Successful Transvenous Extraction of Adult Dirofilaria immitis Parasites from a Naturally Infected Domestic Ferret (Mustela putorius furo).

Heartworm disease caused by Dirofilaria immitis is a serious and underdiagnosed cardiovascular condition in domestic ferrets. Hemodynamic changes caused by parasitization in ferrets cause a potentially fatal syndrome, but its clinical findings and treatment have not yet been standardized. The objective of this study was to describe the clinical case of a successful surgical extraction in a ferret infected by D. immitis. The patient was a 2-year-old, 1.5 kg asymptomatic male domestic ferret. The infection was diagnosed using a commercial test for the detection of D. immitis antigens. Subsequently, their clinical status was evaluated using serological and imaging diagnostic tests, and it was finally decided to perform surgical extraction of the adult worms. The ferret was anesthetized and placed in the left lateral decubitus position to perform a venotomy in the right jugular vein. Endoscopic extraction basket-shaped devices were used in the right atrial cavity under fluoroscopic guidance following the Seldinger endovascular surgery technique. With careful handling, two adult parasites were manually removed. A transthoracic echocardiogram performed after the procedure confirmed the absence of heartworms. The ferret recovered without complications and was discharged within 24 h. In the clinical review, 30 days after surgery, no notable alterations or symptoms were observed. This case report describes the first complete surgical removal of adult D. immitis parasites in a naturally infected ferret.

Tick-associated Trypanosoma species (Apicomplexa: Kinetoplastida) from cattle ticks (Acari: Ixodidae) in Peninsular Malaysia.

A Trypanosoma screening was conducted on 130 pools comprising 1,241 ticks, collected from 674 selected farm ruminants in Peninsular Malaysia. Of these, nine pools were tested positive for Trypanosoma. Subsequent BLAST searches revealed that the 18S rRNA gene sequences were closely related to Trypanosoma rhipicephalis isolate Chaco CB, with percentage similarities ranging from 95.56 % to 99.84 %. Phylogenetic analysis showed that three of the nine sequences formed a clade with Trypanosoma rhipicephalis. The remaining six Trypanosoma sequences formed a distinct clade, separate from T. rhipicephalis and other Trypanosoma species, with genetic distances of 4.34 % and 4.33-4.58 %, respectively. This study marks the first report of tick-associated Trypanosoma in Malaysia and underscores significant research gaps regarding trypanosome interactions with tick hosts in the region.

Lipids from a snail host regulate the multicellular behavior of a predator of parasitic schistosomes.

Transmission of vector-borne diseases can be slowed by symbionts within the secondary hosts that spread disease. Snails spread schistosomiasis, and the snail symbiont Capsaspora owczarzaki kills schistosome larvae. In studying how Capsaspora colonizes its host snail, we discovered that Capsaspora responded to its host by forming multicellular aggregates. We elucidated the chemical cue for aggregation: hemolymph phosphatidylcholines (PCs). Furthermore, we uncovered that Capsaspora cells aggregate to different degrees in sera from different host snails-and these responses correlate with serum concentrations of PCs. Therefore, Capsaspora senses a host factor that can indicate the identity and physiological state of its host. Since cellular aggregation controls microbial motility, feeding, and immune evasion, this response within host tissue may be important for colonization. If so, snail serum PC and Capsaspora aggregation will be molecular and cellular markers to discern which conditions will favor the colonization of snails (and potential exclusion of schistosomes) by Capsaspora.

Autochthonous Human Babesia divergens Infection, England.

We describe a case of autochthonous human Babesia divergens infection in an immunocompetent woman in England. The patient had fever, hemolysis, multiorgan failure, and 18% parasitemia. We confirmed B. divergens by 18S rDNA PCR and sequencing. Clinicians should consider babesiosis as a differential diagnosis in patients with unexplained hemolysis.

Two cases of Angiostrongylus vasorum, a cardiopulmonary nematode, in a wild black bear and coyote of Tennessee.

Angiostrongylus vasorum, commonly known as the French heartworm, is a metastrongyloid parasitic nematode that infects wild and domestic canids. In North America, A. vasorum is endemic to the Canadian island of Newfoundland, but has been expanding to new areas including Nova Scotia, Prince Edward Island, and West Virginia (USA). Two cases of A. vasorum are reported from the state of Tennessee. The first case in a black bear (Ursus americanus) and the second case in a coyote (Canis latrans). The black bear was found dead in Sevier County in November of 2022, while the coyote was trapped and euthanized as part of a predator control program in Campbell County in January of 2023. Histology of the lungs revealed both animals had verminous pneumonia. DNA was extracted from the lungs of both, and PCR was performed using NC1 and NC2 primers. Sequencing results of the PCR products from the bear and coyote samples indicated that they were 95% and 96% similar, respectively, to European strains of A. vasorum. This report marks the first time A. vasorum has been reported in Tennessee as well as only the second and third report of autochthonous A. vasorum infection in the United States and the first report in an ursid. These two cases confirm the spread of A. vasorum further into North America. This nematode is highly pathogenic to wild and domestic canids, and thus these cases represent an emerging threat to both and underscore the need for further surveillance for the parasite.

