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PubMed1992

Moderation of ruminal fermentation by ciliated protozoa in cattle fed a high-grain diet.

The objective of this study was to assess the influence of ciliated protozoa on ruminal fermentation in cattle fed high-grain diets. Six ruminally cannulated steers fed a corn-based grain diet (85% concentrate plus 15% alfalfa hay) at 12-h intervals were assigned randomly to two groups, ciliate free and faunated, in a crossover design. Defaunation was by ruminal emptying, omasal flushing, and treatment with sodium sulfosuccinate. Two to 3 weeks after defaunation, the ruminal contents of all steers were sampled before the morning feeding (0 h) and at 1, 2, 4, 6, 8, and 12 h after feeding to measure pH, analyze fermentation products, and monitor counts of ciliated protozoa and lactic acid-producing and -fermenting bacterial groups. Total numbers of ciliated protozoa in the faunated steers averaged 4.3 x 10(5)/g, and the protozoa consisted of nine genera. Ciliate-free steers had lower (P less than 0.01) ruminal pHs (pH 5.97) than faunated cattle (pH 6.45); however, the treatment-time interaction was not significant. Ruminal lactate and ammonia concentrations were similar in both groups. The total volatile fatty acid concentration was higher (P less than 0.05) in the ciliate-free steers than in the faunated steers and exhibited a treatment-time interaction (P less than 0.05). The acetate-to-propionate ratio was higher (P less than 0.05) in the faunated group than in the ciliate-free group and showed a treatment-time interaction (P less than 0.05). Total anaerobic bacterial counts were about fourfold higher in the ciliate-free group than in the faunated group.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2000

Importance of passive diffusion in the uptake of polychlorinated biphenyls by phagotrophic protozoa.

Unicellular protozoan grazers represent a size class of organisms where a transition in the mechanism of chlorobiphenyl (CB) introduction, from diffusion through surface membranes to ingestion of contaminated prey, could occur. This study compares the relative importance of these two processes in the overall uptake of polychlorinated biphenyls by protists. Uptake rates and steady-state concentrations were compared in laboratory cultures of grazing and nongrazing protozoa. These experiments were conducted with a 10-microm marine scuticociliate (Uronema sp.), bacterial prey (Halomonas halodurans), and a suite of 21 CB congeners spanning a range of aqueous solubilities. The dominant pathway of CB uptake by both grazing and nongrazing protozoa was diffusion. Organic-carbon-normalized CB concentrations (in the protozoan cell) were equivalent in grazing and nongrazing protozoa for all congeners studied. Rate constants for uptake into and loss from the protozoan cell were independently determined by using [3,3',4, 4'-(14)C]tetrachlorobiphenyl (IUPAC no. 77), 0.38 +/- 0.03 min(-1) and (1.1 +/- 0.1) x 10(-5) (g of organic carbon)(-1) min(-1), respectively. Magnitudes of the uptake and loss processes were calculated and compared by using a numerical model. The model result was consistent with data from the bioaccumulation experiment and supported the hypothesis that diffusive uptake is faster than ingestive uptake in phagotrophic unicellular protozoa.

Animals↗
PubMed2005

Exposure to rumen protozoa leads to enhancement of pathogenicity of and invasion by multiple-antibiotic-resistant Salmonella enterica bearing SGI1.

Multiple-antibiotic-resistant Salmonella enterica serotype Typhimurium is a food-borne pathogen that has been purported to be more virulent than antibiotic-sensitive counterparts. The paradigm for this multiresistant/hyperpathogenic phenotype is Salmonella enterica serotype Typhimurium phage type DT104 (DT104). The basis for the multiresistance in DT104 is related to an integron structure designated SGI1, but factors underlying hyperpathogenicity have not been completely identified. Since protozoa have been implicated in the alteration of virulence in Legionella and Mycobacterium spp., we attempted to assess the possibility that protozoa may contribute to the putative hypervirulence of DT104. Our study reveals that DT104 can be more invasive, as determined by a tissue culture invasion assay, after surviving within protozoa originating from the bovine rumen. The enhancement of invasion was correlated with hypervirulence in a bovine infection model in which we observed a more rapid progression of disease and a greater recovery rate for the pathogen. Fewer DT104 cells were recovered from tissues of infected animals when protozoa were lysed by preinfection chemical defaunation of the bovine or ovine rumen. The protozoan-mediated hypervirulence phenotype was observed only in DT104 and other Salmonella strains, including serovars Agona and Infantis, possessing SGI1.

