Long term classical methods for the preservation of xanthan gum-producing Xanthomonas arboricola pathovar pruni strains / Métodos clássicos de longo prazo parapreservação e produção de gomaxantana por cepas de Xanthomonas arborícola patovar pruni

The maintenance of viable and stable Xanthomonascells is crucial for the xanthan reliable research and industrial production. The method, storage and recovery conditions should preserve bothviability and phenotypical and genotypical features. Here, the effectiveness classical methods on the long-term preservation of different Xanthomonas arboricola pathovar pruni strains was to determine.Strains were preserved by monthly sub-culturing in solid medium and lyophilization. After 12 years the viability of the strains, was assessed, as well as their productive capacity and the viscosity of the xanthan gum produced by these strains kept by lyophilization and sub-culturing. Among the lyophilized strains, only those stored at -18 °C were viable after 12 years. The productive capacity of the strains were poorly affected by lyophilization, the passage of the cultures into a solid nutrition medium being sufficient for them to return to their normal metabolism. The viscosity of the synthesized xanthan gum was method-dependent and higher for the lyophilized strains. The work and its findings arenew and original because a work on this topic has never been published before. The results obtained allow the breaking of paradigms regarding the preservation of Xanthomonas.(AU)

A manutenção de células de Xanthomonas viáveis e estáveis é crucial para se obter uma pesquisa confiável e para a produção de xantana industrial.O método, o armazenamento eascondições de recuperação devem preservar tanto a viabilidade quanto as características fenotípicas e genotípicas. O objetivo do estudo foi determinar a eficácia dos métodos clássicos na preservação a longo prazo de diferentes cepas de Xanthomonas arboricolapatovarpruni. As cepas foram preservadas por subcultivo mensal em meio sólido e liofilização. Após 12 anos,avaliou-se a viabilidade das linhagens, bem como a capacidade produtiva e a viscosidade da goma xantana produzida por essas linhagens mantidas por liofilização e subcultivo. Entre as cepas liofilizadas, somente foram viáveis, após 12 anos, as armazenadas a -18°C. A capacidade produtiva das cepas foi pouco afetada pela liofilização, sendo suficiente a passagem das culturas para um meio de cultivosólido para que elas voltassem ao seu metabolismo normal. A viscosidade da goma xantana sintetizada foi dependente do método e maior para as cepas liofilizadas. O estudo e suas descobertas sãonovos e originais porque um trabalho sobre este tópico nunca foi publicado antes. Os resultados obtidos permitem quebrar paradigmas quanto à preservação de Xanthomonas.(AU)

Taiwan cobra envenoming: serum venom concentration before and after specific treatment and relationship with debridement of necrotic wound tissue

Background: Bivalent freeze-dried neurotoxic (FN) antivenom has been the primary treatment since the 1980s for Taiwan cobra (Naja atra) envenomation in Taiwan. However, envenomation-related wound necrosis is a significant problem after cobra snakebites. In the present study, we analyzed the changes in serum venom concentration before and after antivenom administration to discover their clinical implications and the surgical treatment options for wound necrosis. Methods: The patients were divided into limb swelling and wound necrosis groups. The clinical outcome was that swelling started to subside 12 hours after antivenom treatment in the first group. Serum venom concentrations before and after using antivenoms were measured to assess the antivenom's ability to neutralize the circulating cobra venom. The venom levels in wound wet dressing gauzes, blister fluids, and debrided tissues were also investigated to determine their clinical significance. We also observed the evolutional changes of wound necrosis and chose a better wound debridement timing. Results: We prospectively enrolled 15 Taiwan cobra snakebite patients. Males accounted for most of this study population (n = 11, 73%). The wound necrosis group received more antivenom doses than the limb swelling group (4; IQR:2-6 vs 1; IQR:1-2, p = 0.05), and less records of serum venom concentrations changed before/after antivenom use (p = 0.0079). The necrotic wound site may release venom into circulation and cause more severe envenomation symptoms. Antivenom can efficiently diminish limb swelling in cobra bite patients. However, antivenom cannot reduce wound necrosis. Patients with early debridement of wound necrosis had a better limb outcome, while late or without debridement may have long-term hospital stay and distal limb morbidity. Conclusions: Antivenom can efficiently eliminate the circulating cobra venom in limb swelling patients without wound necrosis. Early debridement of the bite site wound and wet dressing management are suggestions for preventing extended tissue necrosis and hospital stay.(AU)

Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients

Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at ­80o C for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at ­20o C for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at ­80o C for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at ­20o C.(AU)

Gestational follow-up in ewes - placentome size and fecal progesterone metabolite levels

