Characterization of Trypanosoma cruzi Sirtuins as Possible Drug Targets for Chagas Disease

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

Correlation between volume of apical periodontitis determined by cone-beam computed tomography analysis and endotoxin levels found in primary root canal infection

This clinical study was conducted to correlate the levels of endotoxins and bacterial counts found in primary endodontic infection with the volume of periapical bone destruction determined by cone-beam computed tomography (CBCT) analysis. Moreover, the levels of bacteria and endotoxins were correlated with the development of clinical features. Twenty-four root canals with primary endodontic disease and apical periodontitis were selected. Clinical features such as pain on palpation, pain on percussion, and previous episode of pain were recorded. The volume (cubic millimeters) of periapical bone destruction was determined by CBCT analysis. Endotoxins and bacterial samplings were collected by using sterile/apyrogenic paper points. Endotoxins were quantified by using limulus amebocyte lysate assay (KQCL test), and bacterial count (colony-forming units [CFU]/mL) was determined by using anaerobic culture techniques. Data were analyzed by Pearson correlation and multiple logistic regression (P < .05). Endotoxins and bacteria were detected in 100% of the root canal samples (24 of 24), with median values of 10.92 endotoxin units (EU)/mL (1.75-128 EU/mL) and 7.5 × 10(5) CFU/mL (3.20 × 10(5)-8.16 × 10(6) CFU/mL), respectively. The median volume of bone destruction determined by CBCT analysis was 100 mm(3) (10-450 mm(3)). The multiple regression analysis revealed a positive correlation between higher levels of endotoxins present in root canal infection and larger volume of bone destruction (P < .05). Moreover, higher levels of endotoxins were also correlated with the presence of previous pain (P < .05). Our findings revealed that the levels of endotoxins found in root canal infection are related to the volume of periapical bone destruction determined by CBCT analysis. Moreover, the levels of endotoxin are related to the presence of previous pain.

Hepatitis C infection: challenges in dental management and diagnosis of extrahepatic manifestations

Hepatitis C is associated with autoimmune diseases, hepatocellular carcinoma,and extrahepatic manifestations that, in conjunction, may seriously compromise the patient's quality of life. We herein describe a case of chronic hepatitis C with oral manifestations and discuss some implications for diagnosis and treatment. A 63-year-old woman complaining of spontaneous bleeding of the oral mucosa presented with bilateral asymmetric ulcers surrounded by white papules and striae on the buccal mucosa. Her medical history revealed leucopenia, thrombocytopenia, and skin lesions associated with chronic hepatitis C. Propranolol and ranitidine had recently been prescribed. Lichen planus, lichenoid reaction, and erythema multiforme were considered in the differential diagnosis. Histopathological analysis revealed lymphocytic infiltrate in a lichenoid pattern. The lesions partially healed after 1 week and completely regressed after 6 months, despite the maintenance of all medications; no recurrence was observed. The final diagnosis was oral lichen planus associated with hepatitis C. Chronic hepatitis C may present oral manifestations, which demand adjustments in dental treatment planning. Medication side effects may interfere with the clinical presentation and course of the disease and should be accounted for in the differential diagnosis. The possibility of spontaneous remission of oral lichen planus should always be considered, especially when putative etiological factors of a lichenoid lesion are withdrawn in an attempt to differentiate oral lichen planus from lichenoid lesions. This case emphasizes the importance of recognizing the extrahepatic manifestations of hepatitis C as a cause of increased morbidity.

Study on Chagas disease occurrence in the municipality of Monte Negro, State of Rondônia, Brazilian Amazon

Introduction: Studies on Chagas disease deal with the perspective of its occurrence in the Amazon region, which is directly correlated to the population growth and the spread of the bug biotope. The state of Rondônia has an immense source of vectors (Triatomine) and reservoirs of Trypanosoma cruzi. Environmental changes brought forth by the deforestation in the region may cause vector behavior changes and bring these vectors to a closer contact with humans, increasing the probability of vector infection. Methods: This study was carried out to check the occurrence of Chagas disease in the municipality of Monte Negro, Rondônia, Brazil, based on a random sampling of the farms and people wherein blood collection from the population and capturing triatomines were done. The blood samples were submitted to serologic tests to detect antibodies of the IgG class against T. cruzi. The triatomines that were collected had their digestive tract checked for the presence of trypanosomatidae with morphology resembling that of the T. cruzi. Results: The population examined was mostly from other states. From the 322 bugs examined on the microscope, 50% showed parasites with morphology compatible with T. cruzi. From the serology of 344 random samples of human blood, 1.2% was found positive, 6% showed inconclusive results, and 92.8% were negative. Conclusions: Monte Negro shows low prevalence of human infection by T. cruzi and none active vector transmission; however, preventive and surveying measures, which are not performed until now, shall be taken due to the abundance of vectors infected by trypanosomatidae.

