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PubMed1987

Relative contributions of ruminal bacteria and protozoa to the degradation of protein in vitro.

Mixed ruminal microorganisms from a cow fed timothy hay and concentrate supplement (50:50) were incubated with various protein sources for 15 h (no carbohydrates or growth), and deamination was studied under enzyme-limiting substrate-excess conditions (n = 3). Addition of amphotericin (10 micrograms/ml) killed protozoa and decreased (P less than .05) ammonia production from killed bacteria but it had no effect (P greater than .05) on casein deamination. Monensin (5 micrograms/ml) also killed protozoa; however, it decreased (P less than .05) casein deamination to a much greater extent than amphotericin. Antibacterial antibiotics (penicillin G, polymixin B, cephalosporin C and streptomycin) greatly reduced (P less than .05) ammonia formation from casein. Isolated bacteria always produced more ammonia than isolated protozoa, but the difference was less with heat-treated, particulate proteins. Heated soybean protein was as soluble as heated casein but it was deaminated (P less than .05) at a faster rate by bacteria. Nonammonia-nonprotein N accumulation was greater (P less than .05) with the protozoa than bacteria. When incubations containing bacteria or protozoa were compared with combinations of protozoa and bacteria, the combinations always caused a synergistic increase in ammonia and decrease (P less than .05) in nonammonia-nonprotein N. These results suggest: soluble proteins were primarily degraded by bacteria; protozoa could contribute to the degradation of insoluble, particulate proteins; protozoa were limited in their ability to assimilate peptides (or amino acids); low molecular weight products could be fermented more readily by bacteria and monensin was toxic to protozoa, but decreases in ammonia were primarily due to action of monensin on bacteria.

Animals↗
PubMed2006

Bacteria removal in septic effluent: influence of biofilm and protozoa.

Numerous biological, physical and chemical parameters are involved in the retention and removal of bacteria in wastewater treatment systems. Biological parameters, such as biofilms and protozoa grazing activity, are often mentioned but few studies provide a better understanding of their influence. In this study, the effect of bacterivorous protozoa on pathogenic indicator bacteria removal was investigated in septic effluent and in the presence of a biofilm coating glass slides. Endogenous bacteria from septic effluent were quantified. First, bacteria removal was compared between septic effluents treated or not with an inhibitor of protozoa (cycloheximide). The mortality rates were 10 times lower in treated effluent (96 CFU mL(-1) d(-1)) than in untreated effluent (1100 CFU mL(-1) d(-1)). Secondly, the efficiency of bacteria removal was studied (i) with a biofilm surface and active protozoa, (ii) with a biofilm surface and inactivated protozoa, (iii) with a clean surface. Protozoa in the presence of a biofilm were responsible for 60% of bacteria removal. Biofilm without protozoa and a clean surface each removed similar quantities of bacteria. Grazing by protozoa could be an important biological mechanism for bacterial elimination in wastewater treatment systems.

Animals↗
PubMed2000

From protozoa to mammalian cells: a new paradigm in the life cycle of intracellular bacterial pathogens.

It is becoming apparent that several intracellular bacterial pathogens of humans can also survive within protozoa. This interaction with protozoa may protect these pathogens from harsh conditions in the extracellular environment and enhance their infectivity in mammals. This relationship has been clearly established in the case of the interaction between Legionella pneumophila and its protozoan hosts. In addition, the adaptation of bacterial pathogens to the intracellular life within the primitive eukaryotic protozoa may have provided them with the means to infect the more evolved mammalian cells. This is evident from the existence of several similarities, at both the phenotypic and the molecular levels, between the infection of mammalian and protozoan cells by L. pneumophila. Thus, protozoa appear to play a central role in the transition of bacteria from the environment to mammals. In essence, protozoa may be viewed as a 'biological gym', within which intracellular bacterial pathogens train for their encounters with the more evolved mammalian cells. Thus, intracellular bacterial pathogens have benefited from the structural and biochemical conservation of cellular processes in eukaryotes. The interaction of intracellular bacterial pathogens and protozoa highlights this conservation and may constitute a simplified model for the study of these pathogens and the evolution of cellular processes in eukaryotes. Furthermore, in addition to being environmental reservoirs for known intracellular pathogens of humans and animals, protozoa may be sources of emerging pathogenic bacteria. It is thus critical to re-examine the relationship between bacteria and protozoa to further our understanding of current human bacterial pathogenesis and, possibly, to predict the appearance of emerging pathogens.

