Pseudoparasitosis due to accidental ingestion of chicken cecum

The mother of an eighteen months baby brought to the laboratory a round worm-like structure and claimed that it had passed in her child's feces. She mentioned that the child had no particular complaint other than difficulty in the passage of this structure through the small anal sphincter. The length of the worm-like structure was 10 cm by about 1.5 cm in its widest blind part and about 0.3 cm in its narrowest patent part [Figures 1-3]. There was a longitudinal line running on one side of its outer surface to which was attached shreds of fatty tissue [Figure 4]. It had a thick muscular wall and a body cavity, but didn't show the unique smooth configuration of a nematode. There was also brownish sand-like material coming out from the opening at the narrow end and sedimented in the bottom of the container. Microscopic examination of the sediment revealed numerous oocysts of coccidian protozoa of different sizes and early degrees of development [Figure 5]. Histopathological examination of paraffin sections prepared from the provided parasite-like structure and stained with haematoxylm and eosin, revealed parasitophorous vacuoles comparable in size containing developing merozoites in epithelial cells of a chicken's cecum [Figures 6 and 7]

Ultrastructural analysis of posterior polar endocytic phagocytosis by leishmania amastigotes

Leishmania spp. go through a complex life cycle. Developmental forms alternate between elongated, flagellated, motile, extracellular-promastigote forms; and aflagellar, ovoid amastigote-bodies. Mechanisms for survival and procurement of nutrients differ according to stage-specific requirements. To present a mechanistic insight into phagocytosis by protists, and the developmental regulation of endocytosis, achieved by investigation of the ultrastructural interaction of two Leishmania isolates from Egypt, one cutaneous and one visceral, with their host cell cytoplasm. Two leishmanial strains were used. One isolate was from a patient's cutaneous lesion acquired in northern Sinai [SH], and the other obtained from pools of livers and spleens of stray dogs from Agamy resort in Alexandria [D3]. Isolation and maintenance of promastigote and amastigote stages was extracellularly in Offut's medium and intracellularly in laboratory out bred male Syrian hamsters. Culture pellets and biopsies 1 mm thick from various hamster tissues were fixed in cold 2.4% gluteraldehyde in 2% Ca cacodylate buffer [pH 7.2]. The specimens were processed for electron microscopy. Ultrathin sections were cut on a LBK Reichard ultra microtome and post stained with uranyl acetate in lead citrate solution. Sections were examined in a Zeiss EM 952 electron microscope. In amastigotes the subpellicular microtubules, which form the cytoskeleton support for the pellicular membrane, showed pronounced free endings at the posterior pole of the parasites, still covered by the plasma membrane. The posterior area deficient of microtubules formed a cup-like invagination of variable depths according to the parasite's endocytic activity. Evidence of phagocytosis by tissue forms of both isolates is exhibited by the similarity of homogenous bodies engulfed in the posterior invaginations and the host cell cytoplasm. The exhibition of endocytic phagocytosis by intracellular Leishmania supports the concept of a common origin of protists from a phagocytic cell ancestor, and is evidence that phagocytosis, as a criterion, is not secondarily lost in the genus

Molecular characterization of Egyptian human and animal echinococcus granulosus isolates by Rapd-PCR technique

Five primers of known, but arbitrary nucleotide sequence [OPH-03, OPH-05, OPH-12, OPH-15 and OPH-18] were used to detect the genetic variability in the Egyptian human, camel and pig E. Granulosus isolates. OPH-03, OPH-05 and OPH-15 proved to be useful genetic markers of strain variation; while, OPH-12 and OPH-18 allowed the distinction at the genus level, i.e. diversified from Cysticercus tenuicollis. OPH-03 was the most effective, giving sharp distinct banding pattern and the least values of similarity coefficients. Some variations were detected within E. granulosus isolates from the same host. The level of heterogeneity was low in three of the human isolates, camel and pig strains. An individual variation was detectable within other three human isolates. Human and camel isolates were the most related pair, having similar patterns and the highest similarity coefficients. The study implied that human cases in Egypt are of the camel/dog strain and camels are important hosts for the transmission of human hydatidosis

Association of some HLA-DRB1 antigens with echinococcus granulosus specific humoral immune response

