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Filariasis is an endemic health issue in India and Southeast Asian countries. The diagnosis is conventionally made by demonstrating microfilaria in a peripheral blood smear. Detection of microfilaria is rarely reported in cytological evaluation, and even more infrequent is the presence of microfilaria in a neoplastic lesion. Here we report a rare case of microfilaria in a routine cytological smear of a liver aspirate, which was diagnosed as hepatocellular carcinoma.

We report an interesting case of Bancroftian filariasis diagnosed on fine needle aspiration cytology in a young female hailing from a non-endemic hilly area of India, who presented with a subcutaneous swelling on the left forearm. This is an unusual extra-lymphatic site to harbor filarial infestation and the cytology smears showed adult worms, eggs, and microfilariae of Wuchereria bancrofti.

ABSTRACT: We report an interesting case of Bancroftian filariasis diagnosed on fine needle aspiration cytology in a young female hailing from a non-endemic hilly area of India, who presented with a subcutaneous swelling on the left forearm. This is an unusual extra-lymphatic site to harbor filarial infestation and the cytology smears showed adult worms, eggs, and microfilariae of Wuchereria bancrofti. [ABSTRACT FROM AUTHOR]

Dirofilariasis, caused by the nematode Dirofilaria spp., poses significant challenges in diagnosis due to its diverse clinical manifestations and complex life cycle. This comprehensive literature review focuses on the evolution of diagnostic methodologies, spanning from traditional morphological analyses to modern emerging techniques in the context of dirofilariasis diagnosis. The review traces the historical progression of diagnostic modalities, encompassing traditional approaches such as microscopic examination, serological tests (including ELISA and IFA), radiographic imaging, ultrasonography, and necropsy, which laid the foundation for subsequent advancements. The integration of molecular diagnostics marks a significant turning point in dirofilariasis diagnosis with the adoption of polymerase chain reaction (PCR) assays and real-time PCR (qPCR) facilitating enhanced sensitivity and specificity. Furthermore, recent strides in nextgeneration sequencing (NGS) technologies, including whole-genome sequencing (WGS), targeted sequencing (TS), metagenomic sequencing (MS), and RNA sequencing (transcriptome sequencing), have revolutionized the landscape of dirofilariasis diagnostics. Emerging techniques such as loop-mediated isothermal amplification (LAMP), digital PCR (dPCR), and digital microfluidics are also explored for their potential to augment diagnostic accuracy. The review addresses challenges associated with standardizing molecular protocols, tackling false positives/negatives, and discusses the advantages and limitations of each technique. By providing a comprehensive overview of dirofilariasis diagnostic strategies, from traditional to cutting-edge methods, this review aims to enhance understanding of the disease's diagnostic landscape. The insights gained have implications for improved disease management and guide future research endeavors toward refining diagnostic protocols and advancing therapeutic interventions. [ABSTRACT FROM AUTHOR]

ABSTRACT: Filariasis is an endemic health issue in India and Southeast Asian countries. The diagnosis is conventionally made by demonstrating microfilaria in a peripheral blood smear. Detection of microfilaria is rarely reported in cytological evaluation, and even more infrequent is the presence of microfilaria in a neoplastic lesion. Here we report a rare case of microfilaria in a routine cytological smear of a liver aspirate, which was diagnosed as hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]

Abstract Dirofilaria immitis is a mosquito-borne parasitic nematode that causes fatal heartworm disease in canids. The microfilariae are essential for research, including drug screening and mosquito-parasite interactions. However, no reliable methods for maintaining microfilaria long-term are currently available. Therefore, we used severe combined immunodeficiency (SCID) mice to develop a reliable method for maintaining D. immitis microfilaria. SCID mice were injected intravenously with microfilariae isolated from a D. immitis-infected dog. Microfilariae were detected in blood collected from the tail vein 218 days post-inoculation (dpi) and via cardiac puncture 296 dpi. Microfilariae maintained in and extracted from SCID mice showed infectivity and matured into third-stage larvae (L3s) in the vector mosquito Aedes aegypti. L3s can develop into the fourth stage larvae in vitro. Microfilariae from SCID mice respond normally to ivermectin in vitro. The microfilariae in SCID mice displayed periodicity in the peripheral circulation. The SCID mouse model aided in the separation of microfilariae from cryopreserved specimens. The use of SCID mice enabled the isolation and sustained cultivation of microfilariae from clinical samples. These findings highlight the usefulness of the SCID mouse model for studying D. immitis microfilaremia in canine heartworm research.

Setaria equina is a common parasitic infection in horses that is transmitted by mosquitoes. The absence of previous reports regarding the presence of Setaria equina microfilariae in Nigerian horses highlights the need to confirm its existence in the region. Verifying the existence of this parasite would enhance our overall understanding of its global distribution, which could then inform the development of more effective control measures. This paper reports a case of Setaria equina microfilariae infection in an adult Argentine polo pony mare in Nigeria. The recently acquired adult Argentine polo pony mare was exhibiting depression and weight loss. Despite normal vital parameters, the horse had a poor body condition and no history of deworming. Laboratory analyses, including wet blood film examination, complete blood count, and serum biochemistry, revealed the presence of Setaria equina microfilariae and macrocytic hypochromic anaemia. Treatment involved a five-day course of intramuscular multivitamin injections and three weeks of subcutaneous ivermectin injections. Follow-up examinations demonstrated the absence of microfilaria and a return to an active and lively state. The case report details the clinical presentation, diagnostic process, and successful treatment of a recently acquired adult Argentine polo pony mare. A consistent deworming regimen is essential for horses, including those recently acquired, to prevent parasitic infections and enhance their overall well-being.

Filariasis is a very important public health problem endemic in tropical and subtropical countries. It is a debilitating disease with significant socioeconomic burden. In 2016, lymphatic filariasis was successfully eliminated from the Maldives, which was certified by WHO. We report a case of filariasis detected by a rapid antigen test while undergoing a screening program conducted by Health Protection Agency, Maldives in December 2023. The patient was asymptomatic and showed no peripheral blood eosinophilia. Conventional blood smears were negative. Nevertheless, buffy coat smear served as a promising test in detection of parasite, thus confirming the re-emergence of the disease in the country. Therefore, our study highlight the significance of buffy coat smears in detection of hemoparasites. We also focus on the importance of surveillance system in order to sustain the disease elimination goal that was already achieved.

Introduction: Filariasis, a parasitic infection caused by several types of nematode worms, is a matter of great concern for public health in India, The disease is caused by microfilaria, which are released into the peripheral blood circulation with periodicity during the night. However, identifying microfilaria in routine peripheral blood smears, Fine Needle Aspiration (FNAC) smears, and body fluids is challenging due to their scarce nature. Our study aimed to underscore the significance of having a heightened suspicion of filarial infection in patients with swelling, particularly those from endemic regions. Furthermore, the study emphasised the importance of using FNAC as an economical and effective diagnostic tool for detecting microfilaria. Materials and Methods: This retrospective study was conducted in the Pathology Department of ESIC Medical College and Hospital Gulbarga over a period of ten years (2013-2023). A 22-23 gauze needle made aspiration smears were stained by Papanicolaou and May-Grünwald Giemsa stain. Results: In this study, 24 cases showed the presence of microfilaria, a type of parasitic worm, despite the lack of clinical suspicion. Of these cases, eight involved breast lumps, eight involved thyroid swelling, and eight involved lymph nodes. In fifteen of these cases, fragments of adult worms were also reported, indicating a potentially more severe infection. These findings suggest the need for more diligent screening and testing to detect and treat parasitic infections before they can cause further harm. Conclusion: Filariasis is a rare disease that requires thorough screening of FNA smears, particularly in asymptomatic patients from endemic regions, to avoid missing incidental findings. Moreover, the study emphasises the cost-effectiveness and efficacy of FNAC in diagnosing microfilaria. [ABSTRACT FROM AUTHOR]

