Rheological effects of high substitution levels of fats by inulin in whole cassava dough: chemical and physical characterization of produced biscuits.

Inulin has been used as a fat substitute in baked goods due to its gelling properties and functionality. However, it usually has been done in low substitution levels. The aim of this study was to evaluate the effect in rheological parameters in cassava dough caused by high substitution levels of fats by inulin. Physical and chemical characterization of biscuits were done for evaluation of dough's potential usage in bakery products. Substitution of mozzarella cheese by inulin showed that up to 12.5 g inulin per 100 g cassava flour led to a lower storage modulus, suggesting bakery potential for bread like products. Higher substitutions (25 g, 37.5 g, and 50 g per 100 g cassava flour) led to a higher storage modulus and suggest a potential for cookies and similar products. Hardness results obtained support rheological ones, suggesting higher substitution levels are suitable for cookies and similar products. This is inferred by observing an increase in hardness ranging from 5.80 N up to 17.47 N. Cassava dough with fat substituted by inulin in high levels, has potential in the development of different baked goods.

Potential functional bakery products as delivery systems for prebiotics and probiotics health enhancers.

Several health benefits have been associated to probiotics and prebiotics, most of these are involved in the regulation of the host's gut microbiome. Their incorporation to diverse food products has been done to develop potential functional foods. In the case of bakery products, their incorporation has been seen to improve several technological parameters such as volume, specific volume, texture along with sensorial parameters such as flavor and aroma. Scientific literature in this topic has been divided in three main research branches: nutrition, physical quality and sensory analyzes, however, studies rarely cover all of them. Due to the harsh thermal stress during baking, sourdough technology along with microencapsulation of probiotics, has been studied as an alternative to enhance its nutritional values and increase cell viability, though in few occasions. The potential functional baked goods have maintained acceptable physical characteristics and sensorial acceptability, while in some cases an improvement is seen due to the effect of probiotics and prebiotics. The results obtained from several studies done, have shown the viability of developing functional bakery products by applying prebiotics or probiotics. This could be used as an encouragement for more research to be done in this topic.

Variation in the cellular localization of host-protective oncospheral antigens in Taenia saginata and Taenia solium.

Immunohistochemistry and immunofluorescence with confocal microscopy were used to localize the host-protective antigens of Taenia saginata (TSA9 and TSA18) and Taenia solium (TSOL16, TSOL18 and TSOL45). In nonactivated oncospheres, TSA9 and TSOL45 antigens were found primarily in the cytoplasm of the penetration gland type one (PG1) cell. A similar pattern of staining was seen for TSOL45 in oncospheres of T. solium that remained within the oncospheral membrane. In addition, there was less intense staining of TSA9 and TSOL45 in the quadri-nucleate penetration gland type 2 (PG2) cell. TSA18, TSOL16 and TSOL18 were predominantly found in the PG2 cell. In activated oncospheres that had escaped the oncospheral membrane, the antigens (other than TSA9) were seen both in the penetration gland cell locations and throughout the oncospheral parenchyma. Co-localization analyses revealed that only TSOL16 and TSOL18 antigens were co-localized in the PG2 cell of oncospheres that had not escaped the oncospheral membrane. However, in activated oncospheres that escaped the oncospheral membrane, all three antigens of T. solium were co-localized as they were present throughout the parenchyma. No positive staining was observed on the surface of nonactivated or recently activated oncospheres of T. saginata or T. solium.

Proteomic study of activated Taenia solium oncospheres.

Taenia solium cysticerci are a major cause of human seizures and epilepsy in the world. In the gastrointestinal tract of infected individuals, taeniid eggs release the oncospheres, which are then activated by intestinal stimuli, getting ready to penetrate the gut wall and reach distant locations where they transform in cysticerci. Information about oncospheral molecules is scarce, and elucidation of the oncosphere proteome could help understanding the host-parasite relationship during the first steps of infection. In this study, using liquid chromatography and tandem mass spectrometry (LC-MS/MS) analysis, we could identify a set of oncospheral proteins involved in adhesion, protein folding, detoxification and proteolysis, among others. In addition, we have characterized one of the identified molecules, the parasite 14-3-3, by immunoblot and immunolocalization. The identification of these oncospheral proteins represents the first step to elucidate their specific roles in the biology of the host-parasite relationship.

Combined human and porcine mass chemotherapy for the control of T. solium.

A combined (human and porcine) mass chemotherapy program was tested in a controlled design in 12 village hamlets in the Peruvian highlands. A single dose of 5 mg of praziquantel was given to eliminate intestinal taeniasis in humans, and two rounds of oxfendazole (30 mg/kg) were administered to all pigs. The total population in the study villages was 5,658 resident individuals, and the porcine population at the beginning of the study was 716 pigs. Human treatment coverage was 75%, ranging from 69% to 80%. There were only a few refusals of owners for porcine treatment of their animals. The effect of the intervention was measured by comparing incidence rates (seroconversion in pigs who were seronegative 4 months before) in treatment versus control villages, before and up to 18 months after treatment. There was a clear effect in decreasing prevalence (odds ratio, 0.51; P < 0.001) and incidence (odds ratio, 0.39; P < 0.013) in the treatment area after the intervention, which did not leave to extinction of the parasite but stabilized in slightly decreased rates persisting along the follow-up period. Mass chemotherapy was effective in decreasing infection pressure in this hyperendemic area. However, the magnitude of the effect was small and did not attain the goal of eliminating transmission.

Taenia solium infection in a rural community in the Peruvian Andes.

An epidemiological study was conducted in a highland, rural community in Peru, to determine the seroprevalences of human and porcine infection with Taenia solium and the risk factors associated with human infection. The seroprevalences, determined using an assay based on enzyme-linked-immuno-electrotransfer blots (EITB), were 21% (66/316) in the humans and 65% (32/49) in the pigs. The human subjects aged <30 years were more likely to be positive for anti-T. solium antibodies than the older subjects (P < 0.001). The risk factors associated with human seropositivity were lack of education beyond the elementary level [odds ratio (OR)=2.69; 95% confidence interval (CI)=1.09-6.65] and pig-raising (OR=1.68; CI=0.96-2.92). Curiously, sheep-raising was inversely associated with human T. solium infection (OR=0.50; CI=0.28-0.90). The study site appears to be a new endemic focus for T. solium in the central Peruvian Andes. Although, in earlier studies, the seroprevalence of T. solium infection has generally been found to increase with age, the opposite trend was observed in the present study. The results of follow-up studies should help determine if the relatively high seroprevalence in the young subjects of the present study is the result of a transient antibody response.

Seroincidence of porcine T. solium infection in the Peruvian highlands.

We performed repeated serological sampling of pigs in an endemic area of the Peruvian highlands (eight villages) to assess the feasibility of detecting incident cases of Taenia solium infection as indicators of ongoing transmission of the parasite. A total of 2245 samples corresponding to 1548 pigs were collected in three sampling rounds (n=716, 926, and 603, respectively). Village-period specific seroprevalences of antibodies by enzyme-linked immunoelectrotransfer blot (EITB) assay varied from 39% (95% CI: 34, 44) to 76% (95% CI: 72, 79). The prevalence of cysticercosis increased with the age of the pigs (similarly for both sexes). Around 40% of pigs were re-sampled at the end of each 4-month period. Crude incidence risks were 48% (57/120, 95% CI: 43-52) and 58% (111/192, 95% CI: 54-61) for each period. A proportion of seropositive animals became seronegative at the end of each period (23 and 15%). Incidence varied by the village, and the exposure period, and was higher in males than females (but did not differ by age).

Protection of pigs with cysticercosis from further infections after treatment with oxfendazole.

Cysticercosis, the infection by the larvae of Taenia solium, is a major cause of acquired epilepsy in the world; it also causes significant economic loss because of contaminated pork. This disease is endemic in most developing countries and no control strategy has yet been proven efficient and sustainable. To further evaluate the full potential of single-dose oxfendazole treatment for pigs as a control measure, 20 pigs with cysticercosis were treated with oxfendazole and later matched with 41 naive pigs and exposed to a natural challenge in a hyperendemic area. New infections were found by serologic testing in 15 of the 32 controls (47%), and by the presence of cysts at necropsy in 12 of them (37%). Only minute residual scars were detected in the carcasses of oxfendazole-treated pigs. Pigs with cysticercosis, once treated with oxfendazole, are protected from new infections for at least three months.

Short report: transient antibody response in Taenia solium infection in field conditions-a major contributor to high seroprevalence.

The discordance between extremely high seroprevalence of Taenia solium antibodies in disease-endemic populations, relatively few symptomatic cases of neurocysticercosis, and high background levels of putatively inactive brain lesions (mainly calcifications) in seronegative controls have confused researchers, clinicians, and epidemiologists in the last decade. We reviewed longitudinal serologic data from general population serosurveys in 3 different disease-endemic areas of Peru and Colombia and found that approximately 40% of seropositive people were seronegative when resampled after 1 year (3 surveys) or after 3 years (1 survey). Transient antibodies may have significant implications for the epidemiology of and immunity to this disease.

Experimental infection model for Taenia solium cysticercosis in swine. Cysticercosis Working Group in Peru.

A novel method for infecting pigs with Taenia solium using an intramuscular innoculum of oncospheres was investigated in a series of five experiments in 18 animals. The model is simple to perform, requires a minimal number of oncospheres, permits multiple infections per animal, and decreases the variation inherent in oral infection models. This intramuscular oncosphere assay (IMOA) may provide a valuable tool to evaluate therapeutic agents or potential vaccines for cysticercosis.

Natural history of infection with Bartonella bacilliformis in a nonendemic population.

An investigation was performed after an outbreak of bartonellosis in a region of Peru nonendemic for this disorder. Symptoms of acute and chronic bartonellosis were recorded. Serological analysis was performed on 55% of the affected population (554 individuals), 77.5% of whom demonstrated previous infection with Bartonella bacilliformis. The attack rate of Oroya fever was 13.8% (123 cases); the case-fatality rate was 0.7%. The attack rate of verruga peruana was 17.6%. A new specific immunostain was developed and used to confirm the presence of B. bacilliformis in the biopsied skin lesions. Most seropositive individuals (56%) were asymptomatic. The symptoms that were associated with prior infection, as determined by Western blot, included fever (37.2% of the seropositive vs. 17.2% of the seronegative population; P<.001), bone and joint pain (27% vs. 9%; P<.001), headache (27% vs. 12.3%; P <.001), and skin lesions described as verruga peruana (26.8% vs. 4.9%; P<.001). Our findings suggest that infection with B. bacilliformis causes a broad spectrum of disease that is significantly milder in severity than that frequently reported.

Sonicated diagnostic immunoblot for bartonellosis.

Two simple Bartonella bacilliformis immunoblot preparation methods were developed. Antigen was prepared by two different methods: sonication of whole organisms or glycine extraction. Both methods were then tested for sensitivity and specificity. Well-defined control sera were utilized in the development of these diagnostic immunoblots, and possible cross-reactions were thoroughly examined. Sera investigated for cross-reaction with these diagnostic antigens were drawn from patients with brucellosis, chlamydiosis, Q fever, and cat scratch disease, all of whom were from regions where bartonellosis is not endemic. While both immunoblots yielded reasonable sensitivity and high specificity, we recommend the use of the sonicated immunoblot, which has a higher sensitivity when used to detect acute disease and produces fewer cross-reactions. The sonicated immunoblot reported here is 94% sensitive to chronic bartonellosis and 70% sensitive to acute bartonellosis. In a healthy group, it is 100% specific. This immunoblot preparation requires a simple sonication protocol for the harvesting of B. bacilliformis antigens and is well suited for use in regions of endemicity.

Differentiating Taenia solium and Taenia saginata infections by simple hematoxylin-eosin staining and PCR-restriction enzyme analysis.

Species-specific identification of human tapeworm infections is important for public health purposes, because prompt identification of Taenia solium carriers may prevent further human cysticercosis infections (a major cause of acquired epilepsy). Two practical methods for the differentiation of cestode proglottids, (i) routine embedding, sectioning, and hematoxylin-eosin (HE) staining and (ii) PCR with restriction enzyme analysis (PCR-REA), were tested on samples from 40 individuals infected with T. solium (n = 34) or Taenia saginata (n = 6). Microscopic examination of HE staining of sections from 24 cases, in which conserved proglottids were recovered, clearly revealed differences in the number of uterine branches. Distinct restriction patterns for T. solium and T. saginata were observed when the PCR products containing the ribosomal 5.8S gene plus internal transcribed spacer regions were digested with either AluI, DdeI, or MboI. Both HE histology and PCR-REA are useful techniques for differentiating T. solium from T. saginata. Importantly, both techniques can be used in zones of endemicity. HE histology is inexpensive and is currently available in most regions of endemicity, and PCR-REA can be performed in most hospital centers already performing PCR without additional equipment or the use of radioactive material.

Human hydatidosis in the central Andes of Peru: evolution of the disease over 3 years.

To document the natural history of Echinococcus granulosus infection and response to treatment of human hydatidosis, we reexamined 28 of 37 subjects with E. granulosus infection diagnosed in an epidemiological study conducted in 1994. Twenty-six (70%) of those 37 subjects underwent abdominal ultrasonography, chest radiography, and enzyme-linked immunoelectrotransfer blot assay in 1997. Medical records from two additional individuals were reviewed. Eight patients had their cysts surgically removed during the 3-year follow-up interval; no surgical complications or recurrences occurred. Among eight patients with cystic disease not treated by surgery, four had cyst-growth ranging from 0.4 to 1.4 cm during the 3-year interval. One patient developed a new cyst and another's simple cyst became septate; two developed new calcifications. Of 12 seropositive subjects with no cysts present in 1994, 10 reverted to seronegative, a finding that suggests a significant proportion of seropositive subjects in echinococcus-endemic regions may have only transient infection without disease. When cysts do develop, their growth rates and time courses are highly variable; over the 3-year period, we observed growth, septation, degeneration, and calcification of cysts.

Magnitude of the disease burden from neurocysticercosis in a developing country.

Cysticercosis contributes to higher epilepsy rates in developing countries than in industrialized ones, yet no estimate exists for the associated burden of disease. We used epidemiological data on neurocysticercosis in Peru to calculate the burden of disease and applied our model to the other countries of Latin America where neurocysticercosis is endemic to determine a regional estimate. Analysis of 12 population-based community studies demonstrated that neurocysticercosis was endemic in highland areas and high jungles, with seroprevalences from 6% to 24%. In one community, the adult seizure disorder rate was 9.1% among seropositive persons versus 4. 6% among seronegative persons; we used this difference for estimates. On the basis of average prevalence rates in areas of endemicity of 6%-10%, we estimated that there are 23,512-39,186 symptomatic neurocysticercosis cases in Peru. In Latin America, an estimated 75 million persons live in areas where cysticercosis is endemic, and approximately 400,000 have symptomatic disease. Cysticercosis contributes substantially to neurological disease in Peru and in all of Latin America.

Persistence of passively transferred antibodies in porcine Taenia solium cysticercosis. Cysticercosis Working Group in Peru.

We evaluated the presence and persistence of anticysticercal antibodies in piglets born to Taenia solium infected sows. Infected sows from a disease-endemic area of Peru were transported to a nondisease-endemic area and impregnated. Serum samples were collected from sows and piglets on Day 2 through Week 35 after birth. Using an immunoblot specific for cysticercosis, Ig isotypes to 7 cyst antigens were measured and quantified. Serum samples from the piglets contained detectable antibodies from Week 1 through Week 35 (27 weeks after weaning). The primary Ig isotype present in both sows and piglets was IgG. Antibodies did not appear in piglet serum samples until after suckling, demonstrating that anti-cysticercal antibodies are transferred solely via colostrum. Our data have shown that maternally transferred antibodies to cyst antigens may persist through much of a pig's life. Therefore, the presence of passively transferred antibodies must be considered in studies that examine the prevalence of cysticercosis in pigs. Furthermore, when designing control strategies for cysticercosis, careful evaluation and selection of sentinel pigs becomes a crucial component of sentinel selection.

Human and porcine Taenia solium infection in a village in the highlands of Cusco, Peru. The Cysticercosis Working Group in Peru.

A serological survey was performed using the enzyme-linked immunoelectrotransfer blot assay (EITB) in a village in the highlands of Peru where there are three distinct but close neighborhoods, to determine if there is a direct relationship between human and porcine Taenia solium infection. One hundred and eight out of 365 individuals were sampled, and 14 were seropositive (human seroprevalence 13%). Most seropositive individuals were neurologically asymptomatic. Thirty-eight out of 89 sampled pigs (43%) were seropositive. There was a clear geographical clustering of cases, and positive correlation between human and porcine seroprevalence found when comparing the three neighborhoods. Cysticercosis is an important cause of neurological morbidity in most developing countries, and control/eradication trials are now being increasingly applied. Porcine serology provides an appropriate indicator of T. solium environmental contamination and should be used to estimate the risk of infection when evaluating control measures.

Development of a serologic assay to detect Taenia solium taeniasis.

We developed a serologic assay to identify adult Taenia solium tapeworm carriers using excretory/secretory (TSES) antigens collected from in vitro cultured T. solium tapeworms. To identify taeniasis-specific antigens we used an immunoblot assay with serum samples from T. solium tapeworm carriers and cysticercosis patients. Antigens were identified that reacted with antibodies present in serum samples from taeniasis cases and not with those from cysticercosis patients. Using serum samples collected from persons with confirmed T. solium tapeworm infections, the test was determined to be 95% (69 of 73) sensitive. Serum samples (n = 193) from persons with other parasitic infections, including T. saginata tapeworm infections, do not contain cross-reacting antibodies to TSES, indicating that the assay is 100% specific. These data suggest that the immunoblot assay using TSES antigens can be used to identify persons with current or recent T. solium tapeworm infections and provides a new, important tool for epidemiologic purposes, including control and prevention strategies.

Field diagnosis of Echinococcus granulosus infection among intermediate and definitive hosts in an endemic focus of human cystic echinococcosis.

Human, canine and ovine echinococcosis prevalence was determined in a highland community located in the central Peruvian Andes during 1997 and 1998. Human echinococcosis was determined using portable ultrasonography, chest X-ray examination, and an enzyme-linked immunoelectrotransfer blot (EITB) assay. Canine echinococcosis was determined using microscopy stool examinations and a coproantigen detection enzyme-immunoassay (EIA) for Echinococcus granulosus. Ovine echinococcosis was determined by an EITB assay for sheep echinococcosis and necropsy examination of viscera from domestic slaughtered animals. An abdominal ultrasound, a chest X-ray examination and an EITB for echinococcosis were performed on 214 subjects (45% of the village population). The frequency of presumptive liver/abdominal, lung and liver-lung hydatid cysts was 5.1% (11/214), 3.7% (8/214) and 0.5% (1/214), respectively. The overall prevalence of human cystic echinococcosis was 9.3% (20/214). The frequency of canine echinococcosis was 46% (23/50) and 32% (16/50) by the coproantigen EIA test and arecoline purging, respectively. The frequency of sheep echinococcosis was 65% (22/34) by the EITB and 38% (13/34) by necropsy. We demonstrated a high prevalence of human and animal echinococcosis in this Peruvian village. In remote areas where echinococcosis is endemic, both the coproantigen EIA and arecoline purging may be used for the study of canine echinococcosis; the EITB is useful in establishing the diagnosis of echinococcosis in sheep prior to necropsy.