In this study, an ELISA system using crude camel hydatid fluid antigen was used to detect specific IgG and IgG1 in the sera of 35 cystic echinococcosis [CE] patients, in whom the distribution of class II HLA-DR3 anti HLA-DR11 was determined. The recorded sensitivities were 88.6% and 94.35% for IgG and IgG1, respectively. In patients with a high humoral immune response, a statistically highly significant increased frequency of HLA-DR3 was recorded for IgG with a high relative risk value [RR = 3.2] and a reasonable etiologic fraction [EF = 0.3], while HLA-DR11 recorded RR = 2.6 and EF = 0.2. For IgG1, both antigens showed a significant increased frequency [RR = 2.95 and 2.79, respectively, and EF = 0.28 and 0.23, respectively]. HLA-DR3 was highly significantly associated with complicated cases [RR = 4.36 and EF = 0.4], in whom the mean antibody units for both IgG and IgG1 were significantly raised. It was advisable to rely on IgG1 for the diagnosis of CE and to consider the genetic disposition of the patient as an important criterion in the outcome of infection

Evaluation of fast-elisa for the diagnosis of experimental trichinosis

In order to compare between FAST-ELISA and ELISA for the diagnosis of experimental trichinosis and study the kinetics of antibody and eosinophilic responses, 6 New Zealand rabbits were infected orally by Trichinella spiralis larvae. Blood was collected every other day for the first 2 weeks, then weekly for 11 weeks post infection. T. spiralis crude larval antigen was prepared for coating of ELISA plates and FAST-ELISA beads. Blood was examined for eosinophilic count and for serum antibody level by ELISA and FAST-ELISA techniques. The burden of infection was assessed by counting encysted larvae in muscle samples of the infected rabbits. By FAST-ELISA antibodies were detected 7 days post infection [PI], while with ELISA technique antibodies were detected after 10 days. Both tests detected maximum antibody levels on the 4th week. The eosinophilic count reached its peak by the 2nd week. There was a significant inverse correlation between the mean eosinophilic count and the mean larval count. FAST- ELISA proved to be more sensitive than ELISA in early detection of infection, besides being a simple, fast and sensitive assay for antibody detection against T. spiralis

Evaluation of an ELISA test in past and present schistosomiasis

An enzyme linked immunosorbent assay [ELISA] was evaluated in relation to an indirect hemagglutination [IHA] test in schistosomiasis patients who were classified by clinical, sonographic and direct methods of diagnosis. Sensitivities of ELISA and IHA, respectively, proved to be 100% and 69.23% in acute simple intestinal schistosomiasis, 95.5% and 90.4% in chronic active schistosomiasis patients, 86.06% and 67.41% in patients with past history of exposure, 80% and 64.28% in patients with hepatosplenomegaly with past history of schistosomiasis, and 96% and 80% in patients with hepatic fibrosis as shown by sonar. It was apparent that ELISA is more sensitive than IHA in acute simple schistosomiasis in patients with past history of exposure, those with bilharzial hepatosplenomegaly and those with hepatic fibrosis. Both tests were nearly equally sensitive in chronic active schistosomiasis

Comparative study of three tests [indirect hemagglutination, direct agglutination, and indirect immunofluorescence] for detection of antibodies to toxoplasma gondii in pregnant women

Sera of 600 asymptomatic pregnant women were tested by IHAT for toxoplasma antibodies. The positive reactors were further tested by IFAT and direct agglutination test [DAT]. The prevalence of toxoplasma antibodies was found to be 27.3% by IHAT, and 58.5% of those found positive were also positive by IFAT, while only 51.8% of them were positive by DAT with 71.34% degree agreement between both tests. So, IFAT was found to be more sensitive than DAT in reference to IHAT and is recommended as a confirmatory test for those found positive on screening by IHAT, DAT, compared to IFAT, gave 69.7% sensitivity and 73.5% specificity. Results support previous findings that IHAT, IFAT, and DAT measure different antibodies of Toxoplasma gondii