Descriptive and molecular epidemiology of leishmaniasis diagnosed from clinical samples in the United States, 2021-2022.

Leishmaniasis is a rare disease in the United States, with an estimated annual incidence of dozens of cases occurring primarily in travelers, migrants, and military personnel. True disease incidence is unknown, since leishmaniasis is not a nationally notifiable condition. Here, we describe the results of molecular leishmaniasis over a 1-year interval (September 2021 to August 2022) when our laboratory served as the primary national reference laboratory for molecular diagnosis of civilian leishmaniasis. We tested 218 specimens submitted from 36 states yielding 94 of the 186 (50.5%) positive cases with species or species complex-level identification and 18 novel mini-exon alleles. Most species belonged to subgenus Viannia (75.6%) and associated with cutaneous or mucocutaneous disease. Cases were associated with recent travel (18.1%), travel timing unspecified (7.4%), migration (7.4%), remote travel (2.1%), military (1.1%), or unknown history (63.8%). These data illustrate the clinical utility of molecular testing for leishmaniasis and provide unique insight into disease epidemiology. IMPORTANCE: Leishmaniasis is a disfiguring, neglected parasitic infection endemic to the Southern United States and the Americas. Despite significant populations at risk-travelers, military and foreign service members, and migrating persons-the epidemiology of the disease in the United States is poorly understood. Moreover, few clinical laboratories in the United States can test for the disease. Here, we present results from 1 year of testing for this disease at a major reference laboratory. These findings are particularly relevant because they coincide with a temporary "pause" on all clinical testing at the CDC. Our findings suggest at least several hundred cases occur each year in the United States. In particular, mucosal leishmaniasis may be more common than previously reported. We also highlight greater genetic diversity in Leishmania species endemic to the Americas than has been previously sampled, with implications for diagnostic specificity.

Preparing eugregarine parasites and their cricket host Acheta domesticus as a model for gregarine infection studies.

The increasing global demand for house crickets (Acheta domesticus) necessitates effective health measures. Despite their abundance, the effects of gregarines on these hosts remain underexplored. We present a method for controlled gregarine infection and maintenance of gregarine-free cricket populations. This system, adapted from prior work, is essential for experiments on gregarine infection dynamics, providing insights into parasite evolution and host-parasite interactions within the Apicomplexa group. This protocol includes rearing and maintenance of gregarine-free cricket populations for experimental purposes, gregarine production for oocyst solution and cricket infection, and gregarine infection assessment.

ABIOTIC AND DEMOGRAPHIC DRIVERS OF FLEA PARASITISM ON DEER MICE IN A RECOVERING MIXED-CONIFER FOREST A DECADE POSTFIRE.

With the intensity and frequency of wildfires increasing rapidly, the need to study the ecological effects of these wildfires is also growing. An understudied aspect of fire ecology is the effect fires have on parasite-host interactions, including ectoparasites that might be pathogen vectors. Although some studies have examined the impacts of fire on ticks, studies on other ectoparasites, including pathogen vectors, are rare. To help address this knowledge gap, we examined the abiotic and biotic factors that predict the likelihood and extent of parasitism of deer mice (Peromyscus maniculatus) by fleas within a landscape of unburned and recovering burned (>9 yr postfire) mixed conifer forests. We sampled 227 individual deer mice across 27 sites within the Jemez Mountains of northern New Mexico in 2022 and quantified measures of parasitism by fleas (primarily Aetheca wagneri). These sites were distributed in both unburned areas (n = 15) and recovering burned areas (n = 12), with the latter derived from 2 large fires, the Las Conchas fire (2011) and the Thompson Ridge fire (2013). Using these data, we tested for differences in prevalence, mean abundance, and mean intensity of fleas on deer mice, focusing on the predictive importance of host sex and fire history. We also created generalized linear mixed-effects models to investigate the best host and environmental predictors of parasitism by fleas. Approximately a decade postfire, we found minimal evidence to suggest that fire history influenced either the presence or intensity of fleas on deer mice. Rather, at the current forest-regeneration stage, the extent of parasitism by fleas was best predicted by measures of host sex, body condition, and the trapline's ability to accumulate water, as measured through topography. As host body condition increased, the probability of males being parasitized increased, whereas the opposite pattern was seen for females. Male mice also had significantly greater flea loads. Among potential abiotic predictors, the topographic wetness index or compound topographic index (a proxy for soil moisture) was positively related to flea intensity, suggesting larger flea populations in burrows with higher relative humidity. In summary, although fire may potentially have short-term impacts on the likelihood and extent of host parasitism by fleas, in this recovering study system, host characteristics and topographic wetness index are the primary predictors of parasitism by fleas.

