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PubMed1987

Occurrence and transfection of a Giardia virus.

The presence of virus-derived RNA was investigated in 38 axenically growing Giardia isolates from different geographic areas. The RNA virus was demonstrated in Giardia strains from humans in the U.S.A., England and the majority of strains from Poland. Two strains isolated respectively from a cat and a cavia also contained it. Giardia strains from humans in Belgium and Israel did not contain this RNA virus. Transfection of the RNA virus was accomplished from English and Polish strains, as well as from the cat isolate to isolates lacking it. Differences were observed both in sensitivity of Giardia strains to transfection and in infectivity of the RNA virus from different Giardia strains. Transfection could be carried out with sonicated Giardia extract as well as with filter sterilized medium in which Giardia strains containing RNA virus had grown. The RNA virus did not replicate in Giardia-free medium. No correlation could be demonstrated between the presence of the RNA virus in Giardia isolates and their in vitro resistance to some antiprotozoal drugs, nor with the fact that the strain originated from symptomatic or asymptomatic carriers. The presence of the RNA virus in Giardia trophozoites did not influence the isoenzyme patterns or restriction endonuclease patterns of repetitive DNA. A correlation may exist with the length of time since the isolation in axenic culture of the strain.

Animals↗

PubMed2006

Opsonic requirements for the respiratory burst of neutrophils against Giardia lamblia trophozoites.

BACKGROUND: Giardia lamblia is an important cause of parasitic diarrheal disease worldwide. Occasionally, polymorphonuclear neutrophils (PMNs) may participate as effector cells against Giardia lamblia. The present study was performed in order to examine the role of specific antibody and complement components in promoting the respiratory burst (RB) of PMNs against Giardia lamblia. METHODS: PMNs from human adult volunteers were incubated with Giardia trophozoites in the presence of non-immune (NS) or hyperimmune (HS) serum (anti-Giardia titer, >1:1024). Adherence was scored visually on coverslide after staining with Giemsa. The ability of Giardia to trigger the oxidative response of PMNs was measured by the anion superoxide (O2(-)) production using a cytochrome C reduction method and by the luminol amplified chemiluminescence (CL) assay. RESULTS: Incubation with NS or HS increased Giardia adherence to PMNs from 6.9 +/- 3.2% (basal adherence of Giardia incubated in buffer) to 39 +/- 18.6% (p <0.01) and 76 +/- 19.5% (p <0.001), respectively. In absence of serum, Giardia failed to trigger an oxidative response of PMNs. Opsonization with NS or HS increased the PMN O2(-) production from 3.9 +/- 0.92 nmol/2.5 x 10(6) PMNs/10 min to 9.04 +/- 1.68 (p <0.05) and 17.9 +/- 1.32 (p <0.001), respectively. A similar enhancement of the CL response was also observed. The inactivation of complement activity by heat as well as the elimination of specific antibodies by absorption produced a significant abrogation of the oxidative response but in the case of HS heat inactivation alone did not abolish the response. Similar findings (variable abrogation of the oxidative PMN response) were observed when PMNs were incubated with monoclonal antibodies directed against complement C3, C3b or the low-affinity Fc receptors (CR1, CR3 or FcRlo). CONCLUSIONS: These results show that complement components and specific antibodies influence in the Giardia-PMN interaction. Although components of complement can contribute to the RB of PMNs, specific antibodies are critical for an optimal oxidative PMN response.

Animals↗

PubMed1998

Anaerobic bacterial metabolism in the ancient eukaryote Giardia duodenalis.

