Malaria is a major public health burden throughout the world. Resistance to the antimalarial drugs has increased the mortality and morbidity rate that is achieved so far through the malaria control program. Monitoring the drug resistance to the available antimalarial drugs helps to implement effective drug policy, through the in vivo efficacy studies, in vitro drug susceptibility tests and detection of molecular markers. It is important to understand the mechanism of the antimalarial drugs, as it is one of the key factors in the emergence and spread of drug resistance. This review summarizes the commonly used antimalarial drugs, their mechanism of action and the genetic markers validated so far for the detection of drug-resistant parasites.

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Leishmaniasis is a vector borne protozoan disease and it remains a major public health problem world-wide. Lack of an effective vaccine and vector control program makes the chemotherapy as the primary tool for leishmaniasis. Antimonials were used as the first line of treatment for many years. Emergence of resistance against this drug has become a major concern. Literatures and studies published on anti-leishmanial drug resistance, newer drug discovery for leishmanial resistance etc., in PubMed, Medline and Google search and reviewed thoroughly. Various newer drugs have been identified but, are in limited use because of high cost, toxicity, resistance etc., Recently, many newer mechanisms of drug resistance have been identified which may boost in future designing and development of drugs.

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Blastocystis , an unusual anaerobic, single-celled stramenopile, is a remarkably successful intestinal parasite of a vast array of host species including humans. Fecal Deoxyribonucleic acid (DNA) analysis by nucleic-acid based methods in particular has led to significant advances in Blastocystis diagnostics and research over the past few years enabling accurate identification of carriers and molecular characterization by high discriminatory power. Moreover, Blastocystis comprises a multitude of subtypes (STs) (arguably species) many of which have been identified only recently and molecular epidemiological studies have revealed a significant difference in the distribution of STs across host species and geographical regions. Having a cosmopolitan distribution, the parasite is a common laboratory finding in the stools of individuals with and without intestinal symptoms across the entire globe and while the parasite remains extremely difficult to eradicate and isolate in culture, appropriate molecular tools are now available to resolve important questions such as whether the clinical outcome of colonization is linked to ST and whether Blastocystis is transmitted zoonotically. This review summarizes some of the recent advances in the molecular diagnosis of Blastocystis and gives an introduction to Blastocystis STs, including a recommendation of subtyping methodology based on recent data and method comparisons. A few suggestions for future directions and research areas are given in the light of relevant technological advances and the availability of mitochondrial and nuclear genomes.

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Toxoplasma gondii is an obligate intracellular protozoan parasite presenting as a zoonotic infection distributed worldwide. In HIV-positive individuals, it causes severe opportunistic infections, which is of major public health concern as it results in physical and psychological disabilities. In healthy immunocompetent individuals, it causes asymptomatic chronic persistent infections, but in immunosuppressed patients, there is reactivation of the parasite if the CD4 counts fall below 200 cells/μl. The seroprevalence rates are variable in different geographic areas. The tissue cyst or oocyst is the infective form which enters by ingestion of contaminated meat and transform into tachyzoites and disseminate into blood stream. In immunocompetent persons due to cell-mediated immunity the parasite is transformed into tissue cyst resulting in life long chronic infection. In HIV-infected people opportunistic infection by T. gondii occurs due to depletion of CD4 cells, decreased production of cytokines and interferon gamma and impaired cytotoxic T-lymphocyte activity resulting in reactivation of latent infection. The diagnosis can be done by clinical, serological, radiological, histological or molecular methods, or by the combination of these. There is various treatment regimen including acute treatment, maintenance therapy should be given as the current anti T. gondii therapy cannot eradicate tissue cysts. In HIV patients, CD4 counts <100; cotrimoxazole, alternately dapsone + pyrimethamine can be given for 6 months. Hence, early diagnosis of T. gondii antibodies is important in all HIV-positive individuals to prevent complications of cerebral toxoplasmosis.