Background: Transabdominal ultrasonography in sheep allows real-time diagnosis and monitoring of pregnancy as well as measurement of the size of placentomes, which are structures that connect the fetal and maternal portions, that increase as pregnancy progresses. Progesterone is involved in the maintenance of gestation, with high levels noted during pregnancy. In this context, it is hypothesized that measurements of placentome and progesterone could help in monitoring pregnancy in sheep. Therefore, the main objectives of this study included evaluating whether placentome size and fecal progesterone levels are associated with gestational age in ewes and whether measurement of the placentome and progesterone could be used as an estimate of gestational age. Materials, Methods & Results: A total of 63 pregnant adult (between 2 and 5 years of age) Santa Inês ewes were monitored in the city of Boa Vista, Roraima. All ewes were kept in the paddock with Panicum maximun cv. tanzania, mineral salt, and water ad libitum, supplemented with 200 g/animal/day of soybean residue. Ewes were subjected to ultrasonography every 21 days from day 42 of pregancy until term to monitor pregnancy. Ultrasonography was performed using a linear probe to measure the placentome, including length × width in centimeters. Also every 21 days, fecal samples were collected for measurement of progesterone level. Feces were collected directly from the rectal ampulla, stored individually in plastic bags, and frozen at -20ºC until processing. In the laboratory, these feces samples were freeze-dried and subjected to hormonal extraction with 80% methanol. Then, the levels of fecal progesterone metabolites were measured by enzyme immunoassay. Data was tabulated and submitted to statistical analyses, including descriptive stastistic, correlation, KruskalWallis test and Tukey's test. The levels of fecal progesterone metabolites increased significantly between the 3rd and 4th months of pregnancy, with a decrease close to delivery. Placentome size increased significantly between the 2nd and 3rd months of pregnancy. Placentome measurements did not correlate with progesterone levels and showed a weak correlation with gestational age. Therefore, we concluded that the measurements of placentome size and a single evaluation of fecal progesterone metabolites were not considered efficient methods for estimating gestational age. Discussion: Measurement of progesterone level in fecal samples is a non-invasive hormone monitoring method that provides values like that of blood levels and is less stressful for the animals at the time of sample collection. The wide physiological variation makes it difficult or even useless to use fecal progesterone, when is measured only once per animal, as an indicator of the presence of pregnancy. Despite this limitation, serial sampling is useful for monitoring of hormones during the pregnancy period and is an important tool for research purposes. Placentome size measurements correlated weakly with gestational age and did not correlate with progesterone level. Thus, the presence of placentomes is indicative of the presence of pregnancy but was considered inefficient for determining gestational age in sheep.

Glaucoma in dogs and cats - management with cyclocryotherapy

Background: The treatment of glaucoma often requires numerous therapeutic modalities to achieve the desired reduction in intraocular pressure (IOP). Cyclodestructive procedures or ciliary body destruction have been performed using techniques with considerable differences in efficacy and complication rates. Among these methods, cyclocryotherapy is non-invasive and simple for the management of uncontrolled glaucoma in dogs and cats. The objective of this case report is to describe the technique of carbon dioxide cyclocryotherapy to reduce intraocular pressure in dogs and cats with glaucoma. Cases: Nine canine patients and one cat with glaucoma were treated with cyclocryotherapy performed under general anaesthesia. Clinical signs patients included blepharospasm, ocular pain, episcleral congestion and ocular hypertension. The patients showed higher levels of IOP, higher than 30 mmHg. Surgical treatment with general anaesthesia was applied. The pre-anaesthesia protocol included acepromazine 0.05 mg/kg with methadone 0.2 mg/kg, followed by intravenous propofol and maintenance with isoflurane and oxygen. An ophthalmological cryocautery unit was used with carbon dioxide as the cryogenic agent and a retinal cryoprobe of 3.2 mm diameter tip for the procedure. The method used was a double cycle of freezing and thawing for 60 s in each site. The cryoprobe was centred approximately 5 mm posterior to the corneoscleral limbus over the ciliary body. The temperature of each cyclocryotherapy spot was between -60°C and -80°C and each site was maintained in place for 60 s; 4 to 6 spots of the double freeze-thaw cycle were made. This technique did not have any serious complications during or after the application of cryotherapy, but chemosis and hyperaemia of the bulbar conjunctiva developed. Subconjunctival anti-inflammatory steroids were injected to minimise swelling and patient discomfort. Satisfactory results were observed; in all cases, the intraocular pressure decreased, with the usual result being a cosmetic and painless eye. Discussion: Even with recent surgical and medical advances, pain and blindness are still common occurrences in glaucoma in human and veterinary practice. The cyclodestructive procedures included cyclodialysis, cyclodiathermy, cyclocryotherapy, and cyclophotocoagulation. The cryosurgery in veterinary ophthalmology has many indications for the treatment of ocular diseases and is effective at decreasing intraocular pressure in patients with persistent uncontrolled glaucoma. Cyclocryotherapy has been shown to reduce intraocular pressure in dogs, cats, rabbits and humans with normotensive and glaucomatous eyes. The application of a cryoprobe over the ciliary processes results in ablating ciliary tissue so that aqueous humour inflow is reduced to acceptable levels. In the clinical cases evaluated, there was a reduction in intraocular pressure reaching acceptable levels, with the usual result being cosmetic and painless eye. Medical therapy remains the predominant method for treating glaucoma in veterinary patients; therefore, cyclocryotherapy is an effective, simple way to lower IOP and is a reasonable treatment option for glaucoma management. Cyclocryotherapy has shown good results, with a low learning curve and it can also be repeated if necessary.

Effect of three storage methods on physical and chemical properties of colostrum from Bos indicus cows / Efeito de três métodos de conservação nas propriedades físicas e químicas do colostro de vacas Bos indicus

Maternal colostrum provides the neonate with immunoglobulins, essential for passive immunity. However, storage and handling of colostrum can alter its physical and nutritional characteristics. The aim of this study was to evaluate the effects of three conservation methods on colostrum density, pH, total antioxidant capacity (TAC), protein (PP) and lipid peroxidation (LP). Colostrum samples were collected from 15 Bos indicus cows, and divided into three aliquots for storage using three methods: refrigeration at 4°C, freezing at -20°C, and lyophilization. For each method, four evaluation times were performed: day (d) 5, 10, 15, and 20 for refrigeration and d 15, 30, 45, and 60 for freezing and lyophilization. pH and density analyses were performed at each evaluation time for each method. On d 0 and 15 of storage, TAC, LP, and PP analyses were performed. A generalized linear model, Tukey's test for means comparisons, and a Pearson correlation analysis were conducted. A decrease in refrigerated colostrum pH was observed on d 15. Density of the lyophilized colostrum decreased, as compared with that of frozen colostrum. Lyophilization exhibited the lower PP values of samples, whereas refrigeration presented the highest values of LP and PP. No differences in colostrum TAC were observed between storage methods. A positive correlation between PP and colostrum density and a negative correlation between colostrum density and TAC were found. It was concluded that both freezing and lyophilization are suitable storage methods for bovine colostrum, as they limit proteins and lipids oxidation, and maintain the TAC of fresh colostrum.