Assessing de novo parasite genomes assembled using only Oxford Nanopore Technologies MinION data.

In this study, we assessed the quality of de novo genome assemblies for three species of parasitic nematodes (Brugia malayi, Trichuris trichiura, and Ancylostoma caninum) generated using only Oxford Nanopore Technologies MinION data. Assemblies were compared to current reference genomes and against additional assemblies that were supplemented with short-read Illumina data through polishing or hybrid assembly approaches. For each species, assemblies generated using only MinION data had similar or superior measures of contiguity, completeness, and gene content. In terms of gene composition, depending on the species, between 88.9 and 97.6% of complete coding sequences predicted in MinION data only assemblies were identical to those predicted in assemblies polished with Illumina data. Polishing MinION data only assemblies with Illumina data therefore improved gene-level accuracy to a degree. Furthermore, modified DNA extraction and library preparation protocols produced sufficient genomic DNA from B. malayi and T. trichiura to generate de novo assemblies from individual specimens.

Correlating Cases of Soil-Transmitted Helminths and Remote Sensing Covariates: A Case Study Utilizing 2015, 2017, and 2019 Metadata of Camarines Sur, Philippines.

INTRODUCTION: Soil-transmitted helminth (STH) is a group of helminths that are considered to be neglected tropical diseases (NTDs) and, globally, affect more than 1.5 to 2.6 million people yearly. Depending on the species, they can be acquired by ingesting embryonated eggs from contaminated matter or by skin penetration. Most species of STH are found in the tropics, such as the Philippines. Despite the Mass Drug Administration (MDA), the cases of STH infection continue to rise in the country. Surveys from the Research Institute of Tropical Medicine (RITM) indicate that a high prevalence of STH (Ascaris lumbricoides, Trichuris trichiura, and Necator americanus) was primarily observed in the provinces of the country, such as in Camarines Sur. OBJECTIVES: To correlate remote sensing covariates such as Normalized Difference Vegetation Index (NDVI) and Normalized Difference Built-up Index (NDBI) - to STH-infected cases of the 37 municipalities of Camarines Sur. METHODOLOGY: The available public health record of STH cases from 2015 to 2019 were calculated using the Quantum Geographic Information System (QGIS)and correlated using Pearson Correlation Coefficient. RESULTS: The results showed that infection was higher in children than adults, and A. lumbricoides caused 60% of infection. No correlation of indices with infection cases during 2015 and 2017 was observed; however, 2019 showed a moderate strength (p = 0.037) in correlation. CONCLUSION: This indicates that infection relied not mainly on vegetation and urbanization but on additional environmental factors and non-environmental variables.

Fish ectoparasite detection, collection and curation.

Fish parasitology is a dynamic and internationally important discipline with numerous biological, ecological and practical applications. We reviewed optimal fish and parasite sampling methods for key ectoparasite phyla (i.e. Ciliophora, Platyhelminthes, Annelida and Arthropoda) as well as recent advances in molecular detection of ectoparasites in aquatic environments. Ideally, fish capture and anaesthesia as well as parasite recovery methods should be validated to eliminate potential sampling bias and inaccuracy in determining ectoparasite population parameters. There are considerable advantages to working with fresh samples and live parasites, when combined with appropriate fixation methods, as sampling using dead or decaying materials can lead to rapid decomposition of soft-bodied parasites and subsequent challenges for identification. Sampling methods differ between target phyla, and sometimes genera, with optimum techniques largely associated with identification of parasite microhabitat and the method of attachment. International advances in fish parasitology can be achieved through the accession of whole specimens and/or molecular voucher specimens (i.e. hologenophores) in curated collections for further study. This approach is now critical for data quality because of the increased application of environmental DNA (eDNA) for the detection and surveillance of parasites in aquatic environments where the whole organism may be unavailable. Optimal fish parasite sampling methods are emphasised to aid repeatability and reliability of parasitological studies that require accurate biodiversity and impact assessments, as well as precise surveillance and diagnostics.

Seek, and you will find: Cryptic diversity of the cardiopulmonary nematode Angiostrongylus vasorum in the Americas.

