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PubMed2003

Staining of intestinal protozoa with Heidenhain's iron hematoxylin.

Due to its unique properties, iron hematoxylin has been traditionally recommended for staining intestinal protozoa. This process can be simplified by reducing the number of steps and periods of permanence of the slides in some of the liquids used, without detriment to the quality of the results. Thus iron hematoxylin becomes adequate for routine use. Hematoxylin is a natural dye extracted from Haematoxylon campechianum, of the family Leguminosae. It must first be 'ripened', i.e. oxidized to hematein, which reacts with ferric ammonium sulphate to produce the ferric lake (iron hematoxylin), a basic dye. Iron hematoxylin most frequently stains regressively, i.e. the slides are first overstained and then differentiated.

Animals↗
PubMed2003

Cell fractionation of parasitic protozoa: a review.

Cell fractionation, a methodological strategy for obtaining purified organelle preparations, has been applied successfully to parasitic protozoa by a number of investigators. Here we present and discuss the work of several groups that have obtained highly purified subcellular fractions from trypanosomatids, Apicomplexa and trichomonads, and whose work have added substantially to our knowledge of the cell biology of these parasites.

Animals↗
PubMed1998

Induction of cell-mediated immunity during early stages of infection with intracellular protozoa.

Toxoplasma gondii and Trypanosoma cruzi are intracellular parasites which, as part of their life cycle, induce a potent cell-mediated immunity (CMI) maintained by Th1 lymphocytes and IFN-gamma. In both cases, induction of a strong CMI is thought to protect the host against rapid parasite multiplication and consequent pathology and lethality during the acute phase of infection. However, the parasitic infection is not eliminated by the immune system and the vertebrate host serve as a parasite reservoir. In contrast, Leishmania sp, which is a slow growing parasite, appears to evade induction of CMI during early stages of infection as a strategy for surviving in a hostile environment (i.e., inside the macrophages which are their obligatory niche in the vertebrate host). Recent reports show that the initiation of IL-12 synthesis by macrophages during these parasitic infections is a key event in regulating CMI and disease outcome. The studies reviewed here indicate that activation/inhibition of distinct signaling pathways and certain macrophage functions by intracellular protozoa are important events in inducing/modulating the immune response of their vertebrate hosts, allowing parasite and host survival and therefore maintaining parasite life cycles.

Animals↗
PubMed1982

A study of intestinal protozoa including non-pathogenic Entamoeba histolytica from patients in a group of mental hospitals.

Twenty-five per cent of 174 patients residing in three mental hospitals were found to be infected with protozoa. Seven per cent of the patients were infected with Entameba histolytica. Isoenzyme characterization of the E. histolytica grown in culture showed that there were no pathogenic zymodemes of this organism in any of the specimens tested. (Am J Public Health 1982;72:178-180.)

Entamoeba histolytica↗
PubMed2005

Comparison of disinfectants for biofilm, protozoa and Legionella control.

The aim of this study was to compare the efficiency of different disinfectants applicable to Legionella control in domestic water systems. A domestic water supply simulation unit that allowed simulation of real-world conditions was developed for this purpose. The system, consisting of seven identical rigs, was used to compare treatment efficiency under equivalent conditions of system design, materials, hydraulics, water quality, temperature and initial contamination. During the study, each of six loops received continuous application of one of the following disinfectants: chlorine, electro-chlorination, chlorine dioxide, monochloramine, ozone, or copper/silver. The seventh loop was used as a control and remained untreated. Performance evaluation of these disinfectants was based on their ability to reduce not only Legionella, but also protozoa and biofilms, which contribute to the establishment and dissemination of these bacteria in water systems, and their resistance to treatments. Regarding these criteria, chlorine dioxide and chlorine (as bleach or obtained by electro-chlorination) were the most effective treatments in this study. However, in comparison with chlorine, chlorine dioxide showed a longer residual activity in the system, which constituted an advantage in the perspective of an application to extensive pipework systems.

Animals↗
PubMed2004

cAMP signalling in the kinetoplastid protozoa.

