Common Protozoan Infection Research Articles

Pyrimidine salvage in Toxoplasma gondii as a target for new treatment.

Toxoplasmosis is a common protozoan infection that can have severe outcomes in the immunocompromised and during pregnancy, but treatment options are limited. Recently, nucleotide metabolism has received much attention as a target for new antiprotozoal agents and here we focus on pyrimidine salvage by Toxoplasma gondii as a drug target. Whereas uptake of [3H]-cytidine and particularly [3H]-thymidine was at most marginal, [3H]-uracil and [3H]-uridine were readily taken up. Kinetic analysis of uridine uptake was consistent with a single transporter with a Km of 3.3 ± 0.8 µM, which was inhibited by uracil with high affinity (Ki = 1.15 ± 0.07 µM) but not by thymidine or 5-methyluridine, showing that the 5-Me group is incompatible with uptake by T. gondii. Conversely, [3H]-uracil transport displayed a Km of 2.05 ± 0.40 µM, not significantly different from the uracil Ki on uridine transport, and was inhibited by uridine with a Ki of 2.44 ± 0.59 µM, also not significantly different from the experimental uridine Km. The reciprocal, complete inhibition, displaying Hill slopes of approximately -1, strongly suggest that uridine and uracil share a single transporter with similarly high affinity for both, and we designate it uridine/uracil transporter 1 (TgUUT1). While TgUUT1 excludes 5-methyl substitutions, the smaller 5F substitution was tolerated, as 5F-uracil inhibited uptake of [3H]-uracil with a Ki of 6.80 ± 2.12 µM (P > 0.05 compared to uracil Km). Indeed, we found that 5F-Uridine, 5F-uracil and 5F,2'-deoxyuridine were all potent antimetabolites against T. gondii with EC50 values well below that of the current first line treatment, sulfadiazine. In vivo evaluation also showed that 5F-uracil and 5F,2'-deoxyuridine were similarly effective as sulfadiazine against acute toxoplasmosis. Our preliminary conclusion is that TgUUT1 mediates potential new anti-toxoplasmosis drugs with activity superior to the current treatment.

Macroscopic and Microscopic Survey of Sarcocystis spp. Infection in Slaughtered Cattle and Sheep in Tabriz, Iran

Background & Aims: Sarcocystis infection is one of the most common protozoan infections between humans and animals, which is caused by different species of Sarcocystis. The study aimed to investigate Sarcocystis infection in cattle and sheep slaughtered in Tabriz slaughterhouse by microscopic and macroscopic methods. Materials amd Methods: Diaphragmatic muscles of 500 cattle and 800 sheep were randomly selected for macroscopic and microscopic Sarcocystis cysts. A naked eye examination was done for macroscopic sarcocysts, while peptic digestion and Daub smear method were used for the microscopic cysts. Results: The overall prevalence of Sarcocystis spp. infection was 44.0% and 68.25% in cattle and sheep, respectively. The results showed that rate of infection of cattle with Sarcocystis spp. was 10.4%, 90.0%, and 22.3% by macroscopic, peptic digestion, and Daub smear methods, respectively. Meanwhile, the percentage of infected sheep was determined by macroscopic, peptic digestion, and Daub smear methods as 30.6%, 100.0%, and 44.1%, respectively (P<0.05). Conclusion: It is concluded that digestion is a perfect method for diagnosing sarcocysts in cattle and sheep. As well as, the high prevalence of microscopic Sarcocystis spp. in cattle and sheep of Tabriz, is suggested that meat should be cooked sufficiently, the people to be trained not to feed their dogs and cats with uncooked meat.

