Immunohistochemical expression of HER - 2 and Ki - 67 in granulosa cell tumor in bitches.

Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia-pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti-HER2 and anti-Ki-67. The aim of this study was to evaluate the expression of immunohistochemical markers HER-2 and Ki-67 in granulosa cell tumour from bitches´ ovaries. In HER-2 immunomarker analysis using the HercepTest® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki-67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti-HER-2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti-Ki-67 and anti-HER-2 antibody immunostaining intensities, probably due to only one case with a high Ki-67 index. With an effective protocol for HER-2 and Ki-67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.

Non-invasive sampling in Itatiaia National Park, Brazil: wild mammal parasite detection.

BACKGROUND: Non-invasive sampling through faecal collection is one of the most cost-effective alternatives for monitoring of free-living wild mammals, as it provides information on animal taxonomy as well as the dynamics of the gastrointestinal parasites that potentially infect these animals. In this context, this study aimed to perform an epidemiological survey of gastrointestinal parasites using non-invasive faecal samples from carnivores and artiodactyls identified by stool macroscopy, guard hair morphology and DNA sequencing in Itatiaia National Park. Between 2017 and 2018, faeces from carnivores and artiodactyls were collected along trails in the park. The host species were identified through macroscopic and trichological examinations and molecular biology. To investigate the parasites, the Faust, Lutz and modified Ritchie and Sheather techniques and enzyme immunoassays to detect Cryptosporidium sp. antigens were used. RESULTS: A total of 244 stool samples were collected. The species identified were Chrysocyon brachyurus, Leopardus guttulus, Canis familiaris, Cerdocyon thous, Puma yagouaroundi, Leopardus pardalis, Puma concolor and Sus scrofa. There were 81.1% samples that were positive for parasites distributed mainly in the high part of the park. Helminths, especially eggs of the family Ascarididae, were more frequently detected in carnivore faeces (70.9%). Protozoa, especially Cryptosporidium sp., represented the highest frequency of infection in artiodactyl faeces (87.1%). This zoonotic protozoon was detected in eight mammalian species, including in a wild boar. High values of structural richness and Shannon and Simpson diversity indices were observed for the parasites, especially in the faeces of C. brachyurus. Significant differences in parasite diversity were observed between wild and domestic animals, such as C. brachyurus and C. familiaris, respectively, and between taxonomically distant species, such as C. brachyurus and S. scrofa. The highest values for parasite similarity were found among the species that frequented similar areas of the park, such as C. brachyurus and L. guttulus. CONCLUSIONS: The animals and parasite infections were identified through the combination of three techniques. High frequency parasite structures were diagnosed. Zoonotic protozoa were found and mainly occurred in samples from introduced species.

Expression of interleukins 6 and 10 and population of inflammatory cells in the equine endometrium: diagnostic implications.

Endometritis consists of an acute or chronic inflammatory process involving the endometrium and together with endometrosis constitute the main causes of infertility in mares. The aim of this study was to associate the histopathological findings with the immunohistochemical markers interleukins 6 (IL-6) and 10 (IL-10) to evaluate the inflammatory changes and progression of uterine tissue lesions of mares in the diestrus phase and their diagnostic implications. Twelve crossbred cyclic mares were used for endometrial biopsy collection. Samples were collected in the diestrus period (6 ± 1 day after ovulation) without previous artificial insemination. In the histopathological analysis the samples were classified according to the type and intensity of inflammation, alterations regarding endometrial fibrosis and biopsy categories (I, IIA, IIB and III). In the immunohistochemical analysis, the markers of IL-6 and IL-10 were evaluated by scores (0, 2, 4, 6) according to the intensity of the immunostaining and inflammatory cells (CD-3, CD-20, CD-68 and MPO antibodies) and were counted according to the number of cells immunostained in brown, in ten random fields. An association (p ≤ 0.05) occurred between low score (2) for IL-6 in the endometrial glandular area and moderate fibrotic nets; and between high scores (4 and 6) for IL-10 in sub-epithelial connective tissue and moderate periglandular fibrosis. In conclusion, immunohistochemical analysis demonstrated an association between interleukins and inflammatory cells with endometrial lesions. In addition, this research may be useful in the future to evaluate the progress of the inflammatory process, contributing to the adequate optimization of the reproductive management of the mares.

