperatura ambiente (média de 28ºC e umidade média de 78%) em glicerina a 98%. As peles foram analisadas a fresco (T0) e com 30 (T1), 60 (T2), 90 (T3) e 120 (T4) dias de conservação. Os dados foram analisados de forma comparativa. Em todos os tempos analisados e em ambos os tratamentos, não houve crescimento bacteriano ou fúngico e a estrutura da pele e o arranjo de colágeno mantiveram-se íntegros. Conclui-se que, ambos os métodos de conservação são eficientes e capazes de manter a estrutura morfológica tecidual, e impedem o crescimento e a proliferação de contaminantes por até 120 dias.

Seroprevalence and influence of bovine leukemia virus on the incidence of mastitis in dairy herds

Background: The enzootic bovine leukosis (EBL) is a viral disease with a chronic aspect and its etiological agent is anoncogenic deltaretrovirus called bovine leukemia virus (VLB). It has a wide distribution in dairy cattle herds, and it maytake several years before the first clinical manifestations occur. Animals seropositive for VLB are more susceptible todeveloping infectious diseases such as mastitis. The objective of this study was to evaluate the seroprevalence of EBL, theprevalence of mastitis, the possible association between leukosis and mastitis, and the risk factors related to leukosis inmunicipalities of the Caparaó region of Espírito Santo, Brazil.Materials, Methods & Results: The prevalence of enzootic bovine leukosis and its association with mastitis were evaluatedin an 854 clinically healthy crossbred (Bos tauros tauros x Bos taurus indicus) dairy cows were used, in different lactationphases, from farms located in the 12 municipalities that compose the Caparaó Capixaba micro-region, in the southernpart of Espírito Santo, Brazil, in the period from February to December 2015. The seroprevalence of EBL was determinedby the agar gel immunodiffusion test, the presence of clinical mastitis was ascertained by the black background mug testand of subclinical mastitis by the CMT. The reuse of needles, type of milking, reproductive management, calf mortality,presence of reproductive problems and milk yield were the risk factors assessed. The association between the variableswas estimated by the odds ratio (95% confidence interval). The significance of the associations was determined by theChi-square test. The prevalence of EBL, clinical and subclinical mastitis was 56.79%, 5.50% and 43.55%, respectively.There was a positive association between the EBL virus and the presence of...

Seroprevalence and influence of bovine leukemia virus on the incidence of mastitis in dairy herds

Background: The enzootic bovine leukosis (EBL) is a viral disease with a chronic aspect and its etiological agent is anoncogenic deltaretrovirus called bovine leukemia virus (VLB). It has a wide distribution in dairy cattle herds, and it maytake several years before the first clinical manifestations occur. Animals seropositive for VLB are more susceptible todeveloping infectious diseases such as mastitis. The objective of this study was to evaluate the seroprevalence of EBL, theprevalence of mastitis, the possible association between leukosis and mastitis, and the risk factors related to leukosis inmunicipalities of the Caparaó region of Espírito Santo, Brazil.Materials, Methods & Results: The prevalence of enzootic bovine leukosis and its association with mastitis were evaluatedin an 854 clinically healthy crossbred (Bos tauros tauros x Bos taurus indicus) dairy cows were used, in different lactationphases, from farms located in the 12 municipalities that compose the Caparaó Capixaba micro-region, in the southernpart of Espírito Santo, Brazil, in the period from February to December 2015. The seroprevalence of EBL was determinedby the agar gel immunodiffusion test, the presence of clinical mastitis was ascertained by the black background mug testand of subclinical mastitis by the CMT. The reuse of needles, type of milking, reproductive management, calf mortality,presence of reproductive problems and milk yield were the risk factors assessed. The association between the variableswas estimated by the odds ratio (95% confidence interval). The significance of the associations was determined by theChi-square test. The prevalence of EBL, clinical and subclinical mastitis was 56.79%, 5.50% and 43.55%, respectively.There was a positive association between the EBL virus and the presence of...(AU)

Pitiose cutânea equina de localização atípica tratada topicamente com solução de anfotericina B e DMSO / Topical treatment using amphotericin B and DMSO for an atypically located equine cutaneous pythiosis