Trichomonas gallinae Identification and Histopathological Study in Pigeon (Columba livia domestica) in Baghdad City, Iraq

Trichomonas gallinae causes avian trichomoniasis, which is one of the most common protozoan infections in birds worldwide. Therefore, this study was conducted to investigate and identify the Trichomonas gallinae in domestic pigeons (Columba livia domestica) by microscopic examination (direct smear and Giemsa stain) and histopathological examination in Baghdad city, Iraq, during the period from beginning of October 2018 to end of March 2019. Giemsa-stained cytoplasm with light purple and nucleus with dark purple, clarification of flagella, nucleus, and cytoplasm very well. Histopathological findings of infected birds showed gross existence of yellowish white caseous necrotic material in the oral cavity and esophagus. The histopathological examination in the larynx, esophagus, trachea, crop, liver, and lung as infiltration of inflammatory cell mainly (heterophils); thickening of mucosa because of extensive infiltration of heterophils and disruption of esophageal gland; the thickness in bronchi wall of lung due to glandular hyperplasia and muscular fibroplasia, in liver focal necrosis of parenchyma with mononuclear cell (MNCs) infiltration and granuloma composed of MNCs and heterophils. The current study may contribute to determining the histopathological changes of esophagus, trachea, crop, and liver of trichomoniasis- infected pigeons.

Molecular characterization of bovine Cryptosporidium isolated from diarrheic calves in the Sudan.

Cryptosporidiosis is a common protozoan infection causing morbidity and mortality in young cattle and may be zoonotically transmitted to humans. So far, there is no data available on the presence of Cryptosporidium spp. in the Sudan. The aim of this study was to isolate, identify, and genotype Cryptosporidium oocysts sampled from diarrheic calves housed at different farms in three different municipalities in Khartoum State (Khartoum, Khartoum North, Omdurman). A total of 149 fecal samples were evaluated microscopically for the presence of Cryptosporidium oocysts using the modified Ziehl-Neelsen staining method and 87 (58.3%) samples tested positive. Positive and negative samples were further analyzed by nested PCR targeting the SSU rRNA region. Positive samples were subjected to restriction enzyme analysis of PCR amplicons (PCR-RFLP). Nested PCR identified Cryptosporidium DNA in 53 samples (35.5%); restriction digestion of the PCR products revealed the presence of C. parvum (73.5%), C. ryanae (13.2%), C. andersoni (7.5%), and C. bovis (1.8%). Species distribution was clearly related to age with C. parvum being the predominant species in dysenteric pre-weaned calves. Sequencing of three genes (SSU rRNA, COWP, and GP60) for three C. parvum isolates originating from the three different municipalities showed that all belong to C. parvum subtype family IId. Based on data obtained by GP60, sequencing the two C. parvum isolates from Khartoum and Omdurman represent subtype IIdA18G1, whereas oocysts isolated in Khartoum North belong to subtype IIdA19G1. The observed genotypes are zoonotic and thus C. parvum in calves is potentially a health risk to humans in Khartoum State, Sudan. To the best of our knowledge, this is the first reported attempt to characterize Cryptosporidium isolated from cattle in the Sudan.

Prevalence and molecular characterization of bovine Cryptosporidium from dairy cows in Northern Thailand.

Cryptosporidiosis is a common protozoan infection in humans and domestic animals. It is the culprit for significant neonatal morbidity in cattle as well as weight loss and delayed growth, which leads to large economic losses in the farming industry. Furthermore, bovine Cryptosporidium is also a principal source of human Cryptosporidium infections. The purpose of this study is to determine prevalence and genotype of Cryptosporidium spp. from feces of dairy cows from the northern parts of Thailand (Chiang Rai, Chiang Mai, and Lumpang provinces). A total of 500 fecal samples were collected directly from the rectum and they were examined for potential presence of Cryptosporidium infection by using tests such as DMSO-modified acid fast stain, immunofluorescence antibody test (IFAT) and polymerase chain reaction (PCR). The prevalence of Cryptosporidium spp. was 5% by DMSO-modified acid fast stain, 7% by IFAT and 7.6% by PCR respectively. The main genotypes of Cryptosporidium spp. identified were Cryptosporidium parvum and Cryptosporidium bovis. Therefore, as a result of this study, it can be said that, due to the potential cross-species transmission of Cryptosporidium parvum, infected dairy cows may pose a potential zoonotic risk to humans.

Cerebral malaria--clinical manifestations and pathogenesis.