The protozoan parasite, Giardia duodenalis, shares many metabolic and genetic attributes of the bacteria, including fermentative energy metabolism which relies heavily on pyrophosphate rather than adenosine triphosphate and as a result contains two typically bacterial glycolytic enzymes which are pyrophosphate dependent. Pyruvate decarboxylation and subsequent electron transport to as yet unidentified anaerobic electron acceptors relies on a eubacterial-like pyruvate:ferredoxin oxidoreductase and an archaebacterial/eubacterial-like ferredoxin. The presence of another 2-ketoacid oxidoreductase (with a preference for alpha-ketobutyrate) and multiple ferredoxins in Giardia is also a trait shared with the anaerobic bacteria. Giardia pyruvate:ferredoxin oxidoreductase is distinct from the pyruvate dehydrogenase multienzyme complex invariably found in mitochondria. This is consistent with a lack of mitochondria, citric acid cycle, oxidative phosphorylation and glutathione in Giardia. Giardia duodenalis actively consumes oxygen and yet lacks the conventional mechanisms of oxidative stress management, including superoxide dismutase, catalase, peroxidase, and glutathione cycling, which are present in most eukaryotes. In their place Giardia contains a prokaryotic H2O-producing NADH oxidase, a membrane-associated NADH peroxidase, a broad-range prokaryotic thioredoxin reductase-like disulphide reductase and the low molecular weight thiols, cysteine, thioglycolate, sulphite and coenzyme A. NADH oxidase is a major component of the electron transport pathway of Giardia which, in conjunction with disulphide reductase, protects oxygen-labile proteins such as ferredoxin and pyruvate:ferredoxin oxidoreductase against oxidative stress by maintaining a reduced intracellular environment. As the terminal oxidase, NADH oxidase provides a means of removing excess H+, thereby enabling continued pyruvate decarboxylation and the resultant production of acetate and adenosine triphosphate. A further example of the bacterial-like metabolism of Giardia is the utilisation of the amino acid arginine as an energy source. Giardia contain the arginine dihydrolase pathway, which occurs in a number of anaerobic prokaryotes, but not in other eukaryotes apart from trichomonads and Chlamydomonas reinhardtii. The pathway includes substrate level phosphorylation and is sufficiently active to make a major contribution to adenosine triphosphate production. Two enzymes of the pathway, arginine deiminase and carbamate kinase, are rare in eukaryotes and do not occur in higher animals. Arginine is transported into the trophozoite via a bacterial-like arginine:ornithine antiport. Together these metabolic pathways in Giardia provide a wide range of potential drug targets for future consideration.

Amino Acids↗

PubMed1997

Giardia and Cryptosporidium in Canadian farm animals.

Giardia intestinalis and Cryptosporidium spp. are commonly identified intestinal pathogens in humans and animals. In light of the clinical disease, production losses and zoonotic potential of both Giardia and Cryptosporidium infections, a study was undertaken to investigate the prevalence of these parasites in cattle, sheep, pigs and horses in Canadian farms at different geographical locations. A total of 104 cattle, 89 sheep, 236 pigs and 35 horses were sampled from 15 different Canadian geographical locations. Fecal samples were examined after concentration and immunofluorescent staining. Giardia and Cryptosporidium were present in cattle and sheep in six out of six sites sampled. In cattle the overall prevalence was 29% for Giardia and 20% for Cryptosporidium. Giardia was identified in 38% of sheep while 23% of sheep were positive for Cryptosporidium. Giardia and Cryptosporidium were identified in four out of six hog operations with an overall prevalence of 9% for Giardia and 11% for Cryptosporidium. All horse sampling locations (4/4) were positive for Giardia with 20% of animals infected. Cryptosporidium was identified in three out of four sampling sites with a prevalence of 17%. The prevalence of Giardia and Cryptosporidium was greater in calves and lambs compared to adults. This study demonstrates that both Giardia and Cryptosporidium appear to be prevalent in farm livestock.

Animals↗

PubMed2006

A likely molecular basis of the susceptibility of Giardia lamblia towards oxygen.

Giardia lamblia is an amitochondrial protozoan susceptible to oxygen, but the molecular basis for it remains unclear. A Giardia NAD(P)H:menadione oxidoreductase (DT-diaphorase) is known to catalyse a single electron transfer reaction with quinones as the likely two-electron acceptor when oxygen is absent. Here we overexpressed this enzyme in Giardia trophozoites and observed a significantly enhanced susceptibility of the cells towards oxygen. A knock-down of this enzyme resulted, however, in more oxygen-tolerant Giardia cells growing equally well under anaerobic and aerobic conditions. The function of DT-diaphorase could be thus a major, if not the only, cause for the oxygen susceptibility of Giardia. Overexpressed DT-diaphorase is accompanied by increased intracellular hydrogen peroxide. An overexpression of Fe-superoxide dismutase in Giardia led also to a similarly heightened sensitivity to oxygen. Thus, generation of H2O2 from superoxide anion likely produced from DT-diaphorase catalysed reaction using oxygen as electron acceptor may constitute the molecular basis for Giardia susceptibility to oxygen. A functional homologue of DT-diaphorase in Giardia, NADH oxidase, uses oxygen as the preferred electron acceptor and reduces it to water. Overexpression of this enzyme in Giardia resulted in significantly enhanced growth under aerobic conditions. Giardia NADH oxidase could be thus an instrumental enzyme for the organism to adapt to and to tolerate an aerobic living environment.

