Detection of CD4+ and CD8+ lymphocytes in the intestine of broiler chicks treated with lactobacillus spp. and challenged with Salmonella enterica serovar enteritidis

The expression of immune response as a leukocytic infiltrate by CD4+ and CD8+ cells in the epithelium and in the intestinal lamina propria of chicks fed Lactobacillus spp or cecal microflora (CM) and experimentally challenged or not with Salmonella enterica serovar Enteritidis (SE) was studied using immunohistochemistry. Three hundred and twenty day-of-hatch broiler chicks were divided into four groups of 80 birds each and orally received L. reuteri, L. salivarius, L. acidophilus, or CM. Each group was subdivided into four subgroups of 20 birds each, classified as follows: a subgroup did not receive any oral treatment (negative control), subgroup treated with L. spp or CM, subgroup treated with L. spp or CM and challenged with SE, and subgroup only challenged with SE (positive control). The results show that the oral treatment with L. reuteri, L. salivarius, L. acidophilus, or CM and challenge or not with SE stimulated bird immune response as determined by the leukocytic infiltrate by CD8+ lymphocytes followed by CD4+ in the epithelium and in the lamina propria of the duodenum, jejunum, and cecum of chicks up to 12 days of age. CD8+ lymphocyte number was significantly higher in the intestine of chicks receiving CM and challenged with SE. The duodenum, followed by the jejunum, were the segments in which the immune response, as shown by T, CD4+ and CD8+ cells, was stimulated with the greatest intensity.

Helicobacter pylori em boca: epidemiologia e associação com doenças bucais

The association of Helicobacter pylori with different gastric diseases has been continuously discussed in the literature, whereas the last 20 years a large body of publications was intended to characterize the ecological niches and habitats of this pathogen being the mouth a possible habitat for that rod. The present study aimed to discuss the occurrence of this organism as part of the supplemental or transient microbiota of the mouth. Thus, an extensive review of literature covering the period 1990-2014 was carried out with different databases, yielding 5024 articles discussing transmission and occurrence of H. pylori, in English or Portuguese. Articles were selected according to the inclusion and exclusion criteria and data were tabulated and analyzed. Despite the significant heterogeneity of the literature, it was found this microaerophilic has a worldwide distribution, even in the mouth, which behaves as a reservoir for gastric reinfection. The role of oral hygiene and periodontal disease in the distribution of this microorganism remains unclear, but most studies support the hypothesis that oral biofilms and inflammation of periodontium may facilitate the installation of this pathogen in the mouth. Studies suggest that this microorganism could collaborate as a predisposing factor for oral cancer.

Ceratoconjuntivite por Encephalitozoon hellem em periquitos agapornis (Agapornis spp.) no Brasil: relato de caso

Relata-se um caso de ceratoconjuntivite causada por Encephalitozoon hellem em agapornis (Agapornis spp.) adultos, provenientes de um criatório comercial. Cinco animais apresentaram sinais clínicos de ceratoconjuntivite, blefaroespasmo e blefaroedema bilateral, com presença de secreção seropurulenta. Amostras fecais foram colhidas e foi realizado exame coproparasitológico, com resultado negativo. Dois animais foram necropsiados, sendo detectados, em impressões de raspado de conjuntiva ocular, esporos e outros estádios evolutivos de Microsporidium. A confirmação do diagnóstico foi feita pela reação em cadeia de polimerase e sequenciamento de fragmentos amplificados, com utilização de primers específicos para o gene da subunidade 18S do rRNA de E. hellem. A análise dos fragmentos amplificados demonstrou 100% de similaridade com outras sequências de E. hellem publicadas no GenBank. Este é primeiro relato de infecção por E. hellem em aves no Brasil.