One of the most common central nervous system diseases in tropical countries is cerebral malaria (CM). Malaria is a common protozoan infection that is responsible for enormous worldwide mortality and economic burden on the society. Episodes of Plasmodium falciparum (Pf) caused CM may be lethal, while survivors are likely to suffer from persistent debilitating neurological deficits, especially common in children. In this review article, we have summarized the various symptoms and manifestations of CM in children and adults, and entailed the molecular basis of the disease. We have also emphasized how pathogenesis of the disease is effected by the parasite and host responses including blood brain barrier (BBB) disruption, endothelial cell activation and apoptosis, nitric oxide bioavailability, platelet activation and apoptosis, and neuroinflammation. Based on a few recent studies carried out in experimental mouse malaria models, we propose a basis for the neurological deficits and sequelae observed in human cerebral malaria, and summarize how existing drugs may improve prognosis in affected individuals.

HIV and intestinal parasite co-infections among a Chinese population: an immunological profile.

BackgroundParasite infections often result in a switch of the human body’s predominant immune reaction from T-helper 1 (Th1)-type to Th2-type. Hence, parasite infections are widely expected to accelerate the progression of human immunodeficiency virus (HIV) infections to acquired immunodeficiency syndrome (AIDS). In the People’s Republic of China, both parasitic diseases and AIDS are epidemic in certain rural areas, and co-infections are relatively common. However, no population-based studies have yet investigated the frequency of HIV and parasite co-infections, and its effects on immune responses. We studied (1) the immune status of an HIV-infected population, and (2) the effect of co-infection of HIV and intestinal parasites on selected parameters of the human immune system.MethodsA total of 309 HIV-infected individuals were recruited and compared to an age-matched and sex-matched control group of 315 local HIV-negative individuals. Questionnaires were administered to all participants to obtain information on sociodemographic characteristics, sanitation habits, family income, and recent clinical manifestations. Two consecutive stool samples and 10 ml samples of venous blood were also collected from each individual for the diagnosis of parasite infections and quantitative measurements of selected cytokines and CD4+ T-lymphocytes, respectively.ResultsDuring the study period, 79 HIV-infected individuals were not under highly active antiretroviral therapy (HAART) and were thus included in our analysis; the prevalence of intestinal helminth infections was 6.3% and that of protozoa was 22.8%. The most common protozoan infections were Blastocystis hominis (B. hominis) (13.9%) and Cryptosporidium spp. (10.1%). The prevalence of Cryptosporidium spp. in HIV-infected individuals was significantly higher than that in HIV negative individuals (P < 0.05). Compared to the non-co-infected population, no significant difference was found for any of the measured immunological indicators (P > 0.05). However, the following trends were observed: IFN-γ levels were lower, but the IL-4 level was higher, in the population co-infected with HIV and helminths. In the population co-infected with HIV and B. hominis, the IL-2 level was higher. The population co-infected with HIV and Cryptosporidium spp. had markedly lower CD4+ T-lymphocyte counts.ConclusionAccording to the immunologic profile, co-infection with helminths is disadvantageous to HIV-infected individuals. It was associated with a shift in the Th1/Th2 balance in the same direction as that caused by the virus itself, which might indicate an acceleration of the progress from an HIV infection to AIDS. Co-infection with Cryptosporidium spp. was not associated with a significant change in immune factors but co-infection with Cryptosporidium spp. was associated with a reduced level of CD4 + T-lymphocytes, confirming the opportunistic nature of such infections. Co-infection with B. hominis, on the other hand, was associated with an antagonistic shift in the immunological profile compared to an HIV infection.

Comparison of microscopy, rapid immunoassay, and molecular techniques for the detection of Giardia lamblia and Cryptosporidium parvum