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The unicellular protist Blastocystis has long been an unsolved puzzle for taxonomists, microbiologists and clinicians. Over the years, the organism has been bounced on and off the different branches of the tree of life due the possession of unique phenotypic characters intermediary to different organisms. The organism is polymorphic with only few of forms such as vacuolar, granular, amoeboid, and the cyst form being commonly known. However it could exist in other forms much more frequently than the widely known forms which could be missed by the unaware observer. Certain older concepts in the life cycle of Blastocystis although has been proven wrong are still being followed in various textbooks and other trustworthy internet sources. The causal role of Blastocystis in human disease has long been a subject of controversy. It is widely believed that certain subtypes of the organism are virulent. But this is not so as other factors are also involved in the clinical outcome of the infection. In these contexts, this review intends to shed light on the past misconceptions and the recent findings on the taxonomy, biology and the virulence of this organism.

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The rearing of Aedes mosquitoes is complex and demanding for several reasons. Aedes larvae are affected by temperature, density and available nutrition, mating is not necessarily accomplished naturally and females need a blood meal to develop eggs. The climate chambers where the mosquitoes are kept are warm and sweaty. Due to these tropical conditions the larvae develop fast and need to be cared for daily. The Laboratory of Entomology in National Institute of Malaria Research Bangalore has cultured different colonies of different vectors successfully. In this paper, we discuss different aspects off the rearing process which affect mosquito fitness and are of importance for the quality of fundamental and applied research.

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Toxoplasmosis in organ transplant patients can be a result of donor-transmitted infection, or reactivation of latent infection, or de novo infection. Solid organ transplants including heart, liver, kidney, pancreas and small bowel, and hematogenous stem cell transplants have been implicated in the risk of acquiring infection. In contrast to a benign course in immunocompetent individuals, the spectrum of illness is severe in transplant recipients. Clinical manifestations usually occur within the first 3 months of transplant and may present as encephalitis, pneumonitis, chorioretinitis, meningitis, and disseminated toxoplasmosis with multi-organ involvement. The diagnosis of toxoplasmosis in organ transplant patients is often difficult and is an integration of clinical, radiological, and microbiological workup. Preventive measures include pretransplant evaluation and chemoprophylaxis in view of rapidly progressing and fatal outcome of toxoplasmosis in immunocompromised individuals.

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Amebiasis is a disease caused by the protozoan parasite Entamoeba histolytica. This ameba can colonize the human intestine and persist as a commensal parasite, similar to Entamoeba dispar, an ameba considered to be non-pathogenic. The similarities between E. histolytica and E. dispar make the latter an attractive model for studies aimed at clarifying the pathogenesis of amebiasis. However, in addition to being an interesting experimental model, this relative of E. histolytica remains poorly understood. In the 1990, it was believed that E. dispar was unable to produce significant experimental lesions. This scenario began to change in 1996, when E. dispar strains were isolated from symptomatic patients in Brazil. These strains were able to produce liver and intestinal lesions that were occasionally indistinguishable from those produced by E. histolytica. These and other findings, such as the detection of E. dispar DNA sequences in samples from patients with amebic liver abscess, have revived the possibility that this species can produce lesions in humans. The present paper presents a series of studies on E. dispar that begin to reveal a new facet of this protozoan.

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Hydatid disease (HD) is endemic in many parts of the world. HD can affect virtually any organ system in body and should be kept as differential diagnosis of cystic lesion. HD is mostly asymptomatic; however, it demonstrates a variety of characteristic imaging findings depending on the site of involvement, stage of growth, mass effect, complications, or hematogenous spread, which helps in diagnosis. Radiography, ultrasonography (USG), computed tomography (CT), and magnetic resonance imaging (MRI) are commonly used imaging modalities. Radiography is helpful in chest and for demonstrating calcification. USG demonstrates characteristic findings such as cystic nature, daughter vesicles, membranes, septa, and hydatid sand. CT and MRI are modalities of choice for number, size, anatomic location, identification of local complications, and systemic spread. CT is, especially helpful for osseous involvement, and MRI is better for biliary and neurological involvement. Knowledge of these imaging findings helps in early diagnosis and timely initiation of appropriate therapy.