O colostro materno fornece ao recém-nascido imunoglobulinas que são essenciais para a imunidade passiva. No entanto, o armazenamento e o manuseio do colostro podem alterar suas características físicas e nutricionais. O objetivo deste estudo foi avaliar os efeitos de três métodos de conservação na densidade, pH, capacidade antioxidante total (CAT), peroxidação protéica (PP) e lipídica (LP) do colostro. Amostras de colostro foram coletadas de 15 vacas Bos indicus, e divididas em três alíquotas para armazenamento usando três métodos: refrigeração a 4 °C, congelamento a -20 °C e liofilização. Em cada método, foram realizados quatro tempos de avaliação: dia (d) 5, 10, 15 e 20 para refrigeração e dia 15, 30, 45 e 60 para congelamento e liofilização. Além disso, as análises de pH e densidade foram realizadas em cada momento de avaliação para cada método. No dia 0 e 15 de conservação foram realizadas análises da CAT, PL e PP. Foi realizado um modelo linear generalizado, teste de Tukey para comparação de médias e análise de correlação de Pearson. Uma diminuição no pH do colostro refrigerado foi encontrada no dia 15. A densidade do colostro liofilizado diminuiu em comparação com o colostro congelado. A liofilização apresentou menor PP das amostras, enquanto a refrigeração apresentou os maiores valores de PL e PP. Não foram observadas diferenças no TAC do colostro entre os métodos de armazenamento. Foi encontrada uma correlação positiva entre PP e densidade do colostro e uma correlação negativa entre densidade do colostro e CAT. Assim, concluiu-se que tanto o congelamento quanto a liofilização são métodos adequados de armazenamento do colostro bovino, pois limitam a oxidação de proteínas e lipídios e mantêm o TAC do colostro fresco.

In-vitro production and pre-validation of lyophilized canine platelet-rich plasma for therapeutic use / Produção e validação de plasma rico em plaquetas canino liofilizado para fins de utilização terapêutica

Platelet-rich plasma (PRP) has been considered a promising therapeutic alternative, since platelets are rich in growth factors that are used in the Regenerative Medicine field. However, fresh PRP cannot be stored for long periods. This study aimed to develop a protocol for obtaining lyophilized canine PRP capable of maintaining viability after its reconstitution. For that purpose, canine PRP extraction and lyophilization protocols were initially tested. Subsequently, assays were carried out to quantify the growth factors VEGF and TGF-β, before and after the lyophilization process, gelation test and the three-dimensional gel structure analysis of the reconstituted lyophilized PRP by electron microscopy, as well as in vitro cell proliferation test in lyophilized PRP gel. Additionally, the immunogenicity test was performed, using allogeneic samples of lyophilized PRP. The results showed that the lyophilized PRP had adequate therapeutic concentrations of growth factors VEGF and TGF-β (9.1pg/mL and 6161.6pg/mL, respectively). The reconstituted PRP gel after lyophilization showed an in vitro durability of 10 days. Its electron microscopy structure was similar to that of fresh PRP. In the cell proliferation test, an intense division process was verified in mesenchymal stem cells (MSCs) through the three-dimensional mesh structure of the lyophilized PRP gel. The immunogenicity test showed no evidence of an immune reaction. The findings were promising, suggesting the possibility of having a lyophilized canine PRP that can be marketed. New in vivo and in vitro studies must be carried out for therapeutic confirmation.(AU)

O plasma rico em plaquetas (PRP) é uma alternativa terapêutica promissora, pois as plaquetas são ricas em fatores de crescimento com ação na regeneração de tecidos. No entanto, o PRP fresco não pode ser armazenado por longos períodos. Esse trabalho teve como objetivo desenvolver um protocolo de obtenção de PRP liofilizado canino capaz de manter a viabilidade pós reconstituição. Portanto, foram testados diversos protocolos de extração e liofilização. Para validação do PRP canino liofilizado foi analisada a concentração dos fatores de crescimento VEGF e TGF-β antes e após o processo de liofilização, a estrutura tridimensional do PRP liofilizado reconstituído em forma de gel por microscopia eletrônica e seu efeito in vitro na proliferação de células-tronco mesenquimais. Os resultados demonstraram que o PRP liofilizado apresentou concentrações terapêuticas adequadas dos fatores de crescimento VEGF e TGF- β (9,1pg/ml e 6161,6pg/ml, respectivamente). O gel de PRP reconstituído após liofilização apresentou uma durabilidade in vitro de 10 dias, sua estrutura tridimensional mostrou-se semelhante ao PRP fresco e proporcionou intensa proliferação de células-tronco mesenquimais durante o cultivo. O teste de imunogenicidade não demonstrou evidências de reação imune. Os achados foram promissores, sugerindo a possibilidade de uso de PRP canino liofilizado para o mercado. Novos estudos in vivo e in vitro deverão ser conduzidos para comprovação terapêutica.(AU)

In-vitro production and pre-validation of lyophilized canine platelet-rich plasma for therapeutic use / Produção e validação de plasma rico em plaquetas canino liofilizado para fins de utilização terapêutica