Elephantiasis, also known as filariasis, is a severe public health issue in India. Microfilariae in the bone marrow smears are an unusual and incidental finding. Every peripheral blood and bone marrow aspirate smear must be screened for microfilariae in endemic areas to identify asymptomatic carriers. Here, we report a case of a 50-year-old man who presented with pyrexia of unknown origin. He had mild pallor and right-side inguinal lymphadenopathy. Peripheral blood showed anemia and leukocytosis with neutrophilia. Other serological tests, radiological examination, and aspiration cytology from the lymph node were inconclusive. The bone marrow aspirate smears incidentally displayed microfilaria, and the inguinal lymph node demonstrated an adult filaria worm. The association of microfilaria infection with pyrexia of unknown origin (PUO) and peripheral blood neutrophilia has been rarely reported in the literature. The patient recovered from fever and weakness with antifilarial medication and hematinics. [ABSTRACT FROM AUTHOR]

Elaeophorosis, infection by the filarial worm Elaeophora schneideri, is a parasitic disease of wild ungulates in North America; however, our understanding of the relevance of E. schneideri to moose (Alces alces) morbidity and mortality is incomplete. Between March 2020 and July 2022, necropsy and histopathology were performed on 61 Shiras moose (Alces alces shirasi) in Idaho, US. Among the 41 adults (greater than 1 yr old), 21 moose were from northern Idaho, and 20 were from southeastern Idaho. Elaeophorosis was diagnosed in 24% (10 of 41). All 10 infected moose were from southeastern Idaho; none of the 21 moose from northern Idaho were infected. No juvenile moose (nine from northern and 11 from southeastern Idaho) were infected. Microfilariae were detected histologically in 9 of 10 infected moose, most consistently in brain tissue associated with lesions indicative of ischemic injury to the neuroparenchyma attributed to occlusion of arterioles and capillaries by microfilariae or fibrin thrombi, including edema, necrosis, and glial nodules. Microfilariae found in other tissues of the head, including the eye, tongue, and pinnae of some animals, as well as in lung, heart, liver, and kidney, typically were associated with inflammation. Three of the 10 infected moose had cropped ears attributed to elaeophorosis, and four exhibited abnormal behavior, which may have been due to neuropathology associated with E. schneideri microfilariae in the brain. [ABSTRACT FROM AUTHOR]

Dirofilaria immitis is a mosquito-borne parasitic nematode that causes fatal heartworm disease in canids. The microfilariae are essential for research, including drug screening and mosquito-parasite interactions. However, no reliable methods for maintaining microfilaria long-term are currently available. Therefore, we used severe combined immunodeficiency (SCID) mice to develop a reliable method for maintaining D. immitis microfilaria. SCID mice were injected intravenously with microfilariae isolated from a D. immitis-infected dog. Microfilariae were detected in blood collected from the tail vein 218 days post-inoculation (dpi) and via cardiac puncture 296 dpi. Microfilariae maintained in and extracted from SCID mice showed infectivity and matured into third-stage larvae (L3s) in the vector mosquito Aedes aegypti. L3s can develop into the fourth stage larvae in vitro. Microfilariae from SCID mice respond normally to ivermectin in vitro. The microfilariae in SCID mice displayed periodicity in the peripheral circulation. The SCID mouse model aided in the separation of microfilariae from cryopreserved specimens. The use of SCID mice enabled the isolation and sustained cultivation of microfilariae from clinical samples. These findings highlight the usefulness of the SCID mouse model for studying D. immitis microfilaremia in canine heartworm research. [ABSTRACT FROM AUTHOR]

Abstract Background Onchocerciasis causes chronic systemic inflammation. Several studies have used markers such as haemato-biochemical indices to predict the occurrence of systemic inflammation. This study assessed the variability and predictability of haemato-biochemical indices and blood composite ratios (BCRs) in microfilariae positive (MF+) and microfilariae negative (MF-) subgroups of onchocercomata participants. Methods One hundred and five (105) MF + and 34 MF- participants were retrospectively recruited into the study. Screening for the presence of O. volvulus microfilariae was done from skin snips taken from the left and right iliac crests of participants using established and approved protocols. Haematological and biochemical indices were measured using standard laboratory automated analyzers. Blood composite ratios (BCRs) were calculated as ratios of the absolute parameters involved. Results A significantly increased total WBC, absolute eosinophil, eosinophil percent and absolute basophil were observed in the MF + participants compared to MF- participants. Reduced gamma-glutamyl transferase (GGT) with increased estimated glomerular filtration rate (eGFR) was significantly associated with MF + participants compared to MF- participants. BCRs were significantly higher for eosinophil-to-neutrophil ratio (ENR), eosinophil-to-monocyte ratio (EMR), eosinophil-to-basophil ratio (EBR) and eosinophil-to-lymphocyte ratio (ELR) in MF + participants compared to MF- participants. After multivariate adjustment, onchocercomata participants with increased eosinophil counts (aOR = 13.86, 95% CI [2.07–92.90], p = 0.007), ENR x10 (aOR = 1.42, 95% CI [1.05–1.93], p = 0.025), EMR (aOR = 2.64, 95% CI [1.25–5.60], p = 0.011), EBR (aOR = 1.07, 95% CI [1.01–1.10], p = 0.020) and ELR x10 (aOR = 1.69, 95% CI [1.14–2.51], p = 0.009) were more likely to have microfilaridermia. Conclusions Elevated eosinophil counts with higher ENR, EMR, EBR and ELR levels are significantly associated with microfilaridermia in onchocercomata participants. Combining BCRs with eosinophil count significantly led to an improvement in the conventional model for predicting microfilaridermia.

Background and Aim: Filarial nematode typically produces a larval stage (microfilariae) in the bloodstream of vertebrate hosts, where microfilariae reside in the blood or subcutaneous tissues. Filarial nematodes cause human diseases, such as river blindness and elephantiasis, which are widely studied. However, in avian species, they are overlooked because they are nonpathogenic. In Thailand, microfilaria can be found in wild birds and domestic chickens. Recently, an increase in the number of blood samples submitted to veterinary diagnostic laboratories may have increased the number of microfilariae. Therefore, knowledge about filarial species and reliable detection methods are important. Therefore, this study aimed to investigate the efficacy of buffy coat smear and polymerase chain reaction (PCR)-based methods for the detection of microfilaria in domestic chickens. In addition, parasites were identified using the sequence of the cytochrome c oxidase subunit 1 (COX1) gene. Materials and Methods: Giemsa-stained buffy coat smears from a previous study were reanalyzed. These available buffy coat smears were prepared from 55 domestic chickens raised as backyard free-ranging in Southern Thailand. Fifty-seven frozen genomic DNA extracted from chicken blood were used to detect the presence of the COX1 gene in Onchocercidae nematodes. The nested PCR protocol for amplification of the OnchoCOI_ R2-OnchoCOI_ R2 fragment of the COX1 gene was applied from a previous report. Sequences of COX1 were analyzed to identify Onchocercidae nematodes and if they were single or mixed infections. We constructed Bayesian phylogenetics to identify parasites and assessment of the relationship between filarial nematodes in avian species and other vertebrate hosts. Results: Buffy coat smears from 15 samples revealed microfilaria. Of these 15 samples, only eight were positive for COX1 nested-PCR amplification. The other two buffy coat-negative samples were also positive for nested-PCR. Sequencing of these 11 nested PCR-positive samples revealed that almost all of them were Onchocercidae nematodes. Bayesian phylogenetic analysis showed that chicken Onchocercidae spp. were grouped with other avian filarial nematodes. However, all chickens Onchocercidae spp. showed a double peak in the sequencing chromatogram, indicating mixed filarial infection (species or haplotypes). Therefore, no chicken Onchocercidae sequence was deposited on National Center for Biotechnology Information, GenBank. Conclusion: Giemsa-stained buffy coat smear was a reliable method for the detection of chicken microfilaria in routine veterinary diagnostic laboratories. Development of a new PCR-based method is necessary. This method may provide greater sensitivity and specificity of detection. In addition, the PCR method allowed us to access the genetic characteristics of nematodes, which helped us maximize our knowledge of nematodes. Further investigations, such as the pathogenicity of filarial nematodes in chickens and their potential vectors, are required.