Background: Cutaneous lesions by Pythium insidiosum infection are commonly observed in horses, especially in those living at flooded environments. Equine pythiosis is characterized by the development of tumoral masses that are frequently located at distal limbs, ventral abdômen, thorax, breast and face. The lesions are usually granulomatous, serosanguineous and ulcerated, most often destroyed by self-mutilation due to the intense pruritus. The proposed treatment includes surgical excision followed by antifungal drugs administration, which can be done systemically or topically. Amphotericin B and dimethyl sulfoxide (DMSO) in association has been successfully used for cutaneous pythiosis topical treatment due to the DMSO property to carry any substance through plasmatic membranes. Case: The present report concerns a 12-year-old mixed breed gelding presenting with self-mutilation of a tumoral mass located at the left flank. The owners reported that the horse had initially presented a small wound that had evolved to a 20-cm in diameter mass in 4 weeks. Tissue samples were collected, processed and stained by the Gomori's methenamine silver (GMS) method. The histopathological analysis revealed Pythium insidiosum hyphae in a granulomatous tissue, especially located at peripheral region, where kunkers were present. Surgical excision of the mass followed by cauterization was indicated as initial treatment, and due to financial reasons, the owners elected only the topical antifungal therapy to control the fungus infection after surgery. Flunixin meglumine was also administrated for five days aiming the control of pain and inflammation. The wound was cleaned with povidone-iodine solution and rinsed with a solution containing 50 mg of amphotericin B in 10 mL of sterile water and 10 mL of DMSO. This procedure was carried out twice a day. The wound healed fast due to an excellent centripetal epithelialization, and the horse was discharged after 64 days showing only 5% of the initial wound area. The owner reported by telephone the complete healing and hair growth 10 days after discharge. Discussion: Despite the atypical location of the tumoral lesion described at the present report, the history and clinical manifestations, especially the intense pruritus, showed similarity with other characteristic reports of equine cutaneous pythiosis. The diagnosis was confirmed by the histopathological examination showing hyphae structures, as described to be evidences of the presence of Pythium insidiosum in the tissue. The surgical procedure was the first step to provide remission of clinical signs, and one day after surgery the pruritus desapeared. After excision of the granulomatous tissue and cauterization, daily topical administration of amphotericin B associated with DMSO was effective in destroying the infectious agent, as observed by the excellent epithelization. A pink granulation tissue grew up providing an ideal surface for epithelial migration and the healing process progressed quickly. Centripetal epithelialization reduced the wound area until 3% of the initial area in 64 days of treatment, when the remaining wound was found almost completely healed and covered with hair. At the present report, the horse presenting pythiosis was only topically treated. The recommended therapy using amphotericin B and DMSO solution was effective, economically viable and low risk, considering that the systemic antifungal therapy usually suggested is expensive and extremely nephrotoxic. The atypical location of the lesion on the left flank shows that any anatomical region can be affected by the fungus, since the conditions for its development were present.

Perfil eletroforético das proteínas séricas e do líquido peritoneal de equinos submetidos à experimental do duodeno, íleo e cólon maior / Electrophoretic profile of serum and peritoneal proteins of horses submitted to experimental obstruction of duodenum, ileum and lerge colon

Com o objetivo de avaliar as alterações no proteinograma sérico e peritoneal de equinos submetidos a um modelo experimental de obstrução intestinal, distribuíram-se vinte e quatro animais em quatro grupos: controle instrumentado (GI), obstrução do duodeno (GII), íleo (GIII) e cólon maior (GIV). Foram colhidas amostras de sangue e de líquido peritoneal antes da intervenção cirúrgica (T0), após três horas de obstrução (Ti) e com 1, 3, 12, 24, 72, 120 e 168 horas de reperfusão (Tr). Após centrifugação e fracionamento das amostras, determinaram-se as proteínas totais pelo método de biureto e separaram-se as frações proteicas por eletroforese em gel de agarose. Foram observadas alterações nas concentrações séricas e peritoneais das α, β e γ-globulinas nos animais ensaiados. As alterações deveram-se ao processo inflamatório intestinal desencadeado pelo modelo de obstrução e ao trauma cirúrgico, associado à laparotomia. Animais submetidos à obstrução de duodeno e íleo apresentaram maiores concentrações séricas e peritoneais de proteínas de fase aguda. O fracionamento eletroforético das proteínas contidas no líquido peritoneal é mais sensível no diagnóstico de processos inflamatórios abdominais, quando comparado ao sérico.

