[Irritable bowel syndrome: frequency and phylogenetic relationship of Blastocystis sp. from Mexican patients]. / Síndrome de intestino irritable: frecuencia y relación filogenética de Blastocystis sp. de pacientes mexicanos.

INTRODUCTION: Recent studies reported increased presence of Blastocystis in patients with Irritable Bowel Syndrome (IBS) and an etiologic role has been proposed. The pathogenic role of Blastocystis is controversial, because it is frequently found not only in individuals with enteric symptoms but also in healthy and asymptomatic subjects. Furthermore, there are few studies of blastocistosis in Mexico. OBJECTIVE: To assess the frequency of Blastocystis sp. in IBS patients using molecular techniques and to describe its phylogenetic relationship with sequences of other countries. METHODS: IBS patients according to Rome III criteria were enrolled. In all patients evaluations included: colonoscopies, coproparasitoscopic studies, coproculture, fecal virus screening. PCR and sequencing for Blastocystis sp. were also performed. RESULTS: We recruited 11 men and 51 women with a mean age of 45.6 (SD ± 15.7) years. Eighty-six percent of the IBS patients presented a normal colonoscopy, 8% showed polyps and 6% diverticular disease. Blastocystis sp. was identified in 25% patients (all of them with normal colonoscopy), while two patients had Endolimax nana and Entamoeba histolytica/E. dispar, respectively. Phylogenetic analysis showed that major sequences of Mexican carriers clustered together with sequences of parasites from Japan and Denmark; furthermore, two sequences from IBS patients were grouped in a single cluster. CONCLUSIONS: Blastocystis sp. was identified in 25% of the IBS patients. Our data support the hypothesis of clonal lineages in distinct geographical areas in the world.

Taenia solium: current understanding of laboratory animal models of taeniosis.

Neurocysticercosis is a public health problem in many developing countries and is the most frequent parasitic disease of the brain. The human tapeworm carrier is the main risk factor for acquiring neurocysticercosis. Since the parasite lodges only in the human intestine, experimental models of Taenia solium taeniosis have been explored. Macaques, pigs, dogs, cats and rabbits are unsuccessful hosts even in immunodepressed status. By contrast, rodents are adequate hosts since tapeworms with mature, pregravid and, in some cases, gravid proglottids develop after infection. In this review, information that has been generated with experimental models of taeniosis due to T. solium is discussed. Initially, the use of the model for immunodiagnosis of human taeniosis and evaluation of intervention measures is summarized. Next, descriptions of tapeworms and comparison of hamsters, gerbils and other mammals as experimental models are discussed, as well as data on the humoral immune response, the inflammatory reaction and the production of cytokines associated to Th1 and Th2 responses in the intestinal mucosa. Finally, evaluation of protection induced against the development of tapeworms by recombinant T. solium calreticulin in hamsters is summarized and compared to other studies.

Distribution of Taenia solium Diagnostic Glycoproteins in the Different Developmental Stages of the Parasite.

Taenia solium is a helminth parasite that causes 2 diseases in humans: cysticercosis and taeniasis. The establishment of T. solium metacestodes in the central nervous system causes neurocysticercosis, while development of the adult tapeworm in the small intestine causes taeniasis. Serological diagnosis of neurocysticercosis is performed by Western blot with an enriched fraction of glycoproteins that has been extensively used for clinical diagnosis and epidemiological surveys. The lectin-bound fraction that is used for this assay contains 7 antigenic glycoproteins. These antigenic proteins are considered to be highly specific for cysticercosis when tested with heterologous parasitic diseases. However, recent studies show that people with taeniasis have cross-reactive antibodies against the neurocysticercosis diagnostic glycoproteins and vice versa. Nevertheless, it is not known if these diagnostic proteins are expressed in the adult stage of the parasite. In this paper, we describe the location of 3 of these glycoproteins in T. solium adults and cysticerci using polyclonal antibodies raised against a synthetic peptide based on the amino acid sequence of TS14, a recombinant protein T24H, and the native GP50. The glycoproteins' distribution was different in invaginated and evaginated cysticerci as well as in adult tapeworms. Specifically, the 3 glycoproteins studied were differentially expressed during embryogenesis. Our findings indicate that expression of the diagnostic glycoproteins is developmentally regulated; this is noteworthy since these glycoproteins are considered specific for the diagnosis of neurocysticercosis but nevertheless are present in different structures throughout the development of T. solium. Here we describe the glycoprotein expression and localization, which can be important in understanding their biological functions. In addition, our results help clarify the cross-reaction observed between people with neurocysticercosis and taeniasis to TS14, T24H, and GP50, which are used as diagnostic antigens for neurocysticercosis.

