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.

Cross-sectional study to investigate the seroprevalence and risk factors of Toxoplasma gondii among women attending the antenatal clinic in Namwala, Zambia.

BACKGROUND: Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii (T. gondii). It has a wide host range and is capable of vertical transmission in pregnant women, which may lead to undesirable pregnancy outcomes such as congenital malformations, miscarriage, premature birth and stillbirth. This study investigated the seroprevalence of T. gondii infection among pregnant women attending the antenatal clinic at Namwala District Hospital in Southern Zambia. METHODS: This was a cross-sectional study where blood was collected, and the serum was tested for Toxoplasma IgG and IgM. A questionnaire was administered to participants on demographic characteristics and risk factors. Data were entered in Microsoft Excel and exported to STATA version 14 for analysis. RESULTS: A total of 401 women were enrolled in the study from 3 March to 5 August 2021. The seroprevalence of Toxoplasma IgG was 4.2% (n=17), while the seroprevalence of Toxoplasma IgM was 0.7% (n=3). The median age was 27 (IQR: 24-30) years, and a larger proportion had primary-level education (n=223, 55.6%). The majority (81.6%) of the women were married. None of the risk factors investigated in this study were significant for T. gondii infection. CONCLUSION: There was a low seroprevalence of T. gondii infection among pregnant women in the Namwala district of Southern Province, Zambia, and regular screening may not be warranted in this population. Continued research on toxoplasmosis is recommended to understand its epidemiology across Zambia.

Imaging Assays to Detect DNA Damage in Trypanosome Parasites Using γH2A.

Diseases caused by trypanosomatid parasites remain a significant unmet medical need for millions of people globally. Trypanosomatid parasites such as Trypanosoma cruzi and subspecies of Trypanosoma brucei cause Chagas disease and human African trypanosomiasis (HAT), respectively. Although efforts to find novel treatments have been successful for HAT, Chagas disease is still treated with decades-old therapies that suffer from long treatment durations and severe safety concerns. We recently described the identification and characterization of the cyanotriazole compound class that kills trypanosomes, in vitro and in vivo, by selective inhibition of the trypanosome nuclear topoisomerase II enzyme. To evaluate whether inhibition of the topoisomerase II enzyme led to parasite death due to lethal double-strand DNA breaks, we developed assays for detecting DNA damage in both intracellular amastigotes of T. cruzi and bloodstream-form T. brucei by using the canonical DNA damage marker γH2A. Herein, this article describes the protocols for detecting DNA damage using an immunofluorescence assessment of γH2A by microscopy in trypanosome parasites. Key features • Immunofluorescence-based assay to detect the γH2A response in T. brucei and T. cruzi parasites. • Robust DNA damage pathway-based cellular assays to evaluate topoisomerase II poisons' ability to cause DNA damage. • A 384-well plate-based T. cruzi protocol allows high-resolution and high-throughput evaluation of compounds that cause DNA damage by measuring γH2A in intracellular parasites. • This assay could be modifiable for evaluation of DNA damage responses in various intracellular and extracellular eukaryotic pathogens.

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.

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.

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.

Post operative abdominal wall mucormycosis infection after laparotomy for bowel perforation.

Mucormycosis is a devastating disease with a high mortality rate, typically affecting immunosuppressed individuals. Postoperative surgical site infections due to mucromycosis are rare, with only a handful of cases reported in the literature. Here, we describe a fatal case of post operative abdominal wound infection caused by mucormycosis in an immunocompetent man in his 70 s, who developed the infection following a laparotomy for bowel perforation. Initially, the growth of fungal species from a superficial wound swab was not considered significant until the patient exhibited signs of worsening sepsis. Limited operative debridement was performed for prognostication, in accordance with the family's wishes. There was evidence of extensive significant invasive fungal infection, marked by necrosis extending into the abdominal wall fat and muscle. The patient was then transitioned to comfort measures and subsequently died. This case emphasizes the importance of maintaining a high level of clinical suspicion for mucormycosis, even in patients with minimal risk factors, and highlights the importance of prompt and aggressive treatment.

Blood Fluke Infection (Spirorchidiasis) and Systemic Granulomatous Inflammation: A Case Study of Green Sea Turtles (Chelonia mydas) on Jeju Island, South Korea.