Leishmania sp. infection in dogs from Florianópolis, Santa Catarina, SC, Brazil

The aim of the present study was to investigate the occurrence of Leishmania sp. infection in dogs (N = 491) living in the municipality of Florianópolis, Santa Catarina (SC), Brazil, which was considered a disease-free region for visceral leishmaniasis until 2011, when autochthonous cases of canine disease were notified. Seroprevalence in this population was assessed by ELISA (0.4%; 2/491) and IFAT (4.09%; 24/491). Only one dog exhibited seroreactivity in both serological methods, comprising a total of 25 (5.3%) seroreagent animals. Leishmania sp. DNA, obtained from a sample of whole blood of this animal, was amplified by both conventional and Real-Time PCR. Sequencing of the amplified DNA and, thereby, determination of the Leishmania species involved, was not possible. Our results suggest the necessity of a thorough epidemiological investigation in Florianópolis. (AU).

Borreliosis in horses: epidemiology, experimental infection and therapeutic

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

Secondary aspergillus fumigatus infection associated with coloidal goiter in a Black-masked Lovebird (Agapornis personata)

Aspergillosis is caused by fungus of Aspergillus genus. Is a multifactorial secondary disease and occurs mainly to immunodeficiency. Goiter is the name to non-inflammatory and non-neoplasic thyroid growth which affecting the animal metabolism. In this report we describe a case of aspergillosis and colloidal goiter in a male Black-masked lovebird (Agapornis personata) diagnosed by post mortem exam. The bird was presented for examination due to severe respiratory signs. An initial palliative treatment was performed in order to relieve the symptoms. Despite this, the patient came to die without performing additional ancillary tests. On gross exam, a pulmonary nodule was observed from which we were able to isolate Aspergillus fumigatus on microbial culture. Histological assessment revealed pulmonary aspergilosis and colloid goiter. Based on histopathological and microbiological assessments we conclude that infection probably was secondary to colloid goiter.

Serum protein profile of hookworm infection in dogs

Hookworms are parasitic nematodes that cause anemia and intestinal infections in dogs, especially with large worm burdens. However, the serum protein profile of this parasitological disease is still poorly understood. The present study was design to evaluate 80 asymptomatic dogs (age; 8 months–2 years) to detect the presence of the hookworm thin-shelled, morulated eggs in faeces using faecal flotation and to evaluate the serum protein fractions determined by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Forty dogs had an elevated quantity of eggs in faeces (+++) (PD) and 40 dogs were healthy (HD). Infected dogs showed significant increases for IgG estimated concentrations (PD 1.79±0.8 g/dL and HD 1.44±0.72 g/dL, p= 0.04), for ceruloplasmin estimated concentrations (PD 19±15 mg/dL and HD 5±3.5 mg/dL, p=0.0001), for alpha 1-acid glycoprotein estimated concentrations (PD 31.4±17.9 mg/dL and HD 13.5±12.1 mg/dL, p=0.0001) and for a non-identified protein of 23 kDa estimated concentrations (PD 641.5±194.9 mg/dL and HD 519.8± 197.9 mg/dL, p=0.007). Dogs with hookworm infection showed significant differences in the serum protein profile when compared to healthy animals.

Animal-side serologic assay for rapid detection of Mycobacterium Bovis infection in multiple species of free-ranging wildlife

Numerous species of mammals are susceptible to Mycobacterium bovis, the causative agent of bovine tuberculosis (TB). Several wildlife hosts have emerged as reservoirs of M. bovis infection for domestic livestock in different countries. In the present study, blood samples were collected from Eurasian badgers (n = 1532), white-tailed deer (n = 463), brushtail possums (n = 129), and wild boar (n = 177) for evaluation of antibody responses to M. bovis infection by a lateral-flow rapid test (RT) and multiantigen print immunoassay (MAPIA). Magnitude of the antibody responses and antigen recognition patterns varied among the animals as determined by MAPIA; however, MPB83 was the most commonly recognized antigen for each host studied. Other seroreactive antigens included ESAT-6, CFP10, and MPB70. The agreement of the RT with culture results varied from 74% for possums to 81% for badgers to 90% for wild boar to 97% for white-tailed deer. Small numbers of wild boar and deer exposed to M. avium infection or paratuberculosis, respectively, did not cross-react in the RT, supporting the high specificity of the assay. In deer, whole blood samples reacted similarly to corresponding serum specimens (97% concordance), demonstrating the potential for field application. As previously demonstrated for badgers and deer, antibody responses to M. bovis infection in wild boar were positively associated with advanced disease. Together, these findings suggest that a rapid TB assay such as the RT may provide a useful screening tool for certain wildlife species that may be implicated in the maintenance and transmission of M. bovis infection to domestic livestock.

