Molecular prevalence of Dirofilaria sp. hongkongensis, among the dog population in Thiruvananthapuram, India.

Human dirofilariasis is an emerging vector-borne zoonotic parasitic disease in India. Humans are accidental hosts. Symptomatic dirofilariasis, although uncommon is typically manifested in humans as pulmonary, ocular, or subcutaneous nodules. The present research reports the prevalence of the species, Dirofilaria sp. hongkongensis among the dog population in the Kani tribal settlements within the Agasthyamala Biosphere Reserve located at the southernmost end of the Western Ghats, Kerala. To study the prevalence, we conducted a random blood survey among the dog population in these tribal settlements in Thiruvananthapuram. Whole blood was collected from stray and domesticated animals. Genomic DNA was extracted and the microfilariae in the blood were characterized using the slide smear and COI-based marker specific to nematodes. After processing the data from the Sanger sequencer using BLAST, the sequences were submitted to GenBank. Over 25 % of dogs were found positive for Dirofilaria sp. hongkongensis, which is genetically identical to the strain causing human dirofilariasis as is shown by the phylogenetic analysis. The study reveals that the Kani tribes, who reside in deep forests, are particularly vulnerable to this strain of Dirofilaria sp. hongkongensis. Human case reports of this specific strain have been recorded in Kerala over the past decade. The effective elimination is ultimately impacted by a lack of knowledge regarding the mosquito vectors. Dirofilariasis in dogs and humans is on the rise, which demands both active disease surveillance and proper treatment.

Report of a new species of sand fly, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), from Western Ghats, India.

BACKGROUND: Western Ghats is a biodiversity treasure trove with reports of indigenous leishmaniasis cases. Hence, systematic sand fly surveillance was carried out among the tribal population. The present study reports a novel sand fly species, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), discovered in the Western Ghats of India. METHODS: A comprehensive sand fly survey was conducted across the Kollam, Thrissur, Idukki, Kasaragod and Malappuram districts of Kerala, India. The survey spanned both indoor and outdoor habitats using standard collection methods over a 3-year, 3-month period. DNA barcoding of samples was performed targeting mitochondrial cytochrome c oxidase subunit I (COI) gene, and the sequence generated was subjected to phylogenetic analysis. RESULTS: Phlebotomus (Anaphlebotomus) ajithii, a new sand fly species, is recorded and described in this communication. The morphological relationship of the new species to other members of the subgenus Anaphlebotomus is discussed. Mitochondrial COI barcode followed by phylogenetic analysis confirmed that specimens of Ph. ajithii belong to the same taxonomic group, while a genetic distance of 11.7% from congeners established it as a distinct species. CONCLUSIONS: The Western Ghats, known for its rich biodiversity, has lacked systematic entomological surveys focusing on sand flies. This study aims to fill this gap and reports and describes a new species of sand fly.

Molecular Identification of Host Blood Meal from Phlebotomine Sandflies (Diptera: Psychodidae) from Kerala, India.

Female phlebotomine sandflies serve as vectors for the transmission of Leishmania parasites, perpetuating an enzootic cycle by disseminating between sylvatic and domestic animals. Humans form a part of this cycle because the sandflies search for a blood source required for egg development. The present study aimed to identify the feeding preferences of different sandfly fauna from six districts of Kerala, India, using molecular tools. An entomological survey was conducted during 2021-2023 in Kollam, Kottayam, Thiruvananthapuram, Thrissur, Malappuram, and Palakkad. Both indoor and outdoor habitats were targeted from sandfly collection using different standard tools and methods. Sandflies were identified using standard taxonomic keys, and DNA was extracted from blood meal collected from sandflies. A total of 7366 sandfly specimens were collected during the study period, which belonged to three different genera and 19 species. Blood source was successfully identified from 119 sandflies revealing that the Sergentomyia genus preferably fed on small reptiles and amphibians, whereas Phlebotomus genus was found to mainly feed on mammalian and avian blood. Sergentomyia zeylanica was an exception, as it primarily fed on mammalian blood sources. Interestingly, humans were the second feeding source for Phlebotomus species, which are the proven vectors. Comprehending the feeding patterns of sandflies is crucial, not just for public health but also for obtaining insights into the ecological dynamics between vectors and hosts, ultimately enabling more efficient strategies for disease control and prevention.

Emergence of imported cutaneous leishmaniasis caused by Leishmania major: a case series from Kerala, India.

