Seroprevalence of Toxoplasmosis at a Tertiary Care Centre in North India from 2004 to 2014

Purpose: This study was carried out to determine the seroprevalence of anti-Toxoplasma gondii antibodies in different groups of patients at a tertiary care hospital in North India. Materials and Methods: Clinical and demographic data such as age and gender of patients who had undergone testing for the presence of anti-T. gondii IgG and IgM antibodies between January 2004 and October 2014 were retrospectively analysed. Results: Amongst the 8397 serum samples, an overall seropositivity of 21% (n = 1763) and IgG and IgM seropositivity of 5.7% (n = 481) and 15.3% (n = 1282) were respectively observed. Compared to the period of 2004–2012 (median seroprevalence: 23.6%), a decline in seropositivity to 9.7% in 2013 and 8.1% in 2014 was noted. A rising seroprevalence with age and a higher seroprevalence in females versus males (29.5%, n = 1179 vs. 13.3%, n = 584) were recorded. The highest seroprevalence was observed in suspected ocular toxoplasmosis (47.2%, n = 47), followed by neurological (26.8%, n = 77), human immunodeficiency virus/acquired immunodeficiency syndrome (18.9%, n = 267), post-transplant (17.1%, n = 12) and congenital (7.2%, n = 144) toxoplasmosis. In patients screened for Toxoplasma exposure, the seropositivity was 47.8% (n = 11) in transplant screening and 44.9% (n = 781) in antenatal screening. Conclusion: Toxoplasma infection is highly prevalent in the population of North India across various clinical categories of patients. Future studies focusing on continuous monitoring of seroprevalence trends and elucidation of the risk factors associated with seropositivity in more defined groups of patients are needed.

Diagnostic performance of mini parasep® solvent-free foecal parasite concentrator for the diagnosis of intestinal parasitic infections

Introduction: Various stool concentration techniques have been used to increase the microscopic detection of parasites. We assessed the enclosed, single-vial, Mini Parasep® technique in comparison to the currently used coprodiagnosis procedures. Materials and Methods: A total of 150 stool samples were collected. Samples concentrated by formol–ether method (FEM) and Mini Parasep® SF foecal concentrator and unconcentrated samples were subjected to wet mount, iodine mount microscopy and smear examination by modified acid-fast staining. Results: Direct wet mount detected 72 positive samples (48.6%), whereas 77 (51.3%) and 80 (53.3%) samples were detected by FEM and Mini Parasep® SF methods, respectively. The sensitivity of detection of parasites was 98.7%, 95% and 90.1% with Mini Parasep®, FEM and direct microscopy, respectively. A clearer background with less foecal debris and a better yield of Hymenolepis nana, Trichuris trichiura, Entamoeba coli and Giardia lamblia were noted with Mini Parasep®. Conclusion: Mini Parasep® SF technique is simple, rapid and less cumbersome than conventional diagnostics, making it suitable for routine use. In addition, it offers higher sensitivity and better background clearance than both direct stool examination and FEM.

Clinical and laboratory characteristics of patients with toxocariasis encountered at a tertiary care centre in North India

Toxocariasis is a neglected soil-transmitted helminthic infection caused by either Toxocara canis or Toxocara cati where humans get infected by accidental ingestion of embryonated eggs, and the definitive hosts are dogs and cats. This study aims to analyse the clinical and laboratory characteristics of Toxocara-infected patients and assessment of response to standard treatment with review of literature. The clinical details of patients with Toxocara serology positive for IgG antibodies by ELISA in 5 years (2013–2017) were retrospectively analysed. A total of 29 patients with clinical features and serology suggestive of Toxocara infection were evaluated. A complete history of 14 patients was available for the analysis. Majority (13/14; 96.5%) of cases were children <15 years, males (79%) and belonged to lower socioeconomic status (64.3%). The most common clinical presentation was fever (78.5%) with respiratory symptoms (57%) followed by gastrointestinal features (35.7%). All the patients had eosinophilia (range 8.3%–85%) and raised total IgE levels. Toxocariasis is more common in children, and the true prevalence may be underestimated. In endemic areas, Toxocara workup should be considered in patients with eosinophilia presenting with respiratory and gastrointestinal features.

Free living amoebae in water sources of critical units in a tertiary care hospital in India.