Parasitic infections associated with malignancy and leprosy

Results of parasitic infections, as revealed by urine and stool examination was significant [P < 0.05] in 43.3% of patients suffering from different malignant diseases and non significant [P > 0.05] in 29.3% of leprosy patients compared to 22% in control subjects. The most prevalent parasites were E. histolytica and G. lamblia. Cryptosporidium occysts were not detected. By stool examination and culture, S.stetcoralis larvae were detected only in, the malignancy group. The most common parasites occuring concomitantly were A. duodenale and S. stercoralis. By the IFAT, strongyloidiasis gave significantly higher positive results in the malignancy group than in the leprosy and control groups, IFAT for toxocariasis, showed highly significant positivity in the leprosy group and significantly positivity in the malignancy group. For toxoplasmosis, it showed highly significant positive results in both leprosy and malignancy groups. Eosinophilia was significantly more prominent among malignancy patients and insignificant among those with leprosy. Parasitic infection detected by urine and stool examination among patients with eosinophilia was found in 76% of the malignancy patients and in 66.7% of the leprosy patients

On employing leishmania promastigotes for systemic lupus erythematosus diagnosis

Based on having definite double stranded DNA determinants in the kinetoplast, it was attempted to use Leishmania promastigotes as an IFAT substrate for the detection of serum anti ds-DNA autoantibodies which are the prominent diagnostic marker of SLE. Serological inspection of different Leishmania preparations revealed that Leishmania is not analogous to Crithidia lucilia as an appropriate ds-DNA substrate. It seems likely that the species-specific molecular characters of the kinetoplast-DNA are influencing the reaction. The issue is an evidence of the distinct differentiation of the K-DNA antigenic determinants in haemoflagellates

Evaluation of a direct agglutination test and methylene blue test in diagnosis of cutaneous leishmaniasis

The usefulness and sensitivity of a direct agglutination test [DAT] in the diagnosis of cutaneous leishmaniasis infection has been investigated. Trypsin treated, formalin fixed and coomassie blue stained Leishmania promastigotes were used as antigens: L. infantum, L. donovani, L. aethiopica. Although the titers of sera from patients with localized cutaneous leishmaniasis were low, sera from lepromatous, tuberculous and toxoplasmic patients gave high titers indicating cross reactivity. Comparable results were obtained when the same sera were tested using freshly prepared antigen or antigens stored for 5 months at 4C, and with addition of 0.78% 2-meracaptoethanol to the diluents

Naturally occurring toxoplasma antibodies in serum and milk of lactating women

Serum and milk of lactating women were tested for toxoplasmosis using specific-IgG IFAT. Apparently healthy, 70 women were selected; 54 from rural and 16 from urban areas. Serum and milk were simultaneously collected from each one. Sera were positive in 22 [31.4%] of the total 70, including 16 [29.6%] and 6 [37.5%] of rural and urban groups, respectively. No statistical significant difference was found for positivity and titer levels between the two groups [P > 0.05]. Milk was positive in 12 [17.1%] of the 70 women, including 10 [18.5%] and 2 [12.5%] from rural and urban groups, respectively, having no statistical significant difference [P >0.05]. Comparing serum and milk for positivity and titer levels, also there was no statistical significant difference [P >0.05]. It is concluded that relatively low antibody levels in serum could be excreted in milk and may be protective for suckling babies. Occurrence of antibodies in serum and milk are homogeneously distributed between rural and urban inhabitants

Investigation of sarcocystis as a causative agent in cardiac disease

Sera from 56 cases with cardiomyopathy and myocarditis and 40 cases with different types of valvular diseases were tested, using the indirect fluorescent antibody test [IFAT] for the detection of antibodies to Sarcocystis. None of the cases showed specific reaction where the IFAT was negative at the cut off titer 1: 8 in all cases

Estimation of toxoplasmia gondii antibodies in patients with cardiomyopathy

The possible involvement of Toxoplasma gondii in the pathogenesis or as a complicating factor in patients with cardiomyopathy has been investigated by the serological detection of specific antibodies by IFAT. Fifty six serum samples were collected from patients who presented with dyspnoea on effort, pericardial pains, palpitations and edema of lower limbs. They were investigated by Echo, ECG and epidemiological history, and diagnosed as dilated, ischemic and restrictive cardiomyopathies and myocarditis. Forty serum samples from patients with valvular lesions were examined as a control group. 12 out of 56 samples were positive at end titer of 1: 16 [21.4%]. Sera with positive titers were found to occur mostly in patients with ischemic cardiomyopathy. All samples of the control group were negative

