Diagnosis of Giardia infections by PCR-based methods in children of an endemic area

The present study was designed to estimate the prevalence of Giardia infection in preschool- and school-aged children living in an endemic area. Fecal samples from 573 children were processed by zinc sulfate centrifugal flotation, centrifugal sedimentation (using a commercial device for fecal concentration - TF-Test kit®) and polymerase chain reaction (PCR)-based methods. Of the stool samples assessed, 277 (48.3 percent) were positive for intestinal parasites and/or commensal protozoa. Centrifugal flotation presented the highest diagnostic sensitivity for Giardia infections. The kappa index revealed that both coproparasitological techniques closely agreed on the Giardia diagnosis (86 percent) versus satisfactory (72 percent) and poor (35 percent) concordances for commensal protozoan and helminth infections, respectively. Concerning Giardia molecular diagnosis, from the 71 microscopy-positive samples, specific amplification of gdh and tpi fragments was noted in 68 (95.7 percent) and 64 (90 percent) samples, respectively. Amplification of gdh and tpi genes was observed, respectively, in 95.7 percent and 90 percent of microscopy-positive Giardia samples. For 144 microscopy-negative samples, gdh and tpi gene amplification products were obtained from 8.3 percent and 35.9 percent samples, respectively. The agreement between these genes was about 40 percent. The centrifuge-flotation based method was the most suitable means of Giardia diagnosis assessed in the present study by combining accuracy and low cost.

Genotyping of Brazilian Giardia duodenalis human axenic isolates

Giardia duodenalis is a complex species that comprises at least seven distinct genetic groups (A to G), but only genotypes A and B are known to infect humans and a wide variety of other mammals. Regardless of biological, biochemical and antigenic analysis, several isolates maintained in vitro were not genetically typed yet. So, in the present study, five Brazilian axenic isolates obtained from asymptomatic and symptomatic patients were typed in order to determine the major genetic groups to which the isolates belonged. DNA was extracted from axenic trophozoites, fragments of glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes were amplified by PCR and the isolate genotyping was carried out using restriction fragment length polymorphism (RFLP) and DNA sequencing for both genes. The results revealed that all isolates were assigned to genotype A at both analyzed loci. Indeed, DNA sequence analysis classified the four isolates obtained from asymptomatic individuals into subtype AII, while the isolate obtained from the symptomatic patient was typed as subtype AI. Despite of the limited number of isolates assessed, the findings presented herein provide interesting insights on the occurrence of Giardia genotypes in Brazil and hold the perspective for future molecular and epidemiological investigations.

Occurrence of Tetrameres confusa (Nematoda, Tetrameridae) in Ara ararauna (Psittacidae)

Relata-se a ocorrência de Tetrameres confusa Travassos 1917 (= T. americana (Cram, 1927) Baylis, 1929) (Nematoda, Tetrameridae) em um novo hospedeiro, Ara ararauna Linnaeus, 1758 (Aves, Psittacidae). Este é o primeiro registro da ocorrência do nematódeo em psitacídeos no Brasil.

Strongyloides venezuelensis infection susceptibility of seven inbred strains of mice

A trial was carried out to investigate the susceptibility of seven strains of mice to Strongyloides venezuelensis primary and secondary experimental infections, in order to provide the basis for genetic studies about resistance. Twelve six-week-old male inbred mice of the A/J, BALB/c, CBA/J, C3H/Hepos, C57BL/6, DBA/2 and NIH strains were infected s.c. with 2000 infective larvae. The mean worm counts (± SD) in the small intestine six days after infection were, in increasing order: 28 (± 19) in NIH; 647 (± 228) in BALB/c; 709 (± 425) in DBA/2; 731 (± 151) in C3H/Hepos, 801 (± 174) in CBA/J; 1024 (± 267) in C57BL/6 and 1313 (± 483) in A/J. C57BL/6 mice showed the highest fecal egg counts and NIH, the lowest. No eggs in fecal exams or nematodes in small intestines were recovered from animals reinfected 14 days after primary infection. NIH strain was highly resistant to primary infection by S. venezuelensis. The most susceptible of the other six strains appeared to be the C57BL/6 strain which presented a high nematode counting in intestine and the highest egg output

Parasitological characteristics and tissue response in the abomasum of sheep infected with Haemonchus spp

A histopathological study was conducted on the abomasal mucosa of 40 Corriedale sheep exposed to natural infection with Haemonchus spp. The sheep were allowed to graze on contaminated pastures for 14 days and, after being housed for 28 days, they were slaughtered. Fecal samples were collected for fecal egg counts (FEC) and abomasum samples were obtained for histopathological examination and eosinophil, mast cell and globule leucocyte counts. The number of Haemonchus spp. present in the abomasum was estimated from a 10 per cent aliquot of its content. The number of eggs per female was estimated for 10 Haemonchus spp. females collected from each animal. The following significant correlation coefficients were obtained between the characteristics analyzed: number of Haemonchus and FEC (r = 0.86), female length and number of eggs per female (r = 0.60), female length and FEC (r = 0.53), number of eosinophils and number of Haemonchus (r = 0.48), number of eosinophils and number of globule leucocytes (r = 0.54), number of mast cells and female length (r = -0.39), and number of mast cells and number of globule leucocyte (r = 0.34). The characteristics that showed the highest correlation with animal load was FEC