Use of animals in research is a highly debatable topic. Though their use has led to several discoveries and understanding of many aspects of science but their use in certain sectors needs to be justified. There are national and international laws which govern the use of animals in research, all of which are based on the principles of the 4Rs - replacement, reduction and refinement and the rehabilitation of the use of animals in research. It is mandatory that all institutions involved in animal research develop and abide by the ethical review processes which promote good animal welfare practices by ensuring that the use of animals at the designated establishment is justified. With the availability of many alternatives, the lives of many animals can now be secure.

Diarrhea is a major cause of morbidity and mortality in human immunodeficiency virus (HIV)-infected individuals. Opportunistic enteric parasitic infections are encountered in 30-60% of HIV seropositive patients in developed countries and in 90% of patients in developing countries. Once the CD4 ⁺ cell count drops below 200 cells/μl, patients are considered to have developed acquired immunodeficiency syndrome (AIDS), with the risk of an AIDS-defining illness or opportunistic infection significantly increasing. Opportunistic enteric parasites encountered in these patients are Cryptosporidium, Isospora, Cyclospora, and microsporidia; as well as those more commonly associated with gastrointestinal disease, for example, Giardia intestinalis, Entamoeba histolytica, Strongyloides stercoralis, and also rarely Balantidium coli. In view of AIDS explosion in India, opportunistic enteric parasites are becoming increasingly important and it has to be identified properly. Apart from wet mounts, concentration methods for stool samples and special staining techniques for identification of these parasites, commercially available fecal immunoassays are widely available for the majority of enteric protozoa. Molecular methods such as polymerase chain reaction (PCR), PCR-restriction fragment length polymorphism, flow cytometry, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), have also come in the pipeline for early diagnosis of these infections. Proper disposal of the feces to prevent contamination of the soil and water, boiling/filtering drinking water along with improved personal hygiene might go a long way in preventing these enteric parasitic infections.

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.

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.

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.

Background: Toxoplasmosis is a parasitic disease which may cause some laboratory symptoms in infected individuals. One of the main ways to transmit this organism is placenta to the fetus pathway. If this transmission occurs in the 3 ^rd month of pregnancy, the abortion, central nerve system and ocular disorder will happen. Because of this issue, the precise technique for the detection of Toxoplasma antibodies such as immunoglobulin G (IgG) and immunoglobulin M IgM are important, as they contain ELISA and ELISA avidity. Materials and Methods : i0 n this survey, the main samples are serum and amniotic fluid that were collected from 48 pregnant women infected with Toxoplasma gondii in Shariaty hospital. This survey is attempted to design ELISA avidity kit in Tarbiat Modates University. Results: The results from this survey show that, in these total pregnant women the infection by T. gondii has occurred and many of them are infected currently. Conclusions : i0 n the simple ELISA technique, the only antibody that can be detected precisely is IgM; however, using this technique the IgG antibody can also be detected. In this new technique or ELISA avidity, in addition to detection of IgG antibody against T. gondii, the month of transmission of Toxoplasma is also interpreted.

Background: Many immunological techniques have been developed over years using the different Fasciola antigens for diagnosis of parasitic infection and to replace the parasitological techniques, which are time consuming and usually lack sensitivity and reproducibility. Materials and Methods: In this study, Fasciola gigantica paramyosin (Pmy) antigen was early detected in cattle sera using sandwich enzyme-linked immunosorbent assay (ELISA), to evaluate the Pmy antigen performance in diagnosis. This work was conducted on 135 cattle blood samples, which were classified according to parasitological investigation into, healthy control (30), fascioliasis (75), and other parasites (30) groups. Results: The sensitivity of Sandwich ELISA was 97.33%, and the specificity was 95%, in comparison with parasitological examination, which recorded 66.66% sensitivity and 100% specificity, respectively. Conclusions: It was clear that the native F. gigantica Pmy is considered as a powerful antigen in early immunodiagnosis of fascioliasis, using a highly sensitive and specific sandwich ELISA technique.

Background: Vivax malaria is the most widely distributed human malaria and is responsible for up to 400 million infections every year. Recently, it has become evident that Plasmodium vivax monoinfection could also result in multiple organ dysfunction and severe life-threatening disease as seen in Plasmodium falciparum infection. Materials and Methods: The aim of this study was to note the different clinical and biochemical profiles of adult patients with the severe vivax malaria with regards to complications and outcome. This was a prospective observational study carried out at a tertiary care hospital in Kolkata over 9 month's period. Detailed history and examination findings were noted in all patients. Their clinical presentations, complications, course in ward until discharge or death was noted. Results: A total of 900 cases of vivax malaria were included in the study. Severe disease was present in 200 (22.2%) cases of malaria. There were 108 (54%) patients with single complication (SC) and 92 (46%) patients with the multiple complications (MC). Patients with SC had jaundice (48.1%) followed by cerebral involvement (25.9%), renal failure (7.4%), and pulmonary involvement (3.7%). The MC was found in various combinations and the majority (47.8%) had constellation of two different complications. The mortality rate of patients with the SC and MC was 7.4% and 34.8%. The overall mortality observed in severe vivax malaria was 20% (40/200). Conclusions: In recent years, the clinical pattern of vivax malaria has changed. Severe vivax malaria is now very common with increasing mortality. Not only the number, but also the type of complication influences the outcome of complicated malaria.

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.

