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.

Fixation of pfcrt chloroquine resistance alleles in Plasmodium falciparum clinical isolates collected from unrest tribal agencies of Pakistan / Fixação de alelos de resistência à cloroquina pfcrt em isolados clínicos de Plasmodium falciparum coletados de agitações de agências tribais do Paquistão

Abstract Plasmodium falciparum resistance to Chloroquine (CQ) is a significant cause of mortality and morbidity worldwide. There is a paucity of documented data on the prevalence of CQ-resistant mutant haplotypes of Pfcrt and Pfmdr1 genes from malaria-endemic war effected Federally Administered Tribal Areas of Pakistan. The objective of this study was to investigate the prevalence of P. falciparum CQ-resistance in this area. Clinical isolates were collected between May 2017 and May 2018 from North Waziristan and South Waziristan agencies of Federally Administrated Trial Area. Subsequently, Giemsa-stained blood smears were examined to detect Plasmodium falciparum. Extraction of malarial DNA was done from microscopy positive P. falciparum samples, and P. falciparum infections were confirmed by nested PCR (targeting Plasmodium small subunit ribosomal ribonucleic acid (ssrRNA) genes). All PCR confirmed P. falciparum samples were sequenced by pyrosequencing to find out mutation in Pfcrt gene at codon K76T and in pfmdr1 at codons N86Y, Y184F, N1042D, and D1246Y. Out of 121 microscopies positive P. falciparum cases, 109 samples were positive for P. falciparum by nested PCR. Pfcrt K76T mutation was found in 96% of isolates, Pfmdr1 N86Y mutation was observed in 20%, and 11% harboured Y184F mutation. All samples were wild type for Pfmdr1 codon N1042D and D1246Y. In the FATA, Pakistan, the frequency of resistant allele 76T remained high despite the removal of CQ. However, current findings of the study suggest complete fixation of P. falciparum CQ-resistant genotype in the study area.

Resumo A resistência do Plasmodium falciparum à cloroquina (CQ) é uma causa significativa de mortalidade e morbidade em todo o mundo. Há uma escassez de dados documentados sobre a prevalência de haplótipos mutantes CQ-resistentes dos genes Pfcrt e Pfmdr1 da guerra endêmica da malária em áreas tribais administradas pelo governo federal do Paquistão. O objetivo deste estudo foi investigar a prevalência de resistência a CQ de P. falciparum nesta área. Isolados clínicos foram coletados entre maio de 2017 e maio de 2018 nas agências do Waziristão do Norte e do Waziristão do Sul da Área de Ensaio Administrada Federalmente. Posteriormente, esfregaços de sangue corados com Giemsa foram examinados para detectar Plasmodium falciparum. A extração do DNA da malária foi feita a partir de amostras de P. falciparum positivas para microscopia, e as infecções por P. falciparum foram confirmadas por nested PCR (visando genes de ácido ribonucleico ribossômico de subunidade pequena de Plasmodium (ssrRNA)). Todas as amostras de P. falciparum confirmadas por PCR foram sequenciadas por pirosequenciamento para descobrir a mutação no gene Pfcrt no códon K76T e em pfmdr1 nos códons N86Y, Y184F, N1042D e D1246Y. De 121 microscopias de casos positivos de P. falciparum, 109 amostras foram positivas para P. falciparum por nested PCR. A mutação Pfcrt K76T foi encontrada em 96% dos isolados, a mutação Pfmdr1 N86Y foi observada em 20% e 11% abrigou a mutação Y184F. Todas as amostras eram do tipo selvagem para o códon N1042D e D1246Y de Pfmdr1. No FATA, Paquistão, a frequência do alelo resistente 76T permaneceu alta apesar da remoção de CQ. No entanto, as descobertas atuais do estudo sugerem a fixação completa do genótipo resistente a CQ de P. falciparum na área de estudo.

CA 15-3, CRP, and LDH correlates with prognostic parameters in canine mammary neoplasms

The identification of putative prognostic factors in canine mammary neoplasms (CMNs) has been focused on tissue-specific biomarkers, but the serum biomarkers, including cancer antigen 15-3 (CA 15-3), c-reactive protein (CRP), and lactate dehydrogenase (LDH) have been demonstrated to display clinical application in cases of CMNs. The aim of the study was to evaluate the levels of these serum biomarkers and their association with well-established prognostic factors in CMNs. Samples from 15 female canines with CMNs and 15 clinically healthy ones were collected. The results were evaluated using the Tukey's, Pearson, or Spearman tests. The cut-off point, sensitivity, specificity, and area under curve (AUC) were evaluated using the receiver operating characteristic (ROC) curve analysis in a logistic regression model (P<0.05). The levels of CA 15-3, CRP and LDH were significantly higher in the serum of female dogs with CMNs compared to the healthy ones. Moreover, these factors were positively correlated with ulceration, tumor size, histopathological grade, metastatic lymph node, and clinical staging. Female dogs with CMNs were found to exhibit highest serum levels of CA 15-3, CRP, and LDH. Therefore, they can be applied to improve the efficacy of the diagnosis and prognostic evaluation in casas of CMNs.(AU)