Cutaneous leishmaniasis (CL) is often considered a 'great imitator' and is the most common form of leishmaniasis. The Leishmania species responsible for CL varies among countries, as these species exhibit specific distribution patterns. The increased mobility of people across countries has resulted in the imported incidences of leishmaniasis caused by non-endemic species of Leishmania. During 2023, we confirmed three CL cases caused by L. major from Kerala, India, and upon detailed investigation, these were identified to be imported from the Middle East and Kazakhstan regions. This is the first report of CL caused by L. major from Kerala. The lesion morphology, detection of anti-rK 39 antibody and Leishmania parasite DNA from the blood samples were the unique observations of these cases. Kerala, being an emerging endemic zone of visceral leishmaniasis (VL) and CL, the imported incidences of leishmaniasis by non-endemic species can pose a significant threat, potentially initiating new transmission cycles of leishmaniasis caused by non-endemic species.

Comparative Analysis of Phlebotomus argentipes Vector of Leishmaniasis in India and Sri Lanka.

Phlebotomus argentipes is the predominant sandfly vector of leishmaniasis in the Indian subcontinent. India and Sri Lanka primarily report visceral and cutaneous leishmaniasis caused by Leishmania donovani. We compared Ph. argentipes from two locations, focusing on its morphological, molecular, and salivary protein characteristics. Sandflies were captured using CDC light traps and cattle-baited net traps. Species identification and morphological comparisons were carried out using standard taxonomic keys. DNA extracted from 12 Sri Lankan sandfly samples was PCR-amplified and sequenced for the variable region of Cytochrome oxidase subunit I. Existing DNA sequences of India from GenBank were utilized for a phylogenetic analysis between Sri Lanka and India. Salivary protein profiles were studied using SDS-PAGE, Western blot, and electrospray ionization/LC/MS/MS. The morphological similarities observed between female Ph. argentipes from India and Sri Lanka suggest the presence of Ph. argentipes var. glaucus. A phylogenetic analysis showed genetic divergence between Ph. argentipes populations, but both shared a similar salivary protein profile. A common, strong 30 kDa immunogenic band comprised PagSP05, PagSP06, and PagSP17 proteins of Ph. argentipes. The similarity between the immunogenic salivary proteins suggests their potential use as common markers for vector exposure or immune response stimulants across regions. The use of multiple samples for each category of serum would improve the comprehensiveness of the immunogenic profiles obtained.

Genetic diversity of Leishmania donovani causing visceral and cutaneous leishmaniasis in the Western Ghats, India.

The visceral and atypical cutaneous leishmaniasis (VL and CL) caused by Leishmania donovani is an emerging infectious disease in the Western Ghats, Kerala, India. In this study, L. donovani specific kinetoplast minicircle DNA (k-DNA) sequence analysis was conducted to ascertain the genetic variability among the L. donovani isolates from the Western Ghats. Out of 23 CL and 5 VL suspected patient samples, 18 CL and 3 VL tested positive for k-DNA diagnostic PCR. Subsequently, 17 CL and 3 VL samples were found positive for L. donovani specific k-DNA PCR. Although the genetic diversity among the VL and CL isolates was low, there was clear variation from the parasites reported from other countries. The parasites characterized from the current study were more related to those reported from East Africa and India.

Identification and molecular characterization of Culex flavivirus in Culex quinquefasciatus (Diptera: Culicidae): first report from India.

BACKGROUND: Culex quinquefasciatus is a notorious vector known to transmit pathogens such as Wuchereria bancrofti (causing Lymphatic filariasis) and flaviviruses such as West Nile virus in India and St. Louis Encephalitis virus in the USA. It is the vector of the Rift Valley Fever virus, also on the African continent. Mosquitoes also harbor other non-pathogenic insect-specific flaviviruses (ISFs), such as Culex flavivirus (CxFV) and Aedes flavivirus. Recent studies have implicated ISFs interfering with the vectorial efficiency of the pathogenic arbo-viruses. METHODS: One hundred specimens of the Cx. quinquefasciatus population in two urban areas in Kerala State, India, were screened to have an understanding of the prevalence of these flaviviruses in this vector species. Viral RNA was extracted from individual specimens and was subjected to RT-PCR towards amplification of the CxFV non-structural protein 5 (NS5) gene. RESULTS: Among the 100 specimens, 7.0% were found to be harboring CxFV infection. The phylogenetic analysis of the gene sequences showed that the virus isolates were genetically related to Kenya, with 98-99% sequence similarities. CONCLUSION: This is the first report on the occurrence of CxFV from Cx. quinquefasciatus from India. The occurrence of these viruses in mosquitoes could play a critical role in disease vector management.

Polylactic acid/chitosan films for packaging of Indian white prawn (Fenneropenaeus indicus).