Background: The occurrence of fetal death is one of the tragedies that confront the physician providing obstetric care. Among the various agents associated with infections of pregnancy, viruses are the most important followed by bacteria and protozoa. Among protozoal infections in pregnancy, toxoplasmosis is reported to have a high incidence, sometimes causing fetal death. The study was intended to observe the seroprevalence of Toxoplasmosis in pregnant women presenting with bad obstetric history (BOH). Materials and Methods: A total of 92 antenatal women were included in the study (80 in the study group and 12 in control group). The study group comprised of antenatal women with BOH in the age group of 20-35 years. Antenatal women with Rh incompatibility, pregnancy induced hypertension, diabetes mellitus, renal disorders and syphilis were not included in the study. The control group included women in reproductive age group without BOH. All the samples were screened by enzyme linked immuno sorbent assay (ELISA) for Toxoplasma specific Immunoglobulin M (IgM) and Immunoglobulin G (IgG) antibodies. Results: Of the 80 antenatal women in the study group, 36 (45%) were seropositive for Toxoplasma specific IgG antibodies (P < 0.005), 16 (20%) were seropositive for Toxoplasma specific IgM antibodies (P < 0.005) and 8 (10%) were seropositive for both IgG and IgM antibodies (P < 0.005). Various predisposing factors for acquiring Toxoplasmosis such as contact with cats, contact with soil, food habits, illiteracy, socio-economic status and residential status were also studied. Conclusions: We conclude that toxoplasmosis during pregnancy causes congenital fetal infection with possible fetal loss. ELISA was found to be a sensitive serological test for diagnosis of Toxoplasmosis in pregnant women with BOH. Major cause of fetal loss in BOH cases in the study group was abortion.

Background: Annual mass drug administration (MDA) with diethyl carbamazine (DEC) and Albendazole is the most cost-effective strategy to control lymphatic filariasis (LF). Materials and Methods: The aim of the present study was to assess the coverage and the compliance of MDA, to elicit factors that influenced compliance, to document side-effects reported and to assess the awareness of the community regarding the disease and MDA program in Bankura district, West Bengal after 2012 round of MDA. Multistage cluster sampling method was adopted. Total four clusters; three villages and one urban municipality ward were selected. In each cluster, minimum 30 families were randomly selected and the head or any responsible family member was interviewed using a pre-designed, pre-tested schedule after taking written informed consent. Data were compiled and analyzed using SPSS 19.0. Results: Total eligible population was 683 among which 98.8% received both the drugs. About 5% of the recipients took none of the drugs. More than two-thirds of the families took unsupervised dose. Drug compliance rate was significantly lower in urban (90.7%) than in the rural clusters (95.7%) (z = 2.46, P < 0.05). Effective coverage rate was significantly lower in urban than in the rural clusters (87.4% vs. 95.3%; z = 3.57, P < 0.01). Coverage compliance gap was higher in urban (5.7%) than in rural cluster (3.9%). Fear of side-effects was the main reason for non-compliance. Reported side-effects were few, mild, and transient. Around 60% of the surveyed families were aware about the MDA program whereas, 67% of them heard about LF. Only 41% families were provided information, education, and communication in last 15 days before MDA. Major sources of information for the surveyed families were leaflets (20.3%) and poster (9.8%). Conclusions: Widespread rural urban variation in performance status, poor social mobilization activities, lack of supervised dosing, and lack of knowledge of the community about the disease and the program are the major areas of concern.

Background: Boophilus microplus is an important ectoparasite of livestock. Apart from transmitting diseases, heavy tick burden can decrease production and damage hides. The synthetic pyrethroids which are advantageous over other acaricides for treatment of this infestation are now losing their efficacy due to development of resistant strains of ticks. Materials and Methods: Boophilus microplus ticks with a previous history of acaricidal treatment especially synthetic pyrethroids (SP) such as cypermethrin, deltamethrin and flumethrin were randomly collected from different pockets of four Southern States of India namely Tamil Nadu, Kerala, Karnataka, Andhra Pradesh, and Puducherry from cattle. Deoxyribonucleic acid extracted from pooled adult B. microplus tick from each State was subjected to polymerase chain reaction-restriction fragment length polymorphism to detect point mutation in carboxyl esterase gene. Results: A product size of 372 bp was obtained for cattle tick samples collected from all over Southern States of India. Conclusions: B. microplus ticks found in Southern part of India are not resistant to commonly used SP.

Human subcutaneous dirofilariasis is caused by filarial worms of the genus Dirofilaria. The parasites are transmitted to man by mosquitoes. We report three cases of human subcutaneous dirofilarias caused by Dirofilaria repens from Dibrugarh, Assam, north east India. The cases presented as subcutaneous nodules, on the chest, cheek and the anterior abdominal wall. Noting the frequency of the cases reported within 6 months, it is emphasized that subcutaneous dirofilariasis is a potentially emerging zoonosis in Assam and should be included in the differential diagnosis of patients presenting with subcutaneous nodules in Assam.

We report an unusual case of multiple pyogenic liver abscesses containing Ascariasis lumbricoides in a 35-year-old post-partum female who had delivered 1 month back. Open drainage of liver abscess along with liver worm was done. Patient did well post-operatively.

Intraocular infestation by live Gnathostoma spinigerum is a rare occurrence in humans. Most of the published reports are from South-East Asia. We report a case of ocular gnathostomiasis from Western Maharashtra, where the worm was removed live from the anterior chamber of the left eye. Identification of the worm was confirmed by light microscopy.