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Medical ethics is a sensible branch of moral philosophy and deals with conflicts in obligations/duties and their potential outcome. Two strands of thought exist in ethics regarding decision-making: deontological and utilitarian. In deontological approach, outcomes/consequences may not just justify the means to achieve it while in utilitarian approach; outcomes determine the means and greatest benefit expected for the greatest number. In brief, deontology is patient-centered, whereas utilitarianism is society-centered. Although these approaches contradict each other, each of them has their own substantiating advantages and disadvantages in medical practice. Over years, a trend has been observed from deontological practice to utilitarian approach leading to frustration and discontentment. Health care system and practitioners need to balance both these ethical arms to bring congruity in medical practice.

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Cryptosporidium spp. is under recognized as an important pathogen causing diarrhea in children and HIV-infected individuals with associated high morbidity and mortality. In endemic areas, most symptomatic infections are in childhood and in immunocompromised adults. The immune status of the host plays a critical role in determining the severity of cryptosporidiosis. Infection is self-limited in immunocompetent hosts, but can be severe and persistent in the immunocompromised such as AIDS patients or malnourished children. Cryptosporidiosis in developing countries is a major cause of acute and persistent diarrhea in children and is associated with subsequent impairment in growth, physical fitness, and cognitive function. Despite recognition of the importance of immune status, the correlates of protective immunity in cryptosporidiosis in humans are poorly understood, and treatment modalities are limited.

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The Kala-azar/visceral leishmaniasis (VL) turns epidemic form once in every 15 years in the endemic regions of Indian subcontinent. The goal of elimination of Kala-azar from India by 2010 was lost despite paramount efforts taken by the Government of India and World Health Organization and Regional Office for South East Asia. The main objective of this review was to elucidate the possible reason for the failure of Kala-azar elimination program and to suggest possible remedial measures to achieve the goal in future. The annual numbers of VL cases and deaths recorded in India since 1977 were plotted on a graph, to see if the temporal trends could be associated with changes in the vector control practices or co-infection with human immunodeficiency virus (HIV) or therapeutic modalities used against VL. The VL cases flares up whenever the effect of dichlorodiphenyltrichloroethane (DDT) diminished after the withdrawal of spray. The fading effectiveness was clearly correlated with an increasing number of VL cases. Therapeutic modalities were found to be highly correlating with VL mortality not with VL morbidity. The diminishing efficacy of first and second line drugs and the introduction of new drugs and drugs combination were responsible for ups and downs in the VL mortality. The VL mortality is constantly declining since 1993, but cases started increasing from 2003 to 2007 and then recently again from 2010 to 2011. This shows a serious lacuna in the vector control practices applied. The extent of HIV co-infection did not show any correlation with number/trend of VL cases or death over the study period. It is concluded that, by strict vector control practices, the VL cases can be reduced and by applying proper therapeutic strategies, the VL mortality can be reduced. HIV-VL co-infection does not seem to be in a worried stage.

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Cryptosporidiosis is a major etiology of persistent and chronic diarrhea in children and immunocompromised patients. In addition, it is also one of the important pathogens causing zoonotic or waterborne outbreaks. A number of conventional and molecular tests are available, but each test is riddled with few limitations. Further, there is a need to develop point of care tests and multiplexed tests for simultaneous detection of multiple pathogens.

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Background and Objectives: Cryptosporidiosis is a very important opportunistic infection and is responsible for significant morbidity and mortality in HIV/AIDS patients. The objective of this study is to evaluate Ziehl-Neelsen staining, auramine phenol staining, antigen detection enzyme linked immunosorbent assay and polymerase chain reaction, for the diagnosis of intestinal cryptosporidiosis. Materials and Methods: The study was designed to determine the efficacy of modified Ziehl-Neelsen (ZN), Auramine-Phenol (AP) staining, antigen detection enzyme linked immunosorbent assay (ELISA) and nested polymerase chain reaction (PCR) for detection of cryptosporidia in 671 HIV-seropositive patients, 353 HIV-seronegative patients including 198 children with diarrhea and 50 apparently healthy adults. Results: Cryptosporidium was detected in 26 (3.9%), 37 (5.5%), 32 (4.8%) and 40 (6%) HIV-seropositive and 8 (2.3%), 10 (2.9%), 9 (2.6%) and 9 (2.6%) HIV-seronegative patients by ZN staining, AP staining, antigen detection ELISA and PCR, respectively. None of the healthy controls were infected with Cryptosporidium. Based on criteria of 'true positive' samples, i.e. positive by any two of the four techniques out of ZN, AP, antigen detection ELISA and PCR, sensitivity of ZN and ELISA was 79.06% and 95.35% respectively. AP and PCR were found to be 100% sensitive. Specificity of ZN and ELISA was 100% while specificity of AP and PCR was 99.59% and 99.39% respectively. Conclusions: Auramine phenol staining is a rapid, sensitive and specific technique for diagnosis of intestinal cryptosporidiosis.