Platelet-rich plasma (PRP) has been considered a promising therapeutic alternative, since platelets are rich in growth factors that are used in the Regenerative Medicine field. However, fresh PRP cannot be stored for long periods. This study aimed to develop a protocol for obtaining lyophilized canine PRP capable of maintaining viability after its reconstitution. For that purpose, canine PRP extraction and lyophilization protocols were initially tested. Subsequently, assays were carried out to quantify the growth factors VEGF and TGF-β, before and after the lyophilization process, gelation test and the three-dimensional gel structure analysis of the reconstituted lyophilized PRP by electron microscopy, as well as in vitro cell proliferation test in lyophilized PRP gel. Additionally, the immunogenicity test was performed, using allogeneic samples of lyophilized PRP. The results showed that the lyophilized PRP had adequate therapeutic concentrations of growth factors VEGF and TGF-β (9.1pg/mL and 6161.6pg/mL, respectively). The reconstituted PRP gel after lyophilization showed an in vitro durability of 10 days. Its electron microscopy structure was similar to that of fresh PRP. In the cell proliferation test, an intense division process was verified in mesenchymal stem cells (MSCs) through the three-dimensional mesh structure of the lyophilized PRP gel. The immunogenicity test showed no evidence of an immune reaction. The findings were promising, suggesting the possibility of having a lyophilized canine PRP that can be marketed. New in vivo and in vitro studies must be carried out for therapeutic confirmation.(AU)

O plasma rico em plaquetas (PRP) é uma alternativa terapêutica promissora, pois as plaquetas são ricas em fatores de crescimento com ação na regeneração de tecidos. No entanto, o PRP fresco não pode ser armazenado por longos períodos. Esse trabalho teve como objetivo desenvolver um protocolo de obtenção de PRP liofilizado canino capaz de manter a viabilidade pós reconstituição. Portanto, foram testados diversos protocolos de extração e liofilização. Para validação do PRP canino liofilizado foi analisada a concentração dos fatores de crescimento VEGF e TGF-β antes e após o processo de liofilização, a estrutura tridimensional do PRP liofilizado reconstituído em forma de gel por microscopia eletrônica e seu efeito in vitro na proliferação de células-tronco mesenquimais. Os resultados demonstraram que o PRP liofilizado apresentou concentrações terapêuticas adequadas dos fatores de crescimento VEGF e TGF- β (9,1pg/ml e 6161,6pg/ml, respectivamente). O gel de PRP reconstituído após liofilização apresentou uma durabilidade in vitro de 10 dias, sua estrutura tridimensional mostrou-se semelhante ao PRP fresco e proporcionou intensa proliferação de células-tronco mesenquimais durante o cultivo. O teste de imunogenicidade não demonstrou evidências de reação imune. Os achados foram promissores, sugerindo a possibilidade de uso de PRP canino liofilizado para o mercado. Novos estudos in vivo e in vitro deverão ser conduzidos para comprovação terapêutica.(AU)

Evaluation of traditional Brazilian sausage (linguiça calabresa) elaborated with oregano and basil extracts as natural antioxidants / Avaliação da linguiça calabresa elaborada com extratos de orégano e manjericão como antioxidantes naturais

This study aimed to evaluate traditional Brazilian sausage (linguiça calabresa) elaborated with oregano and basil extracts as natural antioxidants. Hydroalcoholic extracts of dried oregano and basil leaves were prepared, freeze-dried, and evaluated for antioxidant activity by Folin-Ciocalteu, DPPH, FRAP, and ABTS assays. Freeze-dried oregano extract showed higher (p < 0.05) antioxidant activity than freeze-dried basil extract by all methods. Extracts were used in preparations of seven sausage formulations, as follows: F1, 0.05% oregano extract and 0.05% basil extract; F2, 0.1% oregano extract and 0.05% basil extract; F3, 0.05% oregano extract and 0.1% basil extract; F4, 0.075% oregano extract and 0.075% basil extract, F5, 0.1% oregano extract and 0.1% basil extract; F6 (control), without extract; and F7, 0.01% butylated hydroxytoluene (BHT). Sausage formulations were analyzed for chemical composition, pH, acidity, water activity, lipid oxidation, color, and texture. Formulations showed differences in pH at different storage periods. F1, F2, F3, and F7 had the lowest (p < 0.05) lipid oxidation values at 53 days of storage. F1, F2, F6, and F7 showed higher a* values and lower b* values. Hardness and chewiness decreased (p < 0.05) during storage for all formulations. F1, F2, and F3 had similar efficiency to that with synthetic antioxidant BHT (F7), therefore, oregano and basil extracts have potential application as natural antioxidants in the food industry.(AU)

Este estudo objetivou avaliar linguiça calabresa elaborada com extratos de orégano e manjericão como antioxidantes naturais. Extratos hidroalcoólicos de orégano e manjericão desidratados foram preparados, liofilizados e submetidos a avaliação de sua atividade antioxidante pelos métodos de FolinCiocalteu, DPPH, FRAP e ABTS. O extrato liofilizado de orégano apresentou maior atividade antioxidante (p < 0,05) que o extrato de manjericão para todos os métodos. Os extratos liofilizados foram usados em sete formulações de linguiça calabresa, como segue: F1 com 0,05% de cada extrato, F2 com 0,1% de extrato de orégano e 0,05% de extrato de manjericão, F3 com 0,05% de extrato de orégano e 0,1% de extrato de manjericão, F4 com 0,075% de cada extrato, F5 com 0,1% de cada extrato, F6 (controle), sem adição de extratos e F7 adição de 0,01% de hidroxitolueno butilado (BHT). As linguiças calabresas foram avaliadas, quanto a composição química, pH, acidez, atividade de água, oxidação lipídica, cor e textura. As formulações apresentaram variação de pH nos diferentes períodos de estocagem. As formulações F1, F2, F3 e F7 apresentaram os menores valores de oxidação lipídica (p < 0,05) aos 53 dias de armazenamento. As formulações F1, F2, F6 e F7 apresentaram maiores valores de a* e menores valores de b*. Observouse redução (p < 0,05) de dureza e mastigabilidade ao longo do período de armazenamento para todas as formulações. As formulações F1, F2 e F3 apresentaram eficiência similar à formulação com antioxidante sintético BHT (F7), portanto, os extratos de orégano e manjericão tem potencial como antioxidante natural na indústria de alimentos.(AU)