Lack of postmortem digestion of tapeworms in Golden hamsters experimentally infected with Taenia solium.

Taenia solium causes human neurocysticercosis, a public health problem in Mexico and other developing countries. Surprisingly, tapeworm carriers are very rarely found and in necropsy studies practically no tapeworms have been reported. In this paper we analyze the possibility that, after the death of the host, tapeworms could easily be destroyed in the intestine. Our experiments, performed in the hamster model, suggest that the absence of tapeworms in human intestine during necropsy is not due to postmortem digestion.

Survey of drug resistance of Mycobacterium tuberculosis in 3 Mexican states, 1997.

BACKGROUND: Drug resistance threatens global tuberculosis (TB) control efforts. Population-based estimates of drug resistance are needed to develop strategies for controlling drug-resistant TB in Mexico. OBJECTIVE: To obtain population-based data on Mycobacterium tuberculosis drug resistance in Mexico. METHODS: To obtain drug resistance data, we conducted a population-based study of TB cases in the states of Baja California, Sinaloa, and Oaxaca, Mexico. We performed cultures and drug susceptibility testing on M tuberculosis isolates from patients with newly diagnosed, smear-positive TB from April 1 to October 31, 1997. RESULTS: Mycobacterium tuberculosis was isolated from 460 (75%) of the 614 patients. Levels of resistance in new and retreatment TB cases to 1 or more of the 3 current first-line drugs used in Mexico (isoniazid, rifampin, and pyrazinamide) were 12.9% and 50.5%, respectively; the corresponding levels of multi-drug-resistant TB were 2.4% and 22.4%. Retreatment cases were significantly more likely than new cases to have isolates resistant to 1 or more of the 3 first-line drugs (relative risk [RR], 3.9; 95% confidence interval [CI], 2.8-5.5), to have isoniazid resistance (RR, 3.6; 95% CI, 2.5-5.2), and to have multi-drug-resistant TB (RR, 9.4; 95% CI, 4.3-20.2). CONCLUSIONS: This population-based study of M tuberculosis demonstrates moderately high levels of drug resistance. Important issues to consider in the national strategy to prevent M tuberculosis resistance in Mexico include consideration of the most appropriate initial therapy in patients with TB, the treatment of patients with multiple drug resistance, and surveillance or periodic surveys of resistance among new TB patients to monitor drug resistance trends.

Proposed diagnostic criteria for neurocysticercosis.

Neurocysticercosis is the most common helminthic infection of the CNS but its diagnosis remains difficult. Clinical manifestations are nonspecific, most neuroimaging findings are not pathognomonic, and some serologic tests have low sensitivity and specificity. The authors provide diagnostic criteria for neurocysticercosis based on objective clinical, imaging, immunologic, and epidemiologic data. These include four categories of criteria stratified on the basis of their diagnostic strength, including the following: 1) absolute--histologic demonstration of the parasite from biopsy of a brain or spinal cord lesion, cystic lesions showing the scolex on CT or MRI, and direct visualization of subretinal parasites by funduscopic examination; 2) major--lesions highly suggestive of neurocysticercosis on neuroimaging studies, positive serum enzyme-linked immunoelectrotransfer blot for the detection of anticysticercal antibodies, resolution of intracranial cystic lesions after therapy with albendazole or praziquantel, and spontaneous resolution of small single enhancing lesions; 3) minor--lesions compatible with neurocysticercosis on neuroimaging studies, clinical manifestations suggestive of neurocysticercosis, positive CSF enzyme-linked immunosorbent assay for detection of anticysticercal antibodies or cysticercal antigens, and cysticercosis outside the CNS; and 4) epidemiologic--evidence of a household contact with Taenia solium infection, individuals coming from or living in an area where cysticercosis is endemic, and history of frequent travel to disease-endemic areas. Interpretation of these criteria permits two degrees of diagnostic certainty: 1) definitive diagnosis, in patients who have one absolute criterion or in those who have two major plus one minor and one epidemiologic criterion; and 2) probable diagnosis, in patients who have one major plus two minor criteria, in those who have one major plus one minor and one epidemiologic criterion, and in those who have three minor plus one epidemiologic criterion.