Several species of kinetoplastid protozoa cause major human infectious diseases. Trypanosoma cruzi is responsible for the fatal Chagas disease in large parts of South America, the various species of Leishmania cause a number of different human diseases with millions of patients world-wide, and the African trypanosome Trypanosoma brucei is the agent of human sleeping sickness, a disastrously re-emerging epidemic of fatal infections in Sub-Saharan Africa. Chemotherapy of all of these infections is in a very unsatisfactory state. cAMP signalling pathways in humans have provided interesting drug targets for a number of clinical conditions, from asthma to impotency. Similarly, cAMP signalling in kinetoplastids might offer useful targets for the development of novel antiparasitic drugs, which makes their exploration an urgent need. Current knowledge suggests that cAMP signalling proceeds along very similar pathways in all kinetoplastid pathogens (T. cruzi, the Leishmanias and T. brucei). Their adenylyl cyclases are structurally very different from the human enzymes and appear to function as enzyme-linked cell surface receptors. They might represent the major sensory apparatus of the kinetoplastids, guiding much of their environmental sensing and host/parasite interaction. The cAMP-specific phosphodiesterases of the kinetoplastids are rather similar to those of human cells and might function in similar ways. Essentially nothing is known on downstream effectors of cAMP in the kinetoplastids. Homologues of protein kinase A and its regulatory subunits have been identified, but their biochemical properties seem to be disctinct from that of mammalian protein kinase A.

Adenylyl Cyclases↗
PubMed1997

In vitro metabolism of phenylalanine by ruminal bacteria, protozoa, and their mixture.

An in vitro study was conducted to examine the metabolism of phenylalanine (Phe) by mixed rumen bacteria (B), mixed rumen protozoa (P), and a combination of the two (BP). Rumen microorganisms were collected from fistulated goats fed lucerne cubes (Medicago sativa) and a concentrated mixture twice a day. Microbial suspensions were anaerobically incubated at 39 degrees C for 12 h. Phe and some other related compounds in both supernatants and microbial hydrolysates of the incubations were analysed by HPLC. The net degradation rate (&mgr;mol/g microbial nitrogen) of Phe in B was about 1.5-fold higher than that in P. Phe was converted mainly into phenylacetic acid (PAA) and unknown compound(s) that presumably involved tyrosine in B, P, and BP during the 12 h incubation period. Small amounts of benzoic acid (BZA), and traces of phenylpropionic acid (PPR) and phenyllactic acid (PLA) were also produced from Phe. PAA production in B was found to be higher than that in P, whereas it was significantly higher in BP. Although BZA production was less than one-tenth that of PAA production, it was higher in P than in B and BP. PPR was detected in both B and BP, but not in P. PLA was detected only in B. The production of unknown compound(s) was higher in B than in P and BP.

Journal Article↗
PubMed1997

Synthesis of phenylalanine and production of other related compounds from phenylpyruvic acid and phenylacetic acid by ruminal bacteria, protozoa, and their mixture in vitro.

Phenylalanine (Phe) synthesis and the production of other related compounds by mixed ruminal bacteria (B), protozoa (P), and a combination of the two mixture (BP) in an in vitro system were quantitatively investigated using phenylpyruvic acid (PPY) and phenylacetic acid (PAA) as substrates. Rumen microorganisms were collected from fistulated goats fed lucerne cubes (Medicago sativa) and a concentrated mixture twice a day. Microbial suspensions were anaerobically incubated at 39 degrees C for 12 h. Phe and some other related compounds in both supernatants and microbial hydrolysates of the incubations were analysed by HPLC. A large quantity of Phe was produced from both PPY and PAA not only in B but also in P. In B suspensions, free Phe also accumulated in the medium only when PPY was used as a substrate. The ability of B to synthesize Phe from both PPY and PAA (expressed as unit 'per microbial nitrogen') was 5.1 and 24.8% higher than P, respectively. Phe production from PPY in B and P was 43.5 and 55.2% higher than that from PAA. Large amounts of PAA (17-27%) were produced from PPY in all microbial suspension and production amounts were similar in B and P. Small amounts of benzoic acid (BZA) were produced from PPY and PAA in B, P, and BP, and higher BZA production was observed in P as compared to B. Phenylpropionic acid (PPR) was produced in B from both PPY and PAA, but not in P or BP. A trace amount of phenyllactic acid (PLA) was detected only from PPY in B. Higher concentrations of an unknown compound from PPY and PAA were found to be accumulated in the body protein of B and also in the medium of P, and production of the compound from both PPY and PAA was also higher in B than P.

Journal Article↗
PubMed1998

Effects of salinomycin and vitamin B(6) on in vitro metabolism of phenylalanine and its related compounds by ruminal bacteria, protozoa and their mixture.