Accurate rapid diagnostic tests (RDTs) are needed to diagnose lymphatic filariasis (LF) in global elimination programmes. We evaluated the performance of the new STANDARD Q Filariasis Antigen Test (QFAT) against the Bioline Filariasis Test Strip (FTS) for detecting W. bancrofti antigen (Ag) in laboratory conditions, using serum (n = 195) and plasma (n = 189) from LF-endemic areas (Samoa, American Samoa and Myanmar) and Australian negative controls (n = 46). The prior Ag status of endemic samples (54.9% Ag-positive) was determined by rapid test (ICT or FTS) or Og4C3 ELISA. The proportion of samples testing positive at 10 min was similar for QFAT (44.8%) and FTS (41.3%). Concordance between tests was 93.5% (kappa 0.87, n = 417) at 10 min, and it increased to 98.8% (kappa 0.98) at 24 h. The sensitivities of QFAT and FTS at 10 min compared to the prior results were 92% (95% CI 88.0–96.0) and 86% (95% CI 80.0–90.0), respectively, and they increased to 97% and 99% at 24 h. Specificity was 98% for QFAT and 99% for FTS at 10 min. Both tests showed evidence of cross-reaction with Dirofilaria repens and Onchocerca lupi but not with Acanthochilonema reconditum or Cercopithifilaria bainae. Under laboratory conditions, QFAT is a suitable alternative RDT to FTS.

Microfilarial parasites can obstruct the lymphatic tree giving rise to varying lymphatic and extra-lymphatic symptoms. Renal manifestations can range from asymptomatic proteinuria, chyluria, and nephrotic syndrome, to acute glomerulonephritis. The diagnosis of filariasis is usually made by the demonstration of the parasite in the peripheral blood smear, with or without eosinophilia. The renal involvement by this parasite has been sparsely reported in the literature. We hereby report five cases of filariasis detected on histopathological examination of renal biopsies, performed for other indications, along with a brief report of the additional histological findings. Three native and two graft biopsies were included. All our patients were male, with a mean age of 47 years (range 37 to 66 years). The serum creatinine ranged from 1.2 to 12.9 mg/dL. The mean 24-hour urinary protein was 3.6 gm/day. Peripheral blood eosinophilia was not recorded in any case, however, ESR was raised in all cases. Urine examination revealed varying proteinuria, with hematuria in two cases. Histological examination revealed microfilaria in all five biopsies, along with focal segmental glomerulosclerosis in two cases, combined cellular and humoral rejection, minimal change disease and acute tubular necrosis in one case each respectively. All patients were treated with diethylcarbamazine 6mg/kg/day or 12 days, in addition to the renal medications. Diagnosing the parasite is crucial as the patient is likely to benefit due to the timely treatment of the disease. Reporting this case series highlights an interesting finding in nephropathology. [ABSTRACT FROM AUTHOR]

Background Onchocerciasis causes chronic systemic inflammation. Several studies have used markers such as haemato-biochemical indices to predict the occurrence of systemic inflammation. This study assessed the variability and predictability of haemato-biochemical indices and blood composite ratios (BCRs) in microfilariae positive (MF+) and microfilariae negative (MF-) subgroups of onchocercomata participants. Methods One hundred and five (105) MF + and 34 MF- participants were retrospectively recruited into the study. Screening for the presence of O. volvulus microfilariae was done from skin snips taken from the left and right iliac crests of participants using established and approved protocols. Haematological and biochemical indices were measured using standard laboratory automated analyzers. Blood composite ratios (BCRs) were calculated as ratios of the absolute parameters involved. Results A significantly increased total WBC, absolute eosinophil, eosinophil percent and absolute basophil were observed in the MF + participants compared to MF- participants. Reduced gamma-glutamyl transferase (GGT) with increased estimated glomerular filtration rate (eGFR) was significantly associated with MF + participants compared to MF- participants. BCRs were significantly higher for eosinophil-to-neutrophil ratio (ENR), eosinophil-to-monocyte ratio (EMR), eosinophil-to-basophil ratio (EBR) and eosinophil-to-lymphocyte ratio (ELR) in MF + participants compared to MF- participants. After multivariate adjustment, onchocercomata participants with increased eosinophil counts (aOR = 13.86, 95% CI [2.07–92.90], p = 0.007), ENR x10 (aOR = 1.42, 95% CI [1.05–1.93], p = 0.025), EMR (aOR = 2.64, 95% CI [1.25–5.60], p = 0.011), EBR (aOR = 1.07, 95% CI [1.01–1.10], p = 0.020) and ELR x10 (aOR = 1.69, 95% CI [1.14–2.51], p = 0.009) were more likely to have microfilaridermia. Conclusions Elevated eosinophil counts with higher ENR, EMR, EBR and ELR levels are significantly associated with microfilaridermia in onchocercomata participants. Combining BCRs with eosinophil count significantly led to an improvement in the conventional model for predicting microfilaridermia. [ABSTRACT FROM AUTHOR]

An 11-year-old male dog presenting exophthalmia and perforation of the cornea of the left eye was seen at the veterinary hospital of the State University of Maranhão. Upon physical examination, irregularly shaped granulomas of crumbly appearance were observed on the dog’s tongue. The dog was sent for ocular enucleation surgery. The following tests were requested: complete blood count, serum biochemistry, hemoparasite investigation, chest X-Ray, echocardiogram, electrocardiogram, cytology and nodule histopathology. The blood count revealed neutrophilia, signs of anemia, increased total plasma protein and thrombocytopenia. Serum biochemistry and the imaging examinations showed normal results. The cytological examination on the tongue nodules showed microfilariae and the histopathological examination showed chronic glossitis. Polymerase chain reaction (PCR) and genetic sequencing were performed on a blood sample, which made it possible to identify the species Acanthocheilonema reconditum. This is the first record worldwide of the occurrence of microfilariae of A. reconditum on tongue nodules. This finding serves to alert dermatologists and dentists regarding the need for a differential diagnosis for types of oral cavity lesions of unknown etiological origin.

Lymphatic filariasis is a neglected tropical disease that affects the lymphatic system of humans. The major etiologic agent is a nematode called Wuchereria bancrofti, but Brugia malayi and Brugia timoriare sometimes encountered as causative agents. Mosquitoes are the vectors while humans the definitive hosts respectively. The burden of the disease is heavier in Nigeria than in other endemic countries in Africa. This occurs with increasing morbidity and mortality at different locations within the country, the World Health Organization recommended treatments for lymphatic filariasis include the use of Albendazole (400mg) twice per year in co-endemic areas with loa loa, Ivermectin (200mcg/kg) in combination with Albendazole (400mg) in areas that are coendemic with onchocerciasis, ivermectin (200mcg/kg) with diethylcarbamazine citrate (DEC) (6mg/kg) and albendazole (400mg) in areas without onchocerciasis. This paper covered a systematic review, meta-analysis, and scoping review on lymphatic filariasis in the respective geopolitical zones within the country. The literature used was obtained through online search engines including PubMed and Google Scholar with the heading "lymphatic filariasis in the name of the state", Nigeria. This review revealed an overall prevalence of 11.18% with regional spread of Northwest (1.59%), North Central and North East, (4.52%), South West (1.26%), and South-South with South East (3.81%) prevalence. The disease has been successfully eliminated in Argungu local government areas (LGAs) of Kebbi State, Plateau, and Nasarawa States respectively. Most clinical manifestations (31.12%) include hydrocele, lymphedema, elephantiasis, hernia, and dermatitis. Night blood samples are appropriate for microfilaria investigation. Sustained MDAs, the right testing methods, early treatment of infected cases, and vector control are useful for the elimination of lymphatic filariasis for morbidity management and disability prevention in the country. Regional control strategies, improved quality monitoring of surveys and intervention programs with proper records of morbidity and disability requiring intervention are important approaches for the timely elimination of the disease in Nigeria. [ABSTRACT FROM AUTHOR]

Background: Newer generation ophthalmologists practicing in the developed world are not very familiar with some tropical ocular diseases due to the absence of reports in the ophthalmic literature over the past thirty years. Because of world globalization or due to influx of immigrants from sub-Saharan Africa, exotic retinal diseases are being encountered more often in ophthalmology clinics. Methods: A multicenter case series of chorioretinitis or optic neuritis with obscure etiology that used serial multimodal imaging. Results: Four cases qualified with the diagnosis of presumed ocular onchocerciasis based on their residence near fast rivers in endemic areas, multimodal imaging, long term follow-up showing progressive disease and negative workup for other diseases. Characteristic findings include peripapillary choroiditis with optic neuritis or atrophy, subretinal tracts of the microfilaria, progressive RPE atrophy around heavily pigmented multifocal chorioretinal lesions of varying shapes, subretinal white or crystalline dots, and response to ivermectin. Typical skin findings are often absent in such patients with chorioretinitis rendering the diagnosis more challenging. Conclusions: Familiarity with the myriad ocular findings of onchocerciasis, and a high-degree of suspicion in subjects residing in endemic areas can help in the correct diagnosis and implementation of appropriate therapy. Onchocercal chorioretinitis is a slow, insidious, progressive, and prolonged polymorphous disease. [ABSTRACT FROM AUTHOR]