Animals↗
PubMed1975

Protozoa and the decline of Rhizobium populations added to soil.

A fall in Rhizobium abundance occurred in nonsterile soil inoculated with large numbers of the root-nodule bacteria, but many of the rhizobia still survived. No such decline was evident in sterile soil. Protozoa feeding on these bacteria were isolated from soil and other environments. As the abundance of Rhizobium meliloti and a cowpea Rhizobium strain in soil decreased, the protozoan density increased. The inability of the predators to eliminate their prey from soil was not the result of the presence of organisms feeding on the protozoa because many rhizobia survived in sterile soil inoculated with the prey and cultures of individual protozoa, nor was it the result of the rapid multiplication of the bacteria to replace those consumed because survivors were still numerous in essentially organic matter free soil in which the bacteria did not grow appreciably. The lack of elimination also was not associated with a protective effect of soil particles because survivors were still abundant in solutions inoculated with protozoa and bacteria. It is suggested that the size of the prey population diminishes until a density is attained at which the energy used by the predator in hunting for the survivors equals that obtained from the feeding.

Amoeba↗
PubMed1995

New mechanisms of drug resistance in parasitic protozoa.

The main line of defense now available against parasitic protozoa--which are responsible for major diseases of humans and domestic animals--is chemotherapy. This defense is being eroded by drug resistance and, with few new drugs in the pipeline, prevention and circumvention of resistance are medical and veterinary priorities. Although studies of resistance mechanisms in parasites have lagged behind similar studies in bacteria and cancer cells, the tools to tackle this problem are rapidly improving. Transformation with exogenous DNA is now possible with all major parasitic protozoa of humans. Hence, putative resistance genes can be tested in sensitive protozoa, allowing an unambiguous reconstruction of resistance mechanisms. Gene cloning, the polymerase chain reaction, and monoclonal antibodies against resistance-related proteins have made it possible to analyze potential resistance mechanisms in the few parasites that can be obtained from infected people. Hence, the prospect of applying new knowledge about resistance mechanisms to parasites in patients is good, even though today virtually all knowledge pertains to parasites selected for resistance in the laboratory. Resistance mechanisms highlighted in this review include: 1. Decrease of drug uptake because of the loss of a transporter required for uptake. This decrease contributes to resistance to arsenicals and diamidines in African trypanosomes. 2. The export of drugs from the parasite by P-glycoproteins and other traffic ATPases. This export could potentially be an important mechanism of resistance, as these proteins are richly represented in the few protozoa analyzed. There are indications that such transmembrane transporters can be involved in resistance to emetine in Entamoeba spp., to mefloquine in Plasmodium spp., and to antimonials in Leishmania spp. 3. The possible involvement of the P-glycoprotein encoded by the Plasmodium falciparum pfmdr1 gene in chloroquine resistance. We present the available data that lead to the conclusion that overproduction of the wild-type version of this protein results in chloroquine hypersensitivity rather than resistance. 4. The involvement of the PgpA P-glycoprotein of Leishmania spp. in low-level resistance to arsenite and antimonials. We raise the possibility that this protein transports glutathione conjugates of arsenite and antimonials rather than the compounds themselves. 5. Loss of drug activation as the main mechanism of metronidazole resistance in Trichomonas and Giardia spp. Recent evidence indicates that a decrease of the proximal cellular electron donor for metronidazole activation, ferredoxin, is the main cause of resistance in Trichomonas. 6. Resistance arising through alteration of drug targets. The amino acid substitutions in the dihydrofolate reductase-thymidylate synthase of Plasmodium spp. are good examples of this mechanism.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed2003

Identification of trichomonadid protozoa from the bovine preputial cavity by polymerase chain reaction and restriction fragment length polymorphism typing.

Accurate identification of the bovine pathogen Tritrichomonas foetus is sometimes complicated by the presence of other trichomonadid protozoa in clinical samples. A highly specific and reproducible approach for differentiating 3 common types of bovine trichomonadid protozoa found in the bovine preputial cavity, T. foetus, Pentatrichomonas hominis, and a Tetratrichomonas species, was developed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. Universal trichomonadid protozoa primers, TFR1 and TFR2, were used to amplify the 5.85 rRNA gene and internal transcribed spacer regions (ITSRs), and the products were digested with the restriction enzyme HpyCH4IV. Restriction fragment length polymorphism analysis was performed on 55 trichomonad isolates from bovine preputial washing and scraping samples. The RFLP results correlated 100% with 5.85 rRNA gene and ITSR sequence resultsand PCR results with primers specific for T. foetus. The results of this study demonstrate that PCR and RFLP analysis can be used in lieu of DNA sequencing to identify the specific trichomonadid protozoa isolated from the bovine preputial cavity.