Animals↗

PubMed2004

Genotyping of human giardiasis in relation to anti-Giardia secretory IgA and mucosal histopathology.

Comparative study between the prevalence of pathological grading and Giardia genotypes revealed that, in patients infected with Giardia group I and II, out of patients having Giardia genotype I the prevalence of grade 0 was 13.16%, grade I was 21.05%, grade II was 47.37%, grade III was 13.16% and grade IV was 2.26% in comparison to 0%, 30.77%, 46.15%, 7.69% and 15.38% in genotype II (13 patients) and 10%, 40%, 20%, 20% and 10% in group III (10 patients) also in relation to 25%, 43.75%, 18.75%, 6.25% and 6.25% in mixed genotype infections group (16 patients) and 25%, 25%, 35.71%, 10.71% and 3.57% in undetermined infection group (28 patients) for grade 0, I, II, III & IV pathology respectively. There was no statistically significant difference regarding the prevalence of pathological grading in different Giardia genotypes in Gs I & II (P > 0.05). The mean OD of anti-Giardia secretory IgA in relation to Giardia genotypes in patients infected with Giardia Gs I & II was significantly different in the mean OD values of anti-Giardia secretory IgA in patients with different Giardia genotypes which were 1.091 +/- 0.377, 1.079 +/- 0.474, 1.524 +/- 0.503, 1.292 +/- 0.472 & 1.004 +/- 0.31 groups of genotype I, II, III, mixed genotypes infection and undetermined infection group respecttively (P > 0.05), being more increased in patients infected with Giardia genotype III and in mixed genotype infection.

Adolescent↗

PubMed1990

Longitudinal study of Giardia lamblia infection in a day care center population.

An enzyme-linked immunosorbent assay was used to detect Giardia lamblia in stool specimens collected during a 15-month longitudinal study of diarrhea in 82 children 1 to 24 months old attending a day care center (DCC) in Houston. A total of 2727 stool specimens were collected on a weekly basis from the DCC children and were evaluated for rotavirus and Giardia. For DCC children who developed diarrhea stool specimens were also cultured for bacterial enteropathogens. During the 15-month study period, 48 episodes of Giardia infection were detected in 27 of 82 (33%) DCC children, compared with 57 episodes of rotavirus detected in 37 (45%) of these same DCC children. The duration of Giardia excretion was 2.0 +/- 1.5 weeks (mean +/- SD). Only 6 (7%) of the 82 DCC children, or 6 of the 27 (22%) with infection, developed symptoms attributable to Giardia. Ten of the 27 (37%) DCC children infected with Giardia had 2 or more episodes of infection. Giardia was identified in the DCC in all months except June. Two Giardia outbreaks occurred in 1 of the 6 DCC rooms under study. One outbreak was associated with overcrowding. Neither outbreak was associated with the introduction of a new Giardia-positive child into the involved room. In this study Giardia infection occurred commonly in the DCC throughout the year, was rarely associated with illness and was not associated with introduction of asymptomatic carriers into the DCC rooms.

Child Day Care Centers↗

PubMed1991

DNA fingerprinting of the human intestinal parasite Giardia intestinalis with hypervariable minisatellite sequences.