Vimentin, osteocalcin and osteonectin expression in canine primary bone tumors: diagnostic and prognostic implications.

Canine primary bone tumors have a plastic radiographic image, demanding histopathological confirmation. Bone tumors are characterized by the type and amount of extracellular matrix produced what cannot be easily recognized, especially in biopsy samples. Identifying cellular markers that could aid diagnosis has supported various studies in oncological pathology. This study aimed to evaluate 22 canine primary bone neoplasms, establishing their histopathological diagnosis and evaluated vimentin, osteonectin and osteocalcin expression and their implication in diagnosis and prognosis. There were 12 productive osteoblastic osteosarcomas, six minimally productive osteoblastic osteosarcoma, two chondrosarcomas, one fibrosarcoma and one hemangiosarcoma. Immunostaining was cytoplasmatic in all cases, with average percentage of 87.9% for vimentin, 98.0% for osteonectin and 99.9% for osteocalcin. In this last case, only osteosarcomas were considered. Intensity was higher in vimentin labeling (+++), followed by osteonectin (++) and osteocalcin (+). One osteosarcoma showed negative immunostaining for vimentin and of samples submitted to anti-osteocalcin immunostaining, three osteosarcomas and one fibrosarcoma had negative staining. Besides identifying mesenchymal origin, vimentin elevated expression in canine bone tumors can be related to epithelial-mesenchymal transition, leading to more aggressive tumoral phenotypes and metastasis development. Similarly, high osteonectin expression is implicated in neoplastic cell invasion and is also related to metastasis spread. Decreased osteocalcin expression was found in some osteosarcoma samples and can be related to poor prognosis, as in human osteosarcomas. Our findings suggest that vimentin, osteonectin and osteocalcin not only aid diagnosis but can be related to prognosis in canine primary bone tumors, especially osteosarcomas and its osteoblastic subtype.

Preventive Effect of CPP-ACPF Paste and Fluoride Toothpastes Against Erosion and Erosion Plus Abrasion In Vitro - A 3D Profilometric Analysis.

PURPOSE: To evaluate the effect of CPP-ACPF paste and fluoride toothpastes on enamel subjected to erosion and erosion plus abrasion in vitro. MATERIALS AND METHODS: A total of 220 human enamel blocks were divided into eleven groups (n = 20): CPP-ACPF paste (MPP), potassium nitrate/sodium fluoride toothpaste (PE), sodium fluoride toothpaste (FD), fluoride-free toothpaste (SO) and control (erosion only with no paste or toothpastes; CO) according to the experimental design: erosion or erosion plus abrasion immediately after erosion (ERO+I-ABR) or 30 min after erosion (ERO+30min-ABR). For 5 days, the specimens were subjected to: (1) erosive challenge (EC) (cola drink, 4 x 5 min/day), topical application of the undiluted paste or diluted toothpastes (1:2 w/w) (4 x 1 min/ day) plus 1 h in artificial saliva (AS) between cycles and overnight; or (2) EC plus abrasion (4 x /60 s/day) performed with the diluted toothpastes (no MMP) plus 1 h in AS between cycles and overnight. Erosion depth was quantified through a 3D profilometer. Data were analysed using Kruskal-Wallis, Mann-Whitney and Wilcoxon tests (p = 0.05). RESULTS: CPP-ACPF paste and NaF toothpaste showed lowest enamel wear among groups and reduced tissue loss by 89% in erosion challenge. Abrasion led to higher enamel wear than erosion only (p = 0.030). ERO+30min-ABR had no protective effect when compared to ERO+I-ABR (p > 0.05). CONCLUSION: A high frequency of CPP-ACPF paste application (4x daily) is effective in reducing the effects of erosion. A waiting period before performing toothbrushing does not protect enamel against erosion regardless the composition of the toothpastes.

Hydroxymethylbilane synthase gene mutations and polymorphisms in Brazilian families with acute intermittent porphyria.