Abstract Background In onchocerciasis-endemic areas with high ongoing Onchocerca volvulus transmission, a high prevalence of epilepsy has been reported. This study aimed to determine the prevalence and clinical characteristics of epilepsy in the Bono Region of Ghana following 27 years of implementation of ivermectin mass drug administration (MDA). Methods Between October 2020 and August 2021, cross-sectional surveys were conducted in nine communities in the Tain District and Wenchi Municipality of the Bono Region of Ghana. In the first stage, a random door-to-door approach was used to screen the population for epilepsy using a pre-tested questionnaire. Persons suspected of having epilepsy were invited for a second-stage neurological examination for case verification. Community O. volvulus microfilarial infection status and Ov16 seropositivity were also determined. Ninety-five confidence intervals (95% CI) for prevalence values were calculated using the Wilson Score Interval. Results Of the 971 participants, 500 (51.5%) were females, and the median age (interquartile range) was 26 (15‒43) years. Fourteen participants (1.4%, 95% CI: 1.0‒2.0) were diagnosed as having epilepsy with generalized seizures being the most frequent seizure type (85.7%, 12/14). The overall microfilarial prevalence of O. volvulus was 10.3% (November 2020) and 9.9% (August 2021); the Ov16 seroprevalence was 22.2% (June 2021). Only 63.2% took ivermectin in the last round of MDA distribution in March 2021. Conclusions The 1.4% prevalence of epilepsy in the Bono region is similar to the median epilepsy prevalence in sub-Saharan Africa. However, the persistent microfilarial prevalence and low ivermectin study coverage call for the Ghana Onchocerciasis Elimination Programme to step up its efforts to ensure that the gains achieved are consolidated and improved to achieve the elimination of onchocerciasis by 2030. Graphical Abstract

Key Clinical Message: Skin ulcerations are a significant cause of morbidity and can be challenging to manage. Among the various causes of chronic non‐healing ulcers, lymphedema is also considered a possible diagnosis in countries such as Nepal. Lymphatic filariasis has been a significant public health issue in endemic areas. Wuchereria bancrofti is a common nematode parasite that causes lymphatic filariasis. Excessive retention of lymphatic fluid in the interstitial compartment can cause localized tissue swelling, known as lymphedema, which is caused by impaired lymphatic drainage. Microfilariae can be detected in peripheral blood, body fluids, and needle aspirates. Microfilaria is not commonly found in ulcers on elephantiasis legs. We discuss here a case of 73‐year‐old women with elephantiasis legs with pus discharging ulcers in the thighs. Microscopic examination of pus discharge revealed microfilaria which highlights the importance of pus examination as diagnostic modality in endemic countries. [ABSTRACT FROM AUTHOR]

Background: In onchocerciasis-endemic areas with high ongoing Onchocerca volvulus transmission, a high prevalence of epilepsy has been reported. This study aimed to determine the prevalence and clinical characteristics of epilepsy in the Bono Region of Ghana following 27 years of implementation of ivermectin mass drug administration (MDA). Methods: Between October 2020 and August 2021, cross-sectional surveys were conducted in nine communities in the Tain District and Wenchi Municipality of the Bono Region of Ghana. In the first stage, a random door-to-door approach was used to screen the population for epilepsy using a pre-tested questionnaire. Persons suspected of having epilepsy were invited for a second-stage neurological examination for case verification. Community O. volvulus microfilarial infection status and Ov16 seropositivity were also determined. Ninety-five confidence intervals (95% CI) for prevalence values were calculated using the Wilson Score Interval. Results: Of the 971 participants, 500 (51.5%) were females, and the median age (interquartile range) was 26 (15‒43) years. Fourteen participants (1.4%, 95% CI: 1.0‒2.0) were diagnosed as having epilepsy with generalized seizures being the most frequent seizure type (85.7%, 12/14). The overall microfilarial prevalence of O. volvulus was 10.3% (November 2020) and 9.9% (August 2021); the Ov16 seroprevalence was 22.2% (June 2021). Only 63.2% took ivermectin in the last round of MDA distribution in March 2021. Conclusions: The 1.4% prevalence of epilepsy in the Bono region is similar to the median epilepsy prevalence in sub-Saharan Africa. However, the persistent microfilarial prevalence and low ivermectin study coverage call for the Ghana Onchocerciasis Elimination Programme to step up its efforts to ensure that the gains achieved are consolidated and improved to achieve the elimination of onchocerciasis by 2030. [ABSTRACT FROM AUTHOR]

Background: Filarial infections continue to pose a great challenge in endemic countries. One of the central goals in the fight against human filarial infections is the development of strategies that will lead to the inhibition of microfilariae (mf) transmission. Keeping mf under a certain threshold within endemic populations will stop transmission and eliminate the infection. Method: A narrative review was carried out to identify the possibilities and limitations of exploring the use of eosinophil responses as an anti‐filarial vaccine, and biomarker for the detection of filarial infections. An extensive literature search was performed in online scientific databases including PubMed Central, PubMed, BioMed Central, with the use of predefined search terms. Results: A better understanding of the parasite‐host interactions will lead to the development of improved and better treatment or vaccine strategies that could eliminate filariasis as soon as possible. Highlighted in this review is the explorative use of eosinophil‐producing CLC/Galectin‐10 as a potential biomarker for filarial infections. Also discussed are some genes, and pathways involved in eosinophil recruitments that could be explored for anti‐filarial vaccine development. Conclusion: In this short communication, we discuss how eosinophil‐regulated genes, pathways, and networks could be critical in providing more information on how reliably a front‐line immune player could be exploited for anti‐filarial vaccine development and early infection biomarker. [ABSTRACT FROM AUTHOR]

Abstract Background Filarial infections continue to pose a great challenge in endemic countries. One of the central goals in the fight against human filarial infections is the development of strategies that will lead to the inhibition of microfilariae (mf) transmission. Keeping mf under a certain threshold within endemic populations will stop transmission and eliminate the infection. Method A narrative review was carried out to identify the possibilities and limitations of exploring the use of eosinophil responses as an anti‐filarial vaccine, and biomarker for the detection of filarial infections. An extensive literature search was performed in online scientific databases including PubMed Central, PubMed, BioMed Central, with the use of predefined search terms. Results A better understanding of the parasite‐host interactions will lead to the development of improved and better treatment or vaccine strategies that could eliminate filariasis as soon as possible. Highlighted in this review is the explorative use of eosinophil‐producing CLC/Galectin‐10 as a potential biomarker for filarial infections. Also discussed are some genes, and pathways involved in eosinophil recruitments that could be explored for anti‐filarial vaccine development. Conclusion In this short communication, we discuss how eosinophil‐regulated genes, pathways, and networks could be critical in providing more information on how reliably a front‐line immune player could be exploited for anti‐filarial vaccine development and early infection biomarker.

Purpose: To study clinical and pathological features of parasitic lesions in the ocular adnexa in a tertiary care ophthalmic center in south India. Methods: 43 cases of ocular parasitosis were analysed clinically and correlated with the pathological findings (gross morphology and histopathology) over a period of five years (2015–2020). Results: Among the 43 cases, the age group ranged from 9 months to 78 years (mean age of 41.6 years). Female patients were more common than male patients, with a percentage of 63% (27) and 37% (16) respectively. Cystic lesion in the lid or orbit was seen in 23 cases (53.4%); solid mass lesions were seen in 17 cases (39.5%); subconjunctival worms in three cases; and subretinal parasite in one. Gross examination and histopathologic study showed Dirofilaria in 23 cases (53.5%), followed by Cysticercus in six cases (14%) and Microfilariae in four cases (9.3%). Exact species identification was not possible in ten cases (23.25%). Correlation between the type of lesion and type of inflammatory cells with the specific parasite was done. Conclusion: Our study showed that important clinicopathological correlations can be made from the parasitic lesions in the eye and adnexa, which can aid in definitive diagnosis and prompt identification of the parasite for patient management.