Criopreservação do sêmen suíno: desafios a serem superados que auxiliam a identificação de espermatozoides mais férteis / Boar semen cryopreservation: challenges to be overcome that help the identification of more fertile spermatozoa

A criopreservação é o melhor método de armazenamento de sêmen por período indeterminado, conferindo alta flexibilidade de uso. Além disso, é o principal meio para formação de banco de germoplasma. No entanto, congelar sêmen suíno ainda é um desafio a ser enfrentado. Isso porquê, além das características de membrana espermática que ocasionam maiores efeitos deletérios às células durante os processos de congelação e descongelação, ainda é preciso lidar com a grande variabilidade de resultados, seja entre indivíduos, ejaculados de um mesmo indivíduo ou ainda, frações de um mesmo ejaculado. Frente a isso, diferentes estratégias têm sido investigadas a fim de obter melhores resultados e assim, aumentar a aplicabilidade do sêmen congelado suíno a nível comercial. Além disso, acreditamos que haja a possibilidade de se relacionar a capacidade de criotolerância de um determinado espermatozoide a sua maior ou menor capacidade de fertilizar o oócito.(AU)

Cryopreservation is the best method of storing semen indefinitely, providing high flexibility of use. In addition, it is the primary means for forming a germplasm bank. However, freezing boar semen is still a challenge to be faced because, in addition to the characteristics of the spermatic membrane that cause deleterious effects to the cells during the freezing and thawing processes, it is still necessary to deal with the significant variability of results, whether between individuals, ejaculates from the same individual or even fractions of an even ejaculated. Because of this, different strategies have been investigated to obtain better results and thus increase the applicability of frozen boar semen at a commercial level. Furthermore, we believe it is possible to relate the cryotolerance capacity of a given sperm to its greater or lesser capacity to fertilize the oocyte.(AU)

Depicting latent variables considering different pig genetics and nutritional plans in crossbred pigs

We aimed to reduce the dimensionality of quantitative traits in pigs applying factor analysis. Quantitative variables were collected, and the factor analysis extracted five factors with biological meaning related to performance, carcass quality, carcass yield, meat quality, and initial pH. These factors were posteriorly used as dependent variable to evaluate the effects of genetic groups (Piau, Duroc, and Pietrain crossbreds), nutritional plans (low, medium and high lysine levels) and sex. An interaction effect between genetic group and sex was observed in performance, in which Duroc crossbred showed the greatest scores. The Pietrain genetic group showed greater values of carcass quality compared with Duroc and Piau, while Piau crossbred pigs had greater values for meat quality compared with Pietrain and Duroc. A greater carcass yield was observed in Pietrain crossbred compared with Duroc pigs. Pigs fed under low lysine level had the lowest performance values. Carcass yield was affected by nutritional plans, in which the medium lysine level showed the greatest values. Meat quality was improved by the nutritional plan with high lysine level compared with the low lysine level. Initial pH showed increased means when using nutritional plans with low and medium lysine levels in diet composition. With regard to sex effects, a greater carcass quality was found for gilts compared with barrows. The reduction of the data dimensionality allowed the joint evaluation of genetic group, nutritional plan, and sex based on new latent variables that represented the original dataset under easier framework based on factors' biological interpretation.(AU)

Early puberty in short-haired Guinea pigs kept in laboratory animal facilities

Lab animals, such as Guinea pigs (Cavia porcellus), are crucial for scientific development, as they play an important role in the development and quality control chain of vaccines and drugs distributed by the Brazilian public health system. Investigating their biological and physiological parameters is fundamental to raise and keep these animals, so the handling of the facilities that hold them can be updated whenever new information comes up, with the well-being of the animals and alignment with the 3 Rs in mind. In the search for understanding reproductive aspects of Guinea pigs, the present study had the main goal of studying puberty by means of estrous cycle analysis in short-haired Guinea pigs. Guinea pigs have a vaginal occlusive membrane that covers the vaginal orifice. Its rupture takes place gradually and naturally, moments before labor and during estrus. The present study followed 42 females as for the presentation of the vaginal occlusive membrane. Once the membranes ruptured spontaneously, a swab was collected to study vaginal cytology. Membrane rupture was observed in 39 females; six females showed membrane rupture with less than 21 days of age (17 to 21 days). Twenty-three females were characterized as being in estrus due to cytology showing a prevalence of anucleated superficial cells. One of these females was younger than 21 days old. The opening of the vaginal occlusive membrane took place most frequently in intervals between 17 and 18 days, and the membrane remained open between one and three consecutive days. It was possible to follow three cycles of membrane opening on six females. The present study showed the need to adapt handling guidelines for C. porcellus kept in research animal facilities. The early age of puberty imposes the need of separate the female daughters from their fathers at 16 days old.(AU)

The prognostic value of serum neoplastic biomarkers CA 15-3 and CEA in canine mammary neoplasms: a review / O valor prognóstico dos biomarcadores neoplásicos séricos CA 15-3 e CEA em neoplasias mamárias caninas: uma revisão