Individual isolates of the Giardia duodenalis group of protozoan intestinal parasites were identified by DNA fingerprinting with hypervariable minisatellite sequences. A morphologically identical parasite is found in some forty different animal species. Although the species name intestinalis is reserved for the human isolates, electrophoretic karyotyping suggests that most duodenalis isolates fall into the same species grouping. Distinction based upon morphology, restriction endonuclease cleavage of genomic DNA or isoenzyme analysis has not been adequate to identify individual strains. The successful use of hypervariable sequences in the identification of individual human genomes encouraged us to examine the use of these same sequences for the possible identification of parasite isolates. We initially use as a fingerprinting probe the genome of the bacteriophage M13, which has repeated sequences recognising homologous hypervariable sequences in the human genome. The M13 probe recognises a weakly homologous set of hypervariable sequences in Giardia. The number of informative bands is comparable to those seen in mammals, since the lower molecular weight bands are also useful. There is considerable divergence in the sequences of individual Giardia minisatellites. Some cloned Giardia hypervariable sequences are more homologous to M13 than they are to each other. Similar results were observed with the hypervariable repeat sequences 3' to the human alpha-globin gene when they were used as a probe to distinguish Giardia isolates. The poly(dA-dC).poly(dG-dT) probe which recognises frequent TG tracts in a number of organisms also detects a few variable bands amidst a hybridisation background in the Giardia genome. Thus Giardia isolates which could not be distinguished by restriction endonuclease cleavage, antibody typing or isoenzyme analysis have been identified by DNA fingerprinting procedures. Detailed analysis of strain movement, resurgence, variation, host range and drug resistance is now possible. Similar families of sequences may be widespread in lower eukaryotes and useful for generating individual specific fingerprints. A procedure for detecting individual parasites is also presented. Since Giardia is regarded as the most ancient eukaryote before the occurrence of symbiosis with purple non-sulphur bacteria to generate mitochondria, the identification of hypervariable sequences in the Giardia genome should also aid in understanding the mechanism of generation and evolution of these sequences.

Animals↗

PubMedDate not supplied

Evaluation of specific serum anti-Giardia IgM antibody response in diagnosis of giardiasis in children.

Diagnosis of Giardia intestinalis infection is usually made by examination of stool specimens and/or by more invasive methods such as microscopy of duodenal juice or small bowel mucosal biopsies. Serological diagnostic methods have been developed but have not been evaluated in children. In this study specific anti-Giardia immunoglobulin (Ig) M, IgG and IgA antibody titres were measured by enzyme-linked immunosorbent assay. Giardia parasites were sought in jejunal mucosal biopsies and in faeces from 72 children in The Gambia, West Africa; 50 jejunal biopsies, 271 stool samples and 95 serum samples were examined for evidence of Giardia infection. As a diagnostic test, a raised specific anti-Giardia IgM antibody titre (greater than or equal to 1:800) had a sensitivity of 63% and specificity of 93%, with a positive predictive value of 85% and a negative predictive value of 81%. There was poor correlation between positive microscopical identification of Giardia and elevated specific anti-Giardia IgG or IgA antibody titres in children on admission to the study. Elevated serum anti-Giardia IgM, however, was correlated well with active Giardia infection and may prove useful in epidemiological studies of giardiasis in developing countries.

Animals↗

PubMed2006

An examination of the prevalence of and risk factors for shedding of Cryptosporidium spp. and Giardia spp. in cows and calves from western Canadian cow-calf herds.

The primary objectives of this study were to describe the prevalence and risk factors for Cryptosporidium spp. and Giardia spp. infection in cows and calves during the calving season in western Canadian cow-calf herds. Through the calving season of 2002, fresh fecal samples were collected from 560 beef cows and 605 calves in western Canada. Feces were examined for the presence of Cryptosporidium spp. and Giardia spp. using a quantitative sucrose gradient immunoflourescent antibody test. Samples were collected from mature cows on 59 farms and from calves on 100 farms. Only 1.1% (5/560) of the cows were positive for Cryptosporidium spp. whereas 3.1% (19/605) of the calves were positive. Prevalence for Giardia spp. was much higher; Giardia spp. was detected in 17.0% (95/560) of the cow and 22.6% (137/605) of calf fecal samples. Data describing herd management practices, treatment and disease history, age, gender, breed and fecal consistency were gathered to assess potential risk factors associated with shedding. The association between the risk of shedding and average precipitation from December to June and ecological region were also evaluated. Risk factors for infection with Cryptosporidium spp. in either cows or calves could not be evaluated because the multilevel model would not converge due to the relatively low prevalence of the organism in this sample. The prevalence for Giardia spp. was sufficient to explore potential risk factors in both cows and calves. No risk factors were identified for Giardia spp. in beef cows following calving. After the construction of a multivariable model, the only significant predictors for Giardia spp. presence in beef calves was dam age and calf age. Calves born to 2-year-old heifers were 2.3 (95% CI, 1.09-5.06; P = 0.031) times more likely to be shedding Giardia spp. then calves born to cows that were 4-10 years of age. Calves that were 9-18 days of age and calves that were > 18 days of age were 22.4 (95% CI, 5.88-88.18; P < 0.001) and 150 (95% CI, 39.72-603.19; P < 0.001) times more likely, respectively, to be shedding Giardia spp. than calves < or = 4 days of age.