Uso de placa aquecedora visando a incubação do espermatozoide ovino após criopreservação / Use of heating plate aiming at the ovine sperm incubation after cryopreservation

Objetivou-se avaliar a utilização da placa aquecedora, em substituição ao banho-maria, no teste de termorresistência (TTR) de espermatozoides de carneiros após a criopreservação. Foram coletados 10 ejaculados de três carneiros adultos (n=30), por meio de vagina artificial para ovinos. Em seguida, foram diluídos em TrisGema de ovo, a uma concentração final de 200 x106 sptz/mL, e submetidos á curva de resfriamento, para posterior criopreservação em palhetas em nitrogênio líquido. Após descongelação, as amostras foram analisadas quanto à integridade de membrana, de acrossoma e atividade mitocondrial. O sêmen então foi dividido em dois tubos e submetido ao TTR, um em banho-maria e outro em placa aquecedora, e, ainda, avaliado a cada 30 minutos, do tempo zero (pós-descongelação) até 90 minutos de incubação, a 37 °C, quanto à motilidade espermática e à motilidade progressiva. As variáveis foram submetidas à análise de variância e as médias comparadas pelo teste de Tukey a 5% de probabilidade. Após o processo de congelação/descongelação, os espermatozoides apresentaram baixo percentual de integridade de membrana e elevado percentual de atividade mitocondrial e integridade do acrossoma. Não foi observada diferença na motilidade espermática nem na motilidade progressiva ao longo do TTR, quando comparados os espermatozoides que foram incubados em banho-maria aos incubados em placa aquecedora durante o período de 90 minutos. A placa aquecedora pode ser utilizada como meio de incubação dos espermatozoides de carneiros em substituição ao banho-maria.

The objective was to evaluate the use of the hot plate to replace the water bath in the thermo resistance test of ovine sperm after cryopreservation. Ten ejaculates were also collected from three adult sheep (n=30), by means of artificial sheep vagina. The collected samples were diluted in Tris-Egg Yolk, at a final concentration of 200 x106 sptz/mL and subjected to a cooling curve for subsequent cryopreservation in liquid nitrogen straws. Immediately after thawing, the samples were analyzed for membrane and acrosome integrity, and mitochondrial activity. The semen was then divided into two tubes and submitted to TTR, one in a water bath and the other in a hot plate, and evaluated every 30 minutes, from time zero (post-thaw) until 90 minutes of incubation at 37 °C, for sperm motility and progressive motility. The variables were subjected to analysis of variance and the means were compared using the Tukey test at 5% probability. After the freeze/thawing process, sperm showed a low percentage of membrane integrity, high percentage of mitochondrial activity, in addition to maintaining the integrity of acrossome. No difference was observed in sperm motility nor progressive motility, along the TTR, when comparing the sperm that were incubated in a water bath in relation to those incubated in a hot plate during the period of 90 minutes. The heating plate can be used as a means of incubating sheep sperm in replacement of the water bath.

Mitsuokella Jalaludinii Supplementation Improved Nutrient Utilization of Broilers Fed Low-Available Phosphorus Diet

Phytase enzyme is supplemented to poultry feed to improve phosphorus (P) availability. Mitsuokella jalaludinii, bacteria isolated from the rumen of cattle, has been reported as a cheaper alternative source of phytase. As much nutrients are trapped within the phytate complex, we hypothesized that the supplementation of M. jalaludinii phytase to poultry feed would enhance nutrient utilization by poultry. In the current study, the efficacy of freeze-dried M. jalaludinii cells (Mj) as feed supplement for broilers fed low-available phosphorus (low-aP) diet was evaluated. Day-old male Cobb raised in battery cages were assigned to three treatment groups [normal-available phosphorus diet with heat-deactivated Mj (DMj); low-aP diet with DMj; and low-aP diet with Mj], each consisting of four replicates (10 birds per replicate) for a 3-weeks feeding period. Feed intake was recorded daily from day 1-21, whereas broilers were weighted at day 1, 7, 14, and 21. Total excreta were collected at day 11-13 and 18-20. At day 21, twelve broilers from each treatment group were slaughtered to collect plasma and tibia. The results showed that Mj significantly enhanced broilers live weight and feed conversion ratio compared to the control groups (p 0.05). Supplementation with Mj have also enhanced the level of P, Ca, Mn, Cu, and Zn in the sera; and Ca and Mn in the tibia at day 18-20 sampling period (p 0.05). As Mj supplementation can enhance nutrient utilization particularly in broilers fed with low-aP diet, it could provide the market with another option in improving broilers growth rate at a lower cost.(AU)

Mitsuokella Jalaludinii Supplementation Improved Nutrient Utilization of Broilers Fed Low-Available Phosphorus Diet

Phytase enzyme is supplemented to poultry feed to improve phosphorus (P) availability. Mitsuokella jalaludinii, bacteria isolated from the rumen of cattle, has been reported as a cheaper alternative source of phytase. As much nutrients are trapped within the phytate complex, we hypothesized that the supplementation of M. jalaludinii phytase to poultry feed would enhance nutrient utilization by poultry. In the current study, the efficacy of freeze-dried M. jalaludinii cells (Mj) as feed supplement for broilers fed low-available phosphorus (low-aP) diet was evaluated. Day-old male Cobb raised in battery cages were assigned to three treatment groups [normal-available phosphorus diet with heat-deactivated Mj (DMj); low-aP diet with DMj; and low-aP diet with Mj], each consisting of four replicates (10 birds per replicate) for a 3-weeks feeding period. Feed intake was recorded daily from day 1-21, whereas broilers were weighted at day 1, 7, 14, and 21. Total excreta were collected at day 11-13 and 18-20. At day 21, twelve broilers from each treatment group were slaughtered to collect plasma and tibia. The results showed that Mj significantly enhanced broilers live weight and feed conversion ratio compared to the control groups (p 0.05). Supplementation with Mj have also enhanced the level of P, Ca, Mn, Cu, and Zn in the sera; and Ca and Mn in the tibia at day 18-20 sampling period (p 0.05). As Mj supplementation can enhance nutrient utilization particularly in broilers fed with low-aP diet, it could provide the market with another option in improving broilers growth rate at a lower cost.