As neoplasias mamárias caninas (NMCs) são as mais frequentes em cadelas, com isso, uma grande variedade de técnicas tem sido utilizada para identificar os seus fatores prognósticos. Dentre as possibilidades, os biomarcadores neoplásicos séricos antígeno do câncer (CA 15-3) e o antígeno carcinoembrionário (CEA) são os mais promissores. Biomarcadores neoplásicos são substâncias presentes na neoplasia, sangue ou demais produtos biológicos, produzidos pela neoplasia, ou secundariamente pelo paciente, em resposta à sua presença. Na medicina veterinária, esses marcadores são pouco estudados. Em termos gerais, eles não podem ser usados para o diagnóstico primário de neoplasias mamárias, mas estão relacionados com os fatores prognóstico já bem estabelecidos na literatura e tem como vantagem se tratar de uma análise menos invasiva e que é possível de ser feita de forma seriada. O objetivo desta revisão é descrever o CA 15-3 e o CEA como biomarcadores utilizados nas NMCs, o progresso recente feito na literatura e providenciar um sumário dos principais resultados já obtidos. O CA 15-3 e o CEA apresentam potencial prognóstico nas NMCs. Apesar de existir uma grande variação de resultados para esses biomarcadores na literatura, o seu uso deve ser considerado, visto os resultados obtidos na caracterização e prognóstico das NMC.(AU)

Female canine mammary neoplasms (CMNs) are the most frequent neoplasm in bitches, and a variety of techniques have been used to identify they prognostic factors. Among the possibilities, the serum biomarkers cancer antigen (CA 15-3) and carcinoembryonic antigen (CEA) are the most reliable. Neoplastic biomarkers are substances present in the neoplasia, blood, or other biological products, produced mainly by the neoplasia, or secondarily by the patient, in response to their presence. In veterinary medicine, these biomarkers are poorly studied. In general terms they cannot be used for primary diagnosis of mammary neoplasms but are related to the prognostic factors and have the advantage of being a less invasive analysis and that it is possible to be done in a serial way. The objective of this review is to describe CA 15-3 and CEA use as biomarkers in CMNs, the recent progress made in literature and the main overall results that had been already obtained. CA 15-3 and CEA have prognostic potential for CMNs. There is a wide variation of results for these biomarkers in literature, but despite this, its use must be considered as they provide relevant results in terms of characterization and prognostic in CMNs.(AU)

Development and characterization of poultry collagen-based hybrid hydrogels for bone regeneration

Purpose: Poultry by-products can contribute as an innovative natural source for the development of composites based on polymers and minerals aiming at bone regeneration. The objective of this study was the physicochemical and biological characterization of collagen-based hydrogels crosslinked with ultraviolet (UV)-riboflavin. Methods: Pure hydrogels of 100% collagen (G1) or hybrid hydrogels, 90% collagen:10% apatite (G2), 90% collagen:10% nanokeratin (G3), and 90% collagen:5% apatite:5% nanokeratin (G4) were characterized by scanning electron microscope, Fourier-transform infrared spectroscopy, differential scanning calorimetry, swelling degree and quali-quantitative histological analysis. Ectopic implantation in subcutaneous tissue in mice at one, three and nine weeks allowed to assess the inflammation (neutrophils, lymphocytes, macrophages, and giant cells) and repair (neovascularization, and connective tissue) to determine biocompatibility and the integrity of biomaterials to score their biodegradability. Histomorphometry on critical size defects in rat calvaria at one and three months evaluated the percentage of bone, connective tissue, and biomaterials in all groups. Results: The hydrogels presented porous microstructure, water absorption and physicochemical characteristics compatible with their polymeric and/or mineral composition. All materials exhibited biocompatibility, biodegradability, and low osteoconductivity. G2 showed greater density of new bone and biomaterial than the G1, G3 and G4. Conclusions: The collagen-apatite group formulation suggests potential for development as osteopromoting membrane.

Biochemical changes during heat stress in productive animals with an emphasis on the antioxidant defense system

The problem of free radical pathologies in high temperatures in animal husbandry and veterinary medicine is extremely important scientific and practical. In the process of hyperthermia, the structure of cell membranes, the permeability of membranes, and membrane transport systems change, which cause the oxidation of unsaturated fatty acids and the accumulation of toxic products of a radical nature. This plays an important role in the development of many pathological conditions. The review article describes the problem of the influence of heat stress (HS) on a living organism and the problem of hyperthermia in animal husbandry. Emphasis is placed on the role of the body's antioxidant system in a state of HS and the use of antioxidant drugs as additives into feed for farm animals, aimed at mitigating the course and harmful effects in HS. On the one hand, our goal was to describe in an accessible form the main complex biochemical disorders in the body under HS conditions, which lead to a deterioration in the welfare and loss of productivity of animals. On the other hand, we tried to show the special role of the antioxidant defense system in preventing the development of oxidative stress in its classical representation. Also, we substantiated the choice in favor of using natural and synthetic antioxidants as one of the strategies for the prevention of HS in modern animal husbandry.(AU)