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Blastocystis is a highly controversial protozoan parasite. It has been variably regarded as a commensal and pathogen. Scientists have for decades wondered whether it is truly an enteropathogen and if it is observed in symptomatic patients whether treatment is required because patient recovery and improvement has been noted even without any treatment. Though associated with self-limiting infection, treatment is warranted in many patients due to persistence of symptoms. This particularly holds true for children and adults who are immuno compromised. Several drugs have been used to treat Blastocystis but each one of them has produced widely variable rates of clinical cure and eradication of the parasite from the feces. Based on the studies carried out in vitro and clinical responses obtained in patients, metronidazole appears to be the most effective drug for Blastocystis infection. However, the therapy is complicated due to different dosages and regimens adopted and the unresponsiveness to treatment observed in several sections of the population studied. Recently, the finding of different subsets of Blastocystis exhibiting resistance to metronidazole and associated with variable degrees of symptoms has underscored the importance of typing the subsets of the parasite in order to foretell the clinical response and the need to treat. Till date, the mode of action of the drugs used and the mechanism of resistance is not entirely known and is a topic of speculation. Other drugs with anti Blastocystis activity and used in therapy includes trimethoprim sulfamethoxazole and nitazoxanide. Several other compounds have also been evaluated for the treatment either alone or in combination with the first or second line drugs. A lot of interest has also been generated on the role of probiotics particularly Saccharomyces boularrdii and other natural food compounds on eradication of the parasite. This review provides a comprehensive overview of antimicrobials used to target Blastocystis and discusses the issues pertaining to drug resistance, treatment failure, reinfection, and the current views on treatment modalities.

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Toxoplasmosis is caused by a coccidian parasite, Toxoplasma gondii. The parasite is highly prevalent both in humans and in warm-blooded animals. Cat family animals are definitive host, and these animals excrete the infective oocysts in their feces. Humans, though not definitive host, get infection by consuming water or food contaminated with cat feces. Rarely, infection can also take place through transfusing the infected blood, through transplantation of infected organs, or transplacentally from infected mother to fetus. Transplacental infection can cause congenital infection with varied degree of clinical manifestations, which depend on the age of fetus when infection took place. Diagnosis of congenital toxoplasmosis is difficult to establish until it is suspected and laboratory investigations are carried out. In more than 75% of cases, acute infection is missed due to very mild or unnoticeable clinical symptoms and signs. In India, a prevalence rate of 22.4% (8.8-37.3%) has been reported with an overall IgM positivity of 1.43%. It is estimated that approximately between 56,737 and 176,882 children per year are born in India with a possible risk of congenital toxoplasmosis. The diagnosis of congenital toxoplasmosis can be made by serological methods which are most commonly used. The other methods are parasite isolation by culture and molecular methods. Toxoplasmosis is treatable and transplacental transmission can be prevented by spiramycin, which concentrates in the placenta. However, if infection has done any damage to the fetus or the parasite has passed the placenta, spiramycin cannot reverse the damage. Prevention remains the best remedy.

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Cystic echinococcosis and human cysticercosis have recently been included in the list of "neglected tropical diseases" by the World Health Organization (WHO). Both are zoonoses which are prevalent throughout the world and lead to considerable mortality, morbidity, and economic losses as well. This review deals with the disease burden of these two neglected cestode infections. Diagnostic modalities with their specific advantages and disadvantages have also been discussed. Recent developments in immunodiagnostic assays for the two diseases have been dealt with. Various control strategies including the use of veterinary vaccines have been highlighted.