Despite the precarious state of marine turtles as a highly endangered species, our understanding of their diseases remains limited. This case report presents a detailed pathological investigation of spirorchiidiasis, a blood fluke infection that poses a substantial threat to marine turtles. This retrospective study examined three cases of spirorchiid-infected sea turtles, specifically, green sea turtles stranded on Jeju Island, South Korea. Premortem examination of the three spirorchiid-infected green sea turtles demonstrated nonspecific clinical symptoms; blood analysis revealed dehydration, malnutrition, and anemia. Computed tomography scans provided insights into severe pulmonary and extrapulmonary manifestations, including the mass present in the joint region. Post-mortem examinations consistently indicated severe lung lesions and systemic manifestations, with histopathological examination confirming the presence of spirorchiid ova across various organs. Despite the global prevalence of spirorchiidiasis in sea turtles, disease severity varies regionally. This report provides a detailed demonstration of the pathology of spirorchiidiasis in sea turtles from Northeast Asia.

Urogenital schistosomiasis among adult male population in an endemic area of southern Tanzania: a descriptive cross-sectional study.

BACKGROUND: Urogenital schistosomiasis (UGS) caused by Schistosoma haematobium is endemic in Southern Tanzania. The disease has significant implications for both socioeconomic and public health. Because infections with S. haematobium usually peak in childhood, the majority of studies have concentrated on school-aged children leaving other groups such as males which might be continuous reservoir of infection transmission. However, despite its chronic consequences in the male population, the disease has received insufficient attention, especially in sub-Saharan Africa. This study was conducted to describe the previous and current schistosomiasis status among adult males living in high-endemic areas of southern Tanzania DESIGN, SETTING AND PARTICIPANTS: A descriptive cross-sectional study was employed to gather data on the prevalence of UGS among adult men residing at schistosomiasis endemic in the Mtama District Council. Quantitative methods of data collection which included questionnaire and laboratory procedures were used. RESULTS: Out of 245 participants, macrohaematuria and microhaematuria were found in 12 (4.9%, 95% CI 2.4% to 7.8%) and 66 (26.9%, 95% CI 21.6% to 32.7%) participants, respectively. S. haematobium ova were recovered from the urine samples of 54 (22.0%, 95% CI 16.7% to 27.3%) participants. The median intensity of infection was 20 eggs per 10 mL of urine ranging from 1 to 201 eggs per 10 mL of urine (IQR) 60.5). Out of 245 participants 33 (13.5% 95% CI 9.0% to 17.6%) had light intensity of infection and 21 (38.9%, 95% CI; 25.0% to 52.5%) had heavy intensity of infection. Overall, the prevalence of heavy intensity of infection was 8.6% (95% CI 4.9% to 12.6%). The prevalence and intensity of UGS varied significantly by age, marital status and village of residence. CONCLUSION: This study sheds light on the prevalence of UGS among adult males in endemic areas of southern Tanzania. The results highlight the urgent need for comprehensive intervention strategies to address the burden of the disease.

Zipime-Weka-Schista study protocol: a longitudinal cohort study and economic evaluation of an integrated home-based approach for genital multipathogen screening in women, including female genital schistosomiasis, human papillomavirus, Trichomonas and HIV in Zambia.

INTRODUCTION: Multiplathogen home-based self-sampling offers an opportunity to increase access to screening and treatment in endemic settings with high coinfection prevalence of sexually transmitted (HIV, Trichomonas vaginalis (Tv), human papillomavirus (HPV)) and non-sexually transmitted pathogens (Schistosoma haematobium (Sh)). Chronic coinfections may lead to disability (female genital schistosomiasis) and death (cervical cancer). The Zipime-Weka-Schista (Do self-testing sister!) study aims to evaluate the validity, acceptability, uptake, impact and cost-effectiveness of multipathogen self-sampling for genital infections among women in Zambia. METHODS AND ANALYSIS: This is a longitudinal cohort study aiming to enrol 2500 non-pregnant, sexually active and non-menstruating women aged 15-50 years from two districts in Zambia with 2-year follow-up. During home visits, community health workers offer HIV and Tv self-testing and cervicovaginal self-swabs for (1) HPV by GeneXpert and, (2) Sh DNA detection by conventional (PCR)and isothermal (recombinase polymerase assay) molecular methods. Schistosoma ova and circulating anodic antigen are detected in urine. At a clinic follow-up, midwives perform the same procedures and obtain hand-held colposcopic images. High-risk HPV positive women are referred for a two-quadrant cervical biopsy according to age and HIV status. A cost-effectiveness analysis is conducted in parallel. ETHICS AND DISSEMINATION: The University of Zambia Biomedical Research Ethics Committee (UNZABREC) (reference: 1858-2021), the London School of Hygiene and Tropical Medicine (reference: 25258), Ministry of Health and local superintendents approved the study in September 2021.Written informed consent was obtained from all participants prior to enrolment. Identifiable data collected are stored securely and their confidentiality is protected in accordance with the Data Protection Act 1998.