Aiming at evaluating the electrophoresis profile of serum and peritoneal protein in horses submitted to an experimental modelof intestinal obstruction, 24 animals were divided into four groups: instrumented control (GI), duodenum obstruction (GII), ileum obstruction (GIII) and large colon obstruction (GIV). Blood and peritoneal fluid samples were collected before the surgical procedures (T0), 3h after the obstruction (Ti) and 1, 3, 12, 24, 72, 120 and 168h after reperfusion (Tr). After sample centrifuging and fractioning, total protein were determined by biuret method, and the proteic fractions were separated by electrophoresis in agarose gel. Alterations were observed in serum and peritoneal concentrations of α, β and γ globulins on the studied animals. The changes occurred due to the intestinal inflammatory process caused by the obstruction model and the surgical trauma, associated to laparotomy. Animals submitted to duodenum and ileum obstruction showed higher serum and peritoneal concentrations of acute phase proteins. The electrophoretic separation of proteins included in the peritoneal liquid is more sensible when compared to the serum concentration to the diagnosis of abdominal inflammatory process.

Perfil eletroforético das proteínas séricas e do líquido peritoneal de equinos submetidos à experimental do duodeno, íleo e cólon maior / Electrophoretic profile of serum and peritoneal proteins of horses submitted to experimental obstruction of duodenum, ileum and lerge colon

Com o objetivo de avaliar as alterações no proteinograma sérico e peritoneal de equinos submetidos a um modelo experimental de obstrução intestinal, distribuíram-se vinte e quatro animais em quatro grupos: controle instrumentado (GI), obstrução do duodeno (GII), íleo (GIII) e cólon maior (GIV). Foram colhidas amostras de sangue e de líquido peritoneal antes da intervenção cirúrgica (T0), após três horas de obstrução (Ti) e com 1, 3, 12, 24, 72, 120 e 168 horas de reperfusão (Tr). Após centrifugação e fracionamento das amostras, determinaram-se as proteínas totais pelo método de biureto e separaram-se as frações proteicas por eletroforese em gel de agarose. Foram observadas alterações nas concentrações séricas e peritoneais das α, β e γ-globulinas nos animais ensaiados. As alterações deveram-se ao processo inflamatório intestinal desencadeado pelo modelo de obstrução e ao trauma cirúrgico, associado à laparotomia. Animais submetidos à obstrução de duodeno e íleo apresentaram maiores concentrações séricas e peritoneais de proteínas de fase aguda. O fracionamento eletroforético das proteínas contidas no líquido peritoneal é mais sensível no diagnóstico de processos inflamatórios abdominais, quando comparado ao sérico.(AU)

Aiming at evaluating the electrophoresis profile of serum and peritoneal protein in horses submitted to an experimental modelof intestinal obstruction, 24 animals were divided into four groups: instrumented control (GI), duodenum obstruction (GII), ileum obstruction (GIII) and large colon obstruction (GIV). Blood and peritoneal fluid samples were collected before the surgical procedures (T0), 3h after the obstruction (Ti) and 1, 3, 12, 24, 72, 120 and 168h after reperfusion (Tr). After sample centrifuging and fractioning, total protein were determined by biuret method, and the proteic fractions were separated by electrophoresis in agarose gel. Alterations were observed in serum and peritoneal concentrations of α, β and γ globulins on the studied animals. The changes occurred due to the intestinal inflammatory process caused by the obstruction model and the surgical trauma, associated to laparotomy. Animals submitted to duodenum and ileum obstruction showed higher serum and peritoneal concentrations of acute phase proteins. The electrophoretic separation of proteins included in the peritoneal liquid is more sensible when compared to the serum concentration to the diagnosis of abdominal inflammatory process.(AU)

Hemogasometria de eqüinos submetidos à obstrução experimental do duodeno, íleo e cólon maior / Blood gas analysis of horses submitted to experimental obstruction of duodenum, ileum and large colon