Characterization of Egyptian isolates of trichomonas vaginalis: I-serotyping

Antitrichomonal hyperimmune sera against T. vaginalis stocks isolated from Egyptian female patients were employed for serological differentiation of somatic and soluble antigens in the Ouchterlony gel double immunodiffusion technique. It was concluded that soluble trichomonal antigens present in association with living flagellates are stock-specific reacting with some antitrichomonal hyperimmune sera, while those present in association with dead parasites are common antigens reacting with all the sera. Three stocks [E1, E2 and E3] could be differentiated into two strains using their stock-specific antigens. The somatic antigens of six trichomonal stocks reacted with all the hyperimmune sera, denoting common antigenic make-up

Prevalence of giardia lamblia antibodies in serum and milk in lactating women from different social classes in Egypt

Prevalence and levels of systemic and milk antibodies to G. lamblia in the different social classes of the population were studied using the IFAT and nor-partigen immunoglobulin plates. Blood and milk samples were collected simultaneously from lactating women in urban [Cairo] and rural [Benha] areas. Serum IgG was present in 90% of rural low standard mothers, 58% of urban moderate standard mothers, and 25% of urban high standard mothers [P < 0.01, P < 0.001 and P < 0.01]. Antilog of mean of antibody titers was significantly higher in the low standard rural mothers than in the urban moderate and high standard ones. Specific secretory IgA antibody in milk was found in 71% of rural low standard mothers, 31% of urban moderate standard mothers, and 16.6% of urban high standard mothers [P < 0.001, P < 0.01 and P > 0.05]. The antilog of mean S-IgA titers was also higher in the low standard rural mothers. The titer levels of S-IgA in the three classes did not show any correlation with the quantitative levels of total IgA in milk, while specific IgG showed a positive correlation with the total serum IgG in the low standard rural mothers only [P < 0.05].This study documented the widely different antibody response to G.lamblia in individuals living in different social classes

Preliminary study on cross-reactions with toxocariasis

Indirect immunofluorescent antibody test [IFAT], was used to determine seropositivity for Toxocara canis in association with other helminthic infections. One hundred and thirty eight cases were tested: Group [1] composed of 98 with helminthic infections that included 40 schistosomiasis, 22 fascioliasis, 18 hydatidosis 12 ascariasis and 6 ancylostomiasis; Group [2] composed of 40 parasitologically free individuals included as control. All sera were preabsorbed with Ascaris vitulorum antigen. Positive results for toxocariasis in the infected patients were 74.5% as compared to 5% in control group. Its cross-reaction with the other helminthic parasites ranged from 55 to 100% with a statistical significant difference [P< 0.01]. Comparing the positivity between each helminthic group and control there was a statistically highly significant difference [P <0.001]. To reduce cross-reactions, preabsorption of patients sera with multiple parasitic antigens and raising the screening titre is recommended

Comparative evaluation of T spiralis IFAT antigens for serodiagnosis of trichinosis

A comparative study of three IFAT T. spiralis larval antigens [cuticular, frozen and paraffin sections of muscle larvae] is presented. Hyperimmune anti-Trichinella serum was used to evaluate sensitivity while a battery of human sera assembled from patients with other parasitic infections, non-parasitic eosinophilia, malignant diseases as well as from healthy controls were used to evaluate specificity. Results indicate that frozen sections are more stable in the laboratory [for, at least, one year] and in addition to the ease and rapid preparation, possess higher sensitivity and specificity and so, seem to be more reliable and more promising for serodiagnosis of trichinosis

Assessment of sarcocystis infection in patients with soft tissue rheumatism: preliminary evaluation of the diagnostic reliability of the IFAT in human

In preface to investigate patients with soft tissue rheumatism [myositis] for a probable underlying aetiology of Sarcocystis infection, the IFAT was standardized and evaluated. The specificity of Sarcocystis antigen prepared from a S. fusiformis strain isolated from the oesophagus of naturally infected cattle was tested against sera with +ve rheumatoid, +ve antinuclear and +ve anti - DNA factors, which are most likely to occur in patients with connective or mixed connective tissue diseases who may also present with myositis as well as sera from Toxoplasma infected individuals since both parasites are phylogenetically closely related and antibodies incidence in patients with connective tissue diseases as an opportunistic infection. The results showed absolute preclusion of false positive reactions. For sensitively, patients with different muscle complaints were examined. Three cases out of twenty showed specific antibodies. The present issues obviously indicate the validity of the IFAT for the serodiagnosis of sarcocystosis in human cases. The approach is much promising in clinical applications. By revealing the infection as an aetiology of some cases, which may be misdiagnosed, much hazards of inappropriate treatment will be presumably avoided