Animals↗
PubMed2006

Fatty acid composition of ruminal bacteria and protozoa, with emphasis on conjugated linoleic acid, vaccenic acid, and odd-chain and branched-chain fatty acids.

Knowledge of the fatty acid profile of microbial lipids is of great nutritional importance to the animals and, subsequently, their products. This study was conducted to examine the fatty acid profiles of mixed rumen bacteria and protozoa. Bacterial and protozoal cells were isolated by differential centrifugation of rumen contents. The main fatty acids were palmitic (16:0) and stearic (18:0) in both the bacterial and protozoal fractions. Palmitic acid was 74% greater in the protozoal fatty acids than in the bacterial fatty acids, whereas bacteria had 2.25-times greater stearic acid (18:0) proportions compared with protozoa. The total odd-chain plus branched-chain fatty acids were 16.5% of bacterial fatty acids and 11.0% of protozoal fatty acids. The anteiso-17:0 proportions in bacterial and protozoal fatty acids were 1.4 and 2.9%, respectively. The most abundant trans-18:1 isomer, vaccenic acid (18:1 trans-11), was 6.6% of total fatty acids in protozoa and 2.0% of total fatty acids in bacteria. The cis-9, trans-11 CLA was 8.6-times greater in the protozoal fraction (1.32% of total fatty acids) than in the bacterial fraction (0.15%). These results suggest that the presence of protozoa in the rumen may increase the supply of CLA and other unsaturated fatty acids for lower gut absorption by ruminants.

Animals↗
PubMed1982

Effect of diet on amino and nucleic acids of rumen bacteria and protozoa.

Amino acid composition and nucleic acid content of pure cultures of rumen bacteria (17 species) were analyzed. Amino acid composition between gram-positive and -negative organisms was not different. The total nitrogen content of gram-negative bacteria (10.8%) was significantly higher than gram-positive organisms (9.9%). Deoxyribonucleic acid-nitrogen: total nitrogen (mg/g) differed between gram-positive (8.8) and gram-negative (18.9) bacteria, but there was no significant difference in ratio of ribonucleic acid-nitrogen to total nitrogen. In a second experiment six rumen-fistulated cattle were fed either a high roughage (85% alfalfa hay plus 15% concentrate) or high concentrate diet (15% alfalfa hay and 85% concentrate). Cattle were adapted 14 days and rumen contents sampled on 3 consecutive days. Nitrogen content was higher in protozoa from cattle fed low concentrate (8.4%) than in protozoa from cattle fed high concentrate (7.9%) but was similar in bacteria for both diets. Deoxyribonucleic acid nitrogen: total nitrogen (mg/g in bacteria decreased from 27.2 in cattle fed the low concentrate diet to 20.9 in those fed the high concentrate diet. Differences between sampling days were significant for both bacteria and protozoa for ratio of deoxyribonucleic acid-nitrogen to total nitrogen but were significant only in protozoa for ratio of ribonucleic acid nitrogen to total nitrogen. Ribonucleic acid may serve as a marker for estimating microbial production in the rumen if sources of variation are recognized and corrected adequately.

Amino Acids↗
PubMed1983

Effect of energy concentration and source of nitrogen on numbers and types of rumen protozoa.

Two rumen-fistulated Holstein heifers were fed three protein-free semipurified diets: 70% roughage, 30% concentrate; 50% roughage, 50% concentrate; and 30% roughage, 70% concentrate. Wheat straw was the roughage, and the concentrate was a pelleted mixture of corn starch, dextrose, fat, urea, vitamins, and minerals. Each animal received the three dietary treatments in a randomized complete block design. On days 1, 3, and 5 following a 21-day adaptation, rumen fluid samples were taken before feeding and at 1, 2, 3, 4, and 5 h after feeding. Total protozoa concentrations for low, medium, and high concentrate diets were 1.5, 2.5, 4.1 X 10(5)/ml. Dasytricha concentrations were not affected by diet, but Entodinium, Diplodinium, Isotricha, Epidinium, and Ophryoscolex increased with increases of the concentrate proportion of the diets. Entodinium, the predominant protozoa with all diets, increased as concentrate proportion of the diet increased. In a second experiment the two heifers from Experiment 1 were fed semipurified diets that contained either urea or soybean meal as the nitrogen source. There were larger populations of Dasytricha and Isotricha when the diet contained soybean meal, and larger populations of Charon, Entodinium, and total protozoa when the diet contained urea. In both experiments, holotrichs were highest 1 to 2 h after feeding, and entodiniomorphs were highest before feeding and 4 to 6 h after feeding. An active protozoan population can exist in the rumen of cattle fed diets virtually devoid of natural protein, and protozoa are influenced by energy and nitrogen source.

