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PubMed1989

In vitro metabolism of 2,2'-diaminopimelic acid from gram-positive and gram-negative bacterial cells by ruminal protozoa and bacteria.

Bacillus megaterium GW1 and Escherichia coli W7-M5 were specifically radiolabeled with 2,2'-diamino[G-3H]pimelic acid [( 3H]DAP) as models of gram-positive and gram-negative bacteria, respectively. These radiolabeled bacterial mutants were incubated alone (control) and with mixed ruminal bacteria or protozoa, and the metabolic processes, rates, and patterns of radiolabeled products released from them were studied. Control incubations revealed an inherent difference between the two substrates; gram-positive supernatants consistently contained 5% radioactivity, whereas even at 0 h, those from the gram-negative mutant released 22%. Incubations with ruminal microorganisms showed that the two mutants were metabolized differently and that protozoa were the major effectors of their metabolism. Protozoa exhibited differential rates of engulfment (150 B. megaterium GW1 and 4,290 E. coli W7-M5 organisms per protozoan per h), and they extensively degraded [3H]DAP-labeled B. megaterium GW1 at rates up to nine times greater than those of ruminal bacteria. By contrast, [3H]DAP-labeled E. coli W7-M5 degradation by either ruminal bacteria or ruminal protozoa was more limited. These fundamental differences in the metabolism of the two mutants, especially by ruminal protozoa, were reflected in the patterns and rates of radiolabeled metabolites produced; many were rapidly released from [3H]DAP-labeled B. megaterium GW1, whereas few were slowly released from [3H]DAP-labeled E. coli W7-M5. Most radiolabeled products derived from [3H]DAP-labeled B. megaterium GW1 were peptides of bacterial peptidoglycan origin. The ruminal metabolism of DAP-containing gram-positive and gram-negative bacteria, even with the same peptidoglycan chemotype, is thus likely to be profoundly different.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids, Diamino↗
PubMed1993

Protozoa in subsurface sediments from sites contaminated with aviation gasoline or jet fuel.

Numbers of protozoa in the subsurface of aviation gasoline and jet fuel spill areas at a Coast Guard base at Traverse City, Mich., were determined. Boreholes were drilled in an uncontaminated location, in contaminated but untreated parts of the fuel plumes, and in the aviation gasoline source area undergoing H(2)O(2) biotreatment. Samples were taken from the unsaturated zone to depths slightly below the floating free product in the saturated zone. Protozoa were found to occur in elevated numbers in the unsaturated zone, where fuel vapors mixed with atmospheric oxygen, and below the layer of floating fuel, where uncontaminated groundwater came into contact with fuel. The same trends were noted in the biotreatment area, except that numbers of protozoa were higher. Numbers of protozoa in some contaminated areas equalled or exceeded those found in surface soil. The abundance of protozoa in the biotreatment area was high enough that it would be expected to significantly reduce the bacterial community that was degrading the fuel. Little reduction in hydraulic conductivity was observed, and no bacterial fouling of the aquifer was observed during biotreatment.

Journal Article↗
PubMed2006

Development and validation of a real-time PCR method to quantify rumen protozoa and examination of variability between entodinium populations in sheep offered a hay-based diet.

PCR and real-time PCR primers for the 18S rRNA gene of rumen protozoa (Entodinium and Dasytricha spp.) were designed, and their specificities were tested against a range of rumen microbes and protozoal groups. External standards were prepared from DNA extracts of a rumen matrix containing known numbers and species of protozoa. The efficiency of PCR (epsilon) was calculated following amplification of serial dilutions of each standard and was used to calculate the numbers of protozoa in each sample collected; serial dilutions of DNA were used similarly to calculate PCR efficiency. Species of Entodinium, the most prevalent of the rumen protozoa, were enumerated in rumen samples collected from 100 1-year-old merino wethers by microscopy and real-time PCR. Both the counts developed by the real-time PCR method and microscopic counts were accurate and repeatable, with a strong correlation between them (R2= 0.8), particularly when the PCR efficiency was close to optimal (i.e., two copies per cycle). The advantages and disadvantages of each procedure are discussed. Entodinium represented on average 98% of the total protozoa, and populations within the same sheep were relatively stable, but greater variation occurred between different sheep (10(0) and 10(6) entodinia per gram of rumen contents). With this inherent variability, it was estimated that, to detect a statistically significant (P = 0.05) 20% change in Entodinium populations, 52 sheep per treatment group would be required.