Reducing the Public Health Impact of Bovine Tuberculosis by Controlling Disease Transmission Between Cattle and White-Tailed Deer in Northwestern Minnesota

Bovine tuberculosis, caused by infection with Mycobacterium bovis, is a re-emerging zoonotic disease. It has staged a comeback by establishing infections in wildlife and cattle, creating the potential for human disease in locations where it was thought to be under control. In northwestern Minnesota, infected cattle and white-tailed deer were first discovered in 2005. A major bovine tuberculosis eradication campaign is underway in the state, with multiple efforts employed to control M. bovis infection in both cattle and deer populations. In order to effectively eradicate bovine tuberculosis in Minnesota, there is a need for better understanding of the factors that increase the risk of deer and cattle interacting in a way that facilitates tuberculosis transmission. By reducing the risk of disease transmission within the animal populations, we will also reduce the risk that bovine tuberculosis will again become a common disease in human populations. The purpose of this study is to characterize the risk of interactions between cattle and white-tailed deer in northern Minnesota in order to prevent M. bovis transmission. A survey originally developed to assess deer-cattle interactions in Michigan was modified for use in Minnesota, introducing a scoring method to evaluate the areas of highest priority at risk of potential deer-cattle interaction. The resulting semi-quantitative deer-cattle interaction risk assessment was used at 53 cattle herds located in the region adjacent to the bovine tuberculosis “Core Area”. Two evaluators each scored the farm separately, and then created a management plan for the farm that prioritized the areas of greatest risk for deer-cattle interactions. Herds located within the “Management Zone” were evaluated by Minnesota Board of Animal Health staff, and results from these surveys were used as a point of comparison.

Estimating the extent of spatial association of Mycobacterium bovis infection in badgers in Ireland

Mycobacterium bovis infects the wildlife species badgers Meles meles who are linked with the spread of the associated disease tuberculosis (TB) in cattle. Control of livestock infections depends in part on the spatial and social structure of the wildlife host. Here we describe spatial association of M. bovis infection in a badger population using data from the first year of the Four Area Project in Ireland. Using second-order intensity functions, we show there is strong evidence of clustering of TB cases in each the four areas, i.e. a global tendency for infected cases to occur near other infected cases. Using estimated intensity functions, we identify locations where particular strains of TB cluster. Generalized linear geostatistical models are used to assess the practical range at which spatial correlation occurs and is found to exceed 6 in all areas. The study is of relevance concerning the scale of localized badger culling in the control of the disease in cattle.

The Economics of Managing Wildlife Disease

The spread of infectious disease among and between wild and domesticated animals has become a major problem worldwide. Upon analyzing the dynamics of wildlife growth and infection when the diseased animals cannot be identified separately from healthy wildlife prior to the kill, we find that harvest-based strategies alone have no impact on disease transmission. Other controls that directly influence disease transmission and/or mortality are required. Next, we analyze the socially optimal management of infectious wildlife. The model is applied to the problem of bovine tuberculosis among Michigan white-tailed deer, with non-selective harvests and supplemental feeding being the control variables. Using a two-state linear control model, we find a two-dimensional singular path is optimal (as opposed to a more conventional bang-bang solution) as part of a cycle that results in the disease remaining endemic in the wildlife. This result follows from non-selective harvesting and intermittent wildlife productivity gains from supplemental feeding.

Development of a Model of Natural Infection with Mycobacterium bovis in White-Tailed Deer