How drying methods can influence the characteristics of mucilage obtained from chia seed and psyllium husk / Como os métodos de secagem podem influenciar as características da mucilagem obtida de sementes de chia e casca de psyllium

The influence of drying methods (oven drying at 50 °C, and freeze drying) on the centesimal composition, functional characteristics and rheological properties of mucilage obtained from chia seed and psyllium husk were investigated. Results showed that high temperature of oven drying reduced fiber content, solubility, emulsion activity and emulsion stability of mucilage. All samples showed pseudo plastic behavior, with the best result produced by Heschel-Bulkley and Power Law models of chia and psyllium mucilage, respectively. These results will be helpful in selecting suitable drying methods depending on the functional and rheological properties desired of the chia and psyllium mucilage in a food product.(AU)

Este estudo teve como objetivo avaliar a influência dos métodos de secagem (secagem em estufa a 50 °C e liofilização) sobre a composição centesimal, características funcionais e propriedades reológicas da mucilagem obtida a partir de sementes de chia e casca de psyllium. Os resultados mostraram que a alta temperatura de secagem em estufa reduziu o teor de fibras, a solubilidade, a atividade da emulsão e a estabilidade da emulsão das mucilagens. Todas as amostras apresentaram comportamento pseudoplástico, com o melhor ajuste produzido pelos modelos Heschel-Bulkley e Power Law das mucilagens de chia e psyllium, respectivamente. Estes resultados serão úteis na seleção do método de secagem adequado, dependendo das propriedades funcionais e reológicas desejadas das mucilagens de chia e psyllium.(AU)

Pomegranate (Punica granatum L) peel lyophilized extract delays lipid oxidation in tuscan sausages / Estabilidade oxidativa de linguiça tipo Toscana com extrato liofilizado da casca de romã (Punica granatum L)

Pomegranate (Punica granatum L.) contains a high concentration of antioxidant and phenolic compounds. Pomegranate peel extracts at different concentrations were used as natural antioxidant to increased the lipid stability of fresh Tuscan sausages, comparing with sodium erythorbate (SE). Peels were subjected to ultrasound-assisted aqueous extraction and lyophilization. The extract was previously characterized throught phenol and flavonoids determination. The Tuscan Sausages were prepared, and color, pH, peroxides, and thiobarbituric acid reactive substances were assessed at 1, 15, and 30 days of storage, at a mean temperature of 5 ± 1 ºC and under a 12-hour light cycle/day. Sausages containing 0.05 and 0.1% of peel extract showed results similar to sodium erythorbate in lipid peroxidation. Sausages treated with pomegranate peel extracts showed an adequate global acceptability level in the sensory analysis. Pomegranate peel extracts are; therefore, a promising natural alternative for maintaining the lipid stability of Tuscan sausages, promoting the protection of the meat and in addition, being able to bring beneficial of the pomegranate to the human health.(AU)

A romã (Punica granatum L.) contém uma alta concentração de compostos antioxidantes e fenólicos. Extratos de casca de romã em diferentes concentrações foram utilizados como antioxidante natural para aumentar a estabilidade lipídica de linguiça tipo Toscana, comparados ao eritorbato de sódio. As cascas foram submetidas à extração e liofilização aquosas por ultrassom. O extrato foi caracterizado por determinação de fenol e flavonóides. Os parâmetros cor, pH, peróxidos e ácido tiobarbitúrico foram avaliadas nos dias 1, 15 e 30 pós armazenamento, a temperatura de 5 ± 1 ºC e ciclo de luz de 12 horas/dia. As linguiças contendo 0,05 e 0,1% de extrato de casca apresentaram resultados semelhantes ao eritorbato de sódio quanto a peroxidação lipídica. Também apresentaram aceitabilidade global adequada na análise sensorial. Os extratos de casca de romã são, portanto, uma alternativa natural promissora para manter a estabilidade lipídica das linguiças toscanas, promovendo a proteção e podendo trazer benefícios da romã à saúde humana.(AU)

Ação dos crioprotetores glicose, trealose e quitosana na manutenção da viabilidade de células de Escherichia coli, Saccharomyces cerevisiae após liofilização / Action of cryoprotectors glucose, trealose and quitosan in maintaining viability of Escherichia coli, Saccharomyces cerevisiae cells after lyophilization

A liofilização é considerada uma das técnicas mais seguras para obtenção de estabilidade de cepas de cultura. Para este processo, podem ser utilizados crioprotetores, que são substâncias que protegem as estruturas celulares durante o período de congelamento, descongelamento e desidratação. O objetivo do presente trabalho foi avaliar o desempenho da glicose, trealose e quitosana como crioprotetores para manutenção de cepas de Escherichia coli e Saccharomyces cerevisiae. Para tratamento estatístico, utilizou-se ANOVA e teste de Tukey com o software Bioestat, versão 5.3. Para Escherichia coli, a Trealose apresentou melhores resultados após a liofilização, porém nenhum dos tratamentos mostrou-se eficaz em prolongar a viabilidade até os 60 dias de armazenamento. Para Saccharomyces cerevisiae, todos os tratamentos apresentaram-se satisfatórios ao longo dos 60 dias avaliados.