Acute intermittent porphyria (AIP), an autosomal dominant disorder, is caused by a deficiency of hydroxymethylbilane synthase (HMBS). In the present study, we sought to establish a correlation between HMBS activity with the presence of mutations and polymorphisms. Enzyme activity was measured in red blood cells of four Brazilian unrelated AIP families (n = 124) and in blood donors (n = 80). The HMBS mutations in AIP family members were studied by PCR-SSCP followed by direct sequencing. Six intragenic SNPs (1345 G>A, 1500 T>C, 2377 C>A, 2478 A>G, 3581 A>G, and 7064 C>A) were determined by PCR-RFLP. Abnormal SSCP patterns in exons 7, 9, 12, and 15 were observed. DNA sequencing analysis revealed one nonsense mutation, R149X, two missense mutations, G111R and L338P, and one deletion, CT 730-731. All mutation carriers had lower enzyme activity. All polymorphisms, except 2377 C>A and 7064 C>A, showed no significant differences compared with previous reports. Mutation screening allowed the detection of the missense mutation, L338P, and the 730_731delCT deletion, two as yet unreported mutations in Brazilian AIP patients. Our findings also showed a high frequency of 2478 A>G and 3581 A>G polymorphism combinations suggesting that these polymorphisms contributed to enzymatic activity reduction in our study population.

Balantidium coli and other gastrointestinal parasites in captives non-human primates of the Rio de Janeiro, Brazil.

BACKGROUND: Parasites are agents of diarrhea in captive non-human primates (NHP). To broaden information about those etiological agents in captive animals in Brazil, gastrointestinal parasites in captive NHP were researched in nurturing Centers of Rio de Janeiro State. METHODS: Fecal samples were collected from primates, of which 960 came from the Research Center (Cecal/Fiocruz) and 115 from the Primate Center (CPRJ/Inea). The study involved species of the New World (NW) primates and of the Old World (OW). RESULTS: The estimated prevalence was 56.7%, of which 91.3% presented protozoans and 7.4% presented helminths. Statistical difference between the nurturing centers occurred in the overall value of parasitosis and in the isolated frequency of Balantidium coli and Entamoeba sp., especially in the samples of OW primates living in Cecal. CONCLUSIONS: These results demonstrated the need for implements of sanitation programs in the sites for captive primates nurturing.

ROBOTIC VERSUS LAPAROSCOPIC ROUX-EN-Y-GASTRIC BYPASS: A RETROSPECTIVE STUDY IN A SINGLE CENTER.

BACKGROUND: Bariatric surgery is the best treatment option for patients with obesity. As a result of the advancement of technology, the robotic gastric bypass presents promising results, despite its still high costs. AIMS: The aim of this study was to compare patients submitted to a robotic versus a laparoscopic gastric bypass at a single center by a single surgeon. METHODS: This retrospective study collected data from the medical records of 221 patients (121 laparoscopic procedures versus 100 with daVinci platform). The variables analyzed were sex, age, body mass index, comorbidities, surgical time, length of stay, and complications. RESULTS: The mean surgical time for patients in the robotic group was shorter (102.41±39.44 min versus 113.86±39.03 min, p=0.018). The length of hospital stay in robotic patients was shorter (34.12±20.59 h versus 34.93±11.74 h, p=0.007). There were no serious complications. CONCLUSIONS: The group submitted to the robotic method had a shorter surgical time and a shorter hospital stay. No difference was found regarding strictures, bleeding, or leakage.

Serological Evidence of Hepatitis E Virus Infection in Brazilian Equines.

Hepatitis E virus (HEV) infection has been demonstrated in various animal species; those recognized as potential zoonotic reservoirs pose a considerable risk to public health. In Brazil, HEV-3 is the only genotype identified in humans and swine nationwide, in a colony-breeding cynomolgus monkey and, recently, in bovines and capybara. There is no information regarding HEV exposure in the equine population in Brazil. This study aimed to investigate anti-HEV antibodies and viral RNA in serum samples from horses slaughtered for meat export and those bred for sport/reproduction purposes. We used a commercially available ELISA kit modified to detect species-specific anti-HEV, using an anti-horse IgG-peroxidase conjugate and evaluating different cutoff formulas and assay precision. Serum samples (n = 257) were tested for anti-HEV IgG and HEV RNA by nested RT-PCR and RT-qPCR. The overall anti-HEV seroprevalence was 26.5% (68/257) without the detection of HEV RNA. Most municipalities (53.3%) and farms (58.8%) had positive horses. Animals slaughtered for human consumption had higher risk of HEV exposure (45.5%) than those bred for sports or reproduction (6.4%) (p < 0.0001). The statistical analysis revealed sex and breeding system as possible risk-associated factors. The first serological evidence of HEV circulation in Brazilian equines reinforces the need for the surveillance of HEV host expansion in a one-health approach.