The objective of this study was to develop a suitable experimental model of natural Mycobacterium bovis infection in white-tailed deer (Odocoileus virginianus), describe the distribution and character of tuberculous lesions, and to examine possible routes of disease transmission. In October 1997, 10 mature female white-tailed deer were inoculated by intratonsilar instillation of 2 3 103 (low dose) or 2 3 105 (high dose) colony forming units (CFU) of M. bovis. In January 1998, deer were euthanatized, examined, and tissues were collected 84 to 87 days post inoculation. Possible routes of disease transmission were evaluated by culture of nasal, oral, tonsilar, and rectal swabs at various times during the study. Gross and microscopic lesions consistent with tuberculosis were most commonly seen in medial retropharyngeal lymph nodes and lung in both dosage groups. Other tissues containing tuberculous lesions included tonsil, trachea, liver, and kidney as well as lateral retropharyngeal, mandibular, parotid, tracheobronchial, mediastinal, hepatic, mesenteric, superficial cervical, and iliac lymph nodes. Mycobacterium bovis was isolated from tonsilar swabs from 8 of 9 deer from both dosage groups at least once 14 to 87 days after inoculation. Mycobacterium bovis was isolated from oral swabs 63 and 80 days after inoculation from one of three deer in the low dose group and none of four deer in the high dose group. Similarly, M. bovis was isolated from nasal swabs 80 and 85 days after inoculation in one of three deer from the low dose group and 63 and 80 days after inoculation from two of four deer in the high dose group. Intratonsilar inoculation with M. bovis results in lesions similar to those seen in naturally infected white-tailed deer; therefore, it represents a suitable model of natural infection. These results also indicate that M. bovis persists in tonsilar crypts for prolonged periods and can be shed in saliva and nasal secretions. These infected fluids represent a likely route of disease transmission to other animals or humans.

Milk Containing Mycobacterium bovis as a Source of Infection for White-Tailed Deer Fawns (Odocoileus virginianus)

Setting:White-tailed deer represent the first wildlife reservoir of Mycobacterium bovis in the United States. The behavior of does with nursing fawns provides several potential mechanisms for disease transmission. Little information exists concerning transmission between doe and fawn, specifically transmammary transmission. Objective: Determine if fawns can become infected by ingestion of milk replacer containing M. bovis, thus simulating transmission from doe to fawn through contaminated milk. Design: Seventeen, 21-day-old white-tailed deer fawns were inoculated orally with 2x108 CFU (high dose, n=5), 2.5 x 105 to 2.5 x 106 CFU (medium dose, n=5), and 1x104 CFU (low dose, n=5) of M. bovis in milk replacer. Dosages were divided equally and fed daily over a 5-day period. Positive control fawns (n=2) received 1x105 CFU of M. bovis instilled in the tonsillar crypts. Fawns were euthanized and examined 35-115 days after inoculation and various tissues collected for bacteriologic and microscopic analysis. Results: All fawns in the tonsillar, high oral and medium oral dose groups developed generalized tuberculosis involving numerous organs and tissues by 35-84 days after inoculation. Three of five fawns in the low-dose oral group had tuberculous lesions in the mandibular lymph node, and one of five had lesions in the medial retropharyngeal lymph node when examined 115 days after inoculation. Conclusion: White-tailed deer fawns can become infected through oral exposure to M. bovis. Therefore, the potential exists for fawns to acquire M. bovis while nursing tuberculous does.

Bovine Tuberculosis in Swedish Farmed Deer: Detection and Control of the Disease

Bovine tuberculosis (BTB) was introduced into Swedish farmed deer herds in 1987. Epidemiological investigations showed that 10 deer herds had become infected (July 1994) and a common source of infection, a consignment of 168 imported farmed fallow deer, was identified (I). As trace-back of all imported and in-contact deer was not possible, a control program, based on tuberculin testing, was implemented in July 1994. As Sweden has been free from BTB since 1958, few practicing veterinarians had experience in tuberculin testing. In this test, result relies on the skill, experience and conscientiousness of the testing veterinarian. Deficiencies in performing the test may adversely affect the test results and thereby compromise a control program. Quality indicators may identify possible deficiencies in testing procedures. For that purpose, reference values for measured skin fold thickness (prior to injection of the tuberculin) were established (II) suggested to be used mainly by less experienced veterinarians to identify unexpected measurements. Furthermore, the within-veterinarian variation of the measured skin fold thickness was estimated by fitting general linear models to data (skin fold measurements) (III). The mean square error was used as an estimator of the within-veterinarian variation. Using this method, four (6%) veterinarians were considered to have unexpectedly large variation in measurements. In certain large extensive deer farms, where mustering of all animals was difficult, meat inspection was suggested as an alternative to tuberculin testing. The efficiency of such a control was estimated in paper IV and V. A Reed Frost model was fitted to data from seven BTB-infected deer herds and the spread of infection was estimated (< 0.6 effective contacts per deer and year) (IV). These results were used to model the efficiency of meat inspection in an average extensive Swedish deer herd. Given a 20% annual slaughter and meat inspection, the model predicted that BTB would be either detected or eliminated in most herds (90%) 15 years after introduction of one infected deer. In 2003, an alternative control for BTB in extensive Swedish deer herds, based on the results of paper V, was implemented.