Giardia lamblia and Cryptosporidium parvum are recognized as the most common protozoan infections in Saudi Arabia. Microscopic examination of stool samples, either direct or concentrated, for the recovery of G. lamblia cysts and trophozoites and C. parvum oocysts is still the most commonly used for the diagnosis of both parasites. We compared the conventional parasitological techniques of iodine-stained wet mount for G. lamblia and Kinyoun's acid-fast for C. parvum against ImmunoCard STAT® Cryptosporidium/Giardia and real-time polymerase chain reaction (PCR) detecting the 18S rRNA gene of G. lamblia and conventional PCR detecting the same gene of C. parvum at a tertiary hospital in Dhahran, Saudi Arabia. Out of 148 stool samples, 19 and 12 true positives were identified for G. lamblia and C. parvum, respectively, using a composite reference standard. In this case, true positives and negatives were considered as those with at least two positive or negative results out of the three tests. Both ImmunoCard STAT! and PCR methods were more sensitive than the microscopic tests of a single stool specimen of 85.7% (CI=62.6-96.2%) and 85.7% (CI=56.2-97.5%) for G. lamblia and C. parvum, respectively. However, specificity of microscopic tests was higher than other techniques for both parasites. Although PCR seems to be most sensitive for both G. lamblia and C. parvum, its low specificity may render its superiority over other techniques. When a single stool sample is used for detection of G. lamblia and C. parvum, better results can be obtained when coupled with serological testing. Although PCR is the most sensitive method for the detection of both G. lamblia and C. parvum, its use requires attention in relation to the increased possible false positives.

Opportunistic Parasites among Immunosuppressed Children in Minia District, Egypt

A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children (P<0.0001). The most common protozoan infection in immunosuppressed group was Cryptosporidium parvum (60.2%), followed by Blastocystis hominis (12.1%), Isospora belli (9.7%), and Cyclospora caytenensis (7.8%). On the other hand, Entamoeba histolytica (24.6%) and Giardia lamblia (17.6%) were more common than other protozoans in immunocompetent children.

Risk factors for the initial symptomatic giardia infection in a cohort of young Arab-Bedouin children

Giardiasis is a common protozoan infection with clinical manifestations in children ranging from asymptomatic carriage to persistent diarrhoea with malabsorption. It can lead to growth and developmental retardation. The study evaluated risk factors for the initial symptomatic giardiasis (SG) episode among Arab-Bedouin children in Israel. A community-based, prospective cohort study was conducted in Rahat, a Bedouin township in southern Israel. Infants (n=238) were followed by weekly visits from birth to age 18 months. Giardia infection was identified by antigen detection in faecal specimens. Approximately 26% of children experienced one or more SG episode. Mean (SD) age for first SG episode was 12.3 (3.3) months, with 95% of episodes occurring in children >6 months of age. Risk for the first SG in children >6 months of age was associated with it being spring or summer [odds ratio (OR) 6.16, p<0.001], exposure to livestock (OR 4.89, p=0.002) and prior infection with entero-aggregative Escherichia coli (EAEC) (OR 1.12 for each additional percentage in stool prevalence, p=0.02). Weight-for-age Z-scores at age 6 months were inversely related to SG risk (OR 0.62 for each unit increase in Z-score, p=0.029). Giardiasis is an important cause of diarrhoea in Bedouin children. Increased risk of SG in spring/summer might be linked to environmental conditions or seasonal dietary practices which increase virulence or transmission. SG in those exposed to livestock suggests that there are zoonotic risk factors or that hygiene is a causal factor. The association between EAEC infection and SG warrants further investigation.

Duodenal giardiasis: A diagnosis we should not neglect

Background. Giardiasis is the most common protozoan infection, well known and largely widespread all over the world. Infants and younger children are susceptible to Giardia Intestinalis infection. This condition may be responsible of failure to thrive, malabsorption, diarrhoea and abdominal pain. Prevalence is highest in developing countries, but it is also increasing in the industrialized world because of foreign travels and immigration.

A preliminary survey of gastrointestinal and haemoparasites of beef cattle in the tropical livestock farming system in Nan Province, northern Thailand

Parasitism is a primary cause of production losses in most cattle-producing countries of the world, including Thailand. A cross-sectional study was designed to determine baseline data on the prevalence of gastrointestinal and haemoparasites of beef cattle in Nan Province, northern Thailand and to investigate the factors associated with the prevalence of parasitic infections. A total of 207 faecal and 162 blood samples were collected during the summer of 2005. The basic data of management were recorded, including the number of animals on the farm or in the village, major animal health problems, deworming programme and veterinary service. The overall prevalence of gastrointestinal parasitic infections was 61% (126/207). The most common helminth infections in this study were rumen flukes 28% (58), followed by strongyles 27% (55), and the rest were due to Strongyloides 1% (2) and Trichuris 1% (2). The common protozoan infections were Buxtonella cysts 2% (5) and coccidia oocysts 2% (4). Of the 162 blood samples examined, 50% (81) contained Theileria sp., 2% (4) trypanosome, and 1% (1) microfilariae. The high rate of parasitic infections in these areas might be related to the poor management by the farmers, such as sharing the same grazing pasture.