The rectangular filtering microfluidic chip was invented using microfluidics device fabrication technology and can separate living microfilariae from blood samples without a syringe pump. The diagnostic results are highly effective. The device is based on the principle of separating millions of blood cells from microfilariae using a rectangular filter structure. It disperses fluid evenly into the flow-passage channel, and its rectangular filter structure is the key to success in reducing the pressure and separating blood cells from microfilariae effectively. The flow rate and blood cell concentration were optimized in our study. The chip is intended to be a point-of-care device that can reduce the use of superfluous instrumentation in the field. The technology is designed to be rapid, accurate, and easy-to-use for all users, especially those in remote areas.

Filarial parasites are tissue dwelling worms transmitted by hematophagous vectors. Understanding the mechanisms regulating microfilariae (the parasite offspring) development is a prerequisite for controlling transmission in filarial infections. Th2 immune responses are key for building efficient anti-parasite responses but have been shown to also lead to detrimental tissue damage in the presence of microfilariae. Litomosoides sigmodontis, a rodent filaria residing in the pleural cavity was therefore used to characterize pleuropulmonary pathology and associated immune responses in wild-type and Th2 deficient mice. Wild-type and Th2-deficient mice (Il-4ra-/-/Il-5-/-) were infected with L. sigmodontis and parasite outcome was analyzed during the patent phase (when microfilariae are in the general circulation). Pleuropulmonary manifestations were investigated and pleural and bronchoalveolar cells were characterized by RNA analysis, imaging and/or flow cytometry focusing on macrophages. Il-4ra-/-/Il-5-/- mice were hypermicrofilaremic and showed an enhanced filarial survival but also displayed a drastic reduction of microfilaria-driven pleural cavity pathologies. In parallel, pleural macrophages from Il-4ra-/-/Il-5-/- mice lacked expression of prototypical alternative activation markers RELMa and Chil3 and showed an altered balance of some markers of the arginine metabolic pathway. In addition, monocytes-derived F4/80intermediate macrophages from infected Il- 4ra-/-/Il-5-/- mice failed to mature into resident F4/80high large macrophages. Altogether these data emphasize that the presence of both microfilariae and IL- 4R/IL-5 signaling are critical in the development of the pathology and in the phenotype of macrophages. In Il-4ra-/-/Il-5-/- mice, the balance is in favor of parasite development while limiting the pathology associated with the host immune response. [ABSTRACT FROM AUTHOR]

Filarial parasites are tissue dwelling worms transmitted by hematophagous vectors. Understanding the mechanisms regulating microfilariae (the parasite offspring) development is a prerequisite for controlling transmission in filarial infections. Th2 immune responses are key for building efficient anti-parasite responses but have been shown to also lead to detrimental tissue damage in the presence of microfilariae. Litomosoides sigmodontis, a rodent filaria residing in the pleural cavity was therefore used to characterize pleuropulmonary pathology and associated immune responses in wild-type and Th2 deficient mice. Wild-type and Th2-deficient mice (Il-4rα-/-/Il-5-/-) were infected with L. sigmodontis and parasite outcome was analyzed during the patent phase (when microfilariae are in the general circulation). Pleuropulmonary manifestations were investigated and pleural and bronchoalveolar cells were characterized by RNA analysis, imaging and/or flow cytometry focusing on macrophages. Il-4rα-/-/Il-5-/- mice were hypermicrofilaremic and showed an enhanced filarial survival but also displayed a drastic reduction of microfilaria-driven pleural cavity pathologies. In parallel, pleural macrophages from Il-4rα-/-/Il-5-/- mice lacked expression of prototypical alternative activation markers RELMα and Chil3 and showed an altered balance of some markers of the arginine metabolic pathway. In addition, monocytes-derived F4/80intermediate macrophages from infected Il-4rα-/-/Il-5-/- mice failed to mature into resident F4/80high large macrophages. Altogether these data emphasize that the presence of both microfilariae and IL-4R/IL-5 signaling are critical in the development of the pathology and in the phenotype of macrophages. In Il-4rα-/-/Il-5-/- mice, the balance is in favor of parasite development while limiting the pathology associated with the host immune response.

Background and Aim: Filarial infections can significantly impact the health of both humans and animals. In elephants, filariasis has been associated with cutaneous dermatitis and skin nodules. However, molecular evidence for such infections is limited in Thailand. This study aimed to identify the morphological and molecular characteristics of microfilaria in captive Asian elephants in Thailand. Materials and Methods: Whole blood collected from the ear vein of 129 captive Asian elephants was hematologically analyzed, and the blood parasites were evaluated using three standard techniques: The microcapillary test, thin blood smears, and polymerase chain reaction (PCR). Results: Conventional PCR revealed that approximately 17% (22/129) of the sampled elephants were positive for microfilaria. Microscopy revealed that microfilariae are large, unsheathed, with extended nuclei, a short headspace, and a curved tail tapering at the end. Results of internal transcribed spacer region analysis show that the elephant microfilariae are closely related to Onchocerca spp. All of the elephants positive for microfilaria presented with neither skin lesion nor anemic signs. Microfilaria infection was not associated with age; however, microfilariae were more likely to be detected in male elephants due to differences in management systems. Conclusion: This is the first study to provide both morphological and molecular evidence of microfilaria in Thai elephants. There is an urgent need to investigate the long-term and large-scale effects of microfilaria on the health of elephants.

Filarial parasite infestation not only affects the structure and function of lymphatic vessels but is also associated with extralymphatic pathology and disease. Incidence of renal involvement in microfilaria carriers has led to increased cognizance of extralymphatic presentation. Literature set forth clinical syndromes having extralymphatic manifestation of filaria. The diagnosis of filariasis is done by visualisation of microfilaria in peripheral blood smear, lymphatic tissue. Other modalities of diagnosis are Enzyme linked immunosorbent assay (ELISA), Immunochromatographic test. Diethyl carbamazine (DEC) provocation test usually is done to detect microfilaria in night blood smear due to the nocturnal periodicity of microfilaria. The drug DEC flushes the microfilaria into the peripheral circulation leading to high probability of detection. We present a case of a 59-year-old male who was diagnosed as nephrotic syndrome and after a DEC challenge we detected microfilaria in the peripheral smear confirming microfilaria-induced Nephrotic Syndrome after all other secondary conditions were excluded. [ABSTRACT FROM AUTHOR]

Abstract Background Various heartworm (HW) diagnostic testing modalities detect products of, or reactions to, different life cycle stages of Dirofilaria immitis. Microfilariae (Mf) can be directly visualized in blood, antigen (Ag) from immature and adult heartworms may be detected on commercial assays, and antibody (Ab) tests detect the host immune response to larval stages. Ag and Mf tests are commonly used in dogs, which frequently carry adult HW infections, but Ab tests have only been validated for use in cats. In some HW-infected dogs, Ag is blocked by immune complexing leading to false-negative results. Heat-treatment (HT) to disrupt these complexes can increase the sensitivity of HW Ag tests. The aim of this study was to compare different methods for diagnosing HW infection in dogs at high risk using individual and paired diagnostic tests, including an exploration of using Ab tests designed for cats to test canine samples. Methods One hundred stray adult (≥ 2-year-old) dogs in Florida shelters were tested using Mf, HW Ag, and HW Ab tests (feline HW Ab tests currently not commercially validated/approved for use in dogs); two versions of each test platform were used. Results Fourteen dogs tested positive using point-of-care (POC) Ag tests; an additional 2 dogs tested positive with microtiter well assay, and an additional 12 dogs tested positive using HT Ag testing. For individual tests, Ag test sensitivity/specificity compared to HT Ag was 50–57%/100%, and Ab tests were 46–64%/82–94%. Sensitivity estimates for individual tests were higher when comparing to non-HT Ag. Pairing POC Ag tests with Mf tests improved sensitivity without loss of specificity, while pairing POC Ag and Ab tests modestly increased sensitivity at the expense of specificity. Conclusions Screening dogs for HW infection using both POC Ag and Mf detection, which is recommended by the American Heartworm Society, improved diagnostic performance in this study compared to single Ag test use, but may have missed more than one in four infected dogs. The need to improve access to highly accurate, rapid, and inexpensive large-scale HW testing for dogs in animal shelters remains largely unmet by current testing availability. The development of practical and validated protocols that incorporate heat or chemical treatment to disrupt Ag-Ab complexes in POC testing or decreasing the cost and time required for such testing in reference laboratories might provide solutions to this unmet need. Similar studies performed in countries where the prevalence of parasites such as D. repens or A. vasorum is different to the USA could potentially yield very different positive predictive values for both HT and non-HT Ag tests. Graphic abstract