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Of the so-called nonpathogenic intestinal protozoa, Endolimax nana belongs to the ones least well described. Most data on E. nana have emerged from general surveys of intestinal parasites in selected cohorts and mostly in the absence of any particular focus on Endolimax. Hence, the genus of Endolimax remains largely unexplored in terms of morphology, taxonomy, genetic diversity, host specificity, and epidemiology. In this review, we seek to provide an overview of the work that has been performed on the parasite since the genus Endolimax was described by Kuenen and Swellengrebel in 1917 and suggest activities that may pave the way for a better understanding of E. nana in a clinical and public health context.

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Babesiosis is a tick-borne hemoprotozoan disease of domestic and wild animals. The disease is caused by various species of Babesia and some species of Babesia have also zoonotic significance. The parasite in vertebrate hosts' remains in erythrocytes and the morphology of Babesia spp. is not uniform in all vertebrate hosts. With the advancement of science, particularly the use of molecular techniques made it easy to study the evolution of parasites and thereby reclassifying Babesia spp. as per their phylogeny and to establish the relation of one isolate of Babesia spp. with isolates throughout the world. An attempt also made in this communication to enlighten the readers regarding relationship of one isolate of Babesia spp. of a particular area to another isolate of Babesia spp. of that area or other parts of the world and phylogenetic classification of Babesia spp. was also discussed. It has been concluded that as the study on Babesia is complex in nature so monitoring of the infection with the use of modern techniques is very much needed to control the infection. Second, more research work on phylogenetic relationship of Babesia spp. isolated from different hosts is needed, particularly in India to know the evolution of Babesia spp. of a particular area, as it has great importance to study the trans boundary diseases of animals.

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Background: Malaria is a Protozoal disease caused by infection with parasites of the genus Plasmodium and transmitted to man by certain species of infected female Anopheline mosquito. In 2008 there were 1.52 million cases of malaria in India, out of which 0.76 million case of Plasmodium falciparum, comprising 50% of total malaria cases. There were 924 deaths from malaria. Hematological abnormalities have been observed in patients with malaria, with anemia, and thrombocytopenia being the most common. Materials and Methods: We conducted this study to find out the frequency and the degree of thrombocytopenia in patients with malaria. In our study, 230 patients with malaria positive were investigated with platelet count. Results: In the study group of 230 patients: 130 (56.51%) were positive for Plasmodium vivax, 90 (39.13%) were positive for P. falciparum and 10 (4.34%) had mixed infection with both P. vivax and P. falciparum. Out of 130 cases detected with vivax malaria, 100 cases had thrombocytopenia. Out of 90 cases detected with falciparum malaria, 70 cases had thrombocytopenia. Among 10 cases of mixed infection, 9 cases had thrombocytopenia. Conclusions: Presence of thrombocytopenia in a patient with acute febrile illness in the tropics increases the possibility of malaria. The above finding can have therapeutic implications in context of avoiding unnecessary platelet infusion in malaria patients.

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Parasite cultivation techniques constitute a substantial segment of present-day study of parasites, especially of protozoa. Success in establishing in vitro and in vivo culture of parasites not only allows their physiology, behavior and metabolism to be studied dynamically, but also allows the nature of the antigenic molecules in the excretory and secretory products to be vigorously pursued and analyzed. The complex life-cycles of various parasites having different stages and host species requirements, particularly in the case of parasitic helminths, often make parasite cultivation an uphill assignment. Culturing of parasites depends on the combined expertise of all types of microbiological cultures. Different parasites require different cultivation conditions such as nutrients, temperature and even incubation conditions. Cultivation is an important method for diagnosis of many clinically important parasites, for example, Entamoeba histolytica, Trichomonas vaginalis, Leishmania spp., Strongyloides stercoralis and free-living amoebae. Many commercial systems like InPouch TV for T. vaginalis, microaerophilous stationary phase culture for Babesia bovis and Harada-Mori culture technique for larval-stage nematodes have been developed for the rapid diagnosis of the parasitic infections. Cultivation also has immense utility in the production of vaccines, testing vaccine efficacy, and antigen - production for obtaining serological reagents, detection of drug-resistance, screening of potential therapeutic agents and conducting epidemiological studies. Though in vitro cultivation techniques are used more often compared with in vivo techniques, the in vivo techniques are sometimes used for diagnosing some parasitic infections such as trypanosomiasis and toxoplasmosis. Parasite cultivation continues to be a challenging diagnostic option. This review provides an overview of intricacies of parasitic culture and update on popular methods used for cultivating parasites.