Reactivity in ELISA and dot blot of purified GP24, an immunodominant antigen of Taenia solium, for the diagnosis of human neurocysticercosis.

In this paper we report the purification of GP24, one of the seven specific and highly antigenic Taenia solium glycoproteins previously identified by Western blot (WB) with serum, cerebrospinal fluid (CSF) and saliva samples from patients with neurocysticercosis (NC). GP24 was purified and evaluated in ELISA and dot blot for diagnosis. A lentil-lectin-bound glycoprotein fraction (LL-GP) from T. solium cysticerci was submitted to polyacrylamide gel electrophoresis, and the band that corresponded to GP24 was sliced, minced and electroeluted; an aliquot was used to immunize a rabbit, and the antiserum obtained was analysed by WB against LL-GP fraction; only GP24 was detected. ELISA and dot blot were performed with purified GP24 and serum and CSF samples from patients with NC that were previously positive for GP24 in WB and control samples; the latter were negative, while all NC samples were positive. To test for specificity, purified GP24 was incubated in dot blot against 44 sera from patients with other parasitic diseases; no positive reactions were found. Results indicate that GP24 was adequately purified and retained its reactivity; thus in combination with ELISA or dot blot may facilitate immunodiagnosis of NC.

Effect of praziquantel on the migration and survival of developmental stages of Schistosoma mansoni in mice.

Compressed organ autoradiography has been used to determine whether the anthelmintic drug praziquantel (Pzq) modifies the migration of isotopically labelled Schistosoma mansoni during the first 16 days of infection in CBA/Ca mice. The mice were treated with 500 mg kg-1 body weight of the drug on day 1 or day 6. Treatment caused a marked delay in parasite migration from the skin when the drug was administered intradermally at the site of infection on day 1; migration from the lungs was also delayed after such treatment. Pzq injected either intradermally on day 1 or intramuscularly on day 6 effectively reduced the number of parasites that finally arrived in the lungs and the livers by 41 and 47%, respectively. Intramuscular administration of the drug on day 1 had a negligible effect. Worm recoveries assessed on day 38 by perfusion of the hepatic portal system were greatly reduced when Pzq was administered on day 14. The worms proved less susceptible when the drug was administered on day 21 and were completely resistant following drug delivery on day 28. The influence of drug preparation and route of delivery on parasite migration and survival are discussed.

A Taenia solium oncosphere protein homologous to host-protective Taenia ovis and Taenia saginata 18 kDa antigens.

A Taenia solium cDNA (TSOL-18) encoding a protein with close homology to host protective oncosphere antigens from Taenia ovis (To18) and Taenia saginata (TSA-18) is described here. TSOL-18 was cloned from mRNA obtained from hatched and activated oncospheres of T. solium. The high level of predicted amino acid sequence homology among TSOL-18 and other host protective taeniid antigens suggests that the protein expressed by TSOL-18 may be capable of being used as a vaccine against T. solium infection in the parasite's intermediate hosts.

Vaccination against Taenia solium cysticercosis in pigs using native and recombinant oncosphere antigens.

Pigs were immunised with antigens derived from Taenia solium oncospheres or with a pool of three recombinant antigens from Taenia ovis, and subsequently challenged with T. solium eggs. The native oncosphere antigens induced 83% protection against viable, and 89% protection against the total number of cysticerci established following the challenge infection. Immunisation with the recombinant T. ovis antigens induced 93% protection against the establishment of viable cysticerci, and 74% protection against the total number of cysticerci. These results, and those achieved elsewhere with Taenia saginata and T. ovis, support the possibility of developing a practical vaccine to assist in the control of transmission of T. solium through pigs.

Taenia solium: inhibition of spontaneous evagination of cysticerci by the host inflammatory capsule.