Paraoxonase 1 activity in the sperm-rich portion of boar ejaculates is positively associated with sperm quality

Associations of the activity of the paraoxonase 1 (PON1) enzyme with boar sperm quality still needs to be characterized, since boar ejaculates present distinct portions with differences in sperm concentration and quality. This study evaluated PON1 activity in the serum, in the distinct portions of boar ejaculates and estimated correlations with sperm quality parameters. Ejaculates and blood samples were collected from six boars for three weeks (two per week per boar; n = 36). Serum and post-spermatic portion PON1 activities were positively correlated (P = 0.01) but were both uncorrelated with the PON1 activity in the sperm-rich portion and in the whole ejaculate (P > 0.05). Differences in PON1 activity among boars were only observed in the sperm-rich portion of the ejaculate (P < 0.05). The PON1 activity in the serum and in the post-spermatic portion was generally negatively correlated with parameters of spermatozoa kinetics (P < 0.05). In the sperm-rich portion, PON1 activity was positively correlated with sperm concentration (P < 0.0001), curvilinear distance and velocity (both P < 0.05) and DNA integrity (P < 0.05), but negatively correlated with straightness and linearity (P < 0.05). Thus, boar ejaculates with increased PON1 activity in the sperm-rich portion may present increased concentration and spermatozoa with acceptable curvilinear velocity and distance and DNA integrity, which suggests that PON1 activity may be a biomarker for potential fertility.(AU)

Pseudopterígio em coelho - tratamento com tracolimus / Pseudopterygium in a rabbit - treatment with tracolimus

Background: Pseudopterygium, also known as aberrant conjunctival growth, is poorly described in the literature, althoughit is known that this abnormality is uncommon and affects dwarf rabbits and their crossbreeds. The etiology of this diseaseis unknown, but there are hypotheses that the conjunctival growth cause may have its origins in immunological factors,inflammation, traumatic conditions, or cartilage dysplasias. Thus, this study reports the treatment efficacy applied in arabbit, through the continuous use of tracolimus eye drops, after surgical procedure of conjunctival fold resection, as away of controlling the pseudopterygium in rabbits.Case: This case report discusses the positive results from the surgical and therapeutic conduct of a clinical case attended bythe Ophthalmology and Microsurgical Veterinary Service at the Hospital Veterinário Universitário (HVU) of the UFSM. Thepatient was a male rabbit, sterilized, approximately 2-year-old, crossed with a dwarf rabbit. The owner’s main complaintwas the change in the aspect of the left eye, with progressive worsening in the previous four weeks. In the ophthalmologicalexamination, the animal did not present impaired vision or discomfort, however, a vascularized pink membrane was noted,which consisted of a fold of the bulbar conjunctiva, that grew centripetally and covered 90% of the cornea in 360 degrees.The diagnosis was confirmed through visual inspection and the patient’s history. The eye alteration had a characteristicaspect, described as proliferation of the bulbar conjunctiva over the cornea, in a centripetal manner and without signs ofinflammation. In addition, other ophthalmological alterations were ruled out during the patient’s physical and specificexamination. The patient was referred for anesthetic evaluation and, in addition, pre-surgical blood tests were performed,which were normal, according to the expected ranges for the species...

Pseudopterígio em coelho - tratamento com tracolimus / Pseudopterygium in a rabbit - treatment with tracolimus

Background: Pseudopterygium, also known as aberrant conjunctival growth, is poorly described in the literature, althoughit is known that this abnormality is uncommon and affects dwarf rabbits and their crossbreeds. The etiology of this diseaseis unknown, but there are hypotheses that the conjunctival growth cause may have its origins in immunological factors,inflammation, traumatic conditions, or cartilage dysplasias. Thus, this study reports the treatment efficacy applied in arabbit, through the continuous use of tracolimus eye drops, after surgical procedure of conjunctival fold resection, as away of controlling the pseudopterygium in rabbits.Case: This case report discusses the positive results from the surgical and therapeutic conduct of a clinical case attended bythe Ophthalmology and Microsurgical Veterinary Service at the Hospital Veterinário Universitário (HVU) of the UFSM. Thepatient was a male rabbit, sterilized, approximately 2-year-old, crossed with a dwarf rabbit. The owners main complaintwas the change in the aspect of the left eye, with progressive worsening in the previous four weeks. In the ophthalmologicalexamination, the animal did not present impaired vision or discomfort, however, a vascularized pink membrane was noted,which consisted of a fold of the bulbar conjunctiva, that grew centripetally and covered 90% of the cornea in 360 degrees.The diagnosis was confirmed through visual inspection and the patients history. The eye alteration had a characteristicaspect, described as proliferation of the bulbar conjunctiva over the cornea, in a centripetal manner and without signs ofinflammation. In addition, other ophthalmological alterations were ruled out during the patients physical and specificexamination. The patient was referred for anesthetic evaluation and, in addition, pre-surgical blood tests were performed,which were normal, according to the expected ranges for the species...(AU)