Animals↗
PubMed1988

Role of protozoa in microbial acclimation for mineralization of organic chemicals in sewage.

The role of protozoa in affecting the length of the acclimation period for the mineralization of organic compounds in sewage was investigated. The acclimation periods for the mineralization of 2 ng of p-nitrophenol, 100 ng of 2,4-dichlorophenoxyacetic acid, and 100 ng of 2,4-dichlorophenol per milliliter were markedly shortened or eliminated when the indigenous protozoa were inhibited with cycloheximide and nystatin. The extent of mineralization of the test compounds decreased when the protozoa were suppressed. Inhibition of the protozoa increased the total number of bacteria and the density of organisms able to mineralize 2 ng of p-nitrophenol/mL of sewage. Addition of Tetrahymena thermophila to sewage in which the indigenous protozoa were not active lengthened the acclimation period for the mineralization of p-nitrophenol. The addition of T. thermophila to a culture containing a low density of a p-nitrophenol-degrading Pseudomonas sp. did not affect the acclimation period prior to mineralization of the substrate, but the ciliate increased the acclimation period in the presence of high densities of Enterobacter aerogenes added as an alternative prey species. We suggest that protozoan grazing may be responsible for the acclimation period prior to the mineralization of certain organic compounds in sewage.

Animals↗
PubMed2000

Food-borne protozoa.

Pathogenic protozoa are commonly transmitted to food in developing countries, but food-borne outbreaks of infection are relatively rare in developed countries. The main protozoa of concern in developed countries are Toxoplasma, Cryptosporidium and Giardia, and these can be a problem in immunocompromised people. Other protozoa such as Entamoeba histolytica, Cyclospora cayetanensis and Sarcocystis can be a food-borne problem in non-industrialised countries. C. cayetanensis has emerged as a food-borne pathogen in foods imported into North America from South America. Microsporidia may be food-borne, although evidence for this is not yet available. The measures needed to prevent food-borne protozoa causing disease require clear assessments of the risks of contamination and the effectiveness of processes to inactivate them. The globalisation of food production can allow new routes of transmission, and advances in diagnostic detection methods and surveillance systems have extended the range of protozoa that may be linked to food.

Animals↗
PubMed1999

Influence of sampling site on concentrations and carbohydrate-degrading enzyme activities of protozoa and bacteria in the rumen.

Four ruminally cannulated cows were used to assess the distribution of the concentrations and carbohydrate-degrading enzyme activities of the liquid-associated protozoa (LAP) and solid-associated bacteria (SAB) in the rumen. The cows were fed diets (7 kg of DM/d) of 100% hay (Diet F) or 60% barley plus 40% hay (Diet C) in a 2 x 2 crossover experimental design. Samples of ruminal digesta were collected successively from the dorsal (DS), ventral (VS), and anterior (AS) sacs 1h before and 3 h after the morning feeding on two sampling days with a 2-d interval. Irrespective of diet and sampling time, the greatest proportion of entodiniomorphs, representing the main population of protozoa, was found in the DS (40% mean; P < .05). Low pH values observed in the DS (P < .05) indicated higher fermentative activity in this site than in the other parts of the rumen. Protozoa may contribute more than previously thought to the high digestive potential present at the top of the rumen that has classically been attributed to bacteria. The specific activity of plant cell wall polysaccharide-degrading enzymes in the LAP was correspondingly greater (P < .05) for DS than for VS or AS. For the two diets and sampling times, specific activity of fibrolytic enzymes in the SAB tended to be less in the upper than in the lower parts of the rumen, and less in the VS than in the AS. This tendency became significant (P < .05) for total fibrolytic enzyme activity. Differences in bacterial colonization of particles among the three sampling sites may explain such differences in fibrolytic activity of the SAB. Data suggest a spatio-temporal complementary action of the bacteria and protozoa in ruminal plant cell wall degradation, at least with the barley diet, for which the number of protozoa was highest.

