The Role of Senescent CD8+T Cells in the Pathogenesis of Disseminated Leishmaniasis.

Disseminated leishmaniasis (DL) caused by L. braziliensis is characterized by the presence of 10 to more than 1000 lesions spread on the body. While protection against Leishmania is mediated by macrophages upon activation by IFN-γ produced by CD4+T cells, the pathology of disseminated leishmaniasis (DL) could be mediated by macrophages, NK, and CD8+T cells. Herein, we evaluate the participation of senescent CD8+T cells in the pathogenesis of DL. Methods: Peripheral blood mononuclear cells (PBMCs), biopsies, co-cultures of CD8+T cells with uninfected and infected macrophages (MØ), and PBMC cultures stimulated with soluble L. braziliensis antigen (SLA) for 72 h from patients with cutaneous leishmaniasis (CL) and DL were used to characterize senescent CD8+T cells. Statistical analysis was performed using the Mann-Whitney and Kruskal-Wallis tests, followed by Dunn's. Results: Patients with DL have an increase in the frequency of circulating CD8+T cells that present a memory/senescent phenotype, while lesions from DL patients have an increase in the frequency of infiltrating CD8+T cells with a senescent/degranulation phenotype. In addition, after specific stimuli, DL patients' circulating CD8+T with memory/senescent profile, showing degranulation characteristics, increased upon SLA stimuli, and those specific CD8+T cells from DL patients had an increased degranulation phenotype, causing more apoptosis of infected target cells. Conclusions: DL patients show a higher frequency of cytotoxic senescent CD8+T cells compared to CL patients, and that could promote the lysis of infected cells, although without parasite killing, releasing Leishmania to the extracellular compartment, contributing to the spread of parasites.

A Single-Port, Multiple-Access, Custom-Made Device Used in Laparoscopically Assisted Cryptorchidectomy in Standing Horses-A Preliminary Study.

This study evaluates a new multiport device with single access to the abdominal cavity produced with routine hospital supplies that could be applied to laparoscopically assisted cryptorchidectomy in standing horses. Initially, the new device was evaluated on five cadavers of bovine fetuses (n = 5), placed assisted in a minilaparotomy performed in the flank region. Subsequently, the device was evaluated in four cryptorchid horses treated during the hospital routine. During the evaluation of the new device, the possibilities of exploring the abdominal cavity, inspection, and intra-abdominal manipulation with two Babcock forceps were verified. The possibilities were described, and surgical time data were recorded and analyzed using descriptive statistics. In the cadavers, a wide exploration of the abdominal cavity was possible, with a laparoscopic inspection through the right paralumbar fossa and manipulation of intra-abdominal structures with Babcock forceps inserted by the new device. In cryptorchid horses, laparoscopically assisted cryptorchidectomy with a new device was feasible in two patients, and in the others, it allowed the diagnosis of adhesions and ectopic locations in the inguinal region of testicles retained in the cavity. Therefore, the new device was efficient in exploring the inguinal region of cryptorchid horses in the standing position. The present study is preliminary and can support future studies that aim to improve the developed prototype.

Disseminated Leishmaniasis, a Severe Form of Leishmania braziliensis Infection.

Disseminated leishmaniasis (DL) is an emergent severe disease manifesting with multiple lesions. To determine the relationship between immune response and clinical and therapeutic outcomes, we studied 101 DL and 101 cutaneous leishmaniasis (CL) cases and determined cytokines and chemokines in supernatants of mononuclear cells stimulated with leishmania antigen. Patients were treated with meglumine antimoniate (20 mg/kg) for 20 days (CL) or 30 days (DL); 19 DL patients were instead treated with amphotericin B, miltefosine, or miltefosine and meglumine antimoniate. High levels of chemokine ligand 9 were associated with more severe DL. The cure rate for meglumine antimoniate was low for both DL (44%) and CL (60%), but healing time was longer in DL (p = 0.003). The lowest cure rate (22%) was found in DL patients with >100 lesions. However, meglumine antimoniate/miltefosine treatment cured all DL patients who received it; therefore, that combination should be considered as first choice therapy.

