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Lamanema chavezi is an entero-hepatic strongylid parasite specific to South American camelids. It has been reported only on few occasions outside South America. Due to its hepatic migration, it can cause extensive liver damage, leading to granulomatous and fibrotic hepatitis and manifesting with lethargy, anorexia, and even death. We are reporting the second case of L. chavezi infection in Europe and the first in Switzerland. The patient was a three-year old neutered male llama (Lama glama). Clinical examination revealed bloody mucous discharge from the anus. Fecal sedimentation/flotation revealed strongylid eggs consistent with L. chavezi, which were molecularly confirmed by a PCR targeting the ITS2 plus 5.8S and 28S rDNA flanking regions and amplicon sequencing. Eighteen weeks after administration of a single dose of eprinomectin (0.2 mg/kg i.m.), no further L. chavezi eggs were detected in the feces. The source of infection could not be traced back. The entire herd consisted of llamas bred in Switzerland. L. chavezi has been rarely reported outside South America, but its potential for pathogenicity and establishment should not be underestimated. Fecal sedimentation/flotation techniques should be routinely performed to ensure early detection of the parasite., (© 2024. The Author(s).)

A fifteen-month-old Pembroke Welsh corgi with respiratory distress, exercise intolerance, and moderate regenerative anemia was referred to The Norwegian University of Life Sciences, Small Animal Hospital.Hematology revealed moderate regenerative anemia without evidence of hemolysis. Thoracic radiographs showed a generalized mixed interstitial to alveolar lung pattern and enlarged pulmonary arteries. Changes suggestive of moderate pulmonary hypertension were noted on echocardiography. Baermann fecal diagnostic flotation identified large numbers of Angiostrongylus vasorum larvae, and the AngioDetect serological antigen test was positive. The dog was treated with a two-week course with fenbendazole (51 mg/kg q24h po) and topical imidacloprid/moxidectin (250 mg/62.5 mg) and a one-week course with sildenafil (0.45 mg/kg q12h po). Complete clinical, clinicopathological and echocardiographic resolution was observed after only four weeks. Rapid improvement of echocardiographic abnormalities in cases with suspected pulmonary hypertension is not usually reported in cases with angiostrongylosis.Infection with A. vasorum should be considered in dogs with respiratory signs and bleeding tendencies, even in countries with no endemic history or reported cases., (© 2024. The Author(s).)

Abdominal angiostrongyliasis (AA) is a zoonotic and severe parasitic infection caused by Angiostrongylus costaricensis. AA is currently diagnosed by the observation of A. costaricensis-compatible structures in biopsies or the detection of antibodies in serological tests. However, molecular methods targeting homologous sequences of A. costaricensis have not been designed before, and therefore, an HRM-coupled qPCR was developed to detect the internal transcribed spacer 1 (ITS1) of the parasite. The present assay successfully amplified DNA of A. costaricensis obtained from different hosts and identified slight sequence differences through the HRM analysis. The detection limit of the HRM-qPCR was 0.00036 ng/µL, 1.0 ng/µL, and 0.1 ng/µL when A. costaricensis DNA was diluted in nuclease-free water, whole blood, and sera, respectively, which highlights its potential use for cell-free DNA detection. Moreover, the reaction did not cross-amplify DNA of Angiostrongylus cantonensis, Strongyloides stercoralis, and other nematodes, thus emphasizing its specificity. Additionally, the assay tested positive in formalin-fixed paraffin embedded biopsies with visible A. costaricensis adults or eggs, but not in samples without evident parasites or a low number of larvae, which suggests that the reaction is useful for confirming the presence of the nematode in clinical samples. Finally, DNA of sera from patients with AA was evaluated with the HRM-qPCR but none tested positive, possibly due to long storage periods of the samples which could have led to cfDNA degradation. These results indicate that this assay may be useful in the confirmation of AA and its prospection for cell-free DNA detection protocols., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Purpose: Crenosoma vulpis (Dujardin,1845) is a lungworm which has spread worldwide in canines and is associated with upper respiratory infections. In a majority of cases, the infections are accompanied with chronic cough. Diagnosis of lungworms is often underdiagnosed and can be misinterpreted as other respiratory diseases., Methods: The Small Animal Clinic of the University Veterinary Hospital admitted an 11-month-old dog presented with persistent cough associated with difficulty in breathing and even asphyxia. Based on clinical symptoms, the patient underwent radiological and bronchoscopic examination. Bronchoscopy revealed the presence of lungworms obturating the branches of the tracheobronchial tree. Larvae were collected by bronchoscopic lavage and subjected to parasitological and molecular examination., Results: Microscopic detection and morphological identification of the worms removed during the bronchoscopy confirmed the presence of female adult worms. The subsequent molecular characterisation of the mitochondrial (cytochrome c oxidase subunit I gene (cox1) and 12S ribosomal DNA (rDNA)), nuclear (18S rDNA) genes, as well as the analysis of the second internal transcribed spacer (ITS-2) region of the ribosomal DNA, confirmed the Crenosoma vulpis species. Faecal samples were processed using the Baermann method, which confirmed the presence of the larval stage 1 of C. vulpis. The therapy with fenbendazole at a dose of 50 mg/kg of live weight once daily for the period of 7 days was initiated for the patient., Conclusion: This paper presents the first molecularly confirmed clinical case of a Crenosoma vulpis infection in an 11-month-old female dog of the Miniature Schnauzer breed in Slovakia., (© 2024. The Author(s).)

Background: Angiostrongyliasis cantonensis is a severe yet rare parasitic infection caused by the larvae of Angiostrongylus cantonensis. The primary characteristic feature of this foodborne illness in humans is eosinophilic meningitis. Recently, there has been a gradual increase in reported cases globally. Due to the lack of typical clinical symptoms, signs, and specific laboratory tests, early diagnosis of this disease poses significant challenges. Failure to diagnose and treat this condition promptly can result in fatalities., Methods: We present the case of a 13-year-old male patient who initially presented with fever and headache. The patient was preliminarily diagnosed with bacterial meningitis and received treatment with antibacterial drugs. However, the patient's condition worsened, and he developed progressive consciousness disturbances. Eventually, metagenomic next-generation sequencing (mNGS) testing of cerebrospinal fluid samples indicated Angiostrongylus cantonensis infection. Following treatment with albendazole and prednisone, the patient made a full recovery. We include this case report as part of a literature review to emphasize the potential applications of mNGS in the early diagnosis of Angiostrongyliasis cantonensis., Conclusion: mNGS technology plays a crucial role in the diagnosis of angiostrongyliasis cantonensis. As this technology continues to evolve and be applied, we believe it will play an increasingly important role in diagnosing, treating, and monitoring angiostrongyliasis cantonensis., (© 2024. The Author(s).)

Objective: To describe the clinical presentation, diagnostic findings, and medical management of the first suspected autochthonous case of a dog in the US diagnosed with Angiostrongylus vasorum, the French heartworm., Animal: A 10-month-old Goldendoodle born in Oregon and residing in Washington State., Clinical Presentation, Progression, and Procedures: The dog presented for evaluation of intermittent vomiting and diarrhea. Bloodwork revealed leukocytosis with mild lymphocytosis, monocytosis, eosinophilia, and basophilia. Larvae were detected on a fecal flotation, and fecal PCR confirmed A vasorum., Treatment and Outcome: Administration of milbemycin oxime PO once a week for 4 weeks was initiated with recommendation to continue monthly treatment at label dose. The patient improved over the course of treatment., Clinical Relevance: This case highlights the clinical and diagnostic findings and medical management of A vasorum, also known as the French heartworm, in a dog in the US. Few cases of A vasorum have been reported in wild foxes in North America, mostly in eastern Canada and 1 within the eastern US. Here we report for the first time an autochthonous case of A vasorum in a domestic dog in the US and the first report of any canid in the western US. This case highlights the importance of considering A vasorum as a differential for respiratory disease, gastrointestinal disease, or inexplicable eosinophilia in canine veterinary patients in the US and raises awareness for veterinary practitioners to incorporate appropriate preventative and diagnostic measures for their canine patients.

Background: Angiostrongyliasis is a zoonotic parasitic disease caused by the rat lungworm Angiostrongylus cantonensis. The intermediate hosts of A. cantonensis are gastropods, and snail species such as Pomacea canaliculata play a key role in the transmission of human angiostrongyliasis. Detecting A. cantonensis infection in snails is an important component of epidemiological surveillance and the control of angiostrongyliasis., Methods: In this study, a new method for diagnosing A. cantonensis infection in gastropods was developed by recovering larvae from the buccal cavity of three snail species. The entire buccal cavity of a snail was extracted, and the tissue was pressed between two microscope slides to observe whether A. cantonensis larvae were present. Our new method was compared with traditional pathogenic detection methods of lung microscopy, tissue homogenization, and artificial digestion. We artificially infected 160 P. canaliculata, 160 Cipangopaludina chinensis, and 160 Bellamya aeruginosa snails with A. cantonensis. Then, the four different detection methods were used to diagnose infection in each snail species at 7, 14, 21, and 28 days post exposure., Results: We found no significant difference in the percentages of infected P. canaliculata snails using the four methods to detect A. cantonensis larvae. The radula pressing method had a mean detection rate of 80%, while the lung microscopy (81.3%), tissue homogenization (83.8%), and artificial digestion (85%) methods had slightly greater detection rates. Similarly, the percentages of infected C. chinensis snails that were detected using the radula pressing (80%), tissue homogenization (82.1%), and artificial digestion (83.8%) methods were not significantly different. Finally, the percentages of infected B. aeruginosa snails that were detected using the radula pressing (81.3%), tissue homogenization (81.9%), and artificial digestion (81.4%) methods were not significantly different. These results showed that the radula pressing method had a similar detection rate to traditional lung microscopy, tissue homogenization, or artificial digestion methods., Conclusions: This study demonstrates a new method for the qualitative screening of gastropods that act as intermediate hosts of A. cantonensis (and other Angiostrongylus species), provides technical support for the control of human angiostrongyliasis, and furthers research on A. cantonensis., (© 2024. The Author(s).)

Rat lungworm disease or neuroangiostrongyliasis is a cerebral parasitic infection that affects humans and animals alike. Its clinical signs and symptoms can range from mild self-resolving to serious life-threatening conditions. Studies suggest therapeutic interventions during the early stages of infection to be more effective than in later stages. However, early diagnosis of infection is usually problematic without the knowledge of exposure and/or detection of the parasite's DNA or antibody against the parasite in the cerebrospinal fluid. This requires a lumbar puncture, which is an invasive procedure that generally requires hospitalization. This study evaluates an affordable and less invasive alternative to detect parasitic DNA by PCR from the peripheral blood of potentially infected animals. Blood samples from 58 animals (55 dogs and 3 cats) with clinical suspicion of infection were submitted to our lab between February 2019 and August 2022 by local, licensed veterinarians. DNA was extracted from whole blood, plasma, serum, and/or packed cells using the Qiagen DNeasy Blood & Tissue Kit as per the manufacturer's protocol. All 58 animals were tested by real-time PCR using the AcanITS1 assay and 32 of these animals (31dogs; 1 cat) were also tested using the AcanR3990 assay. The PCR results for both assays were classified into strongly positive > positive > weakly positive > negative, and equivocal for ambiguous results, based on the strength of the signal. The percent infection detected using the AcanITS1 and AcanR3990 assays was 12.72% (7/55) and 20.68% (6/29), respectively. The overall percent infection detected was 34.37% (11/32), with only two animals testing positive by both assays. The three cats involved in this study tested negative by both assays. These results are promising and warrant further investigations to increase sensitivity including variables that might affect detection in the blood, such as parasite load, and laboratory methodologies., (© 2024. The Author(s).)

Background: Aelurostrongylus abstrusus is one of the most important respiratory nematodes of felines. Infections may lead to respiratory clinical signs with varying severity or even death, emphasizing the need for preventive treatment of cats with outdoor access to circumvent patent infections., Methods: Therefore, the preventive efficacy of a spot-on formulation of 280 mg/ml fluralaner and 14 mg/ml moxidectin (Bravecto ® Plus spot-on solution for cats, MSD) against A. abstrusus was evaluated in a negative controlled, randomized and partially blinded efficacy study with 28 purpose-bred cats in a non-terminal design. In three different treatment regimes, the minimum recommended dose of 40 mg fluralaner and 2.0 mg moxidectin/kg bodyweight (BW) was administered once at 12, 8 or 4 weeks (study group G1, G2 and G3, respectively) prior to experimental infection with 300 third-stage A. abstrusus larvae, while G4 served as placebo-treated control., Results: From 30 to 46 days post infection (dpi; SD 114 to 130), faeces were sampled to monitor first-stage larvae (L1) excretion for efficacy determination. Secondary efficacy criteria, including respiratory parameters, serological antibody levels and computed tomography (CT) findings, were assessed once before enrolment (SD -7 to -1) and before infection (SD 75 to 83). After infection, CT evaluation was performed once at 47-50 dpi (SD 131 to 134), and respiratory parameters and antibody levels were regularly assessed twice or once a week, respectively (1 up to 78 dpi, SD 85 up to 162). All animals in the control group excreted L1 by 33-37 dpi and remained positive throughout the study period from 41 to 46 dpi (SD 125 to 130). In the treatment groups, only one animal each of G1 and G2 excreted L1 at two consecutive days, and four cats of G1, two of G2 and three of G3 were positive on single occasions. While the geometric mean (GM) of the maximum number of excreted L1 per 5 g of faeces was 7380.89 in the control group (G4), GMs were significantly lower in the treatment groups with 1.63 in G1, 1.37 in G2 and 0.79 in G3. Thus, based on GMs, the reduction in excreted L1 exceeded 99.9% in all three treatment groups. Based on CT severity scores, all lungs of the animals of the control group showed severe pulmonary changes post infection, whereas lungs of the cats of the treatment groups were either unaltered (4 animals), mildly (11 animals), or moderately altered (5 animals). Moreover, seroconversion was observed in all cats of the control group, but not in those of the treatment groups., Conclusions: The combination of diagnostic methods used in this non-terminal study yielded coherent and reliable results. A single administration of Bravecto ® Plus spot-on solution for cats was well tolerated and effective in the prevention of aelurostrongylosis for at least 12 weeks., (© 2024. The Author(s).)

Objectives: Angiostrongylosis is a significant differential for a diverse range of clinical signs in dogs, many of whom present acutely and sometimes with fatal consequences. Point-of-care diagnostic assays include a commercially available Angiostrongylus vasorum qualitative direct lateral flow assay., Materials and Methods: Case records from one referral centre from dogs with an invalid A. vasorum lateral flow assay, comprising an absent control line alongside a visible test line, were reviewed. As control line failure was hypothesised to be due to antigen excess; where available the A. vasorum lateral flow assay was repeated using dilutions of the original serum., Results: Six dogs had an invalid A. vasorum lateral flow assay result. Five dogs had presented with acute-onset, severe clinical disease consistent with angiostrongylosis, and one dog was a clinically healthy in-contact. Clinical suspicion of angiostrongylosis was confirmed using alternative diagnostic testing and/or response to treatment. Repetition of the A. vasorum lateral flow assay, in four cases, using diluted plasma (10% to 12.5% v/v) resulted in the appearance of a control line alongside the visible test line., Clinical Significance: A heavy burden of A. vasorum infection resulting in angiostrongylosis should be suspected in dogs with compatible clinical signs and an invalid A. vasorum lateral flow assay result due to control failure alongside a visible test line. Repetition of the test with a diluted serum may be considered to account for the hook effect, also known as the postzone phenomenon, as a possible cause., (© 2024 The Authors. Journal of Small Animal Practice published by John Wiley & Sons Ltd on behalf of British Small Animal Veterinary Association.)

Although rare, Angiostrongylus costaricensis infection may be a more prevalent etiology of inflammatory bowel disease than ulcerative colitis and Chron's disease in endemic areas in Central and South America. The present study reviewed the occurrence of A. costaricensis in Brazil, its clinical presentation and pathology; and proposed diagnostic criteria and case definitions for abdominal angiostrongyliasis (AA). Southern and southeastern Brazilian regions are the main endemic areas, and AA affects both genders and all age groups. A review of all 23 published reports of 51 Brazilian patients highlighted the following features that were subsequently classified as minor diagnostic criteria: abdominal pain, palpable mass in the right lower abdominal quadrant, history of exposure, ileocecal tumor, and intestinal perforation with wall thickening. Proposed major criteria include right lower quadrant abdominal pain, blood eosinophilia, positive serology (antibody detection), intense eosinophilic infiltration that involves all strata of the intestinal wall, eosinophilic granulomatous reaction, and eosinophilic vasculitis. In addition to the definitions of suspected and possible cases according to increasing strength of evidence of this infection, demonstration of worms/eggs/larvae in tissues or Angiostrongylus DNA in tissues or serum are required for a confirmed diagnosis. The application of the proposed criteria and definitions may improve patient management, epidemiologic surveillance, and identification of new endemic areas., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

In recent years, Angiostrongylus vasorum, Crenosoma vulpis, Eucoleus aerophilus (syn. Capillaria aerophila) and Eucoleus boehmi (syn. Capillaria boehmi), commonly referred to as canine lungworms, have gained a growing interest worldwide as the result of their geographical expansion. Each of these nematode species differs considerably in its biology and pathogenicity. Despite their impact on dogs' health, these parasites are often underdiagnosed owing to diagnostic challenges. Here, we describe the development and validation of a Taq-Man-based multiplex quantitative PCR (qPCR) for the simultaneous detection of the main species of canine lungworms in faeces of infected dogs. Using 10-fold serial dilutions of synthetic gene block fragments containing individual sequence targets of each lungworm species, the analytical sensitivity of the assay ascertained was 1.84 ng/μl for A. vasorum, 3.08 ng/μl for C. vulpis and 0.79 ng/μl for Eucoleus spp. The sensitivity of the assays and their ability to detect mixed species infections were compared with microscopy-based techniques (faecal floatation and Baermann technique) applied to faecal samples submitted for lungworm testing through an accredited diagnostic laboratory at the Institute of Parasitology, University of Zurich, Switzerland, and from community dogs as part of a research project on canine endoparasites in Cambodia. The multiplex qPCR displayed high diagnostic sensitivity (42/46, 91.3%; 95% Confidence Interval (CI): 79.1-97.1%) and a diagnostic specificity of 100% (45/45, 95% CI: 90.6-100%), and was able to detect 42.9% additional mixed lungworm species infections compared with microscopy-based methods. Kappa statistics showed substantial agreement between the qPCRs and microscopy for mixed infections (κ = 0.72, 95% CI: 0.4-1) and Eucoleus spp. (κ = 0.65, 95% CI: 0.45-0.85) and almost perfect agreement for C. vulpis (κ = 0.85, 95% CI: 0.63-1) and A. vasorum (κ = 0.92, 95% CI: 0.84-1). This multiplex qPCR enables timely, accurate, and sensitive diagnosis of canine lungworm species in faecal samples and can be used to monitor the geographical distribution and emergence of these parasitic species, globally., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This study aimed to prospectively evaluate the risk factors of infection by Aelurostrongylus abstrusus in Brazilian cats with cough and/or radiographic changes, using as diagnostic tools the Baermann method (BM), polymerase chain reaction (PCR) of feces, bronchoalveolar lavage fluid (BALF), and cytology. Forty-three cats that were presented with cough or lung radiographic abnormalities compatible with bronchoalveolar disease were included in the study. After clinical evaluation, feces samples were collected to investigate lungworm parasitism through BM and PCR. BALF was performed to provide samples for cytology, bacteriology, and fungal culture. Stool PCR was considered the gold standard for diagnosis tests, and the other methods were evaluated by their agreement. PCR presented 74% (32/43) of positivity for A. abstrusus , while in the BM, 41% (18/43) were positive. BM showed sensitivity of 56.25% and specificity of 100% when compared with PCR. No larva was found in the cytological evaluation of 21 BALF samples. Lungworm is an important cause of bronchopulmonary disease in domestic cats in Brazil and should be included as a differential diagnosis when a cat is presented with cough or radiographic abnormalities. BM is a sensitive, non-invasive, and cheap technique to diagnose the disease, but it is not as sensitive as PCR.

Angiostrongylus cantonensis is a food-borne zoonotic parasite, and human infection may cause eosinophilic meningitis. Non-coding RNAs (ncRNAs) may regulate physiological and pathological processes at multiple biological levels; however, there are few studies pertaining to the regulatory role of ncRNAs in A. cantonensis infection. Based on publications retrieved from PubMed, Wanfang Data and CNKI, the regulatory role of ncRNAs in A. cantonensis infections mainly includes immune responses, cell apoptosis and signaling transduction, and ncRNAs may serve as biomarkers for diagnosis of angiostrongyliasis. This review summarizes the main roles of ncRNAs in A. cantonensis infections and the underlying mechanisms, so as to provide insights into diagnosis and treatment of angiostrongyliasis.

Angiostrongylus cantonensis is the leading cause of neuroangiostrongyliasis worldwide, and east Hawaii Island is a hotspot for the disease in the United States. A combination of glycoproteins with molecular weight of 31 kDa has been used as antigen to evaluate antibody response in human serum samples in Thailand with high specificity and sensitivity. In a previous pilot study, the Thailand-isolated 31-kDa proteins showed efficacy in dot-blot tests using serum samples from 435 human volunteers on Hawaii Island. However, we hypothesized that native antigen isolated from Hawaii A. cantonensis may exhibit higher specificity than the Thailand-isolated 31-kDa antigen due to potential minor variation in epitopes between isolates. In this study, 31-kDa glycoproteins were isolated by sodium dodecyl-sulfate polyacrylamide gel electrophoresis from adult A. cantonensis nematodes collected from rats captured on east Hawaii Island. The resultant proteins were purified by electroelution, pooled, bioanalyzed, and quantified. A subset of 148 samples from human participants of the original cohort of 435 was consented for this study, including 12 of the original 15 clinically diagnosed participants. Results of ELISA using the Hawaii-isolated 31-kDa antigen were compared with results of the same serum samples previously tested with both crude Hawaii antigen ELISA and Thailand 31-kDa antigen dot blot. This study shows a seroprevalence in the general population of East Hawaii Island of 25.0%, similar to previous findings of 23.8% seroprevalence in this cohort using crude antigen from Hawaii A. cantonensis and 26.5% using Thailand 31-kDa antigen.

Background: The lungworm Aelurostrongylus abstrusus infects wild and domestic feline species worldwide and is considered a primary respiratory parasite of cats. Definitive diagnosis is based on the identification of first-stage larvae (L1s) released in faeces approximately 5 to 6 weeks after infection. More recently, serology has been shown to be a diagnostic alternative for A. abstrusus infection in cats. The present study aimed at evaluating the diagnostic performance of serological antibody detection compared to faecal examination for A. abstrusus infection in a population of cats with known infection status from endemic areas in Italy and to identify factors (larval scores, age, co-infections with other helminths) that may influence test sensitivity and specificity of serology., Methods: All cats resulting positive using the Baermann technique (n = 78) were tested with the A. abstrusus ELISA. An additional 90 serum samples from cats living in three geographical areas with infection prevalence > 10%, but that resulted negative on Baermann, were also tested., Results: Among 78 cats copromicroscopically positive for L1s of A. abstrusus (Group 1), 29 (37.2%) were seropositive in ELISA. Of the 90 cats from Group 2 (cats living in three geographical areas in Italy with A. abstrusus prevalence > than 10%, but negative on Baermann examination), 11 (12.2%) were positive on ELISA. The overall seroprevalence was 23.8%. There was no statistical difference either between average optical density (OD) values of cats excreting > 100 L1s vs. cats excreting < 100 L1s (0.84 vs. 0.66; P value = 0.3247) or comparing the OD values with age of infected cats. Few Baermann-negative cats positive for Toxocara cati or hookworms were seropositive, supporting lack of cross-reactivity to these nematodes., Conclusions: Results from the present study suggest that relying solely on faecal examination may underestimate prevalence of A. abstrusus infection in cats and that field surveys based on antibody detection are useful for establishing true prevalence of infected and/or exposed animals., (© 2023. The Author(s).)

Angiostrongylus cantonensis is an invasive nematode parasite that causes eosinophilic meningitis in many vertebrate hosts, including humans. This parasite is spreading rapidly through the six continents, with Europe being the final frontier. Sentinel surveillance may be a cost-effective strategy to monitor the pathogen's arrival to new geographical regions. Necropsy, followed by tissue digestion, is often used to recover helminth parasites from vertebrate host tissues, however, to detect brain parasites, this protocol is poorly utilized. Our brain digestion protocol is easily performed and 1) reduces false positivity and negativity, 2) provides accurate estimates of parasite burden and 3) helps establish a more precise prevalence. Early detection of A. cantonensis increases the efficacy of prevention, treatment, and disease control strategies for susceptible animal populations and humans., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Background: Angiostrongylus cantonensis (rat lungworm) is recognised as the leading cause of human eosinophilic meningitis, a serious condition observed when nematode larvae migrate through the CNS. Canine Neural Angiostrongyliasis (CNA) is the analogous disease in dogs. Both humans and dogs are accidental hosts, and a rapid diagnosis is warranted. A highly sensitive PCR based assay is available but often not readily accessible in many jurisdictions. An alternative DNA amplification assay that would further improve accessibility is needed. This study aimed to assess the diagnostic utility of a newly designed LAMP assay to detect DNA of globally distributed and invasive A. cantonensis and Angiostrongylus mackerrasae, the other neurotropic Angiostrongylus species, which is native to Australia., Methodology/principal Findings: Cerebrospinal fluid (CSF) from dogs with a presumptive diagnosis of A. cantonensis infection (2020-2022) were received for confirmatory laboratory testing and processed for DNA isolation and ultrasensitive Angiostrongylus qPCR targeting AcanR3390. A newly designed LAMP assay targeting the same gene target was directly compared to the reference ultrasensitive qPCR in a diagnostic laboratory setting to determine the presence of A. cantonensis DNA to diagnose CNA. The LAMP assay (Angie-LAMP) allowed the sensitive detection of A. cantonensis DNA from archived DNA specimens (Kappa = 0.81, 95%CI 0.69-0.92; n = 93) and rapid single-step lysis of archived CSF samples (Kappa = 0.77, 95%CI 0.59-0.94; n = 52). Only A. cantonensis DNA was detected in canine CSF samples, and co-infection with A. mackerrasae using amplicon deep sequencing (ITS-2 rDNA) was not demonstrated. Both SYD.1 and AC13 haplotypes were detected using sequencing of partial cox1., Conclusions/significance: The Angie-LAMP assay is a useful molecular tool for detecting Angiostrongylus DNA in canine CSF and performs comparably to a laboratory Angiostrongylus qPCR. Adaptation of single-step sample lysis improved potential applicability for diagnosis of angiostrongyliasis in a clinical setting for dogs and by extension, to humans., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Baláž et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Abdominal angiostrongyliasis (AA) is a severe parasitic infection caused by the nematode Angiostrongylus costaricensis. This disease is characterized by abdominal pain, a strong inflammatory eosinophilic response in the blood and tissues, and eventually intestinal perforation. Diagnosis of AA is challenging since there are no commercially available serological kits for A. costaricensis, and thus, histopathological analysis remains the gold standard. Herein we provide a decision flowchart for clinicians to improve the diagnosis of AA based on a patient's clinical manifestations, laboratory findings, macroscopic observations of the gut lesions, as well as characteristic microscopic alterations in biopsies. A brief discussion of the available polymerase chain reaction and in-house serological methods is also presented. The aim of this mini-review is to improve the diagnosis of AA, which should lead to prompt detection of cases and better estimates of the epidemiology and geographical distribution of A. costaricensis., (© 2023. The Author(s).)

Angiostrongyliasis cantonensis is an emerging infectious disease in China. Snails are intermediate hosts of Angiostrongylus cantonensis and play a critical role in the transmission of angiostrongyliasis cantonensis. Detection of A. cantonensis in snails is an important part of epidemiological surveys. Currently, the rapid developments in the techniques for detection of A. cantonensis in snails facilitate the surveillance of angiostrongyliasis cantonensis and provide an important support for angiostrongyliasis cantonensis prevention and control. This review summarizes the advances in the techniques for detection of A. cantonensis in snails.

Abdominal angiostrongyliasis (AA) is a zoonotic disease caused by the nematode Angiostrongylus costaricensis, which is endemic in southern Brazil. Humans become infected by ingesting third-stage (L3) larvae and are considered accidental hosts since neither eggs nor first-stage (L1) larvae are found in feces. The definitive diagnosis can be made by histopathologic examination of surgical specimens or intestinal biopsies. The present study assessed the use of PCR to carry out the molecular detection of AA from serum samples. A total of 62 human serum samples were divided into three groups: (i) 28 serum samples from human patients with presumptive histopathological diagnosis of AA; (ii) 23 serum samples from individuals with unknown serology for AA; (iii) 11 serum samples from patients that suffered from different parasitosis were included. The serum samples were initially tested by in-house indirect ELISA and then by PCR. A total of 14 samples were positive by ELISA, and 6 were positive by PCR. Six samples that were negative by ELISA were positive by PCR. Amplicons were sequenced, and Angiostrongylus DNA was confirmed. We conclude that PCR amplification can be used to confirm Angiostrongylus DNA in serum, which is especially important in cases where antibody levels are too low to be detected. It may also serve as a useful target for survey studies., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Angiostrongylus vasorum is a widely distributed cardiopulmonary parasite of canids in Europe. Clinical signs in dogs can be highly variable and diagnostically challenging. A correct and early diagnosis is hence indispensable to adequately manage affected patients. First-stage larvae (L1) are excreted in the faeces of definitive hosts and conventionally identified using the Baermann technique. Moreover, ELISAs for the detection of circulating antigen and specific antibodies have been presented. The current study aimed at i) quantitatively assessing larval migration in the Baermann funnel after 12 h and 24 h; ii) investigating the influence of sample storage at 4 °C over the course of three days on the number of detected L1; iii) evaluating potential associations of adult worm burdens with larval shedding in dogs and ELISA optical density (OD) values for circulating parasite antigen and specific antibodies. Faecal samples were obtained from naturally infected dogs (n = 21) and Baermann funnels were set up in duplicate over the course of four consecutive days (days 0-3) starting with the day of sample collection. Funnels were harvested on days 1-4 after 12 and 24 h, respectively, and the number of L1 per gram faeces (LPG) was determined. The LPG did not differ between larval harvest after 12 h from harvest after 24 h. Storage of faecal samples at 4 °C for two and three days entailed a considerable decrease in LPG. Adult worm burdens and larval excretion data from previous experiments demonstrated a correlation between worm burden and LPG. In contrast, no correlations between worm burden and the level of parasite antigen and specific antibody OD values, respectively, were identified. Thus, OD values of both antigen and antibody ELISA did not allow for conclusions on infection intensity reflected by the number of adult parasites. For the detection of L1 in faeces, 12 or 24 h of larval migration time was not discriminating for A. vasorum positivity. Thus, early processing of faecal samples is essential, since larval detection and hence sensitivity of the approach considerably decreased over the course of three days of storage. Therefore, the common recommendation to collect faecal samples for three consecutive days and to subsequently analyse them needs to be reconsidered. The results of this study can be readily translated into precise recommendations for daily practice to adequately assess A. vasorum infected dogs., Competing Interests: Conflict of interest The authors have no conflict of interest to declare., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

An 11-month-old male infant with ascending paralysis had an unremarkable initial cerebrospinal fluid (CSF) analysis and imaging. Progressive neurological symptoms resulted in repeated CSF sampling, microscopy, and plasma microbial cell-free DNA next-generation sequencing analysis, that in combination with epidemiology, confirmed the diagnosis., (© The Author(s) 2022. Published by Oxford University Press on behalf of The Journal of the Pediatric Infectious Diseases Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Angiostrongylus cantonensis is the etiological agent of neuroangiostrongyliasis in humans, which is developed in gastropods and vertebrate species, mainly rodents. Human transmission occurs through consumption of molluscs and paratenic hosts infected with L3, and the migration of larvae to the central nervous system causes eosinophilic meningitis. Laboratory diagnosis is based on molecular and immunological tests, using young or adult females as a source of antigens. However, these tests give positive results only after several weeks of symptoms onset and also cross-reactions with others parasite infections may occur., Objectives: The purpose of this work was to study different antigenic preparations of distinct evolutionary phases of A. cantonensis, in order to improve serological techniques for disease immunodiagnosis., Methods: For this purpose, antigenic fractions of different evolutionary forms were evaluated by Dot-enzyme-linked immunosorbent assay (Dot-ELISA) and Western blot using serum samples., Findings: All analysed fractions showed reactivity with serum samples from patients with neuroangiostrongyliasis, especially female membrane alkaline (FAM) and female soluble alkaline (FAS) fractions together with female soluble saline (FSS), improving the technique specificity., Main Conclusions: The results point to the possibility of use of raw female antigens in association with alkaline membrane antigens extracted from adult worms to aid in diagnosis and helps initiate neuroangiostrongyliasis surveillance and control actions.

Objective: To identify first-stage nematode larvae (L1) recovered from a red fox scat sample and adult female worms recovered from 2 red fox lungs at necropsy, using published molecular methods to confirm a morphological diagnosis of Angiostrongylus vasorum (French heartworm)., Animal: Red fox (Vulpes vulpes) ., Procedure: Nematode larvae recovered from a Baermann examination survey of wild canid scats ( n = 101) conducted from January 2017 to August 2020, were identified by size and morphology and subjected to PCR and DNA sequencing of the small subunit (SSU) rRNA gene, the large subunit (LSU) rRNA gene, or the second internal transcribed spacer (ITS2). In addition, these techniques were applied to adult female worms recovered from the heart/lungs of 2 red foxes (obtained from PEI trappers and stored frozen at -20°C since December of 2018 and 2020)., Results: Size and morphology of L1 recovered by Baermann examination from a wild canid scat sample (presumed to be red fox) collected near Montague, PEI and adult female worms recovered at necropsy from 2 red fox carcasses were identified as A. vasorum . Molecular analysis confirmed the larvae and adult worms were A. vasorum ., Conclusion: These findings indicated that A. vasorum has become endemic in the red fox population on PEI., Clinical Relevance: Angiostrongylus vasorum infection is potentially fatal in dogs. Veterinarians and regional diagnostic laboratories in the Maritime provinces should consider the possibility of A. vasorum infection in dogs with clinical signs of cardiopulmonary and/or central nervous system disease or bleeding disorders., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

Crenosoma vulpis, the fox lungworm, is a helminth parasite endemic to the fox population of New England. Domestic dogs are susceptible to infection via ingestion of snails and slugs. Two dogs from New England were diagnosed with C. vulpis. The predominant clinical sign in both dogs was a chronic cough. Treatment with steroids and antibiotics only temporarily relieved clinical signs. Thoracic radiographs in both dogs revealed bronchial patterns. Endotracheal washes were performed in each dog revealing marked, mixed inflammation consisting mainly of neutrophils with eosinophils in lesser numbers. Helminth larvae could also be visualized on cytology. A fecal flotation revealed helminth larvae in one dog but failed to identify larvae in the second dog. The diagnosis of C. vulpis was confirmed via PCR analysis and sequencing of samples from both endotracheal washes. One dog was treated with fenbendazole (50 mg/kg PO q24h for 14 days), enrofloxacin (13 mg/kg PO q 24 h for 5 days), and a tapering protocol of prednisone (20 mg PO q12h for 5 days, 20 mg PO q24h for 5 days, then 20 mg PO q48h for 10 days). The second dog was treated with fenbendazole (50 mg/kg PO q24h for 10 days) with an additional 7 days of febantel and two doses of milbemycin, achieving complete resolution of clinical signs. This lungworm is becoming increasingly more prevalent in domestic dogs worldwide and may be more prevalent in New England than previously thought. Veterinary practitioners of New England should include this respiratory helminth as a differential in dogs with respiratory signs, and respiratory washes and Baermann fecal examinations are warranted in dogs presenting with non-specific respiratory clinical signs., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Angiostrongylus cantonensis is a worldwide zoonotic parasite that causes eosinophilic meningoencephalitis in many species of animals including humans. This report describes neuro-angiostrongylosis in a white-eared opossum that showed nervous clinical signs such as circling and depression. At necropsy, no relevant macroscopic lesions were observed. Histologically, eosinophilic meningoencephalitis was associated with multiple sections of nematodes and many intracytoplasmic eosinophilic inclusion bodies within gastric parietal cells. Immunohistochemistry was strongly positive for canine distemper virus in the stomach but there was no immunolabeling in the brain. This study describes a fatal case of eosinophilic meningoencephalitis by A. cantonensis with canine distemper virus concurrent infection in a white-eared opossum in southern Brazil, with histological characterization and molecular confirmation of the parasitism., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A red ruffed lemur (Varecia rubra) from a zoo in Louisiana, USA, was euthanized for worsening paresis. Brain and spinal cord histology identified eosinophilic meningoencephalomyelitis with intralesional adult Angiostrongylus sp. nematodes. PCR and sequencing confirmed A. cantonensis infection, indicating this parasite constitutes an emerging zoonosis in the southeastern United States.

Canine angiostrongylosis is a gastropod-borne parasitosis caused by the cosmopolitan nematode Angiostrongylus vasorum. It is associated with cardiorespiratory disorders and eventually fatal coagulopathies. A 6-months-old female dog previously diagnosed with bronchopneumonia and pneumothorax, treated accordingly without success, was hospitalized for further treatment. Complementary diagnostics confirmed previous findings plus pneumomediastinum, thoracic exudate, leucocytosis, thrombocytosis, hypercoagulability, and hyperglobulinemia. Coprological analysis showed a severe patent A. vasorum infection. After the third daily fenbendazole administration, all A. vasorum larvae were found dead in follow-up coprological analyses while in intensive care. Upon release, the dog was monitored until full clinical recovery 3 months later. Up to date, A. vasorum infection-related pneumothorax was rarely described, particularly with full recovery of such a young patient. Thus, a parasitological aetiology should be considered for dogs with severe bronchopneumonia or pneumothorax, especially in Europe and South America where increasing numbers of canine angiostrongylosis cases call for enhanced awareness of veterinary practitioners., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Angiostrongylus vasorum (Nematoda, Metastrongyloidea) is a vascular nematode that resides in the pulmonary arteries and the right side of the heart of a wide variety of carnivores, with an indirect life cycle using coprophagic gastropods as intermediate hosts. For domestic dogs, the infection with A. vasorum can be asymptomatic, but more frequently, it is associated with a wide range of clinical manifestations like cardio-respiratory signs, bleedings, neurological signs, and ocular problems which can lead to death when not treated accordingly. Angiostrongylosis was confirmed for the first time in Romania in red foxes (Vulpes vulpes) in 2017 and two years later a seroepidemiologic study was conducted among domestic dogs. However, to this date, no clinical canine angiostrongylosis cases were published in Romania. The aim of the present paper was to evaluate the knowledge about canine angiostrongylosis among veterinarians in Romania and to update the distribution of this disease using a national wide anonymous questionnaire., Results: Overall, 147 unique responses were submitted, from 31 out of 42 counties. Twelve veterinarians (8%) from 8 counties (26%) acknowledged diagnosing a case of angiostrongylosis including 5 from the Bucharest and 1 from each of the remaining seven counties. All affected dogs had respiratory distress, 75% suffered cardiopathy, 16% coagulopathies and 8% neurological signs. Case diagnosis was based mostly on larval detection by coprology (67%) and serological antigen detection test (42%)., Conclusions: Romanian veterinarians are aware of canine angiostrongylosis and a significant number have clinical experience with the disease. Epidemiological studies are now needed to assess its distribution in the country, and further efforts are required to improve understanding of the disease, its diagnostic and treatment methods among veterinarians., (© 2021. The Author(s).)

Background: Angiostrongylus cantonensis (Ac), or the rat lungworm, is a major cause of eosinophilic meningitis. Humans are infected by ingesting the 3rd stage larvae from primary hosts, snails, and slugs, or paratenic hosts. The currently used molecular test is a qPCR assay targeting the ITS1 rDNA region (ITS1) of Ac., Methods: In silico design of a more sensitive qPCR assay was performed based on tandem repeats predicted to be the most abundant by the RepeatExplorer algorithm. Genomic DNA (gDNA) of Ac were used to determine the analytical sensitivity and specificity of the best primer/probe combination. This assay was then applied to clinical and environmental samples., Results: The limit of detection of the best performing assay, AcanR3990, was 1 fg (the DNA equivalent of 1/100 000 dilution of a single 3rd stage larvae). Out of 127 CDC archived CSF samples from varied geographic locations, the AcanR3990 qPCR detected the presence of Ac in 49/49 ITS1 confirmed angiostrongyliