Determination of the geometrical diarylpropenamine isomers in feces by high-performance liquid chromatography

Diarylpropenamine derivatives are a class of compounds which have been evaluated as potential drug candidates. Here a specific and reproducible HPLC method for the determination of cis- and trans-isomers of the unsubstituted derivative, 3-(4'-bromo-[1,1'-biphenyl]-4-yl)-3-(4-X-phenyl-N,N-dimethyl-2-propen-1-amine (I, where X=H) in feces is described. The analyte I and internal standard, nitro derivative (II, where X=NO2), were isolated from the basified biological matrix using a liquid-liquid extraction with ethyl acetate followed by a solid-phase procedure performed on a silica cartridge. The organic phase was evaporated to dryness, the residue was reconstituted in mobile phase and injected into the HPLC system. The analytes were eluted with ethyl acetate-hexane-triethylamine (59:40:1) in HPLC column (silica) and detected by UV spectrophotometry at 272 nm. Linearity, precision and accuracy data for feces standards after extraction were acceptable. The method has been applied to analyses of feces samples from rats dosed with I, in which it could be anticipated that fecal excretion is quantitatively the major route for I elimination. Copyright (C) 1999 Elsevier Science B.V.

Parasitoses em lactentes de 0-12 meses atendidos na Faculdade de Medicina de Botucatu - São Paulo

Infestation by protozoa and/or helminths is considered to be extremely rare in infants. We therefore reviewed the records of all infants up to 12 mth of age attending the Paediatric Department from January 1973 to June 1977, in order to assess the frequency of stool examinations for parasites within this age group, the percentage of positive results in these examinations, and some personal characteristics which propitiated parasitic infestation. Some characteristics of the investigated infants were compared to those of a representative sample of the total number of infants attending during the same period. Stool examinations were realized in 11.8% of the 1,162 attending patients and of those, 15.3% were positive. The proportion of positive cases became 24.0% when the infants in whom there was a history of parasites being passed were included. Ascaris lumbricoides and Giardia lamblia were the parasites most frequently found, respectively in 42.8% and 37.1% of the positive cases. The only detected difference between the control group and the group of infants who had parasitologic examinations was the presence of diarrhea, significantly more frequent in the investigated group. The proportion of positive cases was significantly greater in girls, in infants older than 6 mth and when diarrhea was present for up to 15 days, than in boys, in infants up to 6 mth of age and in chronic diarrhea, respectively. Our results show that parasites were investigated in only a small percentage of infants under 1 yr of age, but that in this age group already, parasitic infestation does occur with a certain frequency. As investigated cases were selected, mainly due to the presence of diarrhea, the real prevalence of parasitism in infants under 1 yr of age was impossible to evaluate.

Control of parasitic infections among school children in the peri-urban area of Botucatu, Sao Paulo, Brazil

The prevalence of intestinal parasitosis was investigated in a primary school located in Rubiao Junior, a peri-urban district of Botucatu, Sao Paulo state, Brazil, in order to assess the effect of treatment and practical measures of prophylaxis in the control of parasitic infections among 7-to-18-year-old school children of a low socio-economic status. The first series of parasitological examinations included 219 school children, of which 123 (56.1%) were found to be infected with one or more parasite species. Eighty-four children carrying pathogenic parasites were submitted to various anti-parasitic treatment schedules. We re-evaluated 75 (89%) students after 4 to 6 months postchemotherapy. The results indicate that the combination of treatment with prophylactic measures has been successful in the control of parasitic infections, since reinfection rates were generally low (≤5.3%), except for Giardia lamblia infections (18.6%), and a marked reduction on the prevalence rates was observed with a significant percentage of cure (≤73.1%) in children infected with most parasite species. The reasons for the apparent failure in the control of infections caused by Hymenolepsis nana and Strongyloides stercoralis are discussed.

Metabolismo oxidativo de neutrófilos em ovelhas naturalmente infectadas por nematódeos gastrintestinais e correlação entre nível sérico de cortisol e carga parasitária

The relationships between neutrophils oxidative metabolism, cortisol serum levels and worm burden, estimated by fecal egg count (EPG), were studied in sheep naturally infected by gastrointestinal parasites at the end of pregnancy, during lactation, and after weaning. Twenty-two Suffolk sheep three to four year old, of same parity and season of parturition were used. Serum cortisol was determined by radioimmunoassay and the neutrophils oxidative metabolism by the nitroblue tetrazolium (NBT) reduction test. The highest EPG values were observed on the fifth week of lactation while the highest rates of cortisol and reduction of NBT occurred on the fourth week after weaning. A positive correlation (r = 0.52; P<0.01) was observed between the neutrophils capacity to reduce the NBT and the serum cortisol concentration in the pre-parturition period. Otherwise, the neutrophils oxidative metabolism decreased near to the parturition. A negative correlation (r = -0.39; P<0.01) between EPG and NBT reduction test was observed after weaning, which was coincident with the increase in the neutrophils capacity to reduce NBT, indicating that animals showing better immune response after weaning have neutrophils with higher oxidative metabolism and lower worm burden.

Cryptococcosis outbreak in psittacine birds in Brazil

An outbreak of cryptococcosis occurred in a breeding aviary in São Paulo, Brazil. Seven psittacine birds (of species Charmosyna papou, Lorius lory, Trichoglossus goldiei, Psittacula krameri and Psittacus erithacus) died of disseminated cryptococcosis. Incoordination, progressive paralysis and difficulty in flying were seen in five birds, whereas superficial lesions coincident with respiratory alterations were seen in two birds. Encapsulated yeasts suggestive of Cryptococcus sp. were seen in faecal smears stained with India ink in two cases. Histological examination of the birds showed cryptococcal cells in various tissues, including the beak, choana, sinus, lungs, air sacs, heart, liver, spleen, kidneys, intestines and central nervous system. High titres of cryptococcal antigen were observed in the serum of an affected bird. In this case, titres increased during treatment and the bird eventually died. Yeasts were isolated from the nasal mass, faeces and liver of one bird. Cryptococcus neoformans var. gattii serovar B was identified based on biochemical, physiological and serological tests. These strains were resistant (minimum inhibitory concentration 64 μ g/ml) to fluconazole. This is the first report of C. neoformans var. gattii occurring in psittacine birds in Brazil. © 2004 ISHAM.

Avaliação da eficiência dos métodos Direto, COPROTEST e Rugai no diagnóstico da estrongiloidíase

Strongyloidiasis, a relatively common parasitism in tropical and sub-tropical areas, is the result of the infection by the smaller nematode, the Strongyloides stercoralis. Humans can be infected by this parasite, which has in its vital cycle free-life forms of male and female individuals able to live in the ground, and with another step necessary parasitism in the intestinal wall. The diagnostic of the infection is routinely done by the microscopic observation of the larva in stool samples and the high sensibility of urn method over another one allows an trustable and efficient diagnostic. The efficiency of three methods (Direct, COPROTEST and Rugai) used in the Parasitology Sector of the NAC-LACAL in Araraquara (SP) to diagnosis the strongiloidiasis were evaluated. A number of 2346 samples of stool of patients from NAC-LACAL and Nestor Goulart Reis Hospital were analyzed in the period between August and December of 2002. The Rugai Method with an positivity index of 65 % was elected as the most efficient of thee ones.

Identificação de Enterobacter spp. através de técnicas convencionais e do sistema semi-automatizado Microscan (Autoscan-4)

A hundred and ninety-five (195) strains of Enterobacter spp., isolated from diverse clinical specimens - urine, feces, cateter, blood, wound, tracheal aspirate, vaginal fluid - were submitted to the conventional identification by biochemical tests, and were also submitted to the identification by panels NegCombo 20 of the system automated MicroScan - AutoScan- 4 (Dade Behring Inc., West Sacramento, CA, USA). The samples were from patients of the Clinical Laboratory from the School of Pharmacy and Biochemistry of UNOESTE, Presidente Prudente, SP, and from patients hospitalized at the University Hospital Domingos Leonardo Cerávolo, UNOESTE. Of the total of strains tested, 191 (97.9%) presented agreement between the two approaches utilized and 4 strains (2.1%) presented identification disagreement, that is, the genus identified was different in each approach. By this study, the conclusion is that both the approaches utilized for the identification presented advantages and disadvantages related to the cost, facility of execution, quickness, reliability and some other characteristics. Even so, our results showed that conventional methods represent a reliable tool for Enterobacter identification.

Comparison of hemodynamic, clinicopathologic, and gastrointestinal motility effects and recovery characteristics of anesthesia with isoflurane and halothane in horses undergoing arthroscopic surgery

Objective - To compare hemodynamic, clinicopathologic, and gastrointestinal motility effects and recovery characteristics of halothane and isoflurane in horses undergoing arthroscopic surgery. Animals - 8 healthy adult horses. Procedure - Anesthesia was maintained with isoflurane or halothane (crossover study). At 6 intervals during anesthesia and surgery, cardiopulmonary variables and related derived values were recorded. Recovery from anesthesia was assessed; gastrointestinal tract motility was subjectively monitored for 72 hours after anesthesia. Horses were administered chromium, and fecal chromium concentration was used to assess intestinal transit time. Venous blood samples were collected for clinicopathologic analyses before and 2, 24, and 48 hours after anesthesia. Results - Compared with halothane-anesthetized horses, cardiac index, oxygen delivery, and heart rate were higher and systemic vascular resistance was lower in isoflurane-anesthetized horses. Mean arterial blood pressure and the dobutamine dose required to maintain blood pressure were similar for both treatments. Duration and quality of recovery from anesthesia did not differ between treatments, although the recovery periods were somewhat shorter with isoflurane. After isoflurane anesthesia, gastrointestinal motility normalized earlier and intestinal transit time of chromium was shorter than that detected after halothane anesthesia. Compared with isoflurane, halothane was associated with increases in serum aspartate transaminase and glutamate dehydrogenase activities, but there were no other important differences in clinicopathologic variables between treatments. Conclusions and clinical relevance - Compared with halothane, isoflurane appears to be associated with better hemodynamic stability during anesthesia, less hepatic and muscle damage, and more rapid return of normal intestinal motility after anesthesia in horses undergoing arthroscopic procedures.

Avaliação de diferentes métodos no diagnóstico laboratorial de Giardia lamblia

The protozoan parasite Giardia lamblia is responsible for a common intestinal infection in all regions of the world. In this study, four laboratory tests were evaluated for diagnostic reproducibility in the detection of this infection: Coprotest®, Direct modified method, Faust method and iron-hematoxylin staining. Positive diagnoses were tested for association with factors such as the age group and gender of the subject and the month when the sample was taken. Feces of 200 children in the Araraquara region (SP, Brazil) were examined by all four methods and the results compared. G. lamblia was found to be the most frequent parasite and 8.0% of the children showed giardiasis. There was no apparent correlation with gender, but most of the parasites were found in three- to five-year-olds. The highest frequency of infection occurred in January. The most reliable diagnostic results for G. lamblia were achieved by combining at least two methods of good reproducibility, i.e. Coprotest-Faust, Direct-Faust or Coprotest-Direct (A > 0.81).

Prevention of soil-transmitted helminth infection

Soil-transmitted helminths (STHs) form one of the most important groups of infectious agents and are the cause of serious global health problems. The most important STHs are roundworms (Ascaris lumbricoides), whipworms (Trichuris trichiura) and hookworms (Necator americanus or Ancylostoma duodenale); on a global level, more than a billion people have been infected by at least one species of this group of pathogens. This review explores the general concepts of transmission dynamics and the environment and intensity of infection and morbidity of STHs. The global strategy for the control of soil-transmitted helminthiasis is based on (i) regular anthelminthic treatment, (ii) health education, (iii) sanitation and personal hygiene and (iv) other means of prevention with vaccines and remote sensoring. The reasons for the development of a control strategy based on population intervention rather than on individual treatment are discussed, as well as the costs of the prevention of STHs, although these cannot always be calculated because interventions in health education are difficult to measure. An efficient sanitation infrastructure can reduce the morbidity of STHs and eliminates the underlying cause of most poverty-related diseases and thus supports the economic development of a country.

Amplifiability of mitochondrial, microsatellite and amelogenin DNA loci from fecal samples of red brocket deer Mazama americana (Cetartiodactyla, Cervidae)

We tried to amplify mitochondrial, microsatellite and amelogenin loci in DNA from fecal samples of a wild Mazama americana population. Fifty-two deer fecal samples were collected from a 600-ha seasonal semideciduous forest fragment in a subtropical region of Brazil (21°20′, 47°17′W), with the help of a detection dog; then, stored in ethanol and georeferenced. Among these samples 16 were classified as fresh and 36 as non-fresh. DNA was extracted using the QIAamp® DNA Stool Mini Kit. Mitochondrial loci were amplified in 49 of the 52 samples. Five microsatellite loci were amplified by PCR; success in amplification varied according to locus size and sample age. Successful amplifications were achieved in 10/16 of the fresh and in 13/36 of the non-fresh samples; a negative correlation (R = -0.82) was found between successful amplification and locus size. Amplification of the amelogenin locus was successful in 22 of the 52 samples. The difficulty of amplifying nuclear loci in DNA samples extractedfrom feces collected in the field was evident. Some methodological improvements, including collecting fresh samples, selecting primers for shorter loci and quantifying the extracted DNA by real-time PCR, are suggested to increase amplification success in future studies. © FUNPEC-RP.

Haemonchus contortus: A multiple-resistant Brazilian isolate and the costs for its characterization and maintenance for research use

The aim of this work was to determine the resistance level of Haemonchus contortus isolated from the Santa Inês flock of the Embrapa (Brazilian government's Agricultural Research Company), Southeast Livestock Unit (CPPSE), as well as to determine costs of characterizing and maintaining this isolate in host donors. Forty-two male Santa Inês lambs were experimentally infected with 4000 H. contortus infective larvae of the field isolate of CPPSE, called Embrapa2010, and divided into six treatment groups, which received triclorfon, albendazol plus cobalt sulfate, ivermectin, moxidectin, closantel and levamisole phosphate, as well as a negative control group (water). Egg per gram (EPG) counts were performed at 0, 3, 7, 10 and 14. days post treatment when the animals were slaughtered for parasite count. The data were analyzed using the RESO statistical program, considering anthelmintic resistance under 95% of efficacy. EPG and worm count presented a linear and significant relation with 94% determination coefficient. The susceptibility results obtained by RESO through both criteria (EPG and worm count) were equal, except for closantel, showing that the isolate Embrapa2010 is resistant to benzimidazoles, macrocyclic lactones and imidazothiazoles. The need of a control group did not appear to be essential since the result for susceptibility in the analyses with or without this group was the same. Suppression in egg production after treatment did not occur in the ivermectin and moxidectin groups. In the control group, the establishment percentage was just 12.5 because of the low number of third-stage larvae, resistance (innate and infection immunity) of the animals studied plus good nutrition. Drug classes presented similar efficacy between adults and immature stages. The costs for isolate characterization were calculated for 42 animals during 60. days. The total cost based on local market rates was approximately US$ 8000. The precise identification of Brazilian isolates and their establishment in host donors would be useful for laboratorial anthelmintic resistance diagnoses through in vitro tests, which has an annual cost of approximately US$ 2500 for maintenance in host donors. © 2012 Elsevier B.V.

Resistance against gastrointestinal nematodes in Crioulo Lageano and crossbred Angus cattle in southern Brazil

Gastrointestinal nematode (GIN) infection is a major cause of production losses in cattle. This study was carried out to evaluate the natural resistance against nematode infection in Crioulo Lageano and crossbred Angus male calves. Crioulo Lageano is a local cattle breed in the state of Santa Catarina, in southern Brazil. Ten weaned calves of each breed were grazed together on pasture and naturally infected with nematodes between July 2009 and December 2010. Once every 28 days, we collected fecal and blood samples for parasitological and immunological tests, as well as recording body weights. After 19 samplings, all animals were slaughtered for quantification and identification of GINs. We found that the animals had been infected with the following nematode species, in decreasing order by the mean number of specimens: Trichostrongylus axei, Cooperia punctata, Ostertagia ostertagi, Haemonchus placei, Oesophagostomum radiatum, and Trichuris spp. There were no significant differences between the Crioulo Lageano and crossbred Angus groups in terms of worm burden or nematode fecal egg count, nor in terms of the mean levels of immunoglobulin (G and A) against C. punctata and H. placei antigens, except in IgA mean level in abomasal mucus against H. placei adult worms that was significantly higher in crossbred Angus cattle (p<0.05). At the end of the study, the crossbred Angus cattle were heavier than were the Crioulo Lageano cattle (mean live weight, 507.35 and 390.3. kg, respectively). Comparative parasitological and immunological evaluation revealed no difference between two breeds in terms of their natural resistance against GINs. © 2012 Elsevier B.V.

Resistance of beef cattle of two genetic groups to ectoparasites and gastrointestinal nematodes in the state of São Paulo, Brazil

The resistance to infestations by ectoparasites and infections by gastrointestinal nematodes was studied in 45 animals (males and females) of two genetic groups: purebred Nelore (NI, n=28) and Three-Cross (1/2 Angus+1/4 Canchim+1/4 Nelore - TC, n=17). The animals were monitored for 24months, during which they were left to graze in tropical pastures without receiving treatment for parasites. Each month the animals were examined for infestations by external parasites, to count the numbers of cattle ticks Rhipicephalus microplus with diameter greater than 4.5mm present on the left side, horn flies (Haematobia irritans) present in the lumbar region and botfly larvae (Dermatobia hominis) present on the entire body. The H. irritans counts were performed with the aid of digital photographs. At the time of examination, fecal samples were collected to count the eggs per gram (EPG) and to perform coprocultures, and peripheral blood samples were drawn to determine the packed cell volume (PCV) and to count the eosinophils. For statistical analysis, the count data were transformed into log10 (n+1), where n is the number of parasites. For PCV, significant effects (P<0.05) were found for collection month (CO), genetic group (GG) and gender (SX), with means and respective standard errors of 41.5±0.65% for the NI animals, 39.3±0.83% for the TC, 41.5±0.72% for the females and 39.3±0.77% for the males. Regarding the eosinophil counts, only the effect of sex was significant (P<0.01), with means and respective standard errors of 926.0±46.2/μL, for males and 1088.0±43.8/μL of blood, for females. The NI animals presented lower mean counts for all the external parasites compared to the TC animals (P<0.01). For ticks, the transformed means followed by standard errors for the NI and TC animals were 0.06±0.01 and 0.34±0.02, while for horn flies these were 0.92±0.05 and 1.36±0.06 and for botfly larvae they were 0.05±0.03 and 0.45±0.05, respectively. The average EPG values were only influenced by CO (P<0.01). The coprocultures revealed the presence of the following endoparasites: Haemonchus spp., Cooperia spp., Oesophagostomum spp. and Trichostrongylus spp., the last in smaller proportion. There were no significant differences between the genetic groups for the endoparasite loads, except for Cooperia spp., which were present in greater number (P<0.05) in the NI group. The results obtained in this experiment confirm previous findings of greater susceptibility of the Nelore breed to Cooperia spp. and high resistance to ectoparasites. © 2013 Elsevier B.V.