Cryptosporidium species in non-human animal species in Latin America: Systematic review and meta-analysis.

Cryptosporidiosis is an infection caused by a protozoon that inhabits the gastrointestinal tract. More than forty valid species have been described in the genus Cryptosporidium, infecting a broad range of hosts around the world, some with zoonotic transmission and others with predominant anthroponotic transmission. Prevalence studies conducted in Latin American countries have been specific, without consolidating information on species prevalences. Thus, the aim of this study was to perform a systematic review and meta-analysis addressing the prevalence of Cryptosporidium species in animals in Latin America. The estimated pooled prevalence rate for cryptosporidiosis in animals, by means of meta-analysis with a random-effects model, based on species identification, was 18.0% (95% CI 11.0%-27.0%) with high heterogeneity. The estimated overall prevalence was 20.3% (36/177) in pets, 19.9% (1309/6573) in livestock animals and 23.9% (954/3995) in exotic/captive animals. Evidence of circulation of 16 Cryptosporidium species was found in five Latin American countries: Brazil, Colombia, Chile, Argentina and Mexico. Through meta-analysis with a random-effects model, the pooled prevalence rate for Cryptosporidium parvum was 0.7% (95% CI 0.2%-2.4%). Cryptosporidium felis (8.5%) was the most prevalent species in pets, C. parvum (10.3%) in livestock animals and Cryptosporidium galli (17.6%) in exotic/captive animals. C. parvum was the species with the greatest geographical dispersion, which can be explained by its eurixenic and zoonotic potential. Few studies on cryptosporidiosis in animals in Latin America were found, which shows that there is a need for investment in and expansion of studies on this parasite. The pooled prevalence of C. parvum in Latin America and its wide circulation are similar to what has been observed in other developing regions, which reaffirms the importance of this species as the cause of a neglected, emerging and zoonotic parasitosis.

Balantidiasis in humans: A systematic review and meta-analysis.

Balantioides coli is a protozoan that infects different hosts species, including humans, with zoonotic transmission. The parasite, which lives in the large intestine and in other organs, can lead to serious infections that may culminate in death. Information about human balantidiasis is generally still very scanty. In view of the above, the purpose of this study was to analyze the epidemiological, clinical and laboratory characteristics of human balantidiasis based on a systematic review and meta-analysis. The scientific articles were retrieved from various databases and were subjected to descriptive analyses, chi-squared tests, and summarized on a forest plot and the heterogeneity index (I2). A total of 103 articles were eligible and included in this review. Out of these 103 articles, 75 were clinical case reports and 28 were epidemiological studies, indicating a frequency of 997 (3.98%) people potentially infected with B. coli. The publication dates of the analyzed articles ranged from 1910 to 2020, but the majority (68.9%) were published between 1998 and 2020. A considerable number of these articles were published in South America and Asia, mostly in Brazil and India, respectively. However, in Africa, Ethiopia, was observed the higher number of infected people (47.5%). A significant association (p < 0.05) was identified between proximity to pigs and positivity for B. coli, since more than 16% infected people were in proximity with pigs and/or their excreta. Infection by the protozoan was classified mainly as intestinal, and the predominant symptom was dysentery. Extraintestinal infections were found in 27 individuals, with colonization of the genitourinary tract frequently highlighted. Direct examination (17.2%), followed by an association of direct examination and sedimentation (45.7%), were the most commonly performed parasitological techniques, and the most frequently diagnosed form was trophozoites, corresponding to 22.5% of cases. The most common treatment for parasitized individuals (11.8%) was an association of tetracycline drugs with nitroimidazole derivatives. The articles retrieved, mainly epidemiological ones, used in meta-analysis showed high heterogeneity (I2> 50%, p < 0.05), impairing the retrieval and comparison of results. Some articles were found to provide incomplete information, making it difficult to retrieve and analyze variables. However, this review enabled us to compile and restate factors that appear to be associated with cases of human balantidiasis.

Evaluation of Larval Therapy Compared to Antibiotic Therapy in the Treatment of Skin Wounds in Rabbits.