The effect of the host's inflammatory capsule or of physical restraint on the spontaneous evagination capacity of Taenia solium cysticerci was evaluated. Parasites obtained from infected pig muscle, in which the surrounding inflammatory capsule was not eliminated, were unable to evaginate spontaneously after 12 or 60 h of culture. This phenomenon was not abolished when the capsule was eliminated and the parasites were cultured in semi-solid agar or in liquid medium.

Prevalence and risk factors for Taenia solium taeniasis and cysticercosis in humans and pigs in a village in Morelos, Mexico.

In a Mexican village in which Taenia solium infection was known to be endemic, we selected a cluster sample of 368 households (21% of the total) for demographic, environmental, and diagnostic surveys, and medical histories for taeniasis and cysticercosis. Coproparasitologic studies of 1,531 participants revealed infection by Taenia sp. in four (0.3%) individuals; however, 5.8% of the respondents reported a history of having passed tapeworm proglottids in feces. Of 1,552 human serum specimens, 10.8% tested positive in the cysticercosis immunoblot assay. Seropositivity increased with age and reached a maximum in subjects ages 46-55 years. Risk factors associated with seropositivity included a history of passing tapeworm proglottids, frequent consumption of pork, and poor personal and household hygiene (P less than 0.05). A history of seizures was also significantly associated with seropositivity (P less than 0.05); approximately one-third of persons with such histories were seropositive. Of 571 pigs examined by tongue inspection, 23 (4.0%) had cysticerci; infection rates increased with the age of pigs, and were higher in pigs that habitually ran loose or were fed human feces (P less than 0.05). Goodness of fit analysis confirmed that seropositive persons (but not infected pigs) were significantly clustered within households, particularly, in households in which a member reported a history of having passed tapeworm proglottids. The results of this study have identified community behavioral and environmental practices that must be modified to prevent continued transmission of cysticercosis and taeniasis.

Novel antigens for neurocysticercosis: simple method for preparation and evaluation for serodiagnosis.

Neurocysticercosis (NCC), which is caused by infection with the larval stage of the pork tapeworm (Taenia solium), is now recognized as a major cause of neurologic diseases in countries where the infection is endemic. Migration of persons from these countries is resulting in diagnosis and local transmission in nonendemic countries at increasing rates. In the present study, immunoblotting and an ELISA were carried out using antigens of T. solium cysticerci fractionated by isoelectric focusing and serum samples from patients with NCC, alveolar (AE) or cystic echinococcosis (CE), and other diseases. Immunoblot analysis revealed antigens fractionated by isoelectric focusing (pH 9.2-9.6) either from cyst fluid of T. solium cysticerci or from intact cysts had unique components (glycoproteins) highly specific and sensitive for detection of NCC exclusively. All confirmed NCC serum samples (53 of 53) recognized at least three major bands of 10-26-kD of fractions with pH 9.2-9.6 from either intact cysts or cyst fluid. These bands were not recognized by sera from patients with other parasitic diseases including AE (0 of 34), CE (0 of 36), or other heterologous parasitoses (0 of 77), patients with hepatoma (0 of 19) or sarcoidosis (0 of 11), or sera from healthy controls (0 of 29). The ELISA using the antigens showed the same sensitivity and specificity for differentiation of NCC (53 of 53) from other diseases (0 of 107) or healthy individuals (0 of 29). Both immunoblotting and the ELISA using the fractionated antigens readily differentiated all NCC from AE or CE in a blind test of 29 serum samples of persons with NCC, CE, and AE. Antigens fractionated from cyst fluid of T. solium cysticerci by a simple, single-step isoelectric focusing (pH 9.2-9.6) are highly specific and sensitive for differential serodiagnosis of NCC in immunoblotting and/or an ELISA.

Development and evaluation of a health education intervention against Taenia solium in a rural community in Mexico.