Computed Tomography Evaluation of Frozen or Glycerinated Bradypus variegatus Cadavers: A Comprehensive View with Emphasis on Anatomical Aspects.

Bradypus variegatus has unique anatomical characteristics, and many of its vascular and digestive tract aspects have yet to be clearly understood. This lack of information makes clinical diagnoses and surgical procedures difficult. The aim of this study was to evaluate the anatomical aspects of frozen and glycerinated corpses of B. variegatus using computed tomography (CT), emphasizing vascular and digestive contrast studies. Nine corpses that died during routine hospital were examined via CT in the supine position with scanning in the craniocaudal direction. In frozen cadavers, the contrast was injected into a cephalic vein after thawing and, subsequently, was administered orally. In addition to bone structures, CT allowed the identification of organs, soft tissues, and vascular structures in specimens. Visualization of soft tissues was better after contrast been administered intravenously and orally, even without active vascularization. Furthermore, the surfaces of the organs were highlighted by the glycerination method. With this technique, it was possible to describe part of the vascularization of the brachial, cervical, thoracic, and abdominal regions, in addition to highlighting the esophagus and part of the stomach. CT can be another tool for the evaluation of B. variegatus cadavers by anatomists or pathologists, contributing to the identification of anatomical structures.

MicroRNAs regulating macrophages infected with Leishmania L. (V.) Braziliensis isolated from different clinical forms of American tegumentary leishmaniasis.

Background: Leishmaniasis is an infectious disease caused by protozoa of the genus Leishmania. There are still no vaccines, and therapeutic options are limited, indicating the constant need to understand the fine mechanisms of its pathophysiology. An approach that has been explored in leishmaniasis is the participation of microRNAs (miRNAs), a class of small non-coding RNAs that act, in most cases, to repress gene expression. miRNAs play a role in the complex and plastic interaction between the host and pathogens, either as part of the host's immune response to neutralize infection or as a molecular strategy employed by the pathogen to modulate host pathways to its own benefit. Methods: Monocyte-derived macrophages from healthy subjects were infected with isolates of three clinical forms of L. braziliensis: cutaneous (CL), mucosal (ML), and disseminated (DL) leishmaniasis. We compared the expression of miRNAs that take part in the TLR/NFkB pathways. Correlations with parasite load as well as immune parameters were analyzed. Results: miRNAs -103a-3p, -21-3p, 125a-3p -155-5p, -146a-5p, -132- 5p, and -147a were differentially expressed in the metastatic ML and DL forms, and there was a direct correlation between miRNAs -103a-3p, -21-3p, -155-5p, -146a-5p, -132-5p, and -9-3p and parasite load with ML and DL isolates. We also found a correlation between the expression of miR-21-3p and miR-146a-5p with the antiapoptotic gene BCL2 and the increase of viable cells, whereas miR-147a was indirectly correlated with CXCL-9 levels. Conclusion: The expression of miRNAs is strongly correlated with the parasite load and the inflammatory response, suggesting the participation of these molecules in the pathogenesis of the different clinical forms of L. braziliensis.

Leishmania braziliensis exosomes activate human macrophages to produce proinflammatory mediators.

