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Naegleria fowleri the causative agent of Primary Amoebic Meningoencephalitis, is ubiquitously distributed worldwide in various warm aquatic environments and soil habitats. The present study reports on the presence of Naegleria spp. in various water bodies present in Rohtak and Jhajjar district, of state Haryana, India. A total of 107 water reservoirs were screened from summer till autumn (2012 and 2013). In order to isolate Naegleria spp. from the collected water samples, the water samples were filtered and the trapped debris after processing were transferred to non-nutrient agar plates already seeded with lawn culture of Escherichia coli. Out of total 107 water samples, 43 (40%) samples were positive by culture for free living amoeba after incubation for 14 days at 37°C. To identify the isolates, the ITS1, 5.8SrDNA and ITS2 regions were targeted for PCR assay. Out of total 43 positive samples, 37 isolates were positive for Naegleria spp. using genus specific primers and the most frequently isolated species was Naegleria australiensis. Out of 37 Naegleria spp. positive isolates, 1 isolate was positive for Naegleria fowleri. The sequence analysis revealed that the Naegleria fowleri strain belonged to Type 2.

Background & objectives: Diarrhoea is the main clinical manifestation caused by intestinal parasitic infections in patients, with special reference to transplant recipients who require careful consideration to reduce morbidity and mortality. Further, molecular characterization of some important parasites is necessary to delineate the different modes of transmission to consider appropriate management strategies. We undertook this study to investigate the intestinal parasitic infections in transplant recipients with or without diarrhoea, and the genotypes of the isolated parasites were also determined. Methods: Stool samples from 38 transplant recipients comprising 29 post-renal, two liver and seven bone marrow transplant (BMT) recipients presenting with diarrhoea and 50 transplant recipients (42 post-renal transplant, eight BMT) without diarrhoea were examined for the presence of intestinal parasites by light microscopy using wet mount, modified Ziehl-Neelsen staining for intestinal coccidia and modified trichrome staining for microsporidia. Genotypes of Cryptosporidium species were determined by multilocus genotyping using small subunit ribosomal (SSUrRNA), Cryptosporidium oocyst wall protein (COWP) and dihydrofolate reductase (DHFR) as the target genes. Assemblage study for Giardia lamblia was performed using triose phosphate isomerase (TPI) as the target gene. Samples were also screened for bacterial, fungal and viral pathogens. Results: The parasites that were detected included Cryptosporidium species (21%, 8/38), Cystoisospora (Isospora) belli (8%, 3), Cyclospora cayetanensis (5%, 2), G. lamblia (11%, 4), Hymenolepis nana (11%, 4), Strongyloides stercoralis (3%, 1) and Blastocystis hominis (3%, 1). Multilocus genotyping of Cryptosporidium species at SSUrRNA, COWP and DHFR loci could detect four isolates of C. hominis; two of C. parvum, one of mixed genotype and one could not be genotyped. All the C. hominis isolates were detected in adult post-renal transplant (PRT) recipients, whereas the C. parvum isolates included a child with BMT and an adult with PRT. Clostridium difficle, cytomegalovirus and Candida albicans were found in 2, 3 and 2 patients, respectively. Interpretation & conclusions: In the present study, C. hominis was observed as an important parasite causing intestinal infections in transplant recipients. Multilocus genotyping of Cryptosporidium species could detect four isolates of C. hominis; two of C. parvum, one of mixed genotype and one could not be genotyped. Genotyping of G. lamblia revealed that assemblage B was most common.

BackgroundScrub typhus, once known as tsutsugamushi fever and attributed to Rickettsia, has transformed into a growing public health concern. Despite its increasing incidence in India, comprehensive spatiotemporal analyses of scrub typhus have been lacking.MethodsThis study examines the prevalence of scrub typhus cases reported from 2003 to 2023, using available literature to provide a breakdown of cases by year and state in India, aiming to elucidate the disease’s spatiotemporal dynamics. The aetiological association of Orientia tsutsugamushi and scrub typhus is based on geographical, immunological and molecular genetic studies.FindingsThis analysis identified 47 650 cumulative cases of scrub typhus in India over the past two decades. The case fatality rate was 5% out of 35 243 cases. Variations in transmission dynamics and the Leptotrombidium vector’s competence may influence the disease’s distribution. Nonetheless, there has been a notable increase in infections since 2010, peaking in 2019 and 2022. Curtailing and containing such an upsurge can be daunting and requires an interdisciplinary public health approach. Further, there is heterogeneity in studies on general, gastrointestinal, pulmonary and inflammatory symptoms compared with studies on cardiac, hepatic, neurological and other symptoms. The Weil-Felix test was the most common diagnostic technique used, and doxycycline was the treatment for scrub typhus cases.InterpretationsThis meta-analysis can help policy-makers and researchers in India develop scrub typhus management and control policies.PROSPERO registration numberCRD42024611771.