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Introduction: Intestinal parasitic infestation is a major public health problem in children of developing countries Because of poor socio-economic conditions and lack of good hygienic living. The aims of this study were to measure the prevalence of intestinal parasitic infestations and to identify risk factors associated with parasitic infestations among the school children of Itahari Municipality. Materials and Methods: The cross-sectional study was conducted in Grade VI, VII and VIII in Government and private schools of Itahari Municipality. Stratified random sampling method was applied to choose the schools and the study subjects. Semi-structured questionnaire was administered to the study subjects and microscopic examination of stool was done. The Chi-square test was used to measure the association of risk factors and parasitic infestation. Results: Overall intestinal parasitic infestation was found to be 31.5%. Around 13% of the study population was found to be infested with helminthes and 18.5% of the study population was protozoa infected. Not using soap after defecation, not wearing sandals, habit of nail biting and thumb sucking were found to be significantly associated with parasitic infection. Conclusions: The prevalence of intestinal parasitic infestation was found to be high in school children of Itahari. Poor sanitary condition, lack of clean drinking water supply and education is supposed to play an important role in establishing intestinal parasitic infections.

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Entamoeba histolytica , the causative agent of intestinal and extraintestinal amebiasis, is a common parasitic cause of significant morbidity and mortality in the developing countries. Hence, early detection and differentiation of pathogenic E. histolytica from nonpathogenic/commensal Entamoeba spp (Entamoeba dispar/Entamoeba moshkovskii/Entamoeba bangladeshi) plays a crucial role in clinical management of patients with amebiasis. Most diagnostic tests currently available do not reliably differentiate between the species of Entamoeba and are less sensitive, cumbersome to perform. Molecular-based methods are highly sensitive, easy to perform and differentiates the pathogenic Entamoeba from nonpathogenic species, serving the criteria for an ideal diagnostic test for amebiasis. Recently, microarray technology has been found to be a promising tool for the diagnostic and epidemiological evaluation of amebiasis.

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Granulomatous amebic encephalitis is a life-threatening central nervous system (CNS) infection caused by the free-living amoebae Acanthamoeba spp., Balamuthia mandrillaris and Sappinia pedata. The disease has a subacute to chronic onset affecting commonly the immunocompromised population with high mortality rate. The diagnosis of this disease entity requires high suspicion with appropriate sample collection and testing by the laboratory experts. Radiological investigations are nonspecific and commonly confused with CNS tuberculosis, neurocysticercosis, disseminated encephalomyelitis, viral encephalitis etc., delaying the accurate diagnosis of these cases. Early diagnosis plays a crucial role in the survival of these cases since appropriate management can be initiated. No single drug is effective; hence multiple antibiotics targeting various proteins or receptors are required for successful treatment. A combination of surgical and medical interventions involving multiple specialty experts is required to prevent death and morbidity in survivors.

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Babesiosis is a protozoan parasitic infection affecting humans and animals. These infections are commonly transmitted by various species of Ixodes ticks depending upon the geographical location. They can also be transmitted by packed cell transfusion and transplacental route from mother to child. Various species have been reported to cause human infections, of which Babesia microti is the most common species reported globally. Usually, Babesia infections are asymptomatic or mild, but can be severe/life-threatening in immunosuppressed or splenectomized individuals. A high index of clinical suspicion in residents of endemic areas or individuals who had a recent travel history to such areas, with laboratory confirmation can guide an early institution of appropriate antimicrobial therapy, thereby preventing complications and death.

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