Banho no leito convencional e descartável: estudo microbiológico e de custo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A Pilot Study to Determine the Effectiveness of Probiotic Use in Elderly Patients with Antibiotic-Associated Diarrhea

Long-term care settings have the majority of their patients on multiple antibiotics, and outbreaks of antibiotic-associated diarrhea and Clostridium difficile are common. Probiotics have been used with these patients to reduce these side effects. Probiotics can re-establish the composition of intestinal microflora, enhance immune response, and clear pathogens from the host which may reduce the symptoms of antibiotic-associated diarrhea. Therefore, the goal of this study was to conduct a retrospective study of the effectiveness of using probiotic in elderly patients in a long-term care facility in a Midwestern city who suffered from antibiotic-associated diarrhea. The probiotic, CulturelleTM had been administered once a day to eight males and twelve female patients who were taking antibiotics and stool consistency and number were recorded. Out of the original group, seven of the patients receiving the probiotic appeared to have positive effects while two patients had negative effects on stools. Thirteen patients showed no change in stool consistency and number. It was difficult to determine the effects of the probiotic due to the use by the facility of a bowel movement protocol for preventing constipation and impaction, and the lack of dietary records. Published studies in patients in long-term facilities vary greatly in terms of trial design, type and dose of probiotic and duration of treatment, which may explain why probiotics work for some patients and not for others. Probiotic use is becoming more accepted with antibiotic-associated diarrhea but due to the lack of definitive evidence about efficacy and the safety of probiotic use, more studies need to be conducted. Advisors: Kaye Stanek Krogstrand and Julie Albrecht

In vitro Metabolism of Grandisin, a Lignan with Anti-chagasic Activity

Tetrahydrofuran lignans represent a well-known group of phenolic compounds capable of acting as antiparasitic agents. In the search for new medicines for the treatment of Chagas disease, one promising compound is grandisin which has shown significant activity on trypomastigote forms of Trypanosoma cruzi. In this work, the in vitro metabolism of grandisin was studied in the pig cecum model and by biomimetic phase I reactions, aiming at an ensuing a preclinical pharmacokinetic investigation. Although grandisin exhibited no metabolization by the pig microbiota, one putative metabolite was formed in a biomimetic model using Jacobsen catalyst. The putative metabolite was tested against T. cruzi revealing loss of activity in comparison to grandisin.

Endophthalmitis: A Rare but Devastating Metastatic Bacterial Complication of Hemodialysis Catheter-Related Sepsis

There are many infectious complications related to vascular access in patients undergoing maintenance hemodialysis. We report two cases of endophthalmitis as a metastatic infection associated with a tunneled catheter and a temporary dual lumen catheter. Both patients were diabetic. A 61-year-old female on maintenance hemodialysis by a jugular tunnelized catheter during the past year was receiving parenteral antibiotics for catheter salvage due to fever episodes in the last 3 months. She was admitted to the hospital presenting pain, proptosis, conjunctival hyperemia, corneal infiltrate, and visual acuity of no light perception (NLP). A 51-year-old male recently undergoing hemodialysis by a temporary dual lumen catheter presented fever. His catheter was removed, but he was admitted to the hospital presenting fever, decreased vision, edema, and pain in his left eye. On examination, eyelid edema, conjunctival hyperemia, purulent secretion, hypopyon in the pupils, and visual acuity of NLP were verified. A diagnosis of endogenous endophthalmitis was made in both patients on clinical grounds and computed tomography. Evisceration of the left eye was the first option of treatment for both patients due to poor vision. Cultures of the eviscerated ocular globes showed Staphylococcus hemolyticus and Staphylococcus aureus, respectively. After evisceration, both patients received treatment, had a good outcome, and were discharged to continue their hemodialysis program. Metastatic bacterial endophthalmitis is a rare complication of dialysis catheter-related bacteremia. When suspected, urgent ophthalmologic evaluation and treatment are needed to reduce the risk of losing vision in the affected eye.

Restoration of the rabbit corneal surface after total epithelial debridement and complete limbal excision

How is the corneal epithelium restored when all of it plus the limbus have been eliminated? This investigation explored the possibility that this may be achieved through the conjunctival epithelium. The corneal epithelium of the right eye of 12 rabbits (Oryctolagus cuniculus) was totally scraped followed by surgical excision of the limbus plus 1.0-1.5 mm of the adjacent conjunctiva. Antibiotics and corticosteroids were applied for 1 week after surgery. Histological and immunohistochemical techniques were used to monitor the events taking place on the eye surface 2 weeks and 1, 3 and 6 months thereafter. Initially, the corneal surface was covered by conjunctival-like epithelium. After 1 month and more prominently at 3 and 6 months an epithelium displaying the morphological features of the cornea and reacting with the AE5 antibody was covering the central region. It is likely that the corneal epithelium originated from undifferentiated cells of the conjunctiva interacting with the corneal stroma.