Composite films were developed by embedding nanochitosan (0.5%, 1% and 2%) in a polylactic acid (PLA) matrix using polyethylene glycol as a cross linking agent and polyvinyl alcohol as plasticizer. The mixture was casted into films via solvent casting. The interaction between the polylactic acid/nanochitosan (PLA/NCS) and polyethylene glycol had a significant effect on the tensile strength and the heat sealing properties. Antimicrobial properties of PLA/NCS films have been confirmed against aerobic microorganisms. PLA/NCS solvent casted films were used to pack prawn meat (Fenneropeneaus indicus) which was further stored in chilled condition for 18 days. The study proved that microbial and biochemical quality indices of prawn samples packed in different PLA/NCS film were retained and concluded that PLA/NCS composite films can be used for packing of fresh prawn to increase its shelf life.

Effectiveness of a 3 + 0 pneumococcal conjugate vaccine schedule against invasive pneumococcal disease among a birth cohort of 1.4 million children in Australia.

BACKGROUND: Most studies use indirect cohort or case-control methods to estimate vaccine effectiveness (VE) of 7- and 13-valent pneumococcal conjugate vaccines (PCV7 and PCV13) against invasive pneumococcal disease (IPD). Neither method can measure the benefit vaccination programs afford the unvaccinated and many studies were unable to estimate dose-specific VE. We linked Australia's national immunisation register with health data from two states to calculate IPD incidence by vaccination status and VE for a 3 + 0 PCV schedule (doses at 2, 4, 6 months, no booster) among a cohort of 1.4 million births. METHODS: Births records for 2001-2012 were probabilistically linked to IPD notifications, hospitalisations, deaths, and vaccination history (available until December 2013). IPD rates in vaccinated and unvaccinated children <2 years old were compared using Cox proportional hazards models (adjusting for potential confounders), with VE = (1 - adjusted hazard ratio) × 100. Separate models were performed for all-cause, PCV7, PCV13 and PCV13-non-PCV7 serotype-specific IPD, and for Aboriginal and non-Aboriginal children. RESULTS: Following introduction of universal PCV7 in 2005, rates of PCV7 serotype and all-cause IPD in unvaccinated children declined 89.5% and 61.4%, respectively, to be similar to rates in vaccinated children. Among non-Aboriginal children, VEs for 3 doses were 94.2% (95%CI: 81.9-98.1) for PCV7 serotype-specific IPD, 85.6% (95%CI: 60.5-94.8) for PCV13-non-PCV7 serotype-specific IPD and 80.1% (95%CI: 59.4-90.3) for all-cause IPD. There were no statistically significant differences between the VEs for 3 doses and for 1 or 2 doses against PCV13 and PCV13-non-PCV7 serotype-specific IPD, or between Aboriginal and non-Aboriginal children. CONCLUSION: Our population-based cohort study demonstrates that >90% coverage in the first year of a universal 3 + 0 PCV program provided high population-level protection, predominantly attributable to strong herd effects. The size of the cohort enabled calculation of robust dose-specific VE estimates for important population sub-groups relevant to vaccination policies internationally.

Probabilistic linkage of national immunisation and state-based health records for a cohort of 1.9 million births to evaluate Australia's childhood immunisation program.

INTRODUCTION: Several countries have developed national immunisation registers, but only the Nordic countries have linked their registers to other health data in order to comprehensively evaluate the `real world' effectiveness of vaccines. Nordic countries can link datasets deterministically using the national person identifier, but most countries, including Australia, don't have such an identifier to enable this type of linkage. OBJECTIVES: To describe the process for assembling a linked study cohort that will enable the conduct of population-based studies related to immunisation and immunisation policy. METHODS: National death and immunisation databases along with state health data (notifications of vaccine preventable diseases, perinatal data, hospital admissions and emergency department presentations) up until December 2013 were probabilistically linked (using demographic details) for children born between 1996 and 2012 in two states: Western Australia and New South Wales (42% of Australia's population, combined). RESULTS: After exclusions there were 1.95 million children in the study cohort (live born children with both a birth and perinatal record which represents 97.5% of all live births in the state perinatal data collections - our source population) and 18.0 million person years of follow up (mean: 9.2 years per child). The characteristics of children in the cohort were generally similar to those only included in state perinatal databases and outcome measures were in keeping with expected figures from unlinked data sources. However, the lack of a dynamic national population register meant immigrants could not be included. CONCLUSIONS: We have been able to develop a similarly comprehensive system to the Nordic countries based on probabilistic linkage methods. Our experience should provide encouragement to other countries with national immunisation registers looking to establish similar systems.