Animals↗
PubMed2003

Biodiversity of algae and protozoa in a natural waste stabilization pond: a field study.

A field study was carried out on the biodiversity of protozoa and algae from a natural waste stabilization pond during November, 1996 to April, 1997. The raw waste and pond samples were analysed for physico-chemical and biological parameters. High dissolved oxygen (DO) coinciding with phytoplankton peak was recorded. The algae--Chlorella vulgaris, Scenedesmus acuminatus, Oscillatoria brevis and Nostoc piscinale and Protozoa--Paramecium caudatum, Acanthamoeba sp., Bodo saltans and Oikomonas termo were obvious as dominant species, whereas algae Ochromonas pyriformis and Synura uvella and protozoa, Didinium masutum and Stentor coerulus were noted as rare species. Totally 71 species of algae and 13 species of protozoa were identified.

Animals↗
PubMed1992

[Occurrence of the protozoa, Entamoeba gingivalis and Trichomonas tenax in the mouths of children and adolescents with hyperplastic gingivitis caused by phenytoin].

The oral protozoa Entamoeba gingivalis and Trichomonas tenax do not occur in small children and are rarely found in older ones. In adolescents their occurrence rate keeps increasing with age. They parasitize in an oral cavity changed by inflammation, yet also in a healthy mouth. Their highest occurrence rate has been recorded in adults with periodontosis and atrophy of the periodontium, a somewhat lower one in adults with gingivitis. The authors addressed the question whether the presumed low occurrence rate of oral protozoa in children becomes increased in drug-induced gingivitis after treatment with the antiepileptic 5,5-diphenylhydantoin. Cultivation for oral protozoa was performed in 231 children and adolescents. Of these 59 were epileptics. Drug-induced gingivitis was present in 66% of the epileptics. Drug-induced gingivitis did not increase the occurrence rate of oral protozoa as compared to the findings in the rest of the series studied. (Tab. 9, Ref. 7.).

Adolescent↗
PubMed2005

[Evaluation of the protozoa found in fecal samples using the trichrome staining method.].

The morphological diagnosis of intestinal protozoa is a widely accepted method today. A permanent staining method is preferred since it can visualize the morphological details of protozoal cysts and it allows the stool specimens to be reevaluated later. In this study, 2141 stool specimens sent to the clinical microbiology laboratory of the Gazi University Hospital were examined using the trichrome staining method. We determined 181 parasitic organisms in 165 (7.70%) of these 2141 specimens. Helminthic organisms were detected in seven of these samples. Protozoa were found in 174 specimens evaluated with the trichrome staining method. In our study, trichrome staining was found to be 87.93% successful in the identification of intestinal protozoa. In conclusion, it was seen that the trichrome staining method could make a reliable diagnosis possible when used in addition to the direct examination of the stool specimens in the identification of intestinal pathogenic and nonpathogenic protozoa to the species level.

English Abstract↗
PubMed1981

Interactions between murine macrophages and obligate intracellular protozoa.

The diversity of interactions between obligate intracellular protozoa and murine macrophages is just being elucidated. Protozoa of the genera Toxoplasma, Leishmania, and Trypanosoma all enter and replicate within macrophages. This review describes similarities and differences among these organisms with regard to entry mechanisms, sites of replication in the phagolysosomal system, metabolic requirements, effects on macrophage function, macrophage handling of protozoal antigens, the relationship to genetics of immune response, and the characteristics of lymphokine-induced microbicidal and microbistatic processes. These organisms each enter the macrophage by endocytosis, but they then reside at different sites in relation to the phagolysosomal system. The basis of obligate parasitism remains unknown; however, both the protozoa and the host cell have important effects on the function of the other during parasitism. The macrophage may play a pivotal role in the immunosuppression associated with the early stages of infection by each of these microbes. Genetic influences on the response to infection have been clearly identified in murine models. Lymphocyte products from immune cells have marked effects on the interactions of protozoa and macrophages, under some conditions stimulating protozoacidal mechanisms, and in some protozoastatic responses. The dynamic balance between protozoal parasitism and macrophage response must be further defined in order to determine the potential value of chemotherapeutic or immunologic intervention.