Animals↗
PubMed2000

Ruminal fermentation and duodenal flow following progressive inoculations of fauna-free wethers with major individual species of ciliate protozoa or total fauna.

Naturally fauna-free (FF) wethers, equipped with ruminal and duodenal cannulas, were used in two groups of eight (Group A) and seven (Group B) animals in six consecutive experimental periods, each lasting for 28 d. The objective was to measure ruminal fermentation traits, and flows of nonammonia nitrogen (NAN), total amino acid (TAA), and bacterial nitrogen (BN) from the stomach after inoculation with individual ciliate protozoa species in each period. The wethers in both groups were fed a diet based on corn silage, haylage, and soybean meal, and they remained FF during the first period. At the beginning of each other period, the wethers were progressively inoculated intraruminally with one individual major species of ruminal ciliate protozoa or total fauna (TF). Thus, Group A was progressively inoculated (+) with Dasytricha ruminantium (DS), Polyplastron multivesiculatum (PP), Isotricha intestinalis (IS), Entodinium caudatum (EN) and TF-type A. Also, Group B was progressively inoculated (+) with IS, DS, Epidinium ecaudatum (EP), Eudiplodinium maggi (EU), and EN. Duodenal digesta and ruminal fluid were collected and sampled in each period on d 26 and 28, respectively, and subjected to chemical analyses. A significantly higher (P < .05) pH (6.4) in ruminal fluid of the Group A wethers was obtained when each DS, DS+PP, DS-PP-IS+EN, and TF population was present in the rumen than when the wethers were FF (6.2). In the Group B wethers, pH (6.1) was lower (P < .05) for the population of IS-DS-EP+EU than for other populations (6.2 to 6.3). The concentration of total VFA in ruminal fluid was higher (P < .05) in the Group B wethers when IS, IS+DS, or IS-DS+EP populations were present in the rumen than when the wethers were FF. The flow of NAN, TAA, and BN from the stomach to the intestinal tract was generally lower for different protozoa populations than for the FF period. Largest decreases (P < .05) in the flow of NAN, TAA, and BN occurred when EN was added into the rumen of wethers in the A and B groups, which already contained populations of DS-PP+IS and IS-DS-EP+EU, respectively. Holotrich protozoa had very little effect on the protein metabolism in the rumen, but cellulolytic protozoa (PP, EP, and EU) and EN decreased the efficiency of protein utilization by the ruminant host.

Amino Acids↗
PubMed1986

The role of ciliate protozoa in nutrition of the ruminant.

The effects of ciliate protozoa on the ruminal ecosystem, digestion in different parts of the gut, the nature of nutrients available for absorption and their effects upon the nutrition and productivity of their host are reviewed. Compared with fauna-free ruminants, the presence of ciliate protozoa results in a more stable ruminal fermentation, higher levels of ammonia, reduced numbers of bacteria, as well as changes in dry matter (%), liquid volume and turnover rate of ruminal contents. Associated with these differences in the rumen are higher ruminal and total tract digestion of organic matter and fiber in faunated animals. A reduction in net microbial synthesis and an increase in dietary protein degradation in the rumen results in the flow of protein to the small intestine being lower in faunated ruminants. The major nutritional effect of the ciliate protozoa is to change the ratio of protein to energy in the nutrients absorbed, with faunated animals having lower protein and higher energy availabilities compared with ciliate-free ruminants. Of the nutrients available for absorption, the ciliates have no consistent effect on the proportions of volatile fatty acids or amino acids. However, there is evidence that hydrogenation of lipids is increased, as is the supply of choline, and that the bioavailability of copper is reduced by the presence of ciliates. Defaunation of young growing ruminants that are fed high energy diets, containing low levels of ruminal nondegradable protein, results in increased growth rate and feed efficiency. It is unlikely, with the possible exception of wool growth, that there are other situations in which defaunation will be beneficial; and it is more likely to be detrimental to animal productivity. It remains to be determined whether manipulation of the types of ciliate protozoa in the rumen could improve animal performance. Information for this review was largely derived from comparisons of faunated and fauna-free animals. However, it is indicated that there are large differences in protozoa numbers and types between naturally faunated individuals in the same flock or herd, and that the effects of such variations on their host's nutrition are unknown.