Abstract Background Blood parasites have been the subject of much research, with numerous reports of the presence of microfilariae in the peripheral blood (circulating microfilariae) of birds belonging to many orders. Current limitations in molecular characterization methods and species identification using morphological characters of circulating microfilariae are major obstacles to improving our understanding the biology of Filarioidea species, particularly in wildlife. The aim of this study was to partially fill these gaps, with particular emphasis on morphological features of microfilariae, which are the most readily accessible stages of these pathogens. Methods Peripheral blood samples of 206 birds belonging to genera Acrocephalus (five species) and Sylvia (five species) were examined using the buffy coat method to process the blood samples for the presence of microfilariae. Positive birds were dissected to collect adult nematodes. Microfilariae and adult nematodes were described, and sequences of their mitochondrial cytochrome c oxidase subunit I and nuclear 28S rDNA gene fragments were obtained and used for molecular characterization and Bayesian phylogenetic inferences. Results Overall prevalence of microfilariae was 2.9%. Microfilariae were found in the blood samples from six birds (2 Acrocephalus scirpaceus and 1 each of A. arundinaceus, Sylvia atricapilla, S. borin and S. curruca), which were dissected. All parasite species observed were new. Eufilaria acrocephalusi sp. n. and Eufilaria sylviae sp. n. were present in subcutaneous, peritracheal and periesophageal connective tissues in A. scirpaceus and S. borin, respectively. Splendidofilaria bartletti sp. n. was found in finger joins of S. atricapilla. Illustrations of microfilariae and adult nematodes are shown, and morphological and phylogenetic analyses identified the DNA barcode haplotypes that are associated with these species. Phylogenetic analysis places the parasites of different genera in different closely related clades. Conclusions Adult nematode morphological characters, which have been traditionally used in the taxonomy of Filarioidea species, have a phylogenetic value. Importantly, in our study parasites of different genera were readily distinguishable based on the morphology of their microfilariae. The link between molecular and morphology data requires more study in Filarioidea species research, particularly because this approach provides new knowledge on species identity using only readily accessible blood stages (microfilariae), thereby avoiding host dissection and thus minimizing harm to wildlife during research.

Purpose: To study clinical and pathological features of parasitic lesions in the ocular adnexa in a tertiary care ophthalmic center in south India.Methods: 43 cases of ocular parasitosis were analysed clinically and correlated with the pathological findings (gross morphology and histopathology) over a period of five years (2015-2020).Results: Among the 43 cases, the age group ranged from 9 months to 78 years (mean age of 41.6 years). Female patients were more common than male patients, with a percentage of 63% (27) and 37% (16) respectively. Cystic lesion in the lid or orbit was seen in 23 cases (53.4%); solid mass lesions were seen in 17 cases (39.5%); subconjunctival worms in three cases; and subretinal parasite in one. Gross examination and histopathologic study showed Dirofilaria in 23 cases (53.5%), followed by Cysticercus in six cases (14%) and Microfilariae in four cases (9.3%). Exact species identification was not possible in ten cases (23.25%). Correlation between the type of lesion and type of inflammatory cells with the specific parasite was done.Conclusion: Our study showed that important clinicopathological correlations can be made from the parasitic lesions in the eye and adnexa, which can aid in definitive diagnosis and prompt identification of the parasite for patient management. [ABSTRACT FROM AUTHOR]

Abstract Background Current measures for the prevention of dirofilariasis, caused by the dog heartworm, Dirofilaria immitis, rely on macrocyclic lactones, but evidence of drug-resistant isolates has called for alternative approaches to disease intervention. As microfilariae are known to be in a state of developmental arrest in their mammalian host and then undergo two molts once inside the arthropod, the aim of this study was to look at the developmental regulation of D. immitis microfilariae that occurs in their arthropod host using in vitro approaches and to investigate the role of the ecdysone signaling system in this development regulation. Methods Dirofilaria immitis microfilariae extracted from dog blood were incubated under various culture conditions to identify those most suitable for in vitro culture and development of the microfilariae, and to determine the effects of fetal bovine serum (FBS), mosquito cells, and ecdysteroid on the development of the microfilariae. Transcript levels of the ecdysone signaling pathway components were measured with droplet digital PCR (ddPCR). Results In vitro conditions that best promote early development of D. immitis microfilariae to the “late sausage stage” have been identified, although shedding of the cuticle was not observed. FBS had inhibitory effects on the development and motility of the microfilariae, but media conditioned with Anopheles gambiae cells were favorable to microfilarial growth. The transcript level study using ddPCR also showed that ecdysone signaling system components were upregulated in developing microfilariae and that 20-hydroxyecdysone increased the proportion of larvae developing to the sausage and late sausage stages in vitro. Conclusions The arthropod host environment provides cues required for the rapid development of D. immitis microfilariae, and the ecdysone signaling system may play an important role in filarial nematode developmental transitions. This study contributes to a better understanding of the developmental process of D. immitis microfilariae.

Abstract Background Pulmonary manifestations are regularly reported in both human and animal filariasis. In human filariasis, the main known lung manifestations are the tropical pulmonary eosinophilia syndrome. Its duration and severity are correlated with the presence of microfilariae. Litomosoides sigmodontis is a filarial parasite residing in the pleural cavity of rodents. This model is widely used to understand the immune mechanisms that are established during infection and for the screening of therapeutic molecules. Some pulmonary manifestations during the patent phase of infection with L. sigmodontis have been described in different rodent hosts more or less permissive to infection. Methods Here, the permissive Mongolian gerbil (Meriones unguiculatus) was infected with L. sigmodontis. Prevalence and density of microfilariae and adult parasites were evaluated. Lungs were analyzed for pathological signatures using immunohistochemistry and 3D imaging techniques (two-photon and light sheet microscopy). Results Microfilaremia in gerbils was correlated with parasite load, as amicrofilaremic individuals had fewer parasites in their pleural cavities. Fibrotic polypoid structures were observed on both pleurae of infected gerbils. Polyps were of variable size and developed from the visceral mesothelium over the entire pleura. The larger polyps were vascularized and strongly infiltrated by immune cells such as eosinophils, macrophages or lymphocytes. The formation of these structures was induced by the presence of adult filariae since small and rare polyps were observed before patency, but they were exacerbated by the presence of gravid females and microfilariae. Conclusions Altogether, these data emphasize the role of host-specific factors in the pathogenesis of filarial infections.

We present a case report of periocular Loa loa. The key feature of L. loa distinguishing it from other human filarial parasites are cuticular bosses, which are presented in images from a light microscope and a scanning electron microscope. The cuticular bosses could be divided into three subtypes not previously described.

Abstract Background Adult worms of Setaria equina mainly found in the peritoneal cavity of equine. They were nonpathogenic but might induce varied degrees of peritonitis and might migrate to the eye, brain, lung, and scrotum causing lacrimation, blindness, paraplegia, locomotor, and neurological disturbances. Identification by light microscopy is insufficient to differentiate Setaria species, and so scanning electron microscopy (SEM) is required to observe their ultrastructures. The study was performed on 80 donkeys from May 2018 to January 2019 for the detection of microfilaria in blood and the adult worms in the peritoneal cavity. The blood samples were either stained with Giemsa stain or examined by modified Knott’s technique for the detection of microfilariae. Adult worms were morphologically characterized based on light microscope and scanning electron microscopy (SEM). PCR was performed targeting the 12S rRNA gene followed by sequencing and phylogenetic analysis. Results The current study recorded 21.6% and 16.2% prevalence rates for adult worms and microfilariae, respectively. By using SEM, this study was able to clarify the detailed structure of amphids, predeirids, vulva, arrangement, and number of male caudal papillae. PCR amplified products for 12S rRNA gene (408 bp) for adult worm and microfilaria. Sequence and phylogenetic analysis revealed that S. equina isolated in the current study from donkeys in Egypt (accession no., MH345965) shared 100% identity with isolates from horse and man in Italy and Iran, respectively and clustered in the same clade with S. digitata, S. tundra and S. labiatopapillosa. Conclusions Identification with light microscopy lacked the ability to characterize different Setaria species, and so using scanning electron microscopy is considered a good choice to distinguish the ultrastructures. In addition, performing the phylogenetic analysis was necessary to detect relationships between different filarial worms, which could not detect by the morphological characterization of adult worms.