Exosomes, organelles measuring 30-200nm, are secreted by various cell types. Leishmania exosomes consist of many proteins, including heat shock proteins, annexins, Glycoprotein 63, proteins exerting signaling activity and those containing mRNA and miRNA. Studies have demonstrated that Leishmania donovani exosomes downregulate IFN-γ and inhibit the expression of microbicidal molecules, such as TNF and nitric oxide, thus creating a microenvironment favoring parasite proliferation. Despite lacking immunological memory, data in the literature suggest that, following initial stimulation, mononuclear phagocytes may become "trained" to respond more effectively to subsequent stimuli. Here we characterized the effects of macrophage sensitization using L. braziliensis exosomes prior to infection by the same pathogen. Human macrophages were stimulated with L. braziliensis exosomes and then infected with L. braziliensis. Higher levels of IL-1ß and IL-6 were detected in cultures sensitized prior to infection compared to unstimulated infected cells. Moreover, stimulation with L. braziliensis exosomes induced macrophage production of IL-1ß, IL-6, IL-10 and TNF. Inhibition of exosome secretion by L. braziliensis prior to macrophage infection reduced cytokine production and produced lower infection rates than untreated infected cells. Exosome stimulation also induced the consumption/regulation of NLRP3 inflammasome components in macrophages, while the blockade of NLRP3 resulted in lower levels of IL-6 and IL-1ß. Our results suggest that L. braziliensis exosomes stimulate macrophages, leading to an exacerbated inflammatory state that may be NLRP3-dependent.

Laparoscopy-assisted percutaneous correction of abdominal wall defects in the umbilical region in a cadaveric model of bovine fetus.

Abdominal wall defects in calves are commonly diagnosed and treated via laparotomy. This technique has witnessed several advancements in the management of these disorders. This study aimed to create a study model and evaluate the feasibility of video-assisted percutaneous correction of abdominal wall defects in bovine fetuses (corpses) compared with the conventional technique. Sixteen bovine fetuses from pregnant cows slaughtered in slaughterhouses were included in this study. The fetuses were categorized into the control group (CG, n = 8), which was subjected to umbilical abdominorrhaphy via laparotomy, and the video-surgical group (VG, n = 8), which received video-assisted percutaneous sutures with two lateral accesses on the right flank. An abdominal wall defect was created in the VG group to generate a study model, which was corrected using the laparoscopic technique. The procedures were performed in two steps. The first step consisted of creating an abdominal wall defect in the umbilical region by laparoscopic approach in an iatrogenic manner (Step 1: E1). The second stage consisted of conventional abdominorrhaphy of the umbilical region wall defect in the CG group and video-assisted percutaneous suturing of the edges of the iatrogenic abdominal wall defect in the VG group, until reversal of the laparoscopic accesses (Step 2: E2). Step 1 showed no statistically significant difference between the two groups. However, a significant statistical difference (p < 0.0001) was observed between the two groups in step 2. The surgical time of step 2 was longer in the CG group (33.10 ± 0.43 minutes) than that in the VG group (10.13 ± 0.68 minutes, p < 0.0001), and the total surgical time was also longer in the CG group (38.48 ± 0.35 minutes) than that in the VG group (15.86 ± 0.67 minutes). The proposed laparoscopic technique allowed the creation of a study model for video-assisted percutaneous suturing with two portals and reduced the surgical time compared with the conventional technique. However, this method needs to be studied further in live animals.

Purple acid phosphatases in coffee-genome-wide identification and the transcriptional profile of selected members in roots upon Pi starvation.

Phosphorus (P) availability is determinant for crop productivity and, despite the sufficient amount of this nutrient in arable land, most of it remains insoluble, leading to the need of high fertilizer input. To cope with P scarcity forecasts and also for cropping costs alleviation, genotypes better adapted to promote P solubilization and absorption are required, especially for perennial crops. Coffee is one of these important perennial crops cultivated in soils with low P availability, and thus we aimed to study adaptive strategies to coffee genotypes in acquire phosphorus. In this study, we focused on rhizosphere phosphatase activity, a major characteristic related to P solubilization from organic pools. To explore the genetic basis of this characteristic, we firstly identified 29 Purple Acid Phosphatases (PAP) genes in C. canephora genome and selected five candidates with higher potential to encode secreted PAPs. We found that C. arabica genotypes have diverse profiles of rhizosphere phosphatase activity, as well as microbial biomass carbon, which we measured to explore the impact of the plant on microbiota and also to discriminate the phosphatase activity origin (plant or microorganism-derived). We selected two C. arabica cultivars with contrasting phosphatase activity and found that one PAP gene has a correlated gene expression profile with this characteristic. This work explores coffee adaptative responses to P starvation conditions, and the information provided can further contribute to breeding programs aiming better adapted genotypes for sustainable agriculture in face of current challenges. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03399-6.