An in vitro study was conducted to examine the effects of salinomycin (SL) and vitamin B(6) (B(6)) on the production of phenylalanine (Phe) from phenylpyruvic acid (PPY) and phenylacetic acid (PAA) and of PAA from Phe and PPY by mixed rumen bacteria (B), mixed rumen protozoa (P) and their mixture (BP). Rumen microorganisms were collected from fistulated goats fed lucerne cubes (Medicago sativa) and a concentrate mixture (3 : 1) twice a day. Microbial suspensions were anaerobically incubated at 39 degrees C for 12 h. Phe and some other related compounds in both supernatants and microbial hydrolysates of the incubations were analyzed by HPLC. When PPY was used as a substrate, it completely disappeared without additives and converted mainly to Phe and PAA on the average by 396 and 178, 440 and 189, and 439 and 147 &mgr;M in B, P and BP, respectively, during the 12 h incubation period. The rate of disappearance showed no significant differences between the microbial suspensions with and without SL and B(6) during the incubation period. The production of Phe from PPY with SL was enhanced (p<0.05) by 40, 20 and 19% in B, P and BP, respectively, while PAA production from PPY with SL was inhibited (p<0.05) by 35, 37 and 38% in B, P and BP, respectively, during the 12 h incubation period. On the other hand, with B(6), the production of Phe and PAA from PPY tended to be enhanced by 14 and 17, 9 and 11, and 7 and 22% in B, P and BP, respectively, during the 12 h incubation period. When PAA added as a substrate was incubated in the incubation medium without any additives, it disappeared by 483, 462 and 507 &mgr;M and converted mainly to Phe on the average by 231, 244 and 248 &mgr;M in B, P and BP, respectively. The disappearance of PAA with SL was inhibited (p<0.05) by 16, 15 and 20%, in B, P and BP, respectively, whereas the disappearance of PAA with B6 was almost the same as that without B(6) in B and BP suspensions but tended to be enhanced by more than 9% in P suspensions during the 12 h incubation period. The production of Phe from PAA with SL tended to be inhibited by 12, 11 and 8% in B, P and BP, respectively, during the 6 h incubation period, but the inhibition was weakened during the 12 h incubation period, whereas Phe production from PAA with B(6) tended to be enhanced by 13, 16 and 8% in B, P and BP, respectively. When Phe was added as a substrate, the net Phe disappearance without additives was 549, 365 and 842 &mgr;M and converted mainly to PAA on the average by 254, 205 and 461 &mgr;M in B, P and BP, respectively. The net disappearance of Phe with SL was inhibited (p<0.05) by 38, 28 and 46%, whereas the net disappearance of Phe with B(6) was enhanced (p<0.05) by 9, 8 and 7% in B, P and BP, respectively. The production of PAA from Phe with SL was inhibited (p<0.05) by 73, 54 and 76% in B, P and BP, respectively. On the other hand, with B(6), PAA production from Phe was enhanced (p<0.05) by 19, 18 and 20% in B, P and BP, respectively. Based on these results, it seems that SL inhibited Phe disappearance and enhanced the synthesis of Phe from PPY, though not from PAA, and accumulated free Phe in the medium, whereas B(6) also enhanced Phe synthesis both from PPY and PAA, which could provide additional amino N for animals.

Journal Article↗
PubMed2000

Effects of protozoa on bacterial nitrogen recycling in the rumen.

The effects of protozoa on ruminal NH3-N kinetics and bacterial N recycling were measured in five sheep (57.6+/-7.1 kg BW, x +/- SD) with ruminal and duodenal cannulas in naturally faunated, defaunated, and refaunated periods. The sheep were fed a diet of 239 g of alfalfa haylage and 814 g of barley concentrate per day (DM basis) divided into 12 equal portions and allocated at 2-h intervals. A pulse dose of 300 mg of 15N as [15N]NH4Cl was administered into the rumen (on d 1 and 15) and 300 mg of 15N as [15N]urea was administered intravenously to the blood (d 8). Enrichment of 15N was measured in ruminal NH3-N, bacterial N, and plasma urea N over a period of 35 h. Total collection of urine was made for 5 d and analyzed for purine derivatives to calculate the flow of microbial N. Ruminal parameters and nutrient digestibilities were also measured. Sheep were defaunated using a rumen washing procedure 50 d prior to measurements in the defaunated period. Sheep were refaunated with ruminal contents from a faunated sheep receiving the same diet. Measurements began 26 d following refaunation, at which time protozoal numbers had returned to those in the originally faunated sheep. Data reported in parentheses are for faunated, defaunated, and refaunated sheep, respectively. Total culturable and cellulolytic bacterial numbers were unaffected by defaunation, but there was an increase in flow of microbial N from the rumen (10.8, 17.3, and 11.1 g N/d; P < .05) in the defaunated period. Flux, irreversible loss, and intraruminal recycling of NH3-N and recycling of NH3-N from plasma urea N were not affected by defaunation. Defaunation had no effect on reducing the absolute amount (13.8, 10.0, and 11.3 g N/d; P > .20) of bacterial N recycling and the percentage of N flux through the bacterial N pool. Total-tract digestion was reduced in defaunated compared with faunated sheep by 8, 17, 15, and 32% for OM, N, NDF, and ADF, respectively. In conclusion, defaunation improved ruminal N metabolism through the enhancement of bacterial protein synthesis, and improvement in the flow of microbial protein to the host animal.