The purpose of the research is to study efficiency of drugs for veterinary use "Inspector Quadro C” and "Inspector Quadro K” in the case of dogs’ and cats’ dirofilariosis.Materials and methods. Study was conducted on 24 animals of different breeds, which were naturally infected by Dirofilaria immitis and D. repens, including 12 dogs at the age from 7 weeks to four years with the body weight of 6–20 kg, and 12 cats at the age from 7 weeks to two years with the body weight of 0.7–5 kg. Diagnosis as well as drug efficiency was confirmed in its entirety based on clinical signs and laboratory studies (microscopic evaluation of animals’ blood films). V. B. Yastreb concentration method were used as the method of study. The drug was administered to the animals from experimental group by applying on dry, uninjured skin on places inaccessible for licking off in the dose range of 0.1 ml per 1 kg of animal body weight twice at intervals of 10–14 days which corresponds to the dose for cats 1 mg/kg, for dogs 2.5 mg/kg for moxidectin.Results and discussion. It has been established that drugs "Inspector Quadro C” and "Inspector Quadro K” had demonstrated 100 % efficiency at dirofilariosis disease in dogs and cats. Negative influence of drugs on animals’ organisms was not established.

Filariasis is a common, yet neglected tropical disease and has a wide spectrum of clinical presentations. It is particularly endemic in certain regions of India. We report this case due to the rare isolation of Microfilariae from the bone marrow aspirate preparation. Here, we report a patient with a history of pyrexia of unknown origin (PUO). The relevant investigations done failed to identify the causative organism. After ruling out routine causes of fever, bone marrow analysis was done and it revealed Microfilariae in the aspirate. The surprising aspect of the case is the absence of eosinophilia in the complete blood picture. The patient was treated with anthelmintics and the fever subsided within 48 h. This case emphasizes the importance of bone marrow study in patients with PUO and the presence of microfilaria in the absence of eosinophilia.

Filariasis is one of the common health problems in India. Extranodal filariasis occurs rarely, and the breast is one of the uncommon sites. Here, we report two cases of breast filariasis in the middle-aged females presented with a painless lump in the left breast. These cases were clinically mistaken as a malignancy. Cytology smears showed numerous microfilariae in both coiled and uncoiled forms along with benign ductal cells. Thus, the diagnoses of filariasis breast probably arising from intramammary lymph nodes were made in both cases. The patients were treated conservatively, and the recovery was uneventful. Thus, fine-needle aspiration cytology played an important role to avoid unnecessary surgical intervention in these patients.

Background: Various heartworm (HW) diagnostic testing modalities detect products of, or reactions to, different life cycle stages of Dirofilaria immitis. Microfilariae (Mf) can be directly visualized in blood, antigen (Ag) from immature and adult heartworms may be detected on commercial assays, and antibody (Ab) tests detect the host immune response to larval stages. Ag and Mf tests are commonly used in dogs, which frequently carry adult HW infections, but Ab tests have only been validated for use in cats. In some HW-infected dogs, Ag is blocked by immune complexing leading to false-negative results. Heat-treatment (HT) to disrupt these complexes can increase the sensitivity of HW Ag tests. The aim of this study was to compare different methods for diagnosing HW infection in dogs at high risk using individual and paired diagnostic tests, including an exploration of using Ab tests designed for cats to test canine samples. Methods: One hundred stray adult (≥ 2-year-old) dogs in Florida shelters were tested using Mf, HW Ag, and HW Ab tests (feline HW Ab tests currently not commercially validated/approved for use in dogs); two versions of each test platform were used. Results: Fourteen dogs tested positive using point-of-care (POC) Ag tests; an additional 2 dogs tested positive with microtiter well assay, and an additional 12 dogs tested positive using HT Ag testing. For individual tests, Ag test sensitivity/specificity compared to HT Ag was 50–57%/100%, and Ab tests were 46–64%/82–94%. Sensitivity estimates for individual tests were higher when comparing to non-HT Ag. Pairing POC Ag tests with Mf tests improved sensitivity without loss of specificity, while pairing POC Ag and Ab tests modestly increased sensitivity at the expense of specificity. Conclusions: Screening dogs for HW infection using both POC Ag and Mf detection, which is recommended by the American Heartworm Society, improved diagnostic performance in this study compared to single Ag test use, but may have missed more than one in four infected dogs. The need to improve access to highly accurate, rapid, and inexpensive large-scale HW testing for dogs in animal shelters remains largely unmet by current testing availability. The development of practical and validated protocols that incorporate heat or chemical treatment to disrupt Ag-Ab complexes in POC testing or decreasing the cost and time required for such testing in reference laboratories might provide solutions to this unmet need. Similar studies performed in countries where the prevalence of parasites such as D. repens or A. vasorum is different to the USA could potentially yield very different positive predictive values for both HT and non-HT Ag tests. [ABSTRACT FROM AUTHOR]

Background: Blood parasites have been the subject of much research, with numerous reports of the presence of microfilariae in the peripheral blood (circulating microfilariae) of birds belonging to many orders. Current limitations in molecular characterization methods and species identification using morphological characters of circulating microfilariae are major obstacles to improving our understanding the biology of Filarioidea species, particularly in wildlife. The aim of this study was to partially fill these gaps, with particular emphasis on morphological features of microfilariae, which are the most readily accessible stages of these pathogens. Methods: Peripheral blood samples of 206 birds belonging to genera Acrocephalus (five species) and Sylvia (five species) were examined using the buffy coat method to process the blood samples for the presence of microfilariae. Positive birds were dissected to collect adult nematodes. Microfilariae and adult nematodes were described, and sequences of their mitochondrial cytochrome c oxidase subunit I and nuclear 28S rDNA gene fragments were obtained and used for molecular characterization and Bayesian phylogenetic inferences. Results: Overall prevalence of microfilariae was 2.9%. Microfilariae were found in the blood samples from six birds (2 Acrocephalus scirpaceus and 1 each of A. arundinaceus, Sylvia atricapilla, S. borin and S. curruca), which were dissected. All parasite species observed were new. Eufilaria acrocephalusi sp. n. and Eufilaria sylviae sp. n. were present in subcutaneous, peritracheal and periesophageal connective tissues in A. scirpaceus and S. borin, respectively. Splendidofilaria bartletti sp. n. was found in finger joins of S. atricapilla. Illustrations of microfilariae and adult nematodes are shown, and morphological and phylogenetic analyses identified the DNA barcode haplotypes that are associated with these species. Phylogenetic analysis places the parasites of different genera in different closely related clades. Conclusions: Adult nematode morphological characters, which have been traditionally used in the taxonomy of Filarioidea species, have a phylogenetic value. Importantly, in our study parasites of different genera were readily distinguishable based on the morphology of their microfilariae. The link between molecular and morphology data requires more study in Filarioidea species research, particularly because this approach provides new knowledge on species identity using only readily accessible blood stages (microfilariae), thereby avoiding host dissection and thus minimizing harm to wildlife during research. [ABSTRACT FROM AUTHOR]