Effects of eating disorders on oral fungal diversity

Background. The eating disorders anorexia and bulimia nervosa can cause several systemic and oral alterations related to poor nutrition and induced vomiting; however, the oral microflora of these patients is poorly studied. Objective. The aim of this study was to evaluate fungal microflora in the oral cavity of these patients by culture-dependent and culture-independent methods. Study Design. Oral rinse samples were cultured to assess the prevalence of Candida species, and the isolates were identified by API system. Microorganism counts were compared by the Mann-Whitney test (5%). Ribotyping, a type of molecular analysis, was performed by sequencing the D1/D2 regions of 28S rRNA. Results. Our results demonstrated that the eating disorder group showed higher oral Candida spp. prevalence with culture-dependent methods and higher species diversity with culture-independent methods. Conclusions. Eating disorders can lead to an increased oral Candida carriage. Culture-independent identification found greater fungal diversity than culture-dependent methods. (Oral Surg Oral Med Oral Pathol Oral Radiol 2012;113:512-517)

Hydrogen and Methane Production, Energy Recovery, and Organic Matter Removal from Effluents in a Two-Stage Fermentative Process

This study evaluates the potential for using different effluents for simultaneous H-2 and CH4 production in a two-stage batch fermentation process with mixed microflora. An appreciable amount of H-2 was produced from parboiled rice wastewater (23.9 mL g(-1) chemical oxygen demand [COD]) and vinasse (20.8 mL g(-1) COD), while other effluents supported CH4 generation. The amount of CH4 produced was minimum for sewage (46.3 mL g(-1) COD), followed by parboiled rice wastewater (115.5 mL g(-1) COD) and glycerol (180.1 mL g(-1) COD). The maximum amount of CH4 was observed for vinasse (255.4 mL g(-1) COD). The total energy recovery from vinasse (10.4 kJ g(-1) COD) corresponded to the maximum COD reduction (74.7 %), followed by glycerol (70.38 %, 7.20 kJ g(-1) COD), parboiled rice wastewater (63.91 %, 4.92 kJ g(-1) COD), and sewage (51.11 %, 1.85 kJ g(-1) COD). The relatively high performance of vinasse in such comparisons could be attributed to the elevated concentrations of macronutrients contained in raw vinasse. The observations are based on kinetic parameters of H-2 and CH4 production and global energy recovery of the process. These observations collectively suggest that organic-rich effluents can be deployed for energy recovery with sequential generation of H-2 and CH4.

Subconjunctival Loa loa worm: first case report in Brazil

We report the first case of ocular infestation by Loa loa in Brazil. Loiasis is caused by infestation with Loa loa, a filarial parasite originally found in the rainforests of western and central Africa. It is transmitted by the bite of the fly Chrysops and has been recently described in other places other than Africa, in African immigrants or travellers. Our case is a 33 year-old woman from Cameroon who was living in Sao Paulo, Brazil, for 5 years. She was asymptomatic until one morning she started feeling "something moving" in the left eye. Under topical anesthesia, on the slit lamp, a moving worm was removed from the subconjunctival space, which later was confirmed to be a male Loa loa adult specimen. Blood tests revealed microfilaraemia of 129 mf/mL. The patient was treated with 400 mg oral albendazole for 3 weeks and 60 mg prednisone. This report illustrates an unusual ocular disease, which is extremely rare outside of Africa, but easily diagnosed and treated. Ophthalmologists should be aware of it, in face of an increasingly globalized world.