Lyophilization is considered one of the safest techniques for acquiring stability of culture strains. For this process, cryoprotectants, which are substances that protect cell structures during the period of freezing, thawing and dehydration, may be used. The objective of the present work was to evaluate the performance of glucose, trehalose, and chitosan as cryoprotectants for Escherichia coli e Saccharomyces cerevisiae. For statistical treatment, ANOVA and Tukey’s test were used with the software Bioestat, version 5.3. For Escherichia coli Trealose presented better performance after lyophilization, but none of the treatments proved to be effective in prolonging viability up to 60 days of storage. For Saccharomyces cerevisiae all treatments were satisfactory throughout the 60 days evaluated.

Ação dos crioprotetores glicose, trealose e quitosana na manutenção da viabilidade de células de Escherichia coli, Saccharomyces cerevisiae após liofilização / Action of cryoprotectors glucose, trealose and quitosan in maintaining viability of Escherichia coli, Saccharomyces cerevisiae cells after lyophilization

A liofilização é considerada uma das técnicas mais seguras para obtenção de estabilidade de cepas de cultura. Para este processo, podem ser utilizados crioprotetores, que são substâncias que protegem as estruturas celulares durante o período de congelamento, descongelamento e desidratação. O objetivo do presente trabalho foi avaliar o desempenho da glicose, trealose e quitosana como crioprotetores para manutenção de cepas de Escherichia coli e Saccharomyces cerevisiae. Para tratamento estatístico, utilizou-se ANOVA e teste de Tukey com o software Bioestat, versão 5.3. Para Escherichia coli, a Trealose apresentou melhores resultados após a liofilização, porém nenhum dos tratamentos mostrou-se eficaz em prolongar a viabilidade até os 60 dias de armazenamento. Para Saccharomyces cerevisiae, todos os tratamentos apresentaram-se satisfatórios ao longo dos 60 dias avaliados.(AU)

Lyophilization is considered one of the safest techniques for acquiring stability of culture strains. For this process, cryoprotectants, which are substances that protect cell structures during the period of freezing, thawing and dehydration, may be used. The objective of the present work was to evaluate the performance of glucose, trehalose, and chitosan as cryoprotectants for Escherichia coli e Saccharomyces cerevisiae. For statistical treatment, ANOVA and Tukeys test were used with the software Bioestat, version 5.3. For Escherichia coli Trealose presented better performance after lyophilization, but none of the treatments proved to be effective in prolonging viability up to 60 days of storage. For Saccharomyces cerevisiae all treatments were satisfactory throughout the 60 days evaluated.(AU)

Frozen sections compared with paraffin-embedded sections: a retrospective veterinary autopsy study

Histologic examination of tissues collected at autopsy are typically prepared using conventional methods (formalin fixed, paraffin‐embedded tissue). In this study, twenty-five tissue specimens from twelve animals were collected at autopsy and examined using the frozen section technique. Of the 25 specimens examined, an accurate and specific diagnosis was obtained in 21 specimens; the pathologic process was correctly identified, but a specific diagnosis was not obtained in 1 specimen; the diagnosis was missed in 2 specimens, and in 1 specimen the diagnosis was deferred. Of the two incorrect diagnoses, one was due to a sampling error and one was due to an interpretation error. Overall, the use of postmortem frozen sections allowed for complete agreement with conventional methods in 21 specimens (84%) and the results from this study support the use of frozen section examination of tissue samples collected postmortem.(AU)

Frozen sections compared with paraffin-embedded sections: a retrospective veterinary autopsy study

Histologic examination of tissues collected at autopsy are typically prepared using conventional methods (formalin fixed, paraffin‐embedded tissue). In this study, twenty-five tissue specimens from twelve animals were collected at autopsy and examined using the frozen section technique. Of the 25 specimens examined, an accurate and specific diagnosis was obtained in 21 specimens; the pathologic process was correctly identified, but a specific diagnosis was not obtained in 1 specimen; the diagnosis was missed in 2 specimens, and in 1 specimen the diagnosis was deferred. Of the two incorrect diagnoses, one was due to a sampling error and one was due to an interpretation error. Overall, the use of postmortem frozen sections allowed for complete agreement with conventional methods in 21 specimens (84%) and the results from this study support the use of frozen section examination of tissue samples collected postmortem.

Pharmacological evaluation of the hydro-alcoholic extract of Campomanesia phaea fruits in rats / Avaliação farmacológica do extrato hidroalcoólico dos frutos da Campomanesia phaea em ratos

Campomanesia phaea (Myrtaceae), popularly known as cambuci, is one of several species of plants producing comestible fruits, largely used in human nutrition. Despite its consumption and economic potential, limited scientific research is available on the Campomanesia, especially those related to its therapeutic benefits. It is reported by traditional medicine the use of the plant in the treatment of different disorders, such as cardiovascular and nervous system disturbances. So, the aim of this study was to carry out the pharmacological evaluation of the hydro-alcoholic extract (HAE) of Campomanesia fruits in rats by screening consisting of tests: a) neuropharmacological observation, b) test on the cardiovascular system. The HAE, prepared from the extraction of fruits with water/ethanol, was concentrated and freeze-dried. Behavioral responses in rats were investigated in open field test and the cardiovascular actions were investigated by a register of indirect blood pressure and the register of spontaneous beating rate right atrium. The results revealed that HAE induced grooming, hypotension and bradycardia. So, this study identified an action on the central nervous system, represented by grooming, and a cardiovascular activity of Campomanesia. The hypotension, attributed in part to bradycardia, was not related to a cholinergic effect, discarding a possible cholinomimetic action of the plant that could justify both cardiovascular and central actions.(AU)