Phylogenetic analysis of Ellipsomyxa species (Myxosporea) and description of Ellipsomyxa gordeyi n. sp. from the gall bladder of mullets (Mugiliformes: Mugilidae) in Nha Trang Bay of the East Sea, Vietnam.

The coastal waters of Vietnam are home to a wide diversity of fishes, but the parasite diversity of these potential hosts is much less well characterized. To begin addressing this knowledge gap, we carried out surveys of myxozoan parasites in fishes collected from Nha Trang Bay in Vietnam's East Sea in 2018-2019. Mugilid fishes were collected in March-April 2018, January-February 2019, and November-December 2019, and examined for myxozoans. Myxospores consistent with those of the genus Ellipsomyxa were found in the gall bladder of four mullet species, and we thoroughly characterized those from Planiliza melinoptera. Myxospores were elliptoid and devoid of striation, with a distinct sinuous suture line. Polar capsules were pyriform and oriented toward the poles of the spore. Morphological features were compared to nominal species and this species from Vietnam was distinct. Phylogenetic analysis based on partial small subunit rDNA sequence revealed that broadly, Ellipsomyxa species split into three phylogenetic lineages, and although in some branches there are groupings by host family, habitat or locality, there are no clear phylogenetic patterns. The new species we encountered in P. melinoptera had a close sister relationship with Ellipsomyxa adlardi, with both species part of a larger subclade within the Ellipsomyxa lineage. Despite this phylogenetic similarity, these species were morphologically distinct, and partial large subunit DNA sequences were only 93% similar to each other. A combination of the morphological characteristics and molecular data suggest that this is an undescribed species and we propose the name Ellipsomyxa gordeyi n. sp.

Beyond Tradition: Exploring Cutting-Edge Approaches for Accurate Diagnosis of Human Filariasis.

Filariasis is recognised as a global public health threat, particularly in tropical and subtropical regions. It is caused by infection with a nematode parasite of the superfamily Filarioidea, including Wuchereria bancrofti, Brugia malayi, Onchocerca volvulus, and Onchocerca lupi. Three main types of filariasis have been classified: lymphatic filariasis, subcutaneous filariasis, and serous cavity filariasis. The symptoms exhibited by individuals afflicted with filariasis are diverse and contingent upon several variables, including the species of parasite, the host's health and immune response, and the stage of infection. While many classical parasitological techniques are considered indispensable tools for the diagnosis of parasitic infections in humans, alternative methods are being sought due to their limitations. Novel tests based on host-parasite interactions offer a rapid, simple, sensitive, and specific diagnostic tool in comparison to traditional parasitological methods. This article presents methods developed in the 21st century for the diagnosis of filariasis caused by invasion from W. bancrofti, B. malayi, O. volvulus, and O. lupi, as well as techniques that are currently in use. The development of modern diagnostic methods based on molecular biology constitutes a significant advancement in the fight against filariasis.

A high-dimensional platform for observing neutrophil-parasite interactions.