RISK OF INTESTINAL HELMINTH AND PROTOZOAN INFECTION IN A REFUGEE POPULATION

With continuing emigration from endemic countries, screening for parasitic infections remains a priority in U.S. communities serving refugee and immigrant populations. We report the prevalence of helminths and protozoa as well as demographic risk factors associated with these infections among 533 refugees seen at the Santa Clara County, California, Refugee Clinic between October 2001 and January 2004. Stool parasites were identified from 14% of refugees, including 9% found to have one or more protozoa and 6% found to have at least one helminth. Most common protozoan infections were Giardia lamblia (6%) and Dientamoeba fragilis (3%), and for helminths, hookworm (2%). Protozoa were more frequent in refugees < 18 years of age (OR: 2.2 [1.2-4.2]), whereas helminths were more common in refugees from South Central Asia (OR: 8.0 [2.3-27.7]) and Africa (OR: 5.9 [1.6-21.6]) when compared with refugees from Eastern Europe and the Middle East. Among helminths, Ascaris lumbricoides and hookworm were concentrated among South Central Asians (6 of 7 and 10 of 11 cases, respectively), whereas Strongyloides stercoralis was predominantly found in Africans (5 of 7 cases). Although predeparture empirical treatment programs in Saharan Africa may have helped to reduce prevalence among arriving refugees from this region, parasitic infection is still common among refugees to the United States with helminth infections found in more specific populations. As refugees represent only a fraction of recent immigrants from endemic countries, current studies in nonrefugee groups are also needed.

Effectiveness of peptone-yeast extract (P-Y) medium in the cultivation and isolation of Entamoeba histolytica/Entamoeba dispar in Turkish patients

Amebiasis is a common protozoan infection worldwide, causing serious health problems in both children and adults. Today, almost 10% of the world population is infected with Entamoeba histolytica/Entamoeba dispar. The aims of this study were both the comparison of the reproduction rates and densities of E. histolytica/E. dispar in Robinson, Dobell-Laidlaw and P-Y culture media and isolation of E. histolytica/E. dispar from stool samples in Peptone-Yeast (P-Y) medium. Trophozoites and cysts of E. histolytica/E. dispar, maintained in Robinson medium, and stool samples of patients with amebiasis were inoculated into P-Y, Robinson and Dobell-Laidlaw culture media. Reproduction rates reached their peak levels 48 h after the inoculation in all culture media. Reproduction rates in P-Y and Robinson media were found similar; however, they were higher than the reproduction rate in Dobell-Laidlaw medium ( p < 0.01); there was no statistically significant difference between the reproduction rates of P-Y and Robinson media ( p > 0.05). Twelve isolates from 12 patients were cultivated in P-Y medium and checked for reproduction everyday for 7 days. Twelve of the 12 (100%) isolates were cultivated in P-Y medium, indicating that the P-Y was an effective medium for the isolation of E. histolytica/E. dispar in stool samples. According to these results, P-Y medium could be preferred in immunologic, serologic and molecular studies and, thus the definitive diagnosis of amebiasis due to its low cost and simple formula.

Microbes and microbial toxins: paradigms for microbial-mucosal interactions I. Pathophysiological aspects of enteric infections with the lumen-dwelling protozoan pathogen Giardia lamblia.