Amoeba↗
PubMed2003

Effects of protozoa on methane production in rumen and hindgut of calves around time of weaning.

Effects of the presence or absence of ciliate protozoa on methanogenesis in the rumen and hindgut were investigated in young calves during a 7-week period. Ten Holstein calves, aged 7 days, were divided in two groups (n = 5) and fed an increasing amount of a commercial milk replacer and small amounts of a calves starter. One group was inoculated with ciliate fauna on two occasions, week 5 and 6, while the second remained ciliate-free. The absence of protozoa in the rumen decreased rumen empty weight (-23%, P < 0.01), and rumen pool size of N (-36%, P < 0.01) and crude fat (-37%, P < 0.05). Rumen bacteria of non-faunated calves contained a higher proportion of total amino acid-N per 16 g N (+3%, P < 0.01) and D-alanine-N per 16 g N (+13%, P < 0.05) compared to faunated calves. Further results contain a reference for a higher bacterial mass in the ciliate-free rumen with an increased number of bacteria adherent to rumen mucosa. The CH4 production in the rumen increased exponentially with the increase in protozoa population size (R2 = 0.68). In presence of 46 x 10(4) protozoa per ml rumen fluid, the in vitro CH4 production of rumen fluid per mol total VFA was about 34% higher in faunated than in non-faunated calves (P < 0.001). Hydrogen (2H) recovery of rumen fermentation was positively correlated (R2 = 0.55) to the CH4 production rate. Methanogens were attached on rumen mucosa. Methanogenesis, induced by rumen mucosa attached bacteria, was stimulated by ruminal protozoa. In the absence of protozoa in the rumen, the acetate-propionate ratio and butyrate proportion of VFA were reduced. In vivo, in the absence of protozoa not only the whole animal CH4 production (-30%, P < 0.05) but also the digestibility of carbohydrates (-4%, P < 0.05) was reduced. Thereby no difference was observed in the intake of ME per kg DM between the groups. In conclusion, the methanogenesis in the rumen, but not in hindgut, is associated with the development of the ruminal protozoa population. The level of methanogenesis (mol/mol VFA) in the hindgut amounts to 20% of the ruminal methanogenesis.

Ammonia↗
PubMed1983

Phylogeny of protozoa deduced from 5S rRNA sequences.

The nucleotide sequences of 5S rRNAs from three protozoa, Bresslaua vorax, Euplotes woodruffi and Chlamydomonas sp. have been determined and aligned together with the sequences of 12 protozoa species including unicellular green algae already reported by the authors and others. Using this alignment, a phylogenic tree of the 15 species of protozoa has been constructed. The tree suggests that the ancestor for protozoa evolved at an early time of eukaryotic evolution giving two major groups of organisms. One group, which shares a common ancestor with vascular plants, contains a unicellular green flagellate (Chlamydomonas) and unicellular green algae. The other group, which shares a common ancestor with the multicellular animals, includes various flagellated protozoa (including Euglena), ciliated protozoa and slime molds. Most of these protozoa appear to have separated from one another at a fairly early period of eukaryotic evolution.

Animals↗
PubMed1986

Dynamics of large ciliate protozoa in the rumen of cattle fed on diets of freshly cut grass.