A comprehensive study was undertaken in a rural community in the state of Morelos, Mexico to evaluate health education as an intervention measure against Taenia solium. An educational program was developed to promote recognition and knowledge of the transmission of the parasite and to improve hygienic behavior and sanitary conditions that foster transmission. The effects of educational intervention were evaluated by measuring changes in knowledge and practices and prevalence of human taeniasis and swine cysticercosis before and after the campaign. The health education strategy was implemented with the active participation of the population based on the information obtained from a sociologic study. A questionnaire was designed and used before, immediately after the intervention, and six months later. Statistically significant improvements occurred in knowledge of the parasite, its life cycle, and how it is acquired by humans; however, changes in behavior related to transmission were less dramatic and persistent. The prevalences of cysticercosis in pigs at the start of the education intervention were 2.6% and 5.2% by lingual examination and antibody detection (immunoblot assay), respectively, and approximately one year after the intervention they were 0% and 1.2% (P < 0.05). These changes were accompanied by significant reductions in the reported access of pigs to sources of infection and freedom to roam. We conclude that health education, developed along with community involvement, reduced opportunities for transmission of T. solium in the human-pig cycle.

PIP: Neurocysticercosis is an important health problem in Mexico, as well as in many other countries of Latin America, Asia, and Africa where conditions permit completion of the cestode's life cycle in pigs and humans. A study was conducted in a rural community in the state of Morelos, Mexico, to determine whether health education could be an effective measure against Taenia solium. An educational program was developed with community input to promote recognition and knowledge of the transmission of the parasite and to improve hygienic behavior and sanitary conditions which foster transmission. The effects of the educational intervention were then assessed by measuring changes in knowledge, practices, and the prevalence of human taeniasis and swine cysticercosis before and after the campaign. Statistically significant improvements occurred with regard to knowledge of the parasite, its life cycle, and how it is acquired by humans. However, changes in behavior related to transmission were less marked and persistent. Lingual examination and antibody detection found cysticercosis among 2.6% and 5.2% of pigs, respectively, at the start of the intervention. At approximately 1 year after the intervention, prevalences had declined to 0% and 1.2%. The decline was accompanied by significant reductions in the reported access of pigs to sources of infection and freedom to roam.

Prevalence and risk of cysticercosis and taeniasis in an urban population of soldiers and their relatives.

To determine markers of Taenia solium transmission and risk factors in an urban community, we studied 1,000 soldiers from a military camp in Mexico City and their relatives. Serum samples were used to detect antigens and antibodies and fecal specimens were examined for Taenia coproantigens and helminth eggs. Prevalences of 12.2% and 5.8% for cysticercosis were found among soldiers and their relatives, respectively. Taeniasis was found in 0.5% and none of the groups, respectively. Relatives of soldiers positive for cysticercosis and taeniasis markers ate more pork from street stores than restaurants or markets compared with relatives of soldiers without these indicators of infection. Also, 12.0% of the relatives of positive soldiers had a history of expelling tapeworm proglottids in the feces in contrast to 3.7% of the family members of the control group. Prevalence values and risk factors in this urban population are similar to those of previous studies performed in rural populations.

Heparin bonding of bypass circuits reduces cytokine release during cardiopulmonary bypass.

BACKGROUND: Heparin bonding of the cardiopulmonary bypass (CPB) pump circuit decreases complement activation and fibrinolysis. It is not known whether inflammatory cytokines produced during CPB can also be modulated by the more biocompatible heparin-coated circuit. METHODS: This initial study evaluated the impact of heparin-bonded CPB circuits on production of the cytokines interleukin-1 (IL-1), tumor necrosis factor-alpha (TNF-a), IL-6, and IL-8 in adults undergoing complex cardiac operations with prolonged CPB. Twenty patients had blood samples drawn immediately before and at hourly intervals after the start of CPB using either a conventional oxygenator and circuit (n = 14) or a covalently bonded heparin oxygenator and circuit (n = 6). Levels of IL-1, TNF-a, IL-6, and IL-8 were measured in all serum samples using a "sandwich" enzyme-linked immunosorbent assay. RESULTS: The levels of IL-6 and IL-8 increased in a time-dependent fashion in both groups, but the response was significantly less over time in the heparin-bonded group (p < 0.05) for both IL-6 and IL-8. Levels of IL-1 and TNF-a were not significantly elevated with lengthening bypass interval in either group. CONCLUSIONS: These data indicate that the use of heparin-coated bypass pump circuits results in lower serum levels of the inflammatory cytokines IL-6 and IL-8 than standard circuits. Biocompatible materials that decrease the inflammatory response to CPB may ultimately reduce the morbidity associated with cardiac operations.

Implementation of proficiency testing in conjunction with a rechecking system for external quality assurance in tuberculosis laboratories in Mexico.