Background: Isolation of free‑living amoebae (FLA) is reported sparsely from water taps, ventilators, air conditioners, haemodialysis units and dental irrigation systems of hospitals worldwide. Their prevalence in hospital environment especially in wards having immunocompromised patients may pose a risk to this group of susceptible population as they may cause disease themselves or may carry pathogens inside them. No study from India has performed such surveillance. Objective: To evaluate extent of FLA contamination in water sources of bone marrow transplant (BMT) intensive care unit (ICU), transplant ICU, haemodialysis unit and high dependency unit in a tertiary care hospital in India. Materials and Methods: A total of hundred samples including fifty each of tap water samples and swabs from mouth of taps used for drinking, bathing and hand washing purposes in these units were collected according to standard procedure. Samples were inoculated onto non‑nutrient agar plates at room temperature followed by morphological confirmation. Molecular identification including polymerase chain reaction (PCR) and sequencing was performed in culture positive samples. Results: Four tap water samples and ten swab samples showed growth of trophozoites and cyst formation. Morphologically, four amoebae resembled Acanthamoeba spp. which was further confirmed by PCR and sequencing showed them to be of T3 and T4 genotypes. Conclusion: The presence of these FLA in hospital water sources emphasises the urgent need of implementing effective preventive measures. Further studies are required to estimate the true prevalence of FLA in Indian hospitals by taking larger number of samples.

Emergence of Balamuthia mandrillaris meningoencephalitis in India.

We report two cases of fatal chronic meningoencephalitis caused by Balamuthia mandrillaris in immunocompetent men. Diagnosis of amoebic meningoencephalitis was made ante‑mortem in one case and postmortem in another by histopathological examination and confirmed by demonstration of B. mandrillaris deoxyribonucleic acid (DNA) by polymerase chain reaction (PCR).

Development of touch down‑multiplex PCR for the diagnosis of toxoplasmosis.

Purpose: The diagnosis of toxoplasmosis is challenging since conventional methods like culture and immunofluorescence are not universally available. Serology, which is used regularly might be negative during early phase of infection and in immunosuppressed patients or may remain positive for a long time. Several molecular tests have been used for the diagnosis of toxoplasmosis, but none of them have an internal control which would inform us regarding the presence of polymerase chain reaction (PCR) inhibitors thus, undermining the confidence of a laboratory physician. Materials and Methods: We designed a multiplex PCR containing primers targeting human beta globin gene which would act as internal control and two primers against the B1 gene and 5s gene which aid in sensitive detection of T. gondii. Results: Multiplex PCR had a sensitivity of 83.3% and specificity of 100%. Conclusion: Multiplex PCR may provide a sensitive and specific tool for diagnosis of human toxoplasmosis.

The presence of dsRNA virus in Trichomonas vaginalis isolates from symptomatic and asymptomatic Indian women and its correlation with in vitro metronidazole sensitivity.

Purpose: Trichomonas vaginalis, a protozoan parasite, is the causative agent of human trichomoniasis, the most common non-viral sexually transmitted disease. The infection encompasses from a complete asymptomatic presentation to severe sequelae; yet, the virulence markers have been poorly understood. It is suggested that the presence of Trichomonas vaginalis virus (TVV) in T. vaginalis may have an impact on its virulence, and its relatedness to in vitro metronidazole resistance has been reported. The aim of the study was to assess the presence of TVV in fresh and Long -Term Cultivated ( LTC) maintained T. vaginalis isolates from symptomatic (S) and asymptomatic (AS) Indian women and its relatedness, if any, with symptomatology and in vitro drug sensitivity. Materials and Methods: One thousand women (537 S and 463 AS) were screened for the presence of T. vaginalis by wet smear and culture examination of vaginal swab and urine sample. Fresh and LTC (6 months-2 years) maintained 15 isolates each from 15 S and 15 AS women were subjected to agarose gel electrophoresis following total cellular RNA extraction to evaluate the presence of double stranded (ds) RNA viral infection. The susceptibility of isolates to metronidazole was determined in vitro. Results: On agarose gel electrophoresis, three bands (5.5, 2.5 and 1.5 kb) were observed in all the 30 fresh isolates from 15 S and 15 AS women and only in 7 LTC isolates from 3 S and 4 AS women. All the fresh isolates harbouring TVV were found to be sensitive to metronidazole in vitro irrespective of the symptomatology of subjects, and out of seven LTC isolates harbouring TVV, six were sensitive to metronidazole and one showed borderline resistance. Conclusions: The results suggest that the presence of TVV alone may not be a virulence marker and loss of TVV on LTC appears to be related to drug resistance. The T. vaginalis Indian isolates are sensitive to metronidazole.