Age Factors↗

PubMed1992

Protection against infection with Giardia lamblia by breast-feeding in a cohort of Mexican infants.

To determine whether breast-feeding protects infants against symptomatic and asymptomatic infection by Giardia lamblia, we followed 197 infants in a poor area of Mexico City from birth to 18 months of age; symptoms and feeding status were recorded weekly. Stool specimens were collected every 1 to 2 weeks and tested for Giardia by enzyme-linked immunosorbent assay. A mean of 1.0 Giardia infection per child-year was detected; 94 infants had a total of 139 infections; 17% of infections were symptomatic. Ninety-one percent of infants were breast fed from birth and 38% were breast fed at 1 year of age. Lack of breast-feeding was a significant risk factor for first Giardia infection at all ages. The adjusted incidence rate ratio for first Giardia infection for none versus complete breast-feeding was 5.0 (confidence interval (CI) 1.5 to 16.9; p = 0.009), and for none versus any breast-feeding, 1.8 (CI 1.1 to 2.8; p = 0.013). Symptomatic Giardia infection was also associated with lack of breast-feeding (none vs any: incidence rate ratio = 2.5; CI 0.9 to 6.8; p = 0.077), but breast-feeding did not protect against chronic carriage of Giardia. Other significant risk factors for Giardia infection were presence of animals in the household (p = 0.005) and the use of water or nonmilk liquid for infant feedings (p = 0.035). We conclude that breast-feeding protects infants against Giardia by mechanisms that include preventing the establishment of infection.

Animals↗

PubMed2000

Prevalence and genotypic characterisation of Giardia in dairy calves from Western Australia and Western Canada.

In this study, the prevalence of Giardia duodenalis infections was determined in Western Canadian and Western Australian dairy calves. Faecal samples were collected from Holstein calves located on a commercial dairy near Lethbridge, Alta., Canada (N=28) and from calves located on two commercial dairies located near Perth, WA, Australia (N=36). Faecal samples were examined for the presence of Giardia cysts using sucrose gradient centrifugation, followed by immunofluoresence microscopy. DNA was then extracted from Giardia isolates obtained from positive samples. A PCR based method was employed to amplify and sequence a 292bp region of the 16S-rRNA gene. Genetic sequences obtained from Giardia isolates were compared to each other and to previously sequenced isolates. Following a single faecal sample, 58% of Western Australian calves and 57% of Western Canadian calves were positive for Giardia. Geometric mean cyst counts/g of faeces were 839 for Western Australian calves and 3475 for Western Canadian calves, but these values did not differ significantly. Genetic sequences were obtained from 10 calves from Western Canada, while six sequences were obtained from Western Australian calves. Of the Western Canadian isolates, eight aligned with the proposed 'Hoofed livestock' genotype. Of the five isolates obtained from Western Australian calves, four sequences were identical to the 'Hoofed livestock' genotype. Two isolates from the Western Canadian calves and one isolate from the Western Australian calves had the identical genetic sequence to the Genotype (Assemblage) A sequence, a common human genotype. The results of this study demonstrate, for the first time, that Giardia infections occur in Western Australian calves. Also, calves from different geographical locations appear to be primarily infected with a Giardia genotype unique to hoofed livestock. However, calves can shed Giardia cysts potentially infective for humans. Thus, Giardia infections should be considered important to Australian dairy producers, and infections in calves may pose a risk to public health regardless of geographical location.

Animals↗

PubMed1994

Epidemiology of equine Cryptosporidium and Giardia infections.

Prevalence and infection patterns of Cryptosporidium and Giardia infections in horses were studied by a direct immunofluorescence staining method. Faecal examinations of 222 horses of different age groups revealed Cryptosporidium infection rates of 15-31% in 66 foals surveyed in central Ohio, southern Ohio and central Kentucky, USA. Only 1 of 39 weanlings, 0 of 46 yearlings, and 0 of 71 mares were positive. Giardia infection was found in all age groups, although the infection rates for foals were higher (17-35%). Chronological study of infection in 35 foals showed that foals started to excrete Cryptosporidium oocysts between 4 and 19 weeks and Giardia cysts between 2 and 22 weeks of age. The cumulative infection rates of Cryptosporidium and Giardia in foals were each 71%. Some foals were concurrently infected with both parasites and excretion of oocysts or cysts was intermittent and long-lasting. The longest duration of excretion was 14 weeks for Cryptosporidium and 16 weeks for Giardia. Excretion of Cryptosporidium oocysts stopped before weaning, while excretion of Giardia cysts continued thereafter. Infected foals were considered the major source of Cryptosporidium infection in foals, whereas infected mares were deemed the major source of Giardia infection in foals. The high infection rate of Giardia in nursing mares suggested a periparturient relaxation of immunity. The results indicated that Cryptosporidium and Giardia infections are common in horses.