Angiostrongylus vasorum is a metastrongylid parasite infecting wild canids and domestic dogs. Its patchy distribution, high pathogenicity and taxonomical classification makes the evolutionary history of A. vasorum intriguing and important to study. First larval stages of A. vasorum were recovered from feces of two grey foxes, Urocyon cinereoargenteus, from Costa Rica. Sequencing and phylogenetic and haplotypic analyses of the ITS2, 18S and cytochrome oxidase subunit 1 (cox1) fragments were performed. Then p- and Nei´s genetic distance, nucleotide substitution rates and species delimitation analyses were conducted with cox1 data of the specimens collected herein and other Angiostrongylus spp. Cophylogenetic congruence and coevolutionary events of Angiostrongylus spp. and their hosts were evaluated using patristic and phenetic distances and maximum parsimony reconciliations. Specimens from Costa Rica clustered in a separate branch from European and Brazilian A. vasorum sequences in the phylogenetic and haplotype network analyses using the ITS2 and cox1 data. In addition, cox1 p-distance of the sequences derived from Costa Rica were up to 8.6 % different to the ones from Europe and Brazil, a finding mirrored in Nei´s genetic distance PCoA. Species delimitation analysis supported a separate group with the sequences from Costa Rica, suggesting that these worms may represent cryptic variants of A. vasorum, a new undescribed taxon or Angiocaulus raillieti, a synonym species of A. vasorum described in Brazil. Moreover, nucleotide substitution rates in A. vasorum were up to six times higher than in the congener Angiostrongylus cantonensis. This finding and the long time elapsed since the last common ancestor between both species may explain the larger diversity in A. vasorum. Finally, cophylogenetic congruence was observed between Angiostrongylus spp. and their hosts, with cospeciation events occurring at deeper taxonomic branching of host order. Altogether, our data suggest that the diversity of the genus Angiostrongylus is larger than expected, since additional species may be circulating in wild canids from the Americas.

Evaluation of Seroprevalence and Risk Factors of Heartworm Infection for Dogs in Rio de Janeiro with Access to Veterinary Care.

Heartworm infection is a chronic disease with clinical signs and effects ranging from an asymptomatic condition to severe disease and death. The prevalence of heartworm disease in the state of Rio de Janeiro has been reported to be high (21.3%). The present study was conducted to evaluate the seroprevalence and risk factors of heartworm infection for the canine population with access to veterinary services in different areas of the state of Rio de Janeiro, Brazil. A total of 1787 canine blood samples were obtained from 135 practices across 8 different areas of Rio de Janeiro state (Rio de Janeiro municipality, São Gonçalo municipality, Niterói municipality, Baixada Fluminense, and the northern, southern, eastern, and mountainous areas) and tested for the presence of Dirofilaria immitis antigens and antibodies against several tick-borne disease pathogens using a commercial immunochromatography technique (Vetscan® Flex 4 Rapid Test; Zoetis; NJ USA). Pet owners reported living conditions, physical characteristics, demographics, and clinical signs for evaluation of risk factors for heartworm infection. Only two evaluated risk factors were shown to enhance the risk for D. immitis infection, including having a short hair coat vs. having a medium or long hair coat (OR 2.62) or positive for antibodies to tick-borne disease parasites (OR 3.83). Clinical signs reported for dogs with heartworm disease were typical for that condition. The overall prevalence of heartworm disease in the state was 8.2%, ranging from 2.4% in the mountainous region to 29.4% in the eastern area. It could not be determined if veterinarians were not diligent about dispensing heartworm preventatives or if poor levels of compliance by dog owners were responsible for higher infection rates in some areas of the state.

Current status of immunodeficient mouse models as substitutes to reduce cat and dog use in heartworm preclinical research.

Chemoprophylactic prevention of veterinary heartworm disease in companion animals, caused by the vector-borne nematode parasite Dirofilaria immitis, is a multi-billion-dollar global market. Experimental use of cats and dogs in preclinical heartworm drug testing is increasing due to evolving drug-resistance to frontline macrocyclic lactones and renewed investment in alternative preventative drug research. We and others recently published data demonstrating proof-of-concept of utilising lymphopenic severe-combined immunodeficient (SCID) or Recombination Activating Gene (RAG)2 deficient mice with additional knockout of the IL-2/7 receptor gamma chain (γc) as alternative preventative drug screening research models of dirofilariasis. Here we summarise the current knowledge of candidate immunodeficient mouse models tested, including a comparison of susceptibility using different background strains of mice, different D. immitis isolates, following use of anti-inflammatory treatments to further suppress residual innate immunity, and efficacies achieved against different reference anthelmintics. We supplement this precis with new data on treatment response to the veterinary anthelmintic, oxfendazole, and initial evaluation of D. immitis susceptibility in CB.17 SCID and C57BL/6 RAG2 -/-γc -/- mice. We conclude that in addition to NSG and NXG mice, RAG2 -/-γc -/- mice on either a BALB/c or C57BL/6 background offer an alternative screening model option, widening access to academic and commercial laboratories wishing to pursue initial rapid in vivo drug screening whilst avoiding potentially unnecessary cat or dog testing.