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs / Presença de múltiplos metais e agricultura afetam biomarcadores de estresse oxidativo em caranguejos de água doce (Aegla)

Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams (≤ 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Os metais e agroquímicos estão entre os principais contaminantes aquáticos, podendo desencadear estresse oxidativo em organismos expostos. O objetivo deste trabalho foi avaliar uma possível correlação entre o nível de biomarcadores de estresse oxidativo em Aegla (Crustacea, Anomura) com (i) o conjunto de metais presentes no sedimento e (ii) com os usos da terra, em três bacias hidrográficas distintas. O estudo foi realizado em riachos (≤ 2ª ordem) de bacias hidrográficas do Sul do Brasil (Bacias do Rio Suzana, do Rio Ligeirinho-Leãozinho e do Rio Dourado), as quais foram caracterizadas em função do percentual de usos da terra e do nível de Cu, Cr, Cd, Fe, Mn e Zn no sedimento. As enzimas Catalase (CAT) e Glutationa Redutase (GR), bem como o nível de peroxidação lipídica das membranas (TBARS), foram analisadas em fêmeas adultas. Uma análise de PCA mostrou que a distribuição de metais foi distinta entre as bacias. Cd, Cr e Fe no sedimento correlacionaram positivamente com a CAT e negativamente com TBARS e GR. Entretanto, a bacia do Dourado apresentou os menores níveis destes três metais e os maiores níveis de TBARS, o que pode ser justificado pelo predomínio da agricultura nesta bacia, já que o TBARS correlacionou positivamente com o percentual de uso agrícola. Nesta bacia, a atividade da GR foi mais alta do que nas outras, indicando uma resposta compensatória em relação a inibição da CAT. As bacias do rio Suzana e rio Ligeirinho-Leãozinho apresentaram valores menores de TBARS, o que pode decorrer da indução da CAT em função dos metais acumulados no sedimento. Em síntese, este trabalho indica que nas bacias com maior concentração de metais tóxicos ocorre uma resposta adaptativa de indução da CAT, o que reduz os níveis de TBARS em Aegla. Por outro lado, na bacia com menor contaminação metálica os níveis de TBARS foram primariamente influenciados pelo uso agrícola.

Comparative Analysis on the Duodenal Microbiota Community in Geese Fed with the All-grass or Basal Diet

Geese (Anser cygnoides) possess stronger ability of roughage digestion and utilization than other poultries, hence, it has become the focus of attention of scientists. Duodenal, jejunum and ileum were mainly participated in food digestion and nutrient absorption, while the cecum was responsible for biological fermentation. Effects on the geese's cecal microbiota community by feeding with the all-grass diet have been investigated, however, whether it had an influence on the geese's duodenal microbiota community remains unexplored. To address this problem, geese feeding with the basal diet for 28 days (G1), the basal diet for 28 days and the all-grass diet for the following 14 days (G2), the basal diet for 42 days (G3) were selected, respectively. The duodenal segments of geese were collected and the hypervariable V3-V4 region of the bacterial 16S rRNA gene was sequencing. A total of 4 main phyla and 16 main genera were identified. Moreover, we also successfully identified that two taxa including the Helcococcus and Clostridium could be used as distinguishing biomarkers specific to G2. The functional profiles of the duodenum microbiota were mainly involved in the membrane transport (e.g. ABC transporters), amino acid metabolism, energy metabolism, metabolism of cofactors and vitamins, and cellular processes and signaling pathways in geese feeding with the all-grass diet. In conclusion, the all-grass diet could impact the composition of duodenal microbiota. However, to resolve the underlying mechanism of the fiber digesting and utilization in geese's gut microbiota, the whole intestinal system needs to be assessed by further studies.(AU)

Uso da túnica vaginal canina conservada em glicerina a 98% como curativo biológico no tratamento de feridas cutâneas produzidas em ratos Wistar / Use of canine vaginal tunic preserved in 98% glycerin as biological curative in the treatment of skin produced in wound Wistar rats

Na procura por material alternativo no tratamento de feridas cutâneas, buscou-se com este estudo avaliar o comportamento da túnica vaginal canina conservada em glicerina a 98% como curativo biológico no tratamento de feridas cutâneas provocadas experimentalmente em ratos. Foram utilizados 16 ratos Wistar, nos quais foram induzidas experimentalmente duas lesões cutâneas na região dorsal, sendo uma cranial e a outra caudal. Uma ferida serviu de controle, a qual não recebeu tratamento, a outra recebeu a túnica vaginal recobrindo toda extensão da lesão. A seleção de qual das feridas, cranial ou caudal, iria receber a membrana foi feita de forma aleatória. Os animais foram distribuídos em quatro grupos de igual número para análises clínicas, macroscópicas e histológicas decorridos três, sete, 14 e 21 dias de pós-operatório. Na avaliação clínica e macroscópica pode-se observar que o processo de reparação teve evolução semelhante tanto no grupo controle quanto no grupo membrana. Na análise histopatológica foi constatada grande quantidade de infiltrado inflamatório nos períodos iniciais, no entanto, decorridos 21 dias, a epiderme já se encontrava reparada, e em todos os momentos foi observada maior presença de tecido conjuntivo no grupo membrana, sugerindo maior precocidade na cicatrização. Deste modo, concluiu-se que a túnica vaginal canina pode ser utilizada como nova alternativa no tratamento de feridas cutâneas.