Animal Nutritional Physiological Phenomena↗
PubMed2004

[Bio-indicating function of soil protozoa to environmental pollution].

Due to the abundant species and huge biomass, soil protozoa play an important role in soil ecosystem. As a bio-indicator, soil protozoa have many advantages over other soil animals. Studies on the community structures, quantities, and dynamic variations of biodiversity of soil protozoa could provide powerful means to evaluate natural environmental changes and to monitor the environmental pollution brought by anthropic activities. Based on the current study at home and abroad, this paper gave a review on the function of soil protozoa in ecosystems, their advantages as bio-indicator, and their responses to environmental factors, soil contaminants and the change of atmospheric CO2. The application prospect of soil protozoa in eco-toxicity diagnosis was also discussed.

Animals↗
PubMed1989

[Prevalence of Entamoeba histolytica and other intestinal protozoa among the inhabitants of Poznan province over a period of 30 years].

The aim of the study was to present the past and actual prevalence of Entamoeba histolytica and other intestinal protozoa in Poznań Province. Although in Poland there are only few accurate data concerning the prevalence of intestinal protozoa, the results of some studies were found reliable. In a survey conducted in 1958 in an orphanage in Poznań (Kasprzak and Karlewiczowa) the intestinal protozoa were found with high prevalence (Table 1). After six years this prevalence decreased considerably (Karlewiczowa and Kasprzak, 1964). Because at the same time such a decrease in intestinal protozoa prevalence, especially that in Entamoeba coli, was observed in the whole population of the Poznań Province, long-term surveys were undertaken. The diagnosing of intestinal protozoa in faeces of 4165 persons, performed at more points in time in the period of 1963 to 1986 in the Poznań Province, were conducted by the same highly skilled staff. Decreasing rates of infection were seen for each amoeba species at nearly each point in time, particularly in the first eight years of observation (Table 2). By using the "amoebic index" and "amoebic prevalence rate" in assessing epidemiological factors (acc. WHO, 1969), the considerable reduction in both indices should be accounted for the improvement of the sanitation and socio-economic status in our country. After the Second World War some Polish authors expressed the fear that after returning to the country, the Polish soldiers who had fought in the endemic regions of invasive amoebiasis, infected with E. histolytica, would constitute sources for spreading amoebiasis and that the disease would be a nucleus of a new, hitherto unknown, social calamity in Poland. Fortunately, the fear did not come true. Although in this country the infection with amoebae, also with E. histolytica, decreased considerably after several years, unfortunately, some opinions still prevail, being the source of an amoebo-mania in the medical staff and corresponding amoebophobia in the patients.

Amebiasis↗
PubMed1986

Symbiosis of protozoa with hydrogen-utilizing methanogens.

The symbiotic interaction between bacteria and protozoa is a well-known phenomenon. Endosymbiosis of bacteria in the amoeba Pelomyxa was reported as early as 1902 by Pénard and since then endosymbiosis has been described for a variety of protozoa by many authors. Also episymbiosis of bacteria and protozoa has been observed frequently. However, surprisingly little is known about the physiological interaction of both partners in such a close association. This is mostly because of problems that arise in characterizing the bacteria, and the unknown nature of excretion products of one partner utilized by the other. The discovery of some unique fluorescent cofactors involved in methane biochemistry which are specific for methanogenic bacteria enabled recognition of these microorganisms by fluorescence microscopy. Using this technique, several symbiotic associations between anaerobic protozoa and methanogenic bacteria have been found. The aim of this paper is to review the field and to discuss the possible functions of symbiosis for both bacteria and protozoa.

Animals↗
PubMed1983

Oxygen-dependent microbicidal systems of phagocytes and host defense against intracellular protozoa.