Clinical Treatment of Dry Eye Using 0.03% Tacrolimus Eye Drops

Purpose: To report the clinical outcome of the treatment of dry eyes using 0.03% tacrolimus eye drops (olive oil + tacrolimus 0.03%) (Ophthalmos, Sao Paulo, Brazil). Methods: Sixteen eyes of 8 patients with Sjogren syndrome dry eyes (age, 51.13 +/- 9.45 years) were enrolled in this study (prospective noncontrolled interventional case series). Patients were instructed to use topical 0.03% tacrolimus eye drops twice a day (every 12 hours) in the lower conjunctival sac. Schirmer I test, break-up time, corneal fluorescein, and rose bengal staining score were performed in all patients 1 day before, and 14, 28, and 90 days after treatment with 0.03% tacrolimus eye drops. Results: The average fluorescein staining and rose bengal staining scores improved statistically significantly after 14 days of treatment and improved even more after 28 and 90 days. The average Schirmer I test did not improve statistically significantly after 28 days of treatment, although we did observe a significant improvement after 90 days of treatment with 0.03% tacrolimus eye drops. The average break-up time did not improve statistically after 14 days of treatment, although we observed a significant improvement after 28 and 90 days of treatment with 0.03% tacrolimus eye drops. Conclusions: Topical 0.03% tacrolimus eye drops successfully improved tear stability and ocular surface status in patients with dry eyes.

Nevo de células fusiformes de Reed na conjuntiva

Os autores relatam um caso de nevo de Reed, lesão que apresenta aspecto histológico de malignidade, mas tem evolução benigna. Paciente de 48 anos, masculino, cor parda, apresentava pterígio nasal no olho direito, associado a uma lesão pequena e pouco pigmentada localizada na cabeça do mesmo. Realizou-se exérese de ambos, sem intercorrências, sem sinais de recidiva. O exame histopatológico revelou lesão com bordas definidas, restrita ao epitélio, constituída por células fusiformes perpendiculares à superfície, com pigmentação melânica esparsa. O diagnóstico inicial foi nevo de Spitz, mas, posteriormente, chegou-se à conclusão que se tratava do nevo de células fusiformes de Reed. O presente relato é o segundo na literatura mundial e o primeiro no Brasil.

Dangerous waters: outbreak of eye lesions caused by fresh water sponge spicules

Purpose To describe an extremely uncommon outbreak of eye lesions in a specific area of the Brazilian Amazonia. Methods Prospective noncomparative case series. Fifty-nine patients who developed eye lesions after swimming in the Araguaia river of Tocantins state in Brazil were examined. A team of ophthalmologists equipped with a slit-lamp, gonioscopic lenses, and indirect ophthalmoscopy performed full eye examination. Analysis of the flora and fauna of the river water was undertaken by a group of experts. Results and Conclusions Eighty-three eyes were affected. The most common lesions were corneal opacities seen in 34 eyes and conjunctival nodules diagnosed in 12 eyes. Severe visual acuity loss was detected in seven children with unilateral anterior chamber lesions. Spicules of the sponge species Drulia uruguayensis and Drulia ctenosclera were found inside three blind eyes that have been enucleated for diagnostic purposes. All eye lesions could be attributed to an outbreak of foreign bodies from fresh water sponges. Organic enrichment of the water resulting from the absence of sanitation probably was the key factor, which initiated a cycle of ecological imbalance that provoked human disease.

A quantitative TaqMan PCR assay for the detection of Ureaplasma diversum

Ureaplasma diversum in veterinary studies is an undesirable microbe, which may cause infection in bulls and may result in seminal vesiculitis, balanopostitis, and alterations in spermatozoids, whereas in cows, it may cause placentitis, fetal alveolitis, abortion, and birth of weak calves. U. diversum is released through organic secretions, especially semen, preputial and vaginal mucus, conjunctival secretion, and milk. The aim of the present study was to develop a TaqMan probe, highly sensitive and specific quantitative PCR (qPCR) assay for the detection and quantification of U. diversum from genital swabs of bovines. Primers and probes specific to U. diversum 16S rRNA gene were designed. The specificity, detection limit, intra- and inter-assay variability of qPCR to detect this ureaplasma was compared with the results of the conventional PCR assay (cPCR). Swabs of vaginal mucus from 169 cows were tested. The qPCR assay detected as few as 10 copies of U. diversum and was 100-fold more sensitive than the cPCR. No cross-reactivity with other Mollicutes or eubacteria was observed. U. diversum was detected in 79 swabs (46.42%) by qPCR, while using cPCR it was detected in 42 (25%) samples. The difference in cPCR and qPCR ureaplasma detection between healthy and sick animals was not statistically significant. But the U. diversum load in samples from animals with genital disorders was higher than in healthy animals. The qPCR assay developed herein is highly sensitive and specific for the detection and quantification of U. diversum in vaginal bovine samples.