In the search for alternative material in the treatment of cutaneous wounds, this study aimed to evaluate the behavior of canine vaginal tunic conserved in glycerin 98% as a biological dressing in the treatment of cutaneous wounds provoked experimentally in rats. Sixteen animals were used in which two cutaneous lesions were caused in the dorsal region, one cranial and the other caudal. One wound served as a control which did not receive treatment, the other received the vaginal tunic covering all extension of the lesion. The selection of which of the wounds, cranial or caudal, would receive the membrane was made at random. The animals were divided into four groups of equal numbers for clinical, macroscopic, and histological analyzes after three, seven, 14 and 21 postoperative days. In the clinical and macroscopic evaluation, it can be observed that the healing process had similar evolution in both the control group and the membrane group. In the histopathological analysis, a large amount of inflammatory infiltrate was observed in the initial periods; however, after 21 days the epidermis was already healed, and at all times a greater presence of connective tissue was observed in the membrane group, suggesting a greater precocity in healing. Thus, it was concluded that the canine vaginal tunic can be used as a new alternative in the treatment of cutaneous wounds.

Uso da túnica vaginal canina conservada em glicerina a 98% como curativo biológico no tratamento de feridas cutâneas produzidas em ratos Wistar / Use of canine vaginal tunic preserved in 98% glycerin as biological curative in the treatment of skin produced in wound Wistar rats

Na procura por material alternativo no tratamento de feridas cutâneas, buscou-se com este estudo avaliar o comportamento da túnica vaginal canina conservada em glicerina a 98% como curativo biológico no tratamento de feridas cutâneas provocadas experimentalmente em ratos. Foram utilizados 16 ratos Wistar, nos quais foram induzidas experimentalmente duas lesões cutâneas na região dorsal, sendo uma cranial e a outra caudal. Uma ferida serviu de controle, a qual não recebeu tratamento, a outra recebeu a túnica vaginal recobrindo toda extensão da lesão. A seleção de qual das feridas, cranial ou caudal, iria receber a membrana foi feita de forma aleatória. Os animais foram distribuídos em quatro grupos de igual número para análises clínicas, macroscópicas e histológicas decorridos três, sete, 14 e 21 dias de pós-operatório. Na avaliação clínica e macroscópica pode-se observar que o processo de reparação teve evolução semelhante tanto no grupo controle quanto no grupo membrana. Na análise histopatológica foi constatada grande quantidade de infiltrado inflamatório nos períodos iniciais, no entanto, decorridos 21 dias, a epiderme já se encontrava reparada, e em todos os momentos foi observada maior presença de tecido conjuntivo no grupo membrana, sugerindo maior precocidade na cicatrização. Deste modo, concluiu-se que a túnica vaginal canina pode ser utilizada como nova alternativa no tratamento de feridas cutâneas.(AU)

In the search for alternative material in the treatment of cutaneous wounds, this study aimed to evaluate the behavior of canine vaginal tunic conserved in glycerin 98% as a biological dressing in the treatment of cutaneous wounds provoked experimentally in rats. Sixteen animals were used in which two cutaneous lesions were caused in the dorsal region, one cranial and the other caudal. One wound served as a control which did not receive treatment, the other received the vaginal tunic covering all extension of the lesion. The selection of which of the wounds, cranial or caudal, would receive the membrane was made at random. The animals were divided into four groups of equal numbers for clinical, macroscopic, and histological analyzes after three, seven, 14 and 21 postoperative days. In the clinical and macroscopic evaluation, it can be observed that the healing process had similar evolution in both the control group and the membrane group. In the histopathological analysis, a large amount of inflammatory infiltrate was observed in the initial periods; however, after 21 days the epidermis was already healed, and at all times a greater presence of connective tissue was observed in the membrane group, suggesting a greater precocity in healing. Thus, it was concluded that the canine vaginal tunic can be used as a new alternative in the treatment of cutaneous wounds.(AU)

Uso da túnica vaginal canina conservada em glicerina a 98% como curativo biológico no tratamento de feridas cutâneas produzidas em ratos Wistar / Use of canine vaginal tunic preserved in 98% glycerin as biological curative in the treatment of skin produced in wound Wistar rats