Larval therapy (LT) is a therapeutic modality that uses larvae of necrophagous flies for the treatment of wounds. The use of this therapy presents several benefits, due to the action of the larvae that remove necrotic tissue selectively, exercising antimicrobial action, and promoting healing. There are situations in which LT proves to be the only or the best alternative for the patient, such as wounds infected by multidrug-resistant microorganisms or when treatment difficulties may lead to an indication for amputation. The purpose of this study was to compare the efficiency of LT, using larvae of Chrysomya megacephala, with that of antibiotic therapy in the treatment of cutaneous wounds infected with Pseudomonas aeruginosa. Twenty-four rabbits were used to perform the experiment, distributed in four groups: group 1, induced wound without bacterial infection; group 2, induced wound with bacterial infection; group 3, induced wound with bacterial infection and antibiotic therapy; group 4, induced wound with bacterial infection and LT. The macroscopic, microscopic, and statistical analyses indicated that LT was as effective as antibiotic therapy in wound healing.

Bariatric surgery during the evolution of fatty liver-A randomized clinical trial comparing gastric bypass and sleeve gastrectomy based on transient elastography.

Nonalcoholic fatty liver disease (NAFLD) is closely related to patients with obesity. For patients with NAFLD, bariatric surgery is the best treatment. However, the best technique to patient with severe NAFLD is still unknown. Currently available, the imaging methods for assessing and monitoring NAFLD are of limited use for diagnosing. In contrast, compared with liver biopsy and transient hepatic elastography (THE) has shown good accuracy in individuals with obesity. To prospectively compare the evolution of THE parameters of NAFLD right after the procedures: gastric bypass vs sleeve gastrectomy. Patients with obesity were randomized into two groups: gastric bypass and sleeve gastrectomy in a previous study. Iin a previous study one week before and three months after surgery the patients underwent evaluation by THE. The patients were also analyzed with controlled attenuation parameter (CAP), which assesses the degree of hepatic steatosis using the same device. Sleeve gastrectomy group showed a greater decrease in THE values (from 8.13 to 5.53 kPa) compared to the gastric bypass group (from 9.25 to 8.81 kPa; P = .004). CAP also revealed a greater decrease in sleeve subjects (from 287 to 242 dB/m) compared to gastric bypass subjects (from 290 to 276 dB/m; P < .0001). The absolute values of these differences also had a largest decrease with both methods in sleeve gastrectomy group (P = .013 and P = .005 for THE and CAP, respectively).Sleeve gastrectomy showed a greater decrease in both parameters (THE and CAP) than gastric bypass in the first months.

Detection of Dirofilaria immitis using microscopic, serological and molecular techniques among dogs in Cabo Frio, RJ, Brazil.

Heartworm disease is a health problem for dogs and cats, especially in tropical and subtropical coastal regions of the world. Some studies have compared the efficacy of the diagnostic techniques used to detect this parasitosis. Therefore, the aim of this study was to compare parasitological optical microscopy (POM), serological and molecular techniques for diagnosing canine heartworm infection. Samples were collected between July 2015 and April 2016 from 103 dogs in Cabo Frio, RJ, Brazil. The wet fresh blood, thick smears, thin smears and modified Knott's test were used to detect microfilariae. ELISA (Snap™ 4Dx ® IDEXX) was used to detect antigens and the polymerase chain reaction (PCR) was used to detect DNA and enable sequencing for species differentiation and confirmation. 19.4% of samples were positive according to microscopy. Through PCR, 15.5% of the total were positive. Using ELISA, the positivity rate was 29.1%. Occult heartworm infection was detected in 11.6% of the samples. ELISA sensitivity was shown to be higher than PCR or microscopy (P = 0.001). Sequencing of samples confirmed the presence of Dirofilaria immitis and Acanthocheilonema reconditum . ELISA was more effective for serological diagnosis canine heartworm and should be used in clinical and epidemiological studies.

Occurrence of anti-Toxoplasma gondii antibodies and risk factors associated with infection in equids slaughtered for human consumption in Brazil.

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii. Ingestion of raw/undercooked meat is considering an important route of infection. Consumption of meat from equids is common in European and Asian countries and an increase in Brazil has been observed. The aim of this study was to evaluate occurrences of anti-T. gondii antibodies and risk factors relating to infection in equids slaughtered for human consumption in Minas Gerais, Brazil. Blood samples from 192 horses and 208 donkeys were collected in the exsanguination area during the slaughter. Serum samples were subjected to the indirect fluorescent antibody test (IFAT). Association analysis was performed using Pearson's chi-square test (χ2) or Fisher's exact test, to evaluate risk factors relating to the prevalence of seroreagents. Antibodies against T. gondii were found in 13.5% of the equids, with higher occurrence in horses (18.75%) than in donkeys (8.65%). Associations between seropositivity and the following variables were found (p ≤ 0.05): species, animal origin, purpose of rearing and source of water for animal consumption and contact with cats. Farms need to implement preventive measures to control T. gondii infection in these species and avert transmission of the parasite to the human population that will consume their meat.