Animals↗
PubMed1995

[Viruses of parasitic protozoa].

The authors present the actual review on several publications concerning the molecular characterizations of the viruses found in parasitic protozoa such as Giardia, Trichomonas, Leishmania and Entamoeba histolytica. All of the RNA viruses observed in parasitic protozoa showed several similarities and did not considerably differ from the viruses found in simple eukaryotic cells; they closely correspond to dsRNA viruses of yeast. The supposition that the protozoan symbionts detected in laboratories transfer to their hosts in natural conditions seemed to be rational, though, there are no evidences that these symbionts are potential pathogens. However, the opinion reiterates that intestinal protozoa (e.g. Entamoeba histolytica) may serve as vectors for HIV or cofactors of HIV infection. The authors point out that irrespective of the potential role of viruses as vectors in the transfection system for parasitic protozoa, the observed viral system constitutes an unusual experimental system to solve the problems of gene expression.

Animals↗
PubMed1979

A comparison of the formalin-ether concentration and trichrome-stained smear methods for the recovery and identification of intestinal protozoa.

The detection and identification of intestinal protozoa were compared using the formalin-ether concentration technique and trichrome smears. Using the polyvinyl alcohol (PVA) one-vial collection method, 13,194 outpatient samples were examined by both methods. Twenty-three percent or 3077 specimens were positive for one or more species of intestinal protozoa. Of the pathogenic protozoa identified, 96.3 percent of the trophozoites and 44.2 percent of the cysts were identified only by the trichrome-stained smear. It is recommended that both procedures by included in the examination of fecal sepcimens for protozoa.

Dientamoeba↗
PubMedDate not supplied

[Method for an absolute count of the number of parasitic protozoa in fish].

A new technique is given of the absolute calculation of parasitic Protozoa in fishes. The mucus from the body and gills of fishes was mixed with such volume of water (V) that the drop of mixture (v) was clearly seen under themicroscope. The calculation of Protozoa was done in a sample from three drops of mixture covered with three slides (S). From 50 to 100 viewing fields were seen the them and an average number of Protozoa in one field was calculated. The total number of Protozoa in one fish (N) was equal to an average number of parasites in a viewing field (n) multiplied by the total number of viewing fields (m) which could be obtained at the examination of the whole mixture. The latter was calculated conventionally (see articles).

Animals↗
PubMed1994

Macrophage response to viruses, protozoa, and fungi: secretory and cellular activities induced in resting unprimed bone marrow-derived mononuclear phagocytes.

The secretory (tumor necrosis factor, TNF-alpha; nitrite) and cellular response (mitochondrial respiration, TNF-alpha-independent tumoricidal activity) of a pure, lymphocyte-free population of resting, unprimed rat bone marrow-derived mononuclear phagocytes (BMM phi) to direct interaction with viruses, protozoa, and fungi was assessed and compared with that triggered by bacterial agents and interferon-gamma (IFN-gamma). Viruses (herpes simplex, vaccinia, poliomyelitis, vesicular stomatitis, lymphocytic choriomeningitis, Sendai), protozoa (Trypanosoma brucei, Giardia lamblia), and fungi (Penicillium, Trichosporon, Fusarium, Rhizopus, Aspergillus, Geotrichum species) affected primarily the secretion of TNF-alpha and mitochondrial respiration of BMM phi; their effects on the secretion of nitrite and on tumoricidal activity were at best marginal. Collectively, the macrophage response to viruses, protozoa, and fungi was less varied and less marked than that to bacterial agents (intact organisms, peptidoglycan, lipoteichoic acid, lipopolysaccharide) and IFN-gamma.

Animals↗
PubMed1998

Toxicity and bioaccumulation of lead in marine protozoa communities.

The toxicity and bioaccumulation of lead has been studied using marine protozoa communities developed in laboratory microecosystems. The concentrations tested were 500 and 1000 micrograms.L-1 of lead as lead acetate. The protozoan was able to bioaccumulate 27.02-504 micrograms Pb.g-1 dry weight. Bacteria also bioaccumulated lead, but always to a lesser degree than protozoa. Lead caused a significant reduction in the density of protozoa, which could be an indirect response to the cellular increase of lead. On the other hand, the toxicant did not determine a decrease in the number of bacterial cells; this could be due to their capacity to bioaccumulate a lesser amount of lead, the increase in the number of dead cells, and the elimination of their predators by the toxicant. After 120 h, a recovery of the community was observed.

Animals↗