Campomanesia phaea (Myrtaceae), popularmente conhecida como cambuci, é uma das várias espécies de plantas que produzem frutos comestíveis amplamente utilizados na nutrição humana. Apesar de seu consumo e potencial econômico, há poucas pesquisas científicas sobre a Campomanesia, especialmente aquelas relacionadas aos seus benefícios terapêuticos. É relatado pela medicina tradicional o uso da planta no tratamento de diferentes distúrbios, tais como distúrbios do sistema nervoso e cardiovascular. Assim, o objetivo deste estudo foi realizar a avaliação farmacológica do extrato hidroalcoólico (EHA) dos frutos de Campomanesia phaea em ratos através de triagem composta pelos testes: a) observação neurofarmacológica, b) testes no sistema cardiovascular. O EHA, preparado a partir da extração de frutos com água/etanol, foi concentrado e liofilizado. As respostas comportamentais em ratos foram investigadas em teste de campo aberto e as ações cardiovasculares foram investigadas pelo registro da pressão arterial indireta e o registro da taxa de batimentos cardíacos espontâneos em preparações isoladas de átrio direito. Os resultados revelaram que EHA induziu grooming, hipotensão e bradicardia. Assim, este estudo identificou uma ação da Campomanesia sobre o sistema nervoso central, representada por grooming, e uma atividade cardiovascular. A hipotensão, atribuída em parte à bradicardia, não está relacionada a um efeito colinérgico, descartando uma possível ação colinomimética da planta que pudesse justificar tanto as ações cardiovasculares quanto as centrais.(AU)

Influence of glycerol addition on the quality of cat epididymal sperm during the freeze-thaw process / Influência da adição do glicerol sobre a qualidade dos espermatozoides epididimários de gato durante as etapas da congelação-descongelação

Cryopreservation of epididymal sperm is a useful tool for preserving the genetic potential of valuable animal specimens. The domestic cat is used as a model to study and develop cryogenics for other felines. However, regulation of the entire cryopreservation process is essential for the success of this biotechnology. Thus, our aim was to evaluate the effects of glycerol equilibration time and freezethaw stages on the quality of epididymal sperm obtained from domestic cats. Epididymal sperm were recovered with TRIS and immediately evaluated for total motility (TM), vigor, viability, membrane functionality (HOST), and morphology. Then, TRIS-20% egg yolk was added to the samples, which were equally divided into two 1.5 mL tubes and refrigerated at 4 ºC for 1 hour. Subsequently, glycerol was added at a final concentration of 5%. The samples were incubated with glycerol (equilibration time) for either 5 or 10 minutes (groups G5 and G10, respectively) and then frozen. Thawing occurred at 37 ºC for 30 seconds. The samples were evaluated at all stages. A reduction in TM was observed only after thawing; however, it was higher in G5 (39.00 ± 4.07%) than in G10 (18.50 ± 4.54%). Vigor declined in both groups after thawing; however, they did not differ from each other. Sperm viability was maintained in G5 after glycerolization (53.60 ± 2.59%); in G10, sperm viability decreased in the glycerolized sample (48.80 ± 2.93%) when compared to that in the fresh sample (59.90 ± 1.74%). Post-thaw viability of G5 (33.80 ± 1.89%) was higher than that of G10 (18.80 ± 3.01%). In the HOST, a decrease in viability was only observed after thawing, with no difference between the groups (41.50 ± 2.84% for G5 and 40.20 ± 3.49% for G10). With regard to sperm morphology, normal sperm decreased while sperm with post-thaw secondary defects increased in both groups. In conclusion, a shorter equilibration time for glycerolization preserves epididymal sperm quality better and the freeze-thaw process is the most critical stage of thawing.(AU)

A criopreservação dos espermatozoides epididimários é uma ferramenta útil para preservar o potencial genético de um animal valioso. Além disso, o gato doméstico é modelo eleito para o estudo e desenvolvimento da criogenia para os demais felinos. Contudo, para o sucesso dessa biotécnica é essencial o controle de todo o processo de criopreservação. Assim, objetivou-se avaliar o efeito do tempo de equilíbrio da glicerolização e das etapas da congelação-descongelação sobre a qualidade dos espermatozoides epididimários de gato doméstico. Para tanto, espermatozoides epididimários foram recuperados com TRIS e imediatamente avaliados quanto à motilidade total (MT), vigor, viabilidade, funcionalidade de membrana (HOST) e morfologia. Em seguida, as amostras foram adicionadas de TRIS-gema a 20%, fracionadas igualmente em dois tubos de 1,5 mL, refrigeradas a 4 ºC por 1 hora e, posteriormente, adicionadas de glicerol na concentração final de 5%. As amostras foram incubadas com glicerol (tempo de equilíbrio) por 5 ou 10 minutos (grupos G5 e G10, respectivamente) e depois congeladas. A descongelação ocorreu a 37 ºC por 30 segundos. As amostras foram avaliadas em todas as etapas. Uma redução na MT foi observada apenas na pós-descongelação, no entanto G5 (39,00 ± 4,07%) foi superior ao G10 (18,50 ± 4,54%). O vigor declinou pós-descongelação em ambos os grupos; contudo, não diferiram entre si. A viabilidade espermática foi mantida no G5 pós-glicerolização (53,60 ± 2,59%), diferentemente do observado em G10, em que a amostra glicerolizada (48,80 ± 2,93%) reduziu em relação à fresca (59,90 ± 1,74%). A viabilidade pós-descongelação de G5 (33,80 ± 1,89%) foi superior à de G10 (18,80 ± 3,01%). No HOST, uma redução da viabilidade só foi observada pósdescongelação, não havendo diferença entre os grupos (41,50 ± 2,84% para G5 e 40,20 ± 3,49% para G10). Em relação à morfologia espermática, os espermatozoides normais diminuíram, enquanto os espermatozoides com defeitos secundários pós-descongelação aumentaram em ambos os grupos. Conclui-se que um menor tempo de equilíbrio para a glicerolização preserva melhor a qualidade dos espermatozoides epididimários e a etapa mais crítica do processo de congelação-descongelação é a descongelação.(AU)