Na procura por material alternativo no tratamento de feridas cutâneas, buscou-se com este estudo avaliar o comportamento da túnica vaginal canina conservada em glicerina a 98% como curativo biológico no tratamento de feridas cutâneas provocadas experimentalmente em ratos. Foram utilizados 16 ratos Wistar, nos quais foram induzidas experimentalmente duas lesões cutâneas na região dorsal, sendo uma cranial e a outra caudal. Uma ferida serviu de controle, a qual não recebeu tratamento, a outra recebeu a túnica vaginal recobrindo toda extensão da lesão. A seleção de qual das feridas, cranial ou caudal, iria receber a membrana foi feita de forma aleatória. Os animais foram distribuídos em quatro grupos de igual número para análises clínicas, macroscópicas e histológicas decorridos três, sete, 14 e 21 dias de pós-operatório. Na avaliação clínica e macroscópica pode-se observar que o processo de reparação teve evolução semelhante tanto no grupo controle quanto no grupo membrana. Na análise histopatológica foi constatada grande quantidade de infiltrado inflamatório nos períodos iniciais, no entanto, decorridos 21 dias, a epiderme já se encontrava reparada, e em todos os momentos foi observada maior presença de tecido conjuntivo no grupo membrana, sugerindo maior precocidade na cicatrização. Deste modo, concluiu-se que a túnica vaginal canina pode ser utilizada como nova alternativa no tratamento de feridas cutâneas.

In the search for alternative material in the treatment of cutaneous wounds, this study aimed to evaluate the behavior of canine vaginal tunic conserved in glycerin 98% as a biological dressing in the treatment of cutaneous wounds provoked experimentally in rats. Sixteen animals were used in which two cutaneous lesions were caused in the dorsal region, one cranial and the other caudal. One wound served as a control which did not receive treatment, the other received the vaginal tunic covering all extension of the lesion. The selection of which of the wounds, cranial or caudal, would receive the membrane was made at random. The animals were divided into four groups of equal numbers for clinical, macroscopic, and histological analyzes after three, seven, 14 and 21 postoperative days. In the clinical and macroscopic evaluation, it can be observed that the healing process had similar evolution in both the control group and the membrane group. In the histopathological analysis, a large amount of inflammatory infiltrate was observed in the initial periods; however, after 21 days the epidermis was already healed, and at all times a greater presence of connective tissue was observed in the membrane group, suggesting a greater precocity in healing. Thus, it was concluded that the canine vaginal tunic can be used as a new alternative in the treatment of cutaneous wounds.

Diagnosis and treatment of HEp-2 cells contaminated with mycoplasma / Diagnóstico e tratamento de células HEp-2 contaminadas por micoplasma

Contamination of primary and cell cultures by mycoplasmas is one of the main economic and biological pitfalls in basic research, diagnosis and manufacture of biotechnological products. It is a common issue which may be difficult to conduct surveillance on. Mycoplasma presence may affect several physiological parameters of the cell, besides being considered an important source of inaccurate and/or non-reproducible scientific results. Each cell type presents characteristical symptoms, mainly morphological, that indicate a contamination by mycoplasma. HEp-2 cells originate from carcinoma of the larynx and are, therefore, part of the respiratory tract, which is one of mycoplasma habitats. Despite the importance these cells in several biological research (evaluation of cell proliferation and migration, apoptosis, antiviral and antitumor compounds), the alterations induced by mycoplasma contamination in HEp-2 cells have not yet been described. Here, we describe the progressive morphological alterations in culture of HEp-2 cells infected with mycoplasma, as well as the-diagnosis of the infection and its treatment. Mycoplasma contamination described within this work led to cytoplasm elongation, cell-to-cell spacing, thin plasma membrane projections, cytoplasmic vacuoles, fusion with neighboring cells, and, finally, cell death. Contamination was detected by fluorescence imaging (DAPI) and PCR reactions. The cultures were treated with BM-Cyclin antibiotic to eliminate contamination. The data presented here will be of relevance to researchers whose investigations involve cell culture, especially respiratory and HEp-2 cells.(AU)

A contaminação de culturas primárias e celulares por micoplasmas é uma das principais armadilhas econômicas e biológicas da pesquisa básica, diagnóstico e fabricação de produtos biotecnológicos. Trata-se de uma contaminação rotineira, mas de difícil acompanhamento. A presença de micoplasma pode afetar vários parâmetros fisiológicos da célula, além de ser considerada uma importante fonte de resultados científicos imprecisos e/ou não reprodutíveis. Cada tipo de célula apresenta sintomas característicos, principalmente morfológicos, que indicam uma contaminação por micoplasma. As células HEp-2 são originárias do carcinoma da laringe e, portanto, fazem parte do trato respiratório, um dos habitats do micoplasma. Apesar da importância destas células em diversas pesquisas biológicas (avaliação da proliferação e migração celular, apoptose, compostos antivirais e antitumorais), as alterações decorrentes da contaminação por micoplasma nestas células ainda não foi descrita. Aqui, descrevemos as alterações morfológicas progressivas na cultura de células HEp-2 infectadas por micoplasma, bem como o diagnóstico da infecção e seu tratamento. A contaminação por micoplasma descrita neste trabalho resultou em alongamento citoplasmático, espaçamento entre células, projeções delgadas da membrana plasmática, vacúolos citoplasmáticos, fusão de células vizinhas e, finalmente, morte celular. A contaminação foi detectada por imagens de fluorescência (DAPI) e reações de PCR. As culturas foram tratadas com antibiótico BM-Cyclin para eliminar a contaminação. Os dados aqui apresentados serão de relevância para pesquisadores cujas investigações envolvem cultura celular, principalmente células respiratórias e HEp-2.(AU)