Abstract Background Mice are susceptible to infections with the rodent filarial nematode Litomosoides sigmodontis and develop immune responses that resemble those of human filarial infections. Thus, the L. sigmodontis model is used to study filarial immunomodulation, protective immune responses against filariae and to screen drug candidates for human filarial diseases. While previous studies showed that type 2 immune responses are protective against L. sigmodontis, the present study directly compared the impact of eosinophils, IL-5, and the IL-4R on the outcome of L. sigmodontis infection. Methods Susceptible wildtype (WT) BALB/c mice, BALB/c mice lacking eosinophils (dblGATA mice), IL-5−/− mice, IL-4R−/− mice and IL-4R−/−/IL-5−/− mice were infected with L. sigmodontis. Analyses were performed during the peak of microfilaremia in WT animals (71 dpi) as well as after IL-4R−/−/IL-5−/− mice showed a decline in microfilaremia (119 dpi) and included adult worm counts, peripheral blood microfilariae levels, cytokine production from thoracic cavity lavage, the site of adult worm residence, and quantification of major immune cell types within the thoracic cavity and spleen. Results Our study reveals that thoracic cavity eosinophil numbers correlated negatively with the adult worm burden, whereas correlations of alternatively activated macrophage (AAM) numbers with the adult worm burden (positive correlation) were likely attributed to the accompanied changes in eosinophil numbers. IL-4R−/−/IL-5−/− mice exhibited an enhanced embryogenesis achieving the highest microfilaremia with all animals becoming microfilariae positive and had an increased adult worm burden combined with a prolonged adult worm survival. Conclusions These data indicate that mice deficient for IL-4R−/−/IL-5−/− have the highest susceptibility for L. sigmodontis infection, which resulted in an earlier onset of microfilaremia, development of microfilaremia in all animals with highest microfilariae loads, and an extended adult worm survival.

Abstract Background Lymphatic filariasis (LF) is a mosquito-borne parasitic disease which is targeted for elimination as a public health problem worldwide. Niue is a small self-governing South Pacific island nation with approximately 1600 residents that was formerly LF endemic. Here, we review the progress made towards eliminating LF in Niue since 1999. Methods This study has reviewed all the available literature relating to LF in Niue to assess surveillance efforts and the elimination of transmission. Reviewed documentation included both published and unpublished works including historical reports of LF, WHO PacELF records, and Niue Country Reports of the national LF elimination program. Findings Niue conducted mapping of baseline LF endemicity by testing the total present and consenting population for LF antigen with immunochromatographic test (ICT) in 1999, when circulating filarial antigen prevalence was 3.1% (n = 1794). Five nationwide annual mass drug administration (MDA) rounds with albendazole (400 mg) and diethylcarbamazine citrate (DEC) were undertaken from 2000 to 2004, with coverage reported from distribution records ranging from 78 to 99% of the eligible population, which excluded pregnant women and children under 2 years of age. A further whole population survey using ICT in 2001 found 1.3% positive (n = 1630). In 2004, antigen prevalence had reduced to 0.2% (n = 1285). A similar post-MDA survey in 2009 indicated antigen prevalence to be 0.5% (n = 1378). Seven positive cases were re-tested and re-treated every six months until negative. Conclusions After five rounds of MDA, Niue had reduced the LF antigen population prevalence in all ages from 3.1% to below 1% and maintained this prevalence for a further five years. Due to Niue’s small population, surveillance was done by whole population surveys. Niue’s results support the WHO recommended strategy that five to six rounds of annual MDA with effective population coverage can successfully interrupt the transmission of LF. Niue received official acknowledgement of the validation of elimination of LF as a public health problem by the WHO Director-General and WHO Western Pacific Regional Office (WPRO) Regional Director at the 67th session of the Regional Committee for the Western Pacific held in Manila in October 2016.

India has still not eradicated filariasis, which is still endemic throughout the country. This study focuses on the breast as a non-typical but not uncommon site of breast disease characterized by nematodes causing obstruction of lymphatic vessels caused by filariasis. The case described herein involves a 26-year-old married female who presented with a subcutaneous nodule in her left breast. Clinical interrogation of this case as part of the pathological diagnosis, phlebotomy was performed by venepuncture in order to examine peripheral blood smears under a microscope. Nonetheless, the peripheral blood smear did not show microfilariae of the disease. Furthermore, fine needle aspiration cytology was also performed to identify possible things for the study. It was diagnosed that the fine-needle aspiration cytology smears of various lesions indicate the lump carried microfilariae. Conclusively the lymphatic swelling was confirmed to be caused by Wuchereria bancrofti in fine-needle aspiration cytology examination without evidence in peripheral blood smears. It was shown that the subject had filariasis, and the patient responded well to daily treatment with diethylcarbamazine citrate for 42 days. [ABSTRACT FROM AUTHOR]

Dirofilaria immitis causes life-threatening heart disease in dogs, thus screening of dog populations is important. Lens-free technology (LFT) is a low-cost imaging technique based on light diffraction that allows computerized recognition of small objects in holographic images. We evaluated an algorithm capable of recognizing microfilariae in canine whole blood using the LFT. We examined 3 groups of 10 EDTA blood specimens, from dogs with microfilaremia (group A), healthy dogs (B), and dogs with hematologic modifications other than microfilaremia (C). The LFT analyzer photographed repeated series of 5 images of all samples. The algorithm declared a sample positive if a microfilaria was detected on ≥1, ≥2, or ≥3 of the 5 images of a series. Microfilariae were detected visually in the images in 9 of 10 cases in group A; no microfilariae were seen in the images from groups B and C. Of the 30 cases, there were 14, 4, and only 3 false-positives with the 1 of 5, 2 of 5, and 3 of 5 image cutoffs, respectively. There were no false-negatives, regardless of cutoff. LFT seems useful for detecting microfilaria and could have application in clinical pathology. [ABSTRACT FROM AUTHOR]

Lymphatic filariasis is a neglected parasitic disease that is a leading cause of long-term disability. Information obtained from genome sequencing of filarial worm can help us identify systems in the worm that are likely to be useful for novel drug design. Brugia (B.) malayi is still the only lymphatic-dwelling filarial parasite with a nearly complete, fully annotated, and published genome. However, most previous studies were based on the FR3 strain of B. malayi, which originally was isolated from a human patient, and was adapted to the rodent model, then maintained in laboratories for more than 60 years. It is uncertain whether genetic variation exists, thus, sequencing of clinical isolates of lymphatic dwelling filarial parasites is a high priority. Here, we report for the first time the complete mitochondrial genome of B. malayi microfilariae from clinical isolate. Complete mitochondrial (mt) genome of the microfilariae isolated from a blood sample taken from a Thai subject living in Narathiwat Province, which is an endemic area of brugian filariasis, was assembled with sequencing reads obtained by Illumina sequencing. Gene annotation, phylogenetic analysis and single nucleotide polymorphism (SNP) were deployed. A complete 13,658-bp mt genome of B. malayi microfilaria was obtained, and it shows 68x coverage. Based on gene annotation, the mt genome consists of 12 protein-coding, two rRNA, and 23 tRNA genes. Phylogenetic analysis using all protein sequences of DNA sequences of mt genome or cytochrome c oxidase subunit I (COX1) revealed a close relationship among three lymphatic filariae (i.e., B. timori, zoonotic B. pahangi, and Wuchereria spp.). The SNPs in the COX1 gene can differentiate microfilariae of B. malayi in human from those found in canine. Furthermore, the number, order and transcription, and direction of B. malayi microfilariae mitochondrial genes were the same as those found in the FR3 strain of B. malayi. The comparison on mitochondrial genome of B. malayi could have important implications on the development of a new intervention or vaccine to treat or prevent this disease in endemic areas/regions around the world.

Summary: Eosinophils mediate protection against filarial nematodes. Our results demonstrate that eosinophil extracellular traps (EETosis) are induced by microfilariae and infective L3 larvae of Litomosoides sigmodontis. These extracellular DNA traps inhibit microfilariae motility in a DNA- and contact-dependent manner in vitro. Accordingly, microfilariae-injection triggers DNA release in an eosinophil-dependent manner in vivo and microfilariae covered with DNA traps are cleared more rapidly. Using dectin-1, we identify the required receptor for the microfilariae-induced EETosis, whereas signaling via other C-type lectin receptors, prior priming of eosinophils, and presence of antibodies are not required. The DNA released upon microfilariae-induced EETosis is mainly of mitochondrial origin, but acetylated and citrullinated histones are found within the traps. We further demonstrate that the presented DNA-dependent inhibition of microfilariae motility by eosinophils represents a conserved mechanism, as microfilariae from L. sigmodontis and the canine heartworm Dirofilaria immitis induce ETosis in murine and human eosinophils.