Animals↗
PubMed2005

Cyclic GMP-dependent protein kinases in protozoa.

PKG (cGMP-dependent protein kinase) is a major intracellular receptor for cGMP and mediates a range of downstream physiological functions. Although most studies have concentrated on mammalian isoforms, the advent of whole genome sequencing has lead to identification of orthologues from other diverse species. Many biochemical and regulatory properties and structural features of PKG have been conserved throughout evolution. However, in protozoa there can be variation in the number of cGMP-binding sites and a tendency toward degeneracy of these additional sites. Recent work on the protozoan PKGs is the major focus of this review. Important functional data are emerging for these and other non-mammalian isoforms. Intriguingly, a role for PKG in locomotion is becoming a common functional theme across diverse phyla.

Animals↗
PubMed1998

The cloning and sequencing of ribosomal protein S18 of parasitic protozoa, Entamoeba histolytica.

A cDNA clone encoding ribosomal protein S18 has been isolated from parasitic protozoa, Entamoeba histolytica. The cDNA insert was 495 nucleotides long and an open reading frame (468 nucleotides including the stop codon) coded for 156 amino acid protein which was highly homologous with human, plant and yeast counterparts. PROSITE analysis of the ribosomal protein S18 in eukaryote revealed homology with prokaryotic ribosomal protein S13 signature, which is known to be involved in the initiation of translation.

Amino Acid Sequence↗
PubMed1991

Modification of the silver proteinate impregnation technique for protozoa and cultured nerve cells.

Silver impregnation with silver-protein compounds is widely used for staining tissue sections and cell cultures. Some authors report that the results obtained with these methods have not always been reproducible because the reagent's composition varies according to the manufacturer. To avoid this problem in the method described in this paper, a silver proteinate, produced in our own laboratory is used. Although our method is based on Bodian's, the modifications we have made allows its use for both free-living cells (protozoa) and cells grown in culture (nerve cells). The significant modifications are 1) different fixation, 2) postfixation with Cajal's formol-bromide, 3) changes in the duration of the impregnation steps technique and 4) elimination of metallic copper. The method reported here enables us to use silver proteinate whenever we require it and to control the composition of the silver proteinate. This technique can be used for cells cultured in either plastic or glass.

Animals↗
PubMed1998

Pathology of Piscinoodinium sp. (Protozoa: Dinoflagellida), parasites of the ornamental freshwater catfishes Corydoras spp. and Brochis splendens (Pisces: Callichthyidae).

Piscinoodinium sp. (Protozoa, Dinoflagellida) was commonly found on routine smears of samples of Brochis splendens and Corydoras spp. imported into Britain from South America, and on samples of the same group of fish examined at the exporters' holding facilities in Brazil. Infected fish had trophonts of different sizes on the gills and skin. In histological sections of the skin, the trophonts were found to be attached within depressions of different depths or enclosed by hyperplastic epithelial cells. Such enclosed trophonts have not previously been reported. Since some of the enclosed trophonts were dead, it was thought that enclosure was a result of the deep penetration of the trophont and the host defence mechanism. On the gills the Piscinoodinium infection was commonly associated with epithelial hypertrophy, focal and diffuse hyperplasia, oedema of the respiratory epithelium and lamellar fusion. The presence of this protozoan on different species of fish from the same shipment suggests that the infection was acquired before export. The source of infection and the stages of the export process which expose the fish to the highest risk of infection are discussed.

Animals↗
PubMed1998

Changes of intestinal epithelial structure and cell turnover in carp Cyprinus carpio infected with Goussia carpelli (Protozoa: Apicomplexa).