Data on the genetic diversity of Pneumocystis jirovecii causing Pneumocystis pneumonia (PCP) among children are still limited, and there are no available data from the Indian subcontinent, particularly associations between genotypes and clinical characteristics. A total of 37 children (62 days-12 years [median 5.5 years]) were included in this study. Pneumocystis was diagnosed by microscopy using Grocott-Gomori methenamine silver stain in 12 cases and by nested PCR using mtLSUrRNA in 25 cases. Genotyping was performed using three different genes, mitochondrial large subunit ribosomal RNA ( mtLSUrRNA ), dihydropteroate synthase ( DHPS ) and dihydrofolate reductase ( DHFR ). mtLSUrRNA genotype 3 and novel mutations at the gene target DHFR (401 T > C) and DHPS 96/98 were frequently observed and clinically associated with severe PCP and treatment failure. Phylogenetic analyses revealed 13 unique sequence types (STs). Two STs (i) 3-DHFR 401 T  >  C-DHPS 96/98 – PJ1 and (ii) 3-DHFR 401 T  >  C-DHPS 96 - PJ3 were significantly associated with treatment failure and high mortality among PCP-positive patients. In conclusion, the present study strongly suggests the emergence of virulent P . jirovecii strains or genetic polymorphisms, leading to treatment failure and high mortality. Our study is the first of its kind from the Indian subcontinent and has highlighted the genetic diversity of Pneumocystis jirovecii among children and their clinical outcomes. These findings emphasize the need to focus more on genotypes to better understand the epidemiology of Pneumocystis pneumonia.

A 24-year-old man with a kidney transplant presented with a 3-day history of diarrhea. An examination of a saline wet mount of a stool sample showed an organism moving in a “falling leaf” pattern (shown in a video).

Human cryptosporidiosis is endemic worldwide, and at least eight species have been reported in humans; the most common being Cryptosporidium hominis and C. parvum. Detailed understanding of the epidemiology of Cryptosporidium is increasingly facilitated using standardized universal technique for species differentiation and subtyping. In this study micro- and minisatellite targets in chromosome 6 were used to assess genetic diversity of C. hominis by sequence length polymorphisms along with single nucleotide polymorphisms (SNPs). A total of 84 Cryptosporidium positive stool specimens were subjected to speciation and genotyping using small subunit (SSU) ribosomal RNA (rRNA) as the target gene. Genetic heterogeneity amongst C. hominis isolates was assessed by sequencing minisatellites, microsatellites and polymorphic markers including genes encoding the 60 kDa glycoprotein (GP60), a 47 kDa protein (CP47), a mucin-like protein (Mucin-1), a serine repeat antigen (MSC6-7) and a 56 kDa transmembrane protein (CP56). Of the 84 Cryptosporidium positive stool specimens, 77 (92%) were positive by SSU rRNA gene polymerase chain reaction (PCR) assay. Of these 77 isolates, 54 were identified as C. hominis and 23 as C. parvum. Of all the loci studied by multilocus sequence typing (MLST), GP60 gene could reveal the highest genetic diversity. Population substructure analysis of C. hominis performed by combined sequence length and nucleotide polymorphism showed nine multilocus subtypes, all of which were distinct groups in the study population. MLST, a powerful discriminatory test, demonstrated both variations and distribution pattern of Cryptosporidium species and its subtypes.

Hookworm infestations, endemic in India, are a common cause of iron deficiency anemia. Hemosuccus pancreaticus, a rare clinical condition, is due to passage of blood into the pancreatic duct possibly through a route between an aneurysm of an artery close to the pancreas and/or pancreatic duct, leading to gastrointestinal (GI) bleeding. Portal hypertensive upper GI bleed is also known since long. We report a case of a 38-year-old male with a history of alcoholism who was being investigated for GI bleeding who had concomitant hookworm infestation, hemosuccus pancreaticus as well as portal hypertension. To the best of our knowledge, this is the first report of common occurrence of hemosuccus pancreaticus and portal hypertension with hookworm infection. This case signifies the importance of infectious causes of GI bleeding to be considered even in cases where anatomic malformations or pathophysiological alterations are predominant.

A 42-year-old man presented with complaints of foreign body sensation and redness in his left eye after being reportedly struck by an insect during a wind storm. The larva was retrieved from the eye and its microscopic examination revealed that it had an ovalshaped segmented body with prominent oral hooks connected to the internal cephalopharyngeal skeleton (Figs 1c, 1 d).

Abstract Detection of antibodies to Toxoplasma in sera from patients suffering from recurrent unprovoked seizures were performed using “in-house” indirect hemagglutination assay and by commercially available anti-Toxoplasma immunoglobulin G and immunoglobulin M enzyme-linked immunosorbent assays. Serum antibody to toxoplasmosis were detected in 12.3% and 15.3% by indirect hemagglutination assay and enzyme-linked immunosorbent assays and respectively. Controls showed seropositivity of 5.7% for antibody to Toxoplasma using the same methods. Seropositivity was higher in children compared to adults. No gender differences were noted. Individuals with rural background (living in a relatively unhygienic conditions) were more commonly affected compared to people living in urban areas. (J Pediatr Neurol 2003; 1(2): 95–98).