Age Factors↗

PubMed1997

Evaluation of nine immunoassay kits (enzyme immunoassay and direct fluorescence) for detection of Giardia lamblia and Cryptosporidium parvum in human fecal specimens.

It is well known that Giardia lamblia and Cryptosporidium parvum can cause severe symptoms in humans, particularly those who are immunologically compromised. Immunoassay procedures offer both increased sensitivity and specificity compared to conventional staining methods. These reagents are also helpful when screening large numbers of patients, particularly in an outbreak situation or when screening patients with minimal symptoms. The data obtained by using 9 diagnostic kits were compared: direct fluorescent-antibody assay (DFA) kits (TechLab Giardia/Crypto IF kit, TechLab Crypto IF kit, and Meridian Merifluor Cryptosporidium/Giardia) and enzyme immunoassay (EIA) kits (Alexon ProSpecT Giardia EZ Microplate Assay, Alexon ProSpecT Cryptosporidium Microplate Assay, Cambridge Giardia lamblia Antigen Microwell ELISA, Meridian Premier Giardia lamblia, Meridian Premier Cryptosporidium, TechLab Giardia CELISA, Trend Giardia lamblia EIA). The test with the Meridian Merifluor Cryptosporidium/Giardia kit was used as the reference method. In various combinations, 60 specimens positive for Giardia, 60 specimens positive for Cryptosporidium, 40 specimens positive for a Giardia-Cryptosporidium mix, and 50 negative fecal specimens were tested. Different species (nine protozoa, three coccidia, one microsporidium, five nematodes, three cestodes, and one trematode) were included in the negative specimens. The sensitivity of EIA for Giardia ranged from 94% (Alexon) to 99% (Trend and Cambridge); the specificity was 100% with all EIA kits tested. The sensitivity of EIA for Cryptosporidium ranged from 98% (Alexon) to 99% (Meridian Premier); specificities were 100%. All DFA results were in agreement, with 100% sensitivity and specificity; however, the TechLab reagents resulted in fluorescence intensity that was generally one level below that seen with the reagents used in the reference method. In addition to sensitivity and specificity, factors such as cost, simplicity, ease of interpretation of results (color, intensity of fluorescence), equipment, available personnel, and number of tests ordered are also important considerations prior to kit selection.

Animals↗

PubMed2003

Detection of Giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies.

The present study developed and standardized an enzime-linked immunosorbent assay (ELISA) to detect Giardia antigen in feces using rabbit polyclonal antibodies. Giardia cysts were purified from human fecal samples by sucrose and percoll gradients. Gerbils (Meriones unguiculatus) were infected to obtain trophozoites. Rabbits were inoculated with either cyst or trophozoite antigens of 14 Colombian Giardia isolates to develop antibodies against the respective stages. The IgG anti-Giardia were purified by sequential caprylic acid and ammonium sulfate precipitation. A portion of these polyclonal antibodies was linked to alkaline phosphatase (conjugate). One hundred and ninety six samples of human feces, from different patients, were tested by parasitologic diagnosis: 69 were positive for Giardia cysts, 56 had no Giardia parasites, and 71 revealed parasites other than Giardia. The optimal concentration of polyclonal antibodies for antigen capture was 40 g/ml and the optimal conjugate dilution was 1:100. The absorbance cut-off value was 0.24. The parameters of the ELISA test for Giardia antigen detection were: sensitivity, 100% (95% CI: 93.4-100%); specificity, 95% (95% CI: 88.6-97.6%); positive predictive value, 91% (95% CI: 81.4-95.9%); and negative predictive value, 100% (95% CI: 96.1-100%). This ELISA will improve the diagnosis of Giardia infections in Colombia and will be useful in following patients after treatment.

Animals↗

PubMed1992

Detection of specific anti-Giardia antibodies in the serodiagnosis of symptomatic giardiasis.