Epithelial cell turnover in the intestine of common carp Cyprinus carpio infected with the coccidian parasite Goussia carpelli (Leger & Stankovitch, 1921) was investigated during laboratory infection using histological and electron microscopical techniques. During the development of the parasite an increased number of mitotic enterocytes, identified by bromodeoxyuridine (BRDU) uptake, were observed at the base of infected mucosal folds. During the merogonic and gamogonic development of the parasite, severe damage to infected epithelium occurred, and concomitantly BRDU-positive cells spread along the mucosal folds. These cells exhibited immature characteristics, including a squamous to cuboidal shape, nuclear apolarity, a high number of ribosomes, and short or reduced microvilli. Contact with adjoining cells was formed by tight junctions and desmosomes, indicating the epithelial origin of these cells. These cells covered gut segments with damaged epithelium within a few days, suggesting a high regenerative capacity of the carp intestine, and this could explain the mild clinical symptoms in fish affected by G. carpelli-coccidiosis. Our study for the first time describes epithelial cell responses to injuries caused by enteric protozoa in piscine hosts.

Animals↗
PubMed1984

Intestinal protozoa in homosexual men of the San Francisco Bay area: prevalence and correlates of infection.

To compare the prevalence of intestinal parasitic infection in male homosexuals selected from other sources with the levels previously reported from clinic-derived populations, various homosexual groups of the San Francisco Bay Area were checked by means of stool examination and questionnaires for prevalence of intestinal parasites and for related sexual behavioral patterns during a 3-year period. The prevalence of Entamoeba histolytica (28.6%) was similar to that reported in other studies, whereas that of Giardia lamblia was lower. Infection with E. histolytica was correlated significantly with a prior history of syphilis or gonorrhea (P less than 0.0001), with the number of sexual partners in the preceding 12-month period (P less than 0.0001), and with the reported frequency of oral-anal sexual contact (P less than 0.001). Giardial infection was also significantly related to oral-anal sex (P less than 0.001). No relation was seen between the presence or absence of gastrointestinal symptoms and infection with pathogenic protozoa. Factors associated with parasitic infection differed little from those found in previous studies, although significance levels were considerably higher because of the larger size of our study group.

Adult↗
PubMed1975

Variation in response of channel catfish to Henneguya sp. infections (Protozoa: Myxosporidea).

Infections in channel catfish (Ictalurus punctatus, Rafinesque) induced by the sporozoan Henneguya (Protozoa: Myxosporidea) result in seven known and diverse disease manifestations. Most outstanding is an interlamellar branchial form responsible for significant losses among immature catfish, and a unique papillomatous form. The question of whether or not the species of Henneguya involved in these cases is H. exilis remains to be resolved.

Animals↗
PubMed1985

Life cycle and biology of Eimeria lettyae sp. n. (Protozoa: Eimeriidae) from the northern bobwhite, Colinus virginianus (L.).

A new species of coccidium, Eimeria lettyae sp. n. (Protozoa: Eimeriidae) was recovered from feces of the northern bobwhite, Colinus virginianus (L.), from Pennsylvania and Florida. Oocysts measured 21.1 microns (16.4 to 25.8) by 17.2 microns (14.1 to 21.2); index (L/W ratio) = 1.22. Oocysts lacked a micropyle, residuum, and polar granules. Sporozoites penetrated the upper 1/2 of the villi, then moved to the lamina propria at the base of the villi. There were five asexual generations, all of which developed above the nucleus of the host cell. Meronts measured 9.4 X 7.0 microns, 18.6 X 11.2 microns, 11.8 X 10.1 microns, 7.1 X 6.2 microns, and 20.2 X 12.8 microns, respectively. These matured at 32, 40, 48, 56, and 72 hr postinoculation (PI) and contained 12, 50+, 24 to 36, 12 to 24, and 50+ merozoites, respectively. Infection was most intense in the duodenum although some gamonts were found in the ileum and ceca. The prepatent period was 88 to 91 hr PI. Sporulation time was 18 hr at 25 C. The peak of oocyst production was broad and extended from 4 days PI through 14 days PI. Oocysts were passed for at least 67 to 76 days PI. Eimeria lettyae sp. n. did not infect chickens (Gallus domesticus), domestic turkeys (Meleagris gallopavo), ring-necked pheasants (Phasianus cochicus), chukar partridge (Alectoris graeca), or Japanese quail (Coturnix coturnix). Immunizing bobwhite with E. lettyae sp. n. did not protect against challenge with E. dispersa. Immunizing bobwhites 25 times with 10(2) or 10(3) sporulated oocysts of E. lettyae did not entirely eliminate oocyst production following challenge with the same species.

Animals↗