The role of oxygen-dependent microbicidal systems of leukocytes in the host defense against the major nonerythrocytic intracellular protozoa which infect man--Toxoplasma gondii, Trypanosoma cruzi, and the Leishmania species--is reviewed. The hydrogen peroxide-halide-peroxidase microbicidal system is uniformly cidal to these organisms in vitro. Peroxidase-independent oxygen product(s) toxicity is more variable. Studies to data indicate that phagocytes which contain granule peroxidase and which have the capacity to generate a vigorous respiratory burst; eg, neutrophils and monocytes, possess substantial activity against these protozoa. The absence of granule peroxidase together with the markedly attenuated respiratory burst of resident macrophages leaves these cells with a severe microbicidal defect. These protozoa can enter resident macrophages in the absence of antibody and survive and replicate within the intracellular environment. Enhancement of the antiparasite activity of resident macrophages can be accomplished either by activation of these cells by exposure to sensitized T-cell products, or by the introduction of exogenous peroxidase into the vacuole. Other factors influencing the ability of protozoa to survive intracellularly include the capacity of these organisms to avoid effective triggering of the macrophage respiratory burst and the levels of endogenous scavengers of oxygen products within the parasite.

Animals↗
PubMed1977

Further evidence for the regulation of bacterial populations in soil by protozoa.

After the addition to soil of large numbers of a cowpea Rhizobium strain, the population declined steadily until the numbers reached about 10(7)/g, and the protozoa rose to about 10(4)/g. When indigenous protozoa were suppressed by the addition of actidione to the soil, the density of the test rhizobium did not fall initially, but its abundance declined to about 10(7)/g when actidione-resistant protozoa arose in significant numbers. The addition to actidione-treated soil of an antibiotic-resistant strain of Paramecium led to a rapid decrease in the population of the rhizobium, the density reaching essentially the same value as in soil receiving neither the drug nor the paramecia. The same changes occurred with Xanthomonas campestris as test prey except that its numbers fell to about 10(5)/g of soil. These data provide further evidence for the key role of protozoa in controlling the abundance of populations of certain bacteria introduced into soil.

Animals↗
PubMed2005

Diarrhea, CD4+ cell counts and opportunistic protozoa in Indian HIV-infected patients.

The aim of this study was to examine the relationships among diarrhea, CD4+ cell counts and opportunistic protozoa in HIV-infected patients in North India. In a retrospective study, blood and stool samples of 200 HIV-infected patients from March 2001 until 2003, submitted to the AIDS division of National Institute of Communicable Diseases (NICD), were analyzed. Each patient was examined for opportunistic protozoa, HIV-1 status and CD4+ cell counts, and screened for diarrheal symptoms. The rate of diarrhea was 38% in the stool examination. In HIV-infected patients in the groups CD4+ > 500 cells/microL, 200 cells/microL < CD4+ < 500 cells/muL and in the AIDS patients CD4+ < 200 cells/microL, diarrhea was 14.7, 29.8 and 56.1%, respectively. It is clear that the diarrhea in the AIDS patients was significant compared with the two former groups (P < 0.0005). In the AIDS patients CD4+ < 200 cells/microL with diarrhea, Cryptosporidium infection was, at 56.5%, the highest and statistically significant compared with the other parasites (P = 0.037). Microsporidium was detected in 30.4% of the AIDS patients. Diarrhea was common and most strongly associated in patients with low CD4+ cell counts. The data stress the importance of opportunistic protozoa in the HIV-infected patients, and that opportunistic protozoa should be expected in HIV-infected patients with low CD4+ and diarrhea.

AIDS-Related Opportunistic Infections↗
PubMed2001

Exploring Leeuwenhoek's legacy: the abundance and diversity of protozoa.

Towards the end of the 17th century, Leeuwenhoek built "magnifying glasses" that enabled him to see and describe protozoa for the first time. Continued exploration of the natural history of protozoa during the past 300 years has progressed far beyond simply documenting morphospecies (global total probably <20,000). We now realize that protozoan 'biodiversity' is multi-faceted (e.g. sibling species, variant genotypes and syntrophic consortia). Realization of their extraordinary abundance has secured for protozoa the position of dominant phagotrophs and regenerators of nutrients within microbial food webs. And studies of protozoa in the natural environment have done much to effect a paradigm shift in our understanding of why specific microbes live where they do and how they got there in the first place. In particular, the hypothesis of ubiquitous dispersal of protozoan species does seem to be supported by the evidence provided by morphospecies, sibling species and even individual genotypes.