MicroRNAs expression profiles in early responses to different levels of water deficit in Setaria viridis.

Water deficit is a major constraint for crops of economic importance in almost all agricultural regions. However, plants have an active defense system to adapt to these adverse conditions, acting in the reprogramming of gene expression responsible for encoding microRNAs (miRNAs). These miRNAs promote the regulation to the target gene expression by the post-transcriptional (PTGS) and transcriptional gene silencing (TGS), modulating several pathways including defense response to water deficit. The broader knowledge of the miRNA expression profile and its regulatory networks in response to water deficit can provide evidence for the development of new biotechnological tools for genetic improvement of several important crops. In this study, we used Setaria viridis accession A10.1 as a C4 model plant to widely investigate the miRNA expression profile in early responses to different levels of water deficit. Ecophysiological studies in Setaria viridis under water deficit and after rewatering demonstrated a drought tolerant accession, capable of a rapid recovery from the stress. Deep small RNA sequencing and degradome studies were performed in plants submitted to drought to identify differentially expressed miRNA genes and their predicted targets, using in silico analysis. Our findings showed that several miRNAs were differentially modulated in response to distinctive levels of water deficit and after rewatering. The predicted mRNA targets mainly corresponded to genes related to cell wall remodeling, antioxidant system and drought-related transcription factors, indicating that these genes are rapidly regulated in early responses to drought stress. The implications of these modulations are extensively discussed, and higher-effect miRNAs are suggested as major players for potential use in genetic engineering to improve drought tolerance in economically important crops, such as sugarcane, maize, and sorghum. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01226-z.

New hosts for a snake's helminth: First report of intermediate and definitive hosts naturally infected by Ophidascaris arndti (Ascarididae) in the wild.

We wish to report the occurrence of adult nematodes Ophidascaris arndti (Ascarididae) naturally infecting a new definitive host, the Fonseca's lancehead Bothrops fonsecai (Viperidae), and third-stage larvae of O. arndti parasitizing a new intermediate host, the montane grass mouse Akodon montensis (Cricetidae), both found in the Atlantic Forest of the state of Rio de Janeiro, Brazil. We elucidated the morphological characteristics of both adults and larvae using light and scanning electron microscopy (SEM). Taxonomic affinities between larvae and adult worms were assessed using MT-CO1 gene sequences. Adult and larval gene sequences formed a well-supported clade and had low pairwise p-distances, suggesting that they are conspecific. Our phylogenies also supported the 'arndti', 'filaria', and 'obconica' groups as independent lineages and confirmed the allocation of Ophidascaris within the family Ascarididae, although as an early offshoot. This is the first report of natural infection of this helminth's larvae in a wild intermediate host.

Pioglitazone, a Peroxisome Proliferator-Activated Receptor-γ Agonist, Downregulates the Inflammatory Response in Cutaneous Leishmaniasis Patients Without Interfering in Leishmania braziliensis Killing by Monocytes.

Patients with cutaneous leishmaniasis (CL) due to Leishmania braziliensis infection have an exacerbated inflammatory response associated with tissue damage and ulcer development. An increase in the rate of patients who fail therapy with pentavalent antimony has been documented. An adjuvant therapy with an anti-inflammatory drug with the potential of Leishmania killing would benefit CL patients. The aim of the present study was to investigate the contribution of peroxisome proliferator-activated receptor-γ (PPAR-γ) activation by pioglitazone in the regulation of the inflammatory response and L. braziliensis killing by monocytes. Pioglitazone is an oral drug used in the treatment of diabetes, and its main mechanism of action is through the activation of PPAR-γ, which is expressed in many cell types of the immune response. We found that activation of PPAR-γ by pioglitazone decreases the inflammatory response in CL patients without affecting L. braziliensis killing by monocytes. Our data suggest that pioglitazone may serve as an adjunctive treatment for CL caused by L. braziliensis.