Evaluation of cytotoxic potential of latex of Calotropis procera and Podophyllotoxin in Allum cepa root model

In the present study we have utilized the Allium cepa root tip meristem model to evaluate the cytotoxic and anti-mitotic activities of latex of Calotropis procera (DL) and podophyllotoxin. Standard cyto-toxic drug cyclophosphamide and non-cytotoxic drugs cyproheptadine and aspirin served as controls. Like cyclophosphamide, both DL and podophyllotoxin significantly inhibited the growth of roots and mitotic activity in a dose-dependent manner. However, podophyllotoxin was more potent in this regard and produced root decay. Cyproheptadine and aspirin, on the other hand, showed a marginal effect on the root growth and mitotic activity at much higher concentrations

Pediatric HIV infection.

OBJECTIVE: The present cross sectional study was undertaken to study clinical profile of HIV infection in children in Northern India. METHODS: 64 children from newborn to eighteen years, presenting for confirmation of diagnosis of HIV infection or monitoring of CD4-CD8 counts in confirmed cases, were evaluated. Children were categorized as per CDC classification of Pediatric HIV. The diagnosis was confirmed by serological tests or PCR assay. CD4-CD8 counts were done by FACS Count. RESULTS: Majority of the children were between 18 months to 5 years. Adolescents comprised 24% of the case. 51.5% children were infected through the mode of mother to child transmission. 39% of the case was transfusion-mediated. Unsafe medical injections probably contributed to 6.2% and heterosexual promiscuity led to 3.1% cases. Clubbing, not described in Indian studies so far, was seen in 9.3% cases. CONCLUSIONS: HIV infection is a chronic childhood disease extending into adolescence, and contaminated blood and unsafe medical injections are still important routes of HIV transmission in India.

Value added eco-friendly products from tannery solid wastes.

Chrome shavings are the prominent solid wastes in tanning industry. Since chromium is known for its toxicity, the disposal of chrome shavings has been identified as a serious problem from the environmental point of view. At present, the popular utilization mode for chrome shavings is the manufacture of leather boards and related products. But this does not offer complete utilization of chrome shavings. Moreover, return per ton of chrome shavings is low if used for leather board production. In view of this, two processes have been developed to offer an alternative and better solution for the disposal of chrome shavings.The first process is preparation of parchment like membrane and the second process is related to development of leather like material. These products are analyzed for their mechanical behavior and other physicochemical properties. Parchment membrane can be used in the preparation of lampshades, chandeliers, wall hangers, table tops etc. and leather like material can be used in the preparation of chappal uppers, hand bags, purses, valets etc. The utilization of the chrome shavings in preparation of those two products not only reduces the environmental pollution but at the same time value added products can also be obtained.

Prevalence of parenterally transmitted hepatitis viruses in clinically diagnosed cases of hepatitis.

Hepatitis B virus (HBV) is the most important causative agent of blood borne hepatitis in humans. Hepatitis D Virus (HDV) infection occurs either as a coinfection or superinfection in HBV carriers. Hepatitis C virus (HCV) is the major cause of transfusion non-A, non-B hepatitis and continues to be a major cause of human liver disease throughout the world. The present study was conducted on 70 clinically diagnosed cases of viral hepatitis to study the prevalence of parenterally transmitted viral hepatitis. The serum samples were tested for HBsAg, HBeAg, IgM anti-HBc, anti-HBe, anti-HCV and anti-HDV using separate ELISA kits. Of the 70 serum samples tested, 28 (40%) were positive for HBsAg out of which 3 (4.28%) were positive for HBeAg also. Five (7.1%) of the HBsAg positive cases tested positive for IgM anti-HBc also. HBsAg alone was found in 17 (24.28%) cases. The prevalence of anti-HCV was 3 (4.28%) in 70 cases. Thus early screening of clinically diagnosed cases of viral hepatitis is essential for establishing diagnosis and treatment to prevent long term sequelae.

SEN virus infection--a new hepatitis virus: a review.