Animals↗
PubMed1984

Pathogenic and free-living protozoa cultured from the nasopharyngeal and oral regions of dental patients.

Protozoa of nose, mouth, and pharynx of 30 randomly chosen female caries patients at an odontological clinic of the National Autonomous University of Mexico, were surveyed by culture from swabs. Culture tubes of swabs from each patient were observed every other day during 5 weeks. Pathogenic protozoa found included Entamoeba histolytica Schaudinn, 1903; Naegleria fowleri Carter, 1970; Acanthamoeba castelanii Douglas, 1930; Acanthamoeba culbertsoni Singh & Das, 1970; and Balantidium coli (Malmsten, 1857) Stein, 1862. This isolation of pathogens suggests that healthy patients may be healthy carriers of cysts of protozoa, mainly amoebae, responsible for several diseases, including primary amoebic meningoencephalitis. Small pathogenic free-living amoebae have not been isolated before from females in Mexico. Many species of free-living protozoa were also cultured from swabs from the patients.

Adult↗
PubMed2005

Bacterial-protozoa interactions; an update on the role these phenomena play towards human illness.

The usage of water with poor microbiological quality increases the risk of human illness. This review discusses and updates current thinking on the nature of the interaction between a range of human bacterial pathogens and waterborne protozoa. The importance of protozoa acting as protective environments for pathogenic bacteria from disinfection and of promoting extended survival in otherwise hostile environments is highlighted. The significance of biofilms in water systems, and new relationships between Salmonella and Campylobacter and water-borne protozoa are also discussed. The protection of pathogenic bacteria from disinfection within protozoa and/or biofilms has important implications for water safety.

Animals↗
PubMed1983

Cell-mediated killing of protozoa.

Cell-mediated immunity represents an important host defence mechanism against protozoal infections. The effector cells directly involved are neutrophils, macrophages and, ultimately, activated macrophages. Within this simple scheme there are, however, considerable variations in activity. Effector cells from different animal species, and even from different strains of the same species, may be more or less effective in controlling a certain protozoal infection. Different protozoa differ in their susceptibility to cell-mediated killing according to genus, species, strain and morphological form. The most susceptible morphological form is that which occurs in the insect vector, and which has not yet adapted to protect itself from the vertebrate host. Epimastigotes of Trypanosoma and promastigotes of Leishmania are readily killed by phagocytic cells, while the corresponding trypomastigote and amastigote forms are considerably more resistant. Protozoa which live in macrophages, such as amastigotes of Leishmania, endozoites (tachyzoites) of Toxoplasma and amastigotes of reticulotropic strains of T. cruzi, have developed a remarkable resistance to the microbicidal activity of the host cell. Conversely, amastigotes of myotropic strains of T. cruzi, which live in muscle cells, have not developed this resistance to cell-mediated killing by macrophages. Readily accessible protozoa, such as T. brucei trypomastigotes and Plasmodium merozoites in the bloodstream, while they lack the marked resistance developed by reticulotropic protozoa, have a partial protection since they are attacked by phagocytic cells only when specific antibody is present. Granulocyte-mediated killing can be largely attributed to neutrophils. Eosinophils appear to play only a minor role and compete ineffectually when neutrophils are also present. The only group of protozoal species which may be significantly controlled by eosinophils are the stercorarian species of Trypanosoma. In vitro experiments show that antibody-coated trypomastigotes of T. cruzi can be killed by eosinophils, although there is little evidence that this occurs in vivo. Interestingly, this is the only species that has been reported to be susceptible to the major basic protein of eosinophils, a toxic component of the lysosomal granules which is very active against helminths. Neutrophils are not very active against endozoites of Toxoplasma gondii, Trypanosoma, trypomastigotes of salivarian Trypanosoma, free merozoites of Plasmodium, and promastigotes and amastigotes of Leishmania.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