Population genetic structure and morphological diversity of Cruzia tentaculata (Nematoda: Ascaridida), a parasite of marsupials (Didelphinae), along the Atlantic Forest on the eastern coast of South America.

Cruzia tentaculata is a helminth parasite of marsupials and has a wide geographic distribution from Mexico to Argentina. The aim of this study was to analyse the genetic population structure of this nematode along the Atlantic Forest biome. Cruzia tentaculata specimens were recovered from Didelphis aurita, Didelphis albiventris and Philander quica in 9 localities. Morphological and morphometric data were investigated for phenotypic diversity among localities and hosts using multivariate discriminant analysis of principal components. Phylogenetic relationships of C. tentaculata were determined using maximum likelihood and Bayesian inference. The population structure was analysed by fixation indices, molecular variance analysis, Tajima's D and Fu's Fs neutrality tests, Mantel tests and Bayesian clustering analysis. A higher significant morphometric difference for males was observed between localities. In the haplogroup networks, 2 groups were recovered, separating locations from the north and from the south/southeast. The morphometric variation in C. tentaculata between different localities was compatible with this north and southeast/south pattern, suggesting adaptation to different ecological conditions. Population genetic analyses suggested a pattern of evolutionary processes driven by Pleistocene glacial refugia in the northeast and southeast of the Atlantic Forest based on the distribution of genetic diversity.

CRISPR/Cas- and Topical RNAi-Based Technologies for Crop Management and Improvement: Reviewing the Risk Assessment and Challenges Towards a More Sustainable Agriculture.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated gene (Cas) system and RNA interference (RNAi)-based non-transgenic approaches are powerful technologies capable of revolutionizing plant research and breeding. In recent years, the use of these modern technologies has been explored in various sectors of agriculture, introducing or improving important agronomic traits in plant crops, such as increased yield, nutritional quality, abiotic- and, mostly, biotic-stress resistance. However, the limitations of each technique, public perception, and regulatory aspects are hindering its wide adoption for the development of new crop varieties or products. In an attempt to reverse these mishaps, scientists have been researching alternatives to increase the specificity, uptake, and stability of the CRISPR and RNAi system components in the target organism, as well as to reduce the chance of toxicity in nontarget organisms to minimize environmental risk, health problems, and regulatory issues. In this review, we discuss several aspects related to risk assessment, toxicity, and advances in the use of CRISPR/Cas and topical RNAi-based technologies in crop management and breeding. The present study also highlights the advantages and possible drawbacks of each technology, provides a brief overview of how to circumvent the off-target occurrence, the strategies to increase on-target specificity, the harm/benefits of association with nanotechnology, the public perception of the available techniques, worldwide regulatory frameworks regarding topical RNAi and CRISPR technologies, and, lastly, presents successful case studies of biotechnological solutions derived from both technologies, raising potential challenges to reach the market and being social and environmentally safe.

Population genetic structure and phenotypic diversity of Aspidodera raillieti (Nematoda: Heterakoidea), a parasite of Didelphini marsupials in Brazil's South and Southeast Atlantic Forest.