SETTING: In developing countries, tuberculosis is diagnosed by identification of acid-fast bacilli (AFB) on sputum smears. OBJECTIVE: To evaluate the quality of AFB microscopy, the Mexican Secretary of Health National Reference Laboratory implemented proficiency testing for its network of 637 laboratories. DESIGN: A total of 586 (92%) laboratories were inspected and 430 technicians evaluated by proficiency testing consisting of 10 slides with known numbers of AFB. Results were compared with those of slide rechecking and with proficiency testing performed 2 years later. RESULTS: Of the 430 technicians evaluated by proficiency testing in 1998, 196 (46%) scored less than 80% and received intensive training in 1999. From a previous mean score of 65% their results increased to 90% (P < 0.0001). In 2001, they again underwent proficiency testing, and the mean score was 83%. The main factors affecting proficiency testing results were the type of laboratory in which the microscopists worked and the number of low-positive slides (1-9/100) in the test. Laboratories whose work was rechecked had better scores (P = 0.002). Proficiency testing scores and the estimated sensitivity of the microscopist's laboratory were associated (P = 0.01). CONCLUSION: External quality assessment and training improve diagnostic performance. Rechecking and proficiency testing are both viable measures of laboratory performance.

Comparison of two assays (EIA and EITB) and two samples (saliva and serum) for the diagnosis of neurocysticercosis.

The enzyme immunoassay (EIA), standardized with a crude extract, and the enzyme-linked immunoelectrotransfer blot assay (ETIB) with glycoprotein antigens, were compared by using saliva and serum in the search for specific antibodies against Taenia solium larvae, for the diagnosis of neurocysticercosis. Saliva was slightly more sensitive in EIA (82.1%) than serum (74.1%). In EITB serum was far more sensitive (100%) than saliva (70.4%). The use of EITB with serum is thus an excellent choice for diagnosis of clinical cases of neurocysticercosis, while EIA using saliva represents a useful combination for diagnosis and, especially, epidemiology, because saliva is easily obtained by a painless and non-invasive procedure and the technique is simpler to perform. Furthermore, cross-reactivity of EIA with Echinococcus does not interfere in countries like Mexico where human hydatid disease is not present.

Comparative analysis of human and porcine neurocysticercosis by computed tomography.

Human and porcine cases of neurocysticerosis were compared by computed tomography in regard to the morphological characteristics of the parasites and the impact of the infection on the brain. In pigs and young people many parasites were usually found, but this was seldom the case in adult humans. These parasites were small and located mainly in the parenchyma and the subarachnoid space; no calcified cysticerci were found. In contrast, in adult humans cysticerci were heterogeneous in appearance and distribution. The brain of the pigs showed no evidence of inflammatory reaction while young people had oedema, enhancement and hydrocephalus; in contrast, adults showed diverse tomographic characteristics of brain response. Thus, the disease has at least 2 different forms. It may be mild with short development and parasites of similar appearance, as occurs in young hosts. In contrast, a long lasting disease occurs in adults, with diverse tomographic, neurological and parasitological features.

Mass treatment against human taeniasis for the control of cysticercosis: a population-based intervention study.

An intervention study with mass treatment against taeniasis to prevent neurocysticercosis due to Taenia solium in a rural community in Mexico was performed in 1991-96. Information and biological samples were obtained at the beginning of the study, at 6 months and at 42 months after mass treatment with praziquantel at a single dose of 5 mg/kg. Prevalence rates of taeniasis were measured by the detection of Taenia coproantigens and Taenia eggs in faeces; neurocysticercosis was suggested by clinical data and by serum antibodies in humans and also in swine. A reduction of 53% after 6 months and of 56% after 42 months for human taeniasis was seen after treatment. Late-onset general seizures decreased 70%. Anti-cysticercus antibodies in the human population were reduced by 75% after 42 months. Antibodies in pigs also showed a significant reduction of 55% after 6 months. In conclusion, an impact of mass chemotherapy against taeniasis to control cysticercosis in the short and long term was demonstrated. Praziquantel for tapeworm treatment should not be given at doses lower than 10 mg/kg. Late-onset convulsive crisis and specific antibodies are good indicators of neurocysticercosis and of exposure to the parasite, respectively.