The Bibliometric Analysis of the Postgraduate Theses Written on Medical Parasitology in Türkiye.

Objective: The aim of this study was to analyze the theses prepared in the field of medical parasitology in Türkiye and to reveal the importance given to the science of parasitology in the groves of academe and to raise awareness in this field. Methods: Council of Higher Education's National Thesis Center database has been analyzed postgraduate these documents conducted in the field of medical parasitology from January 1985 to September 2022. Results: As a result of the examining, 393 theses made in the field of medical parasitology were detected. It was determined that 52.9% of the theses prepared were master, 28% of PhD and 19.1% were the thesis of medical specialty thesis and 61.3% of the theses prepared were related to protozoa, 16.5% of helminths, 8.6% arthropods and 12.2% of intestinal parasites (helminth and protozoa). The top five types of parasites in the theses were Toxoplasma gondii, Leishmania spp., Echinococcus spp., Giardia intestinalis and Cryptosporidium spp. respectively. Conclusion: In conclusion, it was of the opinion that more importance should be given to the field of medical parasitology in Türkiye.

Non-conformities in the Pre-analytical Phase at the Parasitology-Mycology Laboratory of the Mohammed VI University Hospital in Oujda.

INTRODUCTION: The Parasitology-Mycology Laboratory's analytical process involves three stages: pre-analytical, analytical, and post-analytical. Our focus is on the pre-analytical phase (PAP). This study addresses managing PAP non-conformities at Mohammed VI University Hospital in Oujda, aligning with quality standards like ISO 15189 and GBEA and aiming to detect and resolve deviations. METHODS: This 84-month retrospective study analyzed specimens at the Parasitology-Mycology lab in the Mohammed VI University Hospital in Oujda. Examination requests were made through the hospital's IT system (HOSIX), and samples were transported pneumatically. After administrative and technical checks, samples were rejected, processed, or retained for correction based on findings. Reports of non-conformities were sent to prescribers via the IT system. Data were analyzed and flowcharts were created using Microsoft Excel (Redmond, USA). RESULTS AND DISCUSSION: During the study period, prescription errors were the most common non-conformities (65.88%; n=56), followed by sample nature errors (29.41%; n=25) and sample packaging errors (4.70%; n=4). Prescription discrepancies, mycological exams for patients on antifungal treatment or carrying Henna, and missing clinical information were the main causes. Outpatient samples accounted for 29.41% of non-conformities, while inpatient samples accounted for 70.59%. The majority of inpatient non-conformities came from the dermatology department (n=42; 49.41%). The pre-analytical phase in Parasitology-Mycology is crucial for ensuring accurate results, involving the coordination of various stages such as staff training, documentation, and non-conformity management. Prescription errors were predominant among non-conformities, followed by sample nature and packaging errors. Outpatient samples had fewer non-conformities compared to inpatient ones, possibly due to supervision by a biologist. Non-conformities lead to therapeutic, prognostic, and economic issues, underscoring the need for their reduction. Corrective actions are crucial, along with establishing policies for error detection and control. Potential causes of non-conformities can be analyzed using methods like the 5M approach. Suggestions for improvement include distributing a validated sampling manual, creating electronic test request forms, staff training, ongoing training programs, and regular meetings for information exchange. CONCLUSION: The pre-analytical phase in Parasitology-Mycology is crucial, demanding a quality-focused approach for strict adherence to procedures and traceability. Mastery of this phase ensures result reliability.

From the bench to the farm and back again.

My path to becoming a scientist has taken many twists and turns. This is perhaps not unusual to hear. Indeed, in discussions with my colleagues it seems that for many of us the path was never a straight one. Certainly, for me there have been moments when my whole world was encompassed by science and at other times, I have felt strongly that my time in science was up. I like to think that as scientists we ask a lot of questions and, for many of us, those questions extend to our very purpose as a scientist. My intention with this article is not to document my career path in detail or to provide very specific advice. Rather, I hope to describe how questions have defined my journey and to inspire others to occasionally pause and ask themselves what a career in science means to them. Today, I am an Assistant Professor at a major Canadian university, and here are the questions I asked along the way.