Diarrheal diseases with infectious etiology remain a major cause of death globally, particularly in low-income countries. Entamoeba histolytica is a pathogenic protozoan parasite that is the causative agent of amebiasis. Amebiasis has a wide presentation in clinical severity with many factors, including the bacterial microbiota, contributing to this variation. The innate immune response also plays a critical role in regulating the severity of E. histolytica infection, with neutrophils reported to have a protective role. Despite this, the precise mechanism of how neutrophils mediate amebic killing is poorly understood. Thus, modern platforms that allow for inquiry of granulocyte-ameba interactions will increase our understanding of this disease. Herein, we describe an assay for neutrophil killing of E. histolytica by utilizing high-dimensional spectral flow cytometry. Neutrophils were isolated from wild-type 5-week-old C57BL/6 mice and co-cultured with E. histolytica at various multiplicity of infections (MOIs). After co-culture, neutrophils and E. histolytica were stained for spectral flow cytometry. Cell populations were identified using surface markers and fluorescence minus one (FMO) controls. We have previously shown that animals colonized with a component of the human microbiota, Clostridium scindens, were protected from E. histolytica. This protection was associated with elevated neutrophil count. Here, we explored amebic killing capacity and observed that neutrophils from animals with C. scindens possessed heightened amebic killing compared with controls. Thus, this study establishes a novel platform that can provide an in-depth analysis of granulocyte-parasite interactions in various contexts, including during alteration of the intestinal microbiota.IMPORTANCEThe tools for studying host immune cell-E. histolytica interactions are limited. Factors, such as parasite heterogeneity, infectivity, and difficulties with culture systems and animal models, make interrogation of these interactions challenging. Thus, Entamoeba researchers can benefit from next-generation models that allow for the analysis of both host and parasite cells. Here, we demonstrate the use of a novel platform that allows for the determination of parasite-host cell interactions and customizable high-dimensional phenotyping of both populations. Indeed, spectral flow cytometry can approach >40 markers on a single panel and can be paired with custom-developed parasite antibodies that can be conjugated to fluorochromes via commercially available kits. This platform affords researchers the capability to test highly precise hypotheses regarding host-parasite interactions.

One Health: opportunities for Defence Engagement (Health).

The One Health concept continues to gain traction as a necessary approach to tackle emerging threats to human, animal and environmental health but has not yet been adopted within Defence Engagement (Health). The health of humans, animals and ecosystems are closely interlinked. The One Health concept recognises this interdependence and seeks to balance and optimise the health of all three through an integrated and unifying approach. With a focus on zoonoses, vectorborne disease, antimicrobial resistance and food safety it encourages collaboration, communication, coordination and capacity building. The UK has made commitments to the One Health approach in the Global Health and Security Agenda, and the Integrated Review. This article explores how the One Health approach could and should be adopted within Defence Engagement (Health) activity to offer the potential for high-impact, low-risk activity while facilitating long-term relationship building.

The Cryptosporidium signaling kinase CDPK5 plays an important role in male gametogenesis and parasite virulence.

The protozoan parasite Cryptosporidium is a leading cause of diarrhea in young children. The parasite's life cycle involves a coordinated and timely progression from asexual to sexual stages, leading to the formation of the transmissible oocyst. Underlying molecular signaling mechanisms orchestrating sexual development are not known. Here, we describe the function of a signaling kinase in Cryptosporidium male gametogenesis. We reveal the expression of Cryptosporidium parvum calcium-dependent protein kinase 5 (CDPK5) during male gamete development and its important role in the egress of mature gametes. Genetic ablation of this kinase results in viable parasites, indicating that this gene is dispensable for parasite survival. Interestingly, cdpk5 deletion decreases parasite virulence and impacts oocyst shedding in immunocompromised mice. Using phosphoproteomics, we identify possible CDPK5 substrates and biological processes regulated by this kinase. Collectively, these findings illuminate parasite cell biology by revealing a mechanism controlling male gamete production and a potential target to block disease transmission.

M-O-M mediated denaturation resistant P2 tetramer on the infected erythrocyte surface of malaria parasite imports serum fatty acids.

In Plasmodium falciparum, DNA replication, and asynchronous nuclear divisions precede cytokinesis during intraerythrocytic schizogony. Regulation of nuclear division through the import of serum components was largely unknown. At the trophozoite stage, P. falciparum ribosomal protein P2 (PfP2) is exported to the infected erythrocyte (IE) cytosol and the surface as a denaturation-resistant tetramer. The inaccessibility of the IE surface exposed PfP2 to its bona fide ligand led to the arrest of nuclear division. Here, we show that at the onset of schizogony, denaturation-resistant PfP2 tetramer on the IE surface imports fatty acids (FAs). Blockage of import reversibly arrested parasite schizogony. In 11Met-O-Met11 mediated denaturation resistant PfP2 tetramer, the 12/53Cys-Cys12/53 redox switch regulates the binding and release of FAs based on oxidized/reduced state of disulfide linkages. This mechanistic insight of FAs import through PfP2 tetramer reveals a unique regulation of nuclear division at the onset of schizogony.