Gastrointestinal Parasites in Captive Animals at the Rio de Janeiro Zoo.

BACKGROUND: Gastrointestinal parasites may determine diarrhea, dysentery or even death in captive mammals. These animals tend to be more susceptible to parasitic infections due to confinement and stress. Purpose To increase the information about these etiological agents in captive animals in Brazil, the gastrointestinal parasites of the captive mammals of the Rio de Janeiro Zoo were investigated. METHODS: From 2016 to 2018, 180 fecal samples were collected from animals housed in the Rio de Janeiro Zoo: 63 from animals of the order Primates, 26 of Carnivora, 78 of Artiodactyla, 9 of Perissodactyla and 4 of the order Rheiformes. The feces were processed by direct examination and by the techniques of Faust et al., Sheather, Ritchie, Lutz, and smears were stained with safranin. Immunoenzymatic assays were also performed to investigate antigens of Giardia duodenalis, Cryptosporidium spp., Entamoeba histolytica/Entamoeba dispar. RESULTS: Parasite positivity was identified in 68.3% of the fecal samples, with a parasite positivity rate of 68.2% among primates, 65.3% among carnivores, 69.2% among artiodactyls, 33.3% among perissodactyls, and 100% among rheiformes. The most frequently detected parasite was Entamoeba histolytica/E. dispar antigens, which showed a statistically significant positivity rate (33.3%; p = 0.000), particularly in the feces of carnivores (30.7%) and artiodactyls (53.8%). A statistically significant positivity rate of Balantioides coli (11.1%; p = 0.001) was also detected in feces from nonhuman primates, tapirs, collared peccaries and rheas. The positivity of Cryptosporidium sp. antigens in feces of the orders Carnivora, Artiodactyla and Primates was also statistically significant (7.2%, p = 0.010). Oocysts compatible with Cryptosporidium spp. were detected in 6.3% from primates. The helminths most frequently detected were thin-shelled eggs of nematodes (17.7%, p = 0.000), nematode larvae (15.5%, p = 0.000) and Trichuris trichiura eggs (6.1%, p = 0.018). CONCLUSION: The positivity rate for gastrointestinal parasites demonstrates the need for a sanitation management program to be implemented in the zoo, including routine diagnostic parasitology tests followed by specific treatment for each parasitosis.

Increasing the Efficiency of Spontaneous Sedimentation for the Diagnosis of Intestinal Protozoa.

The aim of this study was to evaluate the association between a longer sedimentation time, the reading of a larger number of slides, and the collection of multiple samples on the efficiency of the spontaneous sedimentation technique. Twenty-two patients with a previous parasitological exam positive for intestinal protozoa were recruited to collect new fecal samples (3 samples per patient) before the beginning of antiparasitic treatment. All collected fecal samples were used for spontaneous sedimentation and centrifuge-flotation techniques. Of these, all 22 patients were positive based on spontaneous sedimentation, and 59.1% (13/22) based on centrifuge flotation. The number of samples and the number of slides analyzed by spontaneous sedimentation influenced the number of positive cases. The modifications applied to the spontaneous sedimentation technique increased its performance in protozoa diagnosis.

Agreement between upper endoscopy and esophagography in the diagnosis of megaesophagus in Chagas disease.