BACKGROUND: The population genetics of parasites may be influenced by host specificity, life cycle, host geographical range, evolutionary history, and host population structure. The nematode Aspidodera raillieti infects different marsupial and rodent hosts in the Nearctic and Neotropical regions, implying a gene flow among populations. However, niche diversification of the main hosts of A. raillieti in superimposed areas may provide conditions for population genetic structuring within this parasite species. We examined the genetic structuring of A. raillieti infecting three marsupial species co-occurring along the South and Southeast Brazilian Atlantic Forest, a hotspot of biodiversity. METHODS: We employed morphometric analyses and partial mitochondrial cytochrome c oxidase I gene sequences (MT-CO1) to characterize populations via phylogenetic and phylogeographic analyses. RESULTS: Among 175 A. raillieti specimens recovered from the marsupial hosts Didelphis aurita, D. albiventris, and Philander quica, we identified 99 MT-CO1 haplotypes forming four haplogroups and four clades in networks and phylogenetic trees, respectively. Clades I and II encompassed parasites of D. albiventris from the South region, clade III comprised parasites of D. aurita from the South and Southeast regions, and clade IV encompassed parasites of D. aurita and D. albiventris from the South and Southeast regions and parasites of P. quica from the South region. High genetic differentiation between clades, with a high fixation index and greater genetic variation in the analysis of molecular variance (AMOVA), indicated low gene flow between clades. Haplotypes shared among host species revealed a lack of host specificity. A significant correlation in the Mantel test suggested parasite isolation by distance, while there was no evidence of geographical structure between populations. Negative neutrality test values for clades III and IV suggested recent population expansion. Morphometric differentiation between A. raillieti specimens recovered from different host species, as well as from different localities, was more evident in males. CONCLUSION: The genetic structure of A. raillieti populations in the South and Southeast Atlantic Forest resulted from historical events rather than from current geographical distribution or host specificity. We also demonstrate morphometric variation associated with host species and localities, suggesting phenotypic plasticity to host attributes and to spatial variables.

Helminths of sigmodontine rodents in an agroforestry mosaic in the Brazilian Atlantic Forest: Patterns and processes of the metacommunity structure.

Phylogenetically or taxonomically related hosts may harbour similar parasite communities due to phylogenetic conservatism. In addition, host attributes may favour their exposure to parasites. This study aimed to characterize the helminth fauna of sigmodontine rodents in an Atlantic Forest area in northeastern Brazil and determine the pattern of the helminth metacommunity structure. The influence of host attributes and host taxonomy on the metacommunity structure was also investigated. The most abundant helminth species were Raillietina sp. and Hassalstrongylus lauroi. Euryoryzomys russatus was the most infected host species for helminth parasites, as approximately 81% (35/43) of the animals were infected by at least one helminth species. The helminth metacommunity structure was coherent at both the infracommunity and the component community scales, indicating that species responded to the same environmental gradient. A quasi-Clementsian pattern was observed for the infracommunities, indicating the occurrence of compartments of parasite species that were substituted along the environmental gradient, which was formed by host individuals. A quasi-Gleasonian pattern was found at the component community scale, showing random boundary clumping, which is consistent with the individualistic responses of parasite species to each host species. These patterns corroborated the high values of beta-diversity observed, indicating high species turnover among communities at both scales. Host taxonomic distance was the most important variable explaining the patterns of the helminth metacommunity structure.

Factors related to the suppression of the antitumour immune response in female dogs with inflammatory mammary carcinoma.

Inflammatory mammary carcinoma (IMC), a neoplasia affecting women and female dogs, is considered an aggressive cancer with high metastatic potential and a low survival rate. Studies focused on the tumour microenvironment indicate that the aggressive behaviour of this tumour is primarily correlated with immunological factors as well as inflammation. The objective of this study was to analyse the possible strategies used by the tumour cells to suppress the immune response in female dogs with IMC. Forty-six female dogs were divided into three groups: control (C, n = 10), IMC (n = 14) and mammary carcinoma (MC, n = 22). Clinical-pathological evaluations, survival at follow-up, immunophenotyping of leukocytes in peripheral blood and tumours, and immunohistochemical evaluation of CD4+, granzyme B, perforin and FAS-L were performed. Clinical and pathological results showed a higher frequency of the primary form of neoplasia, solid arrays of tumor cells and a lower survival rate in the IMC group (30 days). Morphometric analysis of inflammatory infiltrate revealed more lymphocytes and macrophages in the IMC group. Immunophenotyping analysis of peripheral blood revealed a higher frequency of CD8+ T-cells (p = 0.0017), a lower frequency of CD4+ T-cells (p <0.0001), and significantly higher mean MHCI and MHCII CD14+ fluorescence intensity in the IMC group (p = 0.038 and p = 0.0117, respectively). The immunohistochemical evaluation of tumour sections showed fewer FAS-L-positive inflammatory cells in the IMC group. These results suggest the important contribution of CD8+ T-cells, macrophages and FAS-L in the aggressiveness of IMC.