Giardia lamblia is one of the most important causes of waterborne diarrheal disease worldwide, and giardiasis is the most common protozoan infection of the human small intestine. Symptomatic infection is characterized by diarrhea, abdominal pain, and malabsorption, leading to malnutrition and weight loss, particularly in children. The pathogen resides strictly in the lumen of the small intestine, and infection is typically not accompanied by significant mucosal inflammation. Clinical and experimental studies indicate that B cell-dependent host defenses, particularly IgA, are important for controlling and clearing Giardia infection, although B cell-independent mechanisms also contribute to this outcome. In contrast to antigiardial host defenses, much less is known about the pathophysiological mechanisms underlying the clinical symptoms of giardiasis, partly because of the current lack of suitable model systems. In addition to being an important human enteric pathogen, Giardia is an interesting model organism for gaining basic insights into genetic innovations that led to evolution of eukaryotic cells, since it belongs to the earliest diverging eukaryotic lineage known. The completion of the giardial genome project will increase understanding of the basic biology of the protozoan and will help us to better understand host pathogen-interactions as a basis for developing new vaccination and therapeutic strategies.

Giardia lamblia carriage in Israeli Bedouin infants: risk factors and consequences.

Giardiasis is a common protozoan infection, with varying clinical manifestations. We investigated the associations between Giardia lamblia carriage and environmental, family, illness, and growth characteristics. Bedouin infants (n=234) were followed from birth to age 18-23 months. At monthly home visits, stool samples were obtained, history of illness was determined, and an environmental assessment was done. The comparisons presented are between 4 groups defined by length of carriage of G. lamblia. Study children had a mean+/-SD of 4.1+/-2.9 diarrhea episodes. No illness, environmental, or family characteristics were associated with length of carriage. Significant differences were found in weight-for-age and weight-for-height z scores between the never-positive-for-G. lamblia group and all other carriage groups combined. Faltering growth was shown to be subsequent to G. lamblia infection rather than preceding it. Our findings confirm that G. lamblia carriage is not associated with diarrhea. However, the effect on growth deserves further investigation.

Parasitic diseases of the heart II: Toxoplasmosis and other protozoan and helminthic diseases

Toxoplasmosis Toxoplusma gondii is an apicomplexan obligate intracellular parasite with worldwide distribution, capable of causing several syndromes. It is one of the most common protozoan infections of man, In the United States the seroprevalence of T. gondii infection in adults varies from 20% to 70% depending on geographic area. Although most cases of acute toxoplasmosis are asymptomatic, a small number of acute infections result in clinically evident disseminated disease involving multiple organs (1). If a pregnant women is infected for the first time with T. gondii during her pregnancy, her child may acquire congenital toxoplasmosis (2). Iatrogenie immunosuppression associated with chemotherapy and transplantation has resulted in reactivation of latent toxoplasmosis with the development of clinical disease (3). With the advent of the HIV epidemic, there has been a dramatic increase in the number of patients with Toxoplasma encephalitis that may also be associated with dissemination to other organs (4,s). T. gondii has three morphologic forms: tachyzoite, bradyzoite (tissue cyst), and the oocyst. The definitive hosts for T. go&i are members of the cat family (6). Gametogony occurs only in the small intestines of cats resulting in the production of infectious oocysts. Humans and other animals become infected by ingesting oocysts contaminating food or

Evaluation of two serological tests for Trichomonas vaginalis infection.

Trichomonas vaginalis is a widely prevalent, sexually transmitted protozoan infecting both males and females. Despite its prevalence, little is known about its contribution to the morbidity rates for urogenital-tract infections. Currently accepted diagnostic methods are limited to the demonstration of the organism in fresh material, identification in stained material, or in vitro cultivation of organisms from the urogenital tract. We have evaluated the indirect hemagglutination test and the gel diffusion test for efficacy in detecting antibodies in serum samples drawn from two population groups. Sera from patients attending a vaginitis clinic had a seropositivity rate of 69% by indirect hemagglutination and 34% by gel diffusion. Seropositivity rates among culture-positive patients were 78% with indirect hemagglutination and 43% with gel diffusion. A group of normal female hospital employees showed seropositivity rates of 30% by indirect hemagglutination and 3% by gel diffusion. Absorption of reactive sera with Trichomonas antigens reduced or abolished the serological reactivity, confirming the specificity of the test. Serological methods can provide a rapid, sensitive, and economical tool to study the epidemiology of this common protozoan infection.