INTRODUCTION: The diagnosis and classification of megaesophagus can be challenging in patients with Chagas disease. The present study aimed to evaluate the agreement between upper endoscopies and esophagographies for the diagnosis and classification of megaesophagus in Chagas disease. METHODS: A cross-sectional study of 50 patients with Chagas disease with upper digestive symptoms was undertaken. Esophagography and upper endoscopy exams were performed to compare diagnoses. Statistical analysis included sensitivity and specificity used to evaluate the diagnostic accuracy of upper endoscopies, and measures of agreement: linearly weighted Kappa (κw) and Cohen`s classical Kappa (κ) coefficients with 95% confidence intervals (95% CI). RESULTS: Twenty-three patients (46%) were diagnosed with megaesophagus by esophagography. The upper endoscopy sensitivity and specificity for megaesophagus diagnosis were 100% and 33.3%, respectively. Regarding megaesophagus classifications, there was a substantial agreement between the two exams (κw = 0.622; 95% CI: 0.498 to 0.746). Within megaesophagus groups, agreement for group I was slight (κ = 0.096; 95% CI: 0.000 to 0.403); for group II, substantial (κ = 0.703; 95% CI: 0.456 to 0.950); and for groups III and IV, inconclusive (κ = 0.457; 95% CI: 0.000 to 0.967; κ = 0.540; 95% CI: 0.035 to 1.000, respectively). CONCLUSIONS: Upper endoscopy has a high sensitivity, but a low specificity to diagnose megaesophagus. Agreement between the two exams varies depending on the megaesophagus grade. Thus, upper endoscopy can be used in the diagnostic workup of a possible Chagas disease megaesophagus, but all identified cases should undergo esophagography.

Epidermal growth factor receptor 2 immunoexpression in gastric cells of domestic cats with H. heilmannii infection.

The aim of this study was to evaluate the immunoexpression of HER-2 in gastric cells of cats infected with Non H. pylori Helicobacter (NHPH) and to investigate an association with the presence of inflammatory infiltrate. Forty-eight paraffin-embedded gastric samples were retrieved from the archives of the Veterinary Anatomic Pathology Laboratory that had previously been shown to be positive for NHPH with the rapid urease test and cytology. Infection by NHPH was confirmed by histopathology using the Warthin-Starry staining. Hematoxylin-eosin stained sections were reviewed to evaluate inflammatory cell infiltrates. Immunohistochemical analysis was done using anti- H. pylori antibody and anti-HER-2 antibody. Molecular analysis was performed by PCR to confirm the presence of Helicobacter. Statistical analysis was performed to determine whether there was an association between the presence of H. Heilmannii and HER-2 expression in gastric samples. All samples were positive for NHPH, by immunohistochemistry, and confirmed by PCR as H. Heilmannii. On histopathologic analysis, 56,3% of the samples had lymphocytes and plasma cells infiltrates, 52,1% of which were mild and 4,2% moderate. The intensity of the inflammatory infiltrate in the gastric mucosa was significantly greater in the complete plasma membrane of parietal cells of gastric glands that had greater HER-2 immunoexpression (p = 0.0001). A statistically significant association (p = 0.007) between the H. Heilmannii infection score and the expression of HER-2 in the lateral membrane of gastric surface cells was observed. HER-2 expression may be increased in feline gastric cells infected by H. Heilmannii and in parietal cells of gastric glands with an increased inflammatory infiltrate.

Comparison of Four Parasitological Techniques for Laboratory Diagnosis of Eggs from Spirometra spp. in Wild Mammal Fecal Samples.

PURPOSE: This study aimed to compare the performance of four different microscopic coproparasitological techniques in relation to egg recovering and the frequency of alterations in the eggs observed through each technique. METHODS: A total of 213 fecal samples from free-living carnivorous mammals were collected between 2017 and 2018 in Itatiaia National Park, RJ, Brazil. Faust and modified Sheather floatation techniques as well as Lutz and modified Ritchie sedimentation techniques were applied. RESULTS: The total positivity rate for Spirometra spp. eggs was 24.4%, and these were detected mainly through Lutz and modified Ritchie sedimentation techniques, with substantial agreement (kappa = 0.73; p = 0.00 < 0.05; McNemar p value = 1.0; Fisher's exact test p = 0.616). Faust and modified Sheather flotation techniques did not present good egg recovery, with frequencies of 6.6% and 7.5%, respectively. Eggs with morphological alterations were mostly observed through Faust (17.3%) and modified Sheather (13.5%). Both flotation techniques presented statistically significant frequencies of deformed eggs, in comparison with the sedimentation techniques (p = 0.00). Low frequencies of deformed eggs were observed when the samples were analyzed through modified Ritchie and Lutz sedimentation techniques. CONCLUSIONS: From these results, sedimentation techniques such as modified Ritchie and Lutz methods were more efficient for diagnosing the eggs of this helminth in fecal material from free-living carnivores and should always be used when analyzing fecal samples from hosts of different species.