Approximately 10% of transfusion associated hepatitis and 20% of community acquired hepatitis cases do not have a defined etiology, suggesting the existence of an additional causative agent. The recent agents which have been detected in blood were Hepatitis G virus (HGV) and TT virus (TTV), members of the family Circoviridae. These viruses were initially thought to be associated with post-transfusion hepatitis but later studies negated this causal effect. Recently, a novel DNA virus at the moment designated as SEN virus (SEN-V) was discovered and is thought to be associated with post-transfusion hepatitis. This virus is related to the above two agents and belongs to the same family. Whether, like HGV and TTV causal relationship with hepatitis is finally confirmed only time will tell, but, at the moment the association appears to present. The present review summarizes the present available data on this new agent of hepatitis.

Screening by VDRL test to detect hidden cases of syphilis.

A total of 59,450 sera from January 1996 to December 2000 were subjected to VDRL testing. Overall VDRL positivity rate was 3.2% and downward trend was observed in the recent years, 1999 and 2000. Majority of the samples were from Gynaecology department, out of which 1.57% were VDRL positive. Out of 30,045 samples from antenatal females, 517(1.47%) were positive, while 304(1.8%) were positive out of 16,980 samples obtained from couples. Out of 304 samples from couples found positive, 17.4% wives had titre >R16; 27.9% wives had titre R1 to R8, out of which 15.3% husbands had titre of >R16. Also, 166 wives with nonreactive VDRL had 19.3% husbands with titre > R16. Thus, couple VDRL test plays an important role in detection of hidden cases of syphilis in the community and early detection and treatment of such cases will further reduce the perinatal morbidity and mortality.

Naegleria meningitis: a rare survival.

Acute amebic meningoencephalitis caused by free-living amebae naegleria fowleri is extremely rare and uniformly fatal with only seven survivals reported till date. An interesting case of naegleria meningitis diagnosed by wet mount cytology of cerebrospinal fluid (CSF) and treated with amphoterecin B, rifampicin and ornidazole with complete recovery is presented. In cases of suspected pyogenic meningitis, if CSF staining, antigen detection or culture is negative for bacteria, a wet mount cytology of CSF for naegleria is suggested. Early treatment with amphoterecin B and rifampicin may improve survival.

Acanthamoeba adherence to soft contact lens and human corneal stroma.

In the present study, the adhesibility of Acanthamoeba castellani trophozoites and cysts to hydrogel contact lenses and to human cornea was investigated. Segments of unworn contact lenses were prepared (120 segments in total). In addition, 8 corneal buttons were obtained from 8 enucleated eyes. And each cornea cut into 8 radial segments. To these lens and corneal segments, 1.2 x 106/ml and 1.3 x 106/ml of cysts and trophozoites respectively were added under varying conditions. The adhesion was checked at 0, 3 and 24 hours after the exposure. Adhesion analysis showed that the trophozoites adhered equally well to lens or cornea. There is an increase in the number of trophozoites adhering to contact lenses as well as cornea. This difference is more significant for contact lenses. Washing of contact lenses significantly decreased the adherence of the trophozoites after 3 hours of incubation. When the comparison of adhesion was done between the unwashed worn and unwashed unworn contact lenses, it was observed that there was a significant difference in adherence to new lenses where the adherence was much lower. The study shows that washing of contact lenses does decrease the chances of colonization by Acantamoeba catellani and also that older lenses have higher chances of getting colonized probably due to the occurrence of scratches et. on the surface which may help in colonization.

Hepatitis G virus (HGV): current perspectives.

Five viruses are usually associated with hepatitis in humans: A-E. In addition to these viruses as aetiological agents of hepatitis, there remain a number of patients with hepatitis in whom no virus could be identified. It was therefore postulated that there may be other agents which may be causing hepatitis. Recently, two viruses have been associated with hepatitis: hepatitis G virus (HGV), and transfusion transmissible virus (TTV). Hepatitis G virus (HGV) is a single stranded RNA virus which represents a newly discovered virus belonging to the flavivirus family. HGV is distinct from hepatitis C virus (HCV) and the newly discovered GBV-A and GBV-B agents, while GBV-C represents an isolate of HGV. The structure of the HGV genome resembles that of HCV. HGV replicates in peripheral blood cells, while replication in liver cells has not been observed till date. Diagnosis of HGV infection is mainly by use of polymerase chain reaction (PCR), as serological techniques are still being developed. Epidemiological data indicate that the virus is prevalent throughout the world, including India and is transmitted via blood/blood products, sexually and vertically from infected mothers to children. The relationship between infection with the virus and presence of liver pathology is controversial and has not been proven beyond doubt, as majority of patients with HGV have no detectable evidence of disease.