Animal Model of Video-Assisted Cecum and Ileum Instrumentation for Equine Visceral Pain Study.

This study aimed to describe an animal model for studying equine visceral pain using minimally invasive and video-assisted cecum and ileum instrumentation. The access to the cecum and ileum was affected because of a previous typhlostomy. For video-assisted distention of the cecum and ileum, a distention device, which we developed using an endotracheal Rusch probe, was used, adapted, and coupled to a cuffometer to inflate and measure the pressure of the cuff attached to its distal portion. In a video-assisted manner, the distal portion of the device was introduced into the cecum and ileum, which contained the cuff in its distal portion, properly positioning it in the lumen. The cuff of the distension device was insufflated after the measurement of baseline physiological parameters of the animals and video-assisted confirmation of its right placement in the cecum and ileum lumen (M0). Was performed in one moment through two simultaneous cuff compressions and 1 minute of animal observation to evaluate the degree of abdominal discomfort manifestations (M1). To cease these stimuli, the cuff was deflated by disconnecting the extensor of the distension device attached to its proximal portion (M2). The procedure was easily performed in most cases. Slow and progressive insufflation allowed subjective adjustment of the intensity of the pain stimulus based on behavioral manifestations. Even with a low rate of complications, the model is feasible and reproducible for studies on visceral pain and the evaluation of analgesic effects.

Surgical Description of Laparoscopic Ovum Pick-Up in Buffalo Calves.

The technique of laparoscopic oocyte aspiration has been increasingly used in animals; however, there are few records of its use in buffaloes. To describe this technique, six suckling Murrah buffaloes aged between 3 and 5 months were used. Three laparoscopic ovum pick-ups were performed in each animal, with intervals of 15 days between surgeries, completing a total of 18 procedures. The technique used three surgical ports with optics and a high-definition video camera. The introduction of the first portal and insufflation of the abdomen was performed through the open technique, with aspiration using a 20 G needle transabdominally and a vacuum pump calibrated at 50 mmHg. The mean complete surgical time from anesthesia to the removal of the animal from the litter was 49 ± 9.8 min. There were 27.8% cases of insufflation on the wrong side of the omentum. The oocyte recovery rate of 60.3% remained within the normal range. However, the rate of viable oocytes recovered was low, with only 40.8% of those recovered undergoing in vitro embryo production (IVEP). These data demonstrate that this simple, minimally invasive technique is an excellent reproductive tool for the genetic improvement of buffalo species.

Knowledge about the disease and adherence to treatment in patients with diabetes

Introduction: Glycemic decompensation in diabetes is one of the major factors for the development of chronic disease complications. Factors involved in the adequate control of diabetes include adherence to pharmacological treatment and knowledge about the disease.Methods: Cross-sectional study on the factors associated with adherence to drug treatment and knowledge about diabetes in diabetic patients treated at Hospital Universitário de Santa Maria between 2018 and 2019, based on the validated Morisky-Green test and on the Diabetes Knowledge Questionnaire.Results: A total of 201 patients diagnosed with diabetes were included, the majority (85.6%) of which had type 2 diabetes and were white (75.6%), with a mean age of 59.4 years. An association between insufficient knowledge about diabetes and patients with type 2 diabetes was observed. An association was found between patients with type 2 diabetes using insulin and non-adherence to drug treatment compared with patients with type 2 diabetes who did use insulin. The research also showed that non-adherence to drug treatment was associated with higher occurrence of hypoglycemia compared with patients who adhered to drug treatment.Conclusion: The data obtained in our study allows us to conclude that non-adherence to pharmacological treatment makes diabetes therapy more complicated and worsens the prognosis.