Identification and Characterization of the ATG8, a Marker of Eimeria tenella Autophagy / Identificação e Caracterização do ATG8, um marcador de autofagia em Eimeria tenella

Autophagy plays an important role in maintaining cell homeostasis through degradation of denatured proteins and other biological macromolecules. In recent years, many researchers focus on mechanism of autophagy in apicomplexan parasites, but little was known about this process in avian coccidia. In our present study. The cloning, sequencing and characterization of autophagy-related gene (Etatg8) were investigated by quantitative real-time PCR (RT-qPCR), western blotting (WB), indirect immunofluorescence assays (IFAs) and transmission electron microscopy (TEM), respectively. The results have shown 375-bp ORF of Etatg8, encoding a protein of 124 amino acids in E. tenella, the protein structure and properties are similar to other apicomplexan parasites. RT-qPCR revealed Etatg8 gene expression during four developmental stages in E. tenella, but their transcriptional levels were significantly higher at the unsporulated oocysts stage. WB and IFA showed that EtATG8 was lipidated to bind the autophagosome membrane under starvation or rapamycin conditions, and aggregated in the cytoplasm of sporozoites and merozoites, however, the process of autophagosome membrane production can be inhibited by 3-methyladenine. In conclusion, we found that E. tenella has a conserved autophagy mechanism like other apicomplexan parasites, and EtATG8 can be used as a marker for future research on autophagy targeting avian coccidia.(AU)

A autofagia desempenha um papel importante na manutenção da homeostase celular através da degradação de proteínas desnaturadas e outras macromoléculas biológicas. Nos últimos anos, muitos pesquisadores se concentraram no mecanismo da autofagia em parasitas apicomplexos, mas pouco se sabe sobre esse processo na coccidia aviária. No presente estudo, a clonagem, sequenciamento e caracterização de gene relacionado à autofagia Etatg8 foram investigados pela PCR quantitativa em tempo real (RT-qPCR), mancha ocidental (WB), ensaios indiretos de imunofluorescência (IFAs) e microscopia eletrônica de transmissão (TEM), respectivamente. Os resultados mostraram que o gene Etatg8 de E. tenella possui uma ORF de 375 bp, codificando uma proteína de 124 aminoácidos com estrutura e propriedades semelhantes à de outros apicomplexos. RT-qPCR revelou que Etatg8 é expresso durante os quatro estágios de desenvolvimento de E. tenella. Entretanto, seus níveis transcricionais foram significativamente mais elevados na fase de oocisto não esporulados. Os ensaios de manchas ocidental (WB) e de imunofluorescência (IFA) mostraram que a proteína EtATG8 foi lipidada para ligar-se à membrana do autofagossomo sob condições de deficiência nutritiva (em presença de rapamicina) e se agregar no citoplasma de esporozoítas e merozoítas. No entanto, o processo de produção de membrana do autofagossomo pode ser inibido por um inibidor de autofagia (3-meetiladeninatiladenina, 3-MA). Em conclusão, foi demonstrado que E. tenella tem um mecanismo de autofagia conservado, semelhante ao de outros parasitas apicomplexos, e que EtATG8 pode ser usado como um marcador para futuras pesquisas sobre autofagia direcionada à coccidiose aviária.(AU)

Hérnia perineal em cães / Perineal hernia in dogs

Background: Perineal hernia is a serious disease characterized by weakening or atrophy and separation of the muscles andfasciae of the perineal musculature, followed by the caudal displacement of pelvic and abdominal organs to the perineumregion. Treatment is invariably surgical and several approaches have been proposed, but complication and recurrence ratesremain high. This study aimed to evaluate 120 cases of perineal hernia treated at the Veterinary Hospital of Uberaba (HVU)from 2005 to 2020, addressing the clinical and surgical aspects and the postoperative period, seeking to identify the mostrelevant factors to improve care and treatment of future patients affected by this disease.Materials, Methods & Results: The medical records of dogs with a definitive diagnosis of perineal hernia were collected.The prevalence of perineal hernia was calculated. Data were obtained regarding sex, being castrated or not, age, bodymass, race, clinical signs, affected side, possibility of hernia reduction, treatments used, associated pathologies. Of the 120cases, only 69 underwent surgery at the HVU and from these cases data were obtained regarding hernia content, surgicaltechniques, surgical wires used, recurrences and postoperative complications. Fisher’s exact test was applied to assess theinfluence of the type of surgical treatment and surgical thread on the occurrence of complications and recurrences. Perineal hernia was the second most frequently observed hernia. Mixed breed, male, non-neutered and geriatric dogs were themost affected. The most frequently observed clinical signs were related to the digestive and urinary tract, consistent withthe most commonly found hernia contents (bladder, prostate and rectum). The simple herniorrhaphy technique was themost used and showed a high number of complications. The second most used technique was...