The roles of circulating anti-Giardia IgM and IgG antibodies were assayed by the Enzyme linked immunosorbent assay (ELISA) using axenic Giardia lamblia whole trophozoites as antigen for routine serodiagnosis of Giardia infection. Detection of anti-Giardia IgM antibody was found to be useful in the early detection of symptomatic giardiasis, whereas IgG detection was not, since the latter could not discriminate between present infection and previous exposure. A mean optical density (OD) +/- 2 standard deviation (SD) value of 0.088 at 1:100 serum dilution was found optimal for the detection of anti-Giardia IgM antibodies as it completely separated out the acute Giardia cases from the Giardia-free controls. The same sera did not reveal any marked differences when tested for anti-Giardia IgG antibodies. Detection of IgM antibodies correlated with high sensitivity and specificity (96%) in an evaluation of a large number of sera collected from Giardia cases and controls.

Animals↗

PubMed1992

Guanine nucleotide-binding proteins in the intestinal parasite Giardia lamblia. Isolation of a gene encoding an approximately 20-kDa ADP-ribosylation factor.

Giardia lamblia is a protozoan intestinal parasite that has characteristics of both eukaryotes and prokaryotes. To determine whether genes for guanine nucleotide-binding proteins are present in Giardia, genomic DNA and cDNA libraries were screened by polymerase chain reaction and by hybridization with mixed oligonucleotide probes complementary to sequences encoding conserved GTP-binding domains. A gene with a high degree of sequence identity with mammalian ADP-ribosylation factors (ARFs), believed to be important in vesicular transport, was identified. The Giardia ARF gene had a 573-base open reading frame encoding 191 amino acids which are 63-70% identical with known mammalian and yeast ARFs. Sequence conservation among ARFs was greatest in putative GTP-binding domains. A single ARF mRNA species of approximately 750 bases was found in two different Giardia isolates. Primer extension and RNA sequencing of the Giardia ARF transcript revealed a short (6-base) 5'-untranslated region similar in size to those found in other Giardia transcripts. Giardia extracts contained ARF activity, as shown by stimulation of cholera toxin-catalyzed ADP-ribosylation and a Giardia ARF expressed in Escherichia coli as a fusion protein likewise exhibited biochemical activity. Its presence in Giardia is consistent with the view that ARF emerged before the divergence of this protozoan from other eukaryotes (approximately 1.5 billion years ago), and that an ARF-like protein may have been the ancestor of several other classes of signal-transducing guanine nucleotide-binding proteins, including the alpha subunits of the heterotrimeric G proteins.

ADP-Ribosylation Factors↗

PubMed2005

[Experimental infection of the gerbil (Meriones unguiculatus) by Colombian isolates of Giardia duodenalis].

INTRODUCTION: Natural and experimental Giardia infections have been reported from bovines, equines, goats, canines, felines and rodents such as mice, rats and gerbils. The latter have provided successful animal models for Giardia duodenalis and Giardia muris experimental infections. OBJECTIVE: The gerbil model was used to establish the pattern of infection of Colombian Giardia human isolates. MATERIALS AND METHODS: Giardia cysts were obtained from stool specimens of symptomatic giardiasis patients by means of sucrose-percoll gradients. Animal inoculation was performed by gastric intubation and injection with 5 x 10(3) Giardia cysts. The course of infection was established by counting cysts every day and trophozoites weekly throughout a period of 30 days. RESULTS: The pattern of cyst excretion was found to be intermittent. Cysts were released during the second and third weeks of infection but not during the first or fourth weeks. The mean minimal number of cysts released per 2-hr collection period was 79 and the mean maximum number was 17,943. Colonization of the small intestine by trophozoites was observed with a mean number ranging from 15,000 to 6,577,778 trophozoites/ml. DISCUSSION AND CONCLUSIONS: Gerbils inoculated with G. duodenalis isolates obtained from geographical areas outside Colombia resolved the infection between 86 and 114 days after infection, whereas gerbils infected with Colombian G. duodenalis isolates resolved the infection at 30 days. The gerbil proved to be a good animal model for experimental infection with Colombian isolates of G. duodenalis. Experimental Giardia infection of gerbils permit a sufficient yield of cysts and trophozoites to be used as antigens for the immunization of other animals and to obtain Giardia antibodies that could be used for Giardia antigen detection assays in stool specimens.

Animals↗