Visando estudar os parâmetros do equilíbrio ácido-base de eqüinos submetidos a um modelo experimental de obstrução intestinal, 24 animais foram distribuídos em quatro grupos, controle instrumentado (GI), obstrução do duodeno (GII), íleo (GIII) e cólon maior (GIV). As amostras de sangue venoso foram coletadas antes das cirurgias (T0), durante as obstruções (T30ob-T180ob) e após as desobstruções (T60des-T180des). Os eqüinos do GIV, no T30ob, e os eqüinos do GII, nos T60ob, T90ob e T120ob, apresentaram aumento do pH(v), da cHCO-3(vP) e da cBase(v) que, acrescidos do aumento da pCO2(v) e da ctCO2(v), caracterizou a alcalose metabólica com compensação respiratória. Nos T90ob e T120ob, nos animais do GII, e no T180ob, nos animais do GIII, a pO2(v) e a sO2(v) tiveram comportamento semelhante. Os valores baixos apresentados pelos animais do GII foram associados à hipercapnia ou à hipoventilação, desencadeadas para a correção da alcalose metabólica. Entretanto, a hipoxemia apresentada pelos animais do GIII foi associada à hipovolemia presente neste período. As alterações ácido-base observadas constituem-se de alterações leves e temporárias, as quais não são capazes de predizer o diagnóstico das obstruções intestinais específicas em eqüinos com cólica. Entretanto, por relacionarem-se diretamente com a precocidade do distúrbio gastrintestinal, elas auxiliam o pesquisador no prognóstico.(AU)

This study aimed to evaluate parameters of acid-base balance in horses submitted to an experimental model of intestinal obstruction. Twenty-four animals were divided in four groups: instrumented control (GI), duodenum obstruction (GII), ileum obstruction (GIII) and large colon obstruction (GIV). Venous blood samples were collected before surgery (T0), during the obstruction (T30ob-T180ob) and after unblocking procedures (T60des-T180des). Animals from GIV, at T30ob and animals from GII, at T60ob, T90ob and T120ob, presented higher values for pH(v), cHCO-3(vP) and cBase(v) which, added to the increase of pCO2(v) and ctCO2(v), characterized the metabolic alkalosis with respiratory compensation. At T90ob and T120ob, in GII animals, and at T180ob, in GIII animals the pO2(v) and sO2(v) had similar response. Low values presented by GII animals were associated to hypercapnia or hypoventilation triggered for metabolic alkalosis correction. However, a hypoxemia presented by GIII animals was associated to hypovolemia at that period. There were light and temporary acid-base alterations that are not capable to predict specific intestinal obstruction diagnosis in equines with colic. However they help in prognosis since both have a direct relation with the precociousness of gastrointestinal disturbance.(AU)

Equilíbrio ácido-base e hidroeletrolítico em eqüinos com cólica / Acid-base and hidroelectrolytic balance in colic horses

Foram utilizados setenta eqüinos distribuídos em três grupos experimentais, G1 (vinte eqüinos hígidos), G2 (vinte e cinco eqüinos com cólica, os quais passaram por tratamento clínico ou cirúrgico e sobreviveram) e G3 (vinte e cinco eqüinos com cólica, os quais passaram por tratamento clínico ou cirúrgico e foram a óbito ou foram sacrificados). Amostras de sangue foram obtidas em dez diferentes momentos, mediante punção da jugular, para estudo do equilíbrio ácido-base e hidroeletrolítico. Os eqüinos com cólica apresentaram diminuição (P<0,05) nos valores do pH(v), principalmente os animais do G3, a qual, associada com a diminuição da pCO2(v), cBase(v) e cHCO-3(vP), confirmou a acidose metabólica, que teve como origem a produção aumentada de lactato durante a glicólise anaeróbica decorrente da hipovolemia. Adicionalmente apresentaram hiponatremia, hipocalemia e hipercloremia. A presença e a magnitude dos desequilíbrios contribuíram substancialmente com o prognóstico dos animais com cólica.(AU)

Seventy horses were distributed into three experimental groups ad follows: G1, twenty healthy animals; G2, twenty-five colic horses that survived after clinical or surgical treatment; and G3, twenty-five colic horses that were sacrificed or died after clinical or surgical treatment. Blood samples were taken from the jugular vein to assess acid-base balance and water-electrolyte balance. Colic horses had lower pH(v) (P<0.05), mainly G3 animals. Acidosis was confirmed by the lower pH(v) associated with decreased pCO2(v), cBase(v) and cHCO-3(vP), which was probably due to an increased production of lactate during anaerobic glycolysis as a result from hypovolemia. Furthermore, hyponatremia, hypokalemia and hyperchloremia were detected. The presence and the extent of the imbalances contributed substantially to the prognosis of colic horses.(AU)