1. The dynamics of large ciliate (holotrich) protozoa (Isotricha and Dasytricha spp.) in the rumen of cattle given cut, fresh ryegrass (Lolium multiflorium Lam) were studied by means of a single intrarumen injection of 14C-labelled protozoa prepared in vitro by adding [Me14C]choline to rumen fluid containing protozoa and incubating at 39 degrees for 2 h. 2. An indication of the lysis rate of protozoa in the rumen was obtained from the radioactivity apparently lost through the methane pool. 3. The turnover time of the holotrich protozoa indicates that these protozoa were extensively retained in the rumen and that only a small proportion of those produced in the rumen flowed out in the digesta. This was supported by the estimation of the rate of lysis which was approximately 85% of the turnover rate in the rumen. 4. The apparent production rate of the larger protozoa indicates that they contribute only about 9% of the predicted net microbial protein synthesis in the rumen.

Animals↗
PubMed1980

A simple method for plating and cloning ciliates and other protozoa.

A simple method is described for plating and cloning ciliates and other protozoa, based on a principle differing from that traditionally used for plating and cloning bacteria and other microorganisms. This procedure, referred to as te silicone-oil-plating-procedure (SOPP), involves vortexing small volumes of culture medium containing protozoa with larger volumes of a non-toxic silicone oil and plating the resulting unstable emulsion in small plastic petri plates. Discrete microdroplets of culture medium form containing protozoa entrapped and immobilized between the hydrophobic surfaces of teh plastic petri dish and the oil. Protozoa, isolated by this method grow, divide, and multiply to form clones. The procedure may be used for plating and cloning protozoa in bacterized and axenic culture. Variations of te basic method may be applied to isolating protozoa from the wild, washing protozoa to remove microorganisms, screening for potential mutants, and for replica plating.

Animals↗
PubMedDate not supplied

Effects of altered temperature and precipitation on desert protozoa associated with biological soil crusts.

Biological soil crusts are diverse assemblages of bacteria, cyanobacteria, algae, fungi, lichens, and mosses that cover much of arid land soils. The objective of this study was to quantify protozoa associated with biological soil crusts and test the response of protozoa to increased temperature and precipitation as is predicted by some global climate models. Protozoa were more abundant when associated with cyanobacteria/lichen crusts than with cyanobacteria crusts alone. Amoebae, flagellates, and ciliates originating from the Colorado Plateau desert (cool desert, primarily winter precipitation) declined 50-, 10-, and 100-fold, respectively, when moved in field mesocosms to the Chihuahuan Desert (hot desert, primarily summer rain). However, this was not observed in protozoa collected from the Chihuahuan Desert and moved to the Sonoran desert (hot desert, also summer rain, but warmer than Chihuahuan Desert). Protozoa in culture began to encyst at 37 degrees C. Cysts survived the upper end of daily temperatures (37-55 degrees C), and could be stimulated to excyst if temperatures were reduced to 15 degrees C or lower. Results from this study suggest that cool desert protozoa are influenced negatively by increased summer precipitation during excessive summer temperatures, and that desert protozoa may be adapted to a specific desert's temperature and precipitation regime.

Animals↗
PubMed1973

Effects of changes in feed level, starvation, and level of feed after starvation upon the concentration of rumen protozoa in the ovine.

Four rumen fistulated sheep were used in five experiments to investigate the effect of feed level upon the concentration of rumen ciliate protozoa. The sheep were fed once daily 650 g of a pelleted diet composed of corn cobs, 45%; alfalfa meal, 35%; oats, 12.5%; cane molasses, 5%; urea, 0.4%; and vitamins and minerals, 2%. The concentration of protozoa reached minimum and maximum values at 5 and 22.5 h after feeding, respectively. Thus, to estimate apparent generation rates, concentrations of protozoa were determined at 5 and 20 h postfeeding. Apparent generation rate/h = natural log of ([concentration of protozoa at 20 h divided by concentration at 5 h] divided by the time interval, [T20 to T5]). Alteration of the feed to protozoa ratio by starvation and by changing the level of feed (200 to 900 g/day) showed that as the ratio of feed to protozoa increased, generation rate increased. Measurements of liquid turnover rates in the rumen showed that turnover rate decreased as feed level decreased. Turnover rate was near zero when the sheep were starved. Small quantities of soluble substrates, added directly to the rumen of starved sheep, maintained the protozoal population when rumen turnover was minimal. Furthermore, as rumen turnover rate increased with increased levels of feed, the effect of substrate on maintaining the protozoal population was negated. Thus, at high feed levels, turnover rate may be the dominant factor controlling the establishment and concentration of protozoa in the rumen.

Animals↗
PubMed1999

Intestinal protozoa in homosexual men in Edinburgh.

Our objective was to determine the prevalence of intestinal protozoa in homosexual men attending a sexually transmitted diseases (STD) clinic, to compare it with the prevalence in a previous study from the same clinic, and to examine the relationship, if any, between the presence of protozoa and sexual practices. Men who consecutively attended the clinic and who gave a history of having had recent sexual contact with another male were invited to provide a stool sample for protozoological examination. Diarrhoeal samples were examined by direct microscopy for trophozoites and cysts and, after staining, for Cryptosporidium spp. In all cases, a modified formol-ether method was used to concentrate protozoal cysts before microscopy. One hundred and seventy-five of the 257 men invited to participate in the study provided a stool sample. At least one species of protozoan was found in 99 (57%) men. Giardia intestinalis was identified in only 5 (3%) men. Blastocystis hominis was found in 46 (26%) men, but the presence of this protozoan was not associated with diarrhoea. Other than the prevalence of Entamoeba dispar/histolytica and G. intestinalis, which remained unchanged, the proportion of men who harboured the non-pathogenic protozoa was significantly increased from 1981/82. A correlation between oral-anal sex or peno-insertive or peno-receptive anal intercourse and the prevalence of protozoa was not found. There was also no correlation between the number of species of protozoa carried and these sexual practices. The presence of protozoa was not associated with a history of foreign travel or sexual contact with a man who had recently travelled outside the UK. The study showed that, although the prevalence of infection is low, giardiasis is still transmissible amongst homosexual men. A correlation between individual sexual practices and the prevalence of intestinal protozoa was not found.

Animals↗
PubMed1990

Ruminal ciliated protozoa in cattle fed finishing diets with or without supplemental fat.

Ruminal samples were collected at slaughter from 364 unfasted steers fed different finishing diets to obtain information on numbers and species distribution of ciliated protozoa in feedlot cattle. Total numbers of protozoa averaged 1.59 X 10(5)/g of ruminal contents. A total of 47 steers (12.9%) were defaunated, but 4.1% of the steers possessed numbers of protozoa greater than 10(6)/g. Entodinium species did not always dominate the protozoan populations; 41 faunated steers (11.2%) were devoid of entodinia, and 79 additional steers (21.7%) possessed populations dominated (greater than 50%) by other genera. Isotricha was the most commonly occurring genus supplanting Entodinium, but Polyplastron and Epidinium were frequently present in high concentrations. Tallow and soybean soapstock supplementation reduced (P less than .05) numbers of protozoa in steers consuming wheat diets. However, yellow grease supplementation did not affect numbers of protozoa in steers fed either sorghum or corn diets. Average ruminal pH was 6.20 on the wheat diet, 6.05 on the corn diet, and 5.69 and 6.23 for the two sorghum diets, respectively. We found no correlation between ruminal pH and numbers of protozoa on any diet. The presence of relatively high protozoan concentrations and few defaunated animals in feedlot cattle necessitates reevaluation of the role that ciliated protozoa play in ruminal metabolism of animals fed processed, high-concentrate diets.

Animal Feed↗
PubMed1995

[Underestimation of intestinal protozoa as a cause of diarrhea in family practice].

OBJECTIVE: To assess the frequency of intestinal protozoa in stool samples of patients with diarrhoea in general practice. SETTING: General practitioners' laboratory in Haarlem, Netherlands. DESIGN: Descriptive study. METHOD: During one year (1 February 1992 to 31 January 1993) all stool samples from patients with diarrhoea visiting a general practitioner were examined according to a standard protocol consisting of bacterial and protozoal examination. RESULTS: Among 1703 stool examinations requested by general practitioners and performed according to the protocol, pathogenic protozoa were found in 10.8% and pathogenic bacteria in 8.6%. Of the 184 patients who tested positive for pathogenic protozoa 156 harboured Giardia lamblia, 22 Entamoeba histolytica and 6 Cryptosporidium spp. Pathogenic protozoa were predominantly found in patients with diarrhoea persisting for longer than 1 week and in cases with intermittent diarrhoea. In patients with acute diarrhoea (duration < 1 week) we predominantly found pathogenic bacteria (Campylobacyter jejuni). If the search for protozoa in the stool samples would not have been performed routinely, 34% of the pathogenic protozoa (Giardia lamblia) would not have been found. CONCLUSION: Intestinal infections with protozoa are not rare in general practice. It seems worthwhile to perform protozoal examination of the stool samples in case of persistent diarrhoea.

Adolescent↗
PubMed2000

Symbiosis therapy: the potential of using human protozoa for molecular therapy.

The concept of symbiosis is proposed as a disease therapy model. It is hypothesized that protozoa that live naturally in human tissues can be genetically modified for the production and delivery of therapeutic proteins. Approximately 30 identified species of protozoa live in a variety of human tissues, both intracellularly and extracellularly. Leishmania, one species of human protozoa, has been genetically altered for conditional auxotrophy and has shown no pathology in both mouse and nonhuman primate safety tests. Several species of protozoa have been transfected with a variety of genes and have successfully manufactured active foreign proteins. Protozoa have biochemical mechanisms to glycosylate proteins. Human protozoa have evolved sophisticated mechanisms for evading immune rejection and can sometimes persist for the lifetime of the host. Symbiosis therapy does not involve genetic alteration of the host and is potentially fully reversible. Research on treating genetic diseases is currently ongoing. For example, there is a group of more than 40 genetic diseases, lysosomal storage diseases, which result from defects in lysosomal enzymes primarily in macrophages. Leishmania specifically targets the lysosomal compartment of the macrophage and therefore may be the optimal vector for treatment of many of these diseases.

Animals↗
PubMed1998

Growth and survival of protozoa isolated from a tannery effluent.

Industrial effluent from a tannery was used for the growth of algae in a medium containing various inorganic salts. Growth of algal cells became visible after 7 d. Two species of protozoa were observed to proliferate in the algal culture containing no organic supplement in the medium. The culture was kept bacteria-free by the use of antibiotics and was perpetuated for at least 150 d with no decline in the protozoan population. Efficient growth of protozoa in a culture of algae elucidated new modes of nutrition in protozoa. Cr(VI) was added to the medium to check the resistance of algae and protozoa against this heavy metal. Protozoa showed different degrees of resistance. The results indicate the importance of algae and protozoa in the process of bioremediation.

Animals↗
PubMed2000

Effect of dichromate on population and growth of various protozoa isolated from industrial effluents.

Three protozoa belonging to genera Euglena, Vorticella and Stylonychia collected from industrial wastes were cultured in a medium containing inorganic salts, basically meant for the growth of algae. Protozoa showed rapid growth in the medium. Hexavalent chromium (K2Cr2O7) at a concentration of 5 micrograms/L in the medium adversely affected the growth of protozoa. At the end of eight days of Cr administration, the population of Euglena, Vorticella and Stylonychia increased 8-, 4.5- and 10-fold, respectively, as against 30-, 6.75- and 50-fold increase in the control cultures. No apparent death phase and no change in activity or morphology of protozoa was observed at this Cr concentration. The protozoa were also exposed to different metal ions, viz. Pb (2.42 mmol/L), Cr (0.48 mmol/L), Cd (0.36 mmol/L), administered in the culture medium for a period of 2 years. The metal tolerance for S. mytilus and V. microstoma was Pb > Cr > Cd. E. proxima could not tolerate any of the long-term metal treatments. Because of the ability of these protozoa to tolerate high concentrations of heavy metals, their potential role in remediation of heavy metals from industrial wastewater is considered.

Animals↗
PubMed1995

Simulation of the effects of diet on the contribution of rumen protozoa to degradation of fibre in the rumen.

A previously described mathematical model, that stimulates the metabolic activities of rumen bacteria and protozoa, was used to examine the contribution of protozoa to neutral-detergent fibre (NDF) degradation in the rumen of cattle. Comparisons between predicted and experimentally observed NDF degradation showed general agreement. Further simulations were performed with diets containing variable proportions of concentrate (between 0 and 1 kg/kg diet DM) and at intake levels ranging between 5.3 and 21.0 kg DM/d. The simulated protozoal contribution to NDF degradation was 17-21% at the lowest intake level. Except for the all-concentrate diets, raising the feed intake level reduced this contribution to 5-13% at the highest intake level. The changes in contribution of protozoa to NDF degradation were related to variations in the fibrolytic bacteria: protozoa value and the NDF-degrading activities of protozoa predicted by the model. In simulations where dietary NDF levels were reduced and starch and sugar levels were increased independently, protozoal contribution to NDF degradation generally increased. These differences were reflected also in the generally increased protozoal contribution to NDF degradation predicted in response to a decreased roughage:concentrate value. The contribution of protozoa also generally declined in response to added N. These changes in predicted protozoal contribution to NDF degradation resulting from dietary variations provided possible explanations for the differences in rumen NDF degradation observed when animals are defaunated.

Animal Feed↗
PubMed1996

Effect of rumen protozoa on nitrogen utilization by ruminants.

Results obtained during the past decade indicate clearly that protozoa are actively involved in the degradation of dietary and microbial proteins in the rumen. Because of the great ability of protozoa to ingest the particulate matter suspended in the rumen, protozoa are more active in degrading insoluble than soluble proteins. This indicates that studies carried out using lysed and sonicated protozoa are not appropriate for quantifying the actual contribution of protozoa to protein degradation in the rumen. In vivo trials have confirmed that duodenal flow of both undegraded dietary protein plus bacterial protein generally is increased by defaunation. The decrease in ruminal ammonia concentration consistently observed after defaunation accounts for the lower urinary nitrogen (N) excretion found in defaunated animals, whereas the increase in fecal N excretion in the same animals probably results from a shift of plant cell wall digestion from the rumen to the large intestine. Total N excretion is not altered significantly by defaunation. A summary of literature data indicates there are contradictory effects of defaunation on ruminant performance. This implies that animal response to defaunation may depend on the specific nutrient-limiting performance on the one hand and on the modifications of digestion and metabolism resulting from defaunation on the other. Different methods are proposed to either eliminate or decrease the numbers of ruminal protozoa or to alter their makeup. However, none of these approaches has been tested under practical feeding conditions.

Animals↗
PubMed2005

A lack of predatory interaction between rumen ciliate protozoa and Shiga-toxin producing Escherichia coli.

AIMS: To investigate interactions between rumen protozoa and Shiga toxin-producing Escherichia coli (STEC) and to ascertain whether it is likely that rumen protozoa act as ruminant hosts for STEC. METHODS AND RESULTS: The presence of stx genes in different microbial fractions recovered from cattle and sheep rumen contents and faeces was examined using PCR. In animals shedding faecal STEC, stx genes were not detected in the rumen bacterial or rumen protozoal fractions. Direct interactions between ruminal protozoa and STEC were investigated by in vitro co-incubation. Rumen protozoa did not appear to ingest STEC, a STEC lysogen or non-STEC E. coli populations when co-incubated. CONCLUSIONS: The ruminal environment is unlikely to be a preferred habitat for STEC. Bacterial grazing by rumen protozoa appears to have little, if any, effect on STEC populations. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that ruminal protozoa are unlikely to be a major factor in the survival of STEC in ruminants. They appear as neither a host that protects STEC from the ruminal environment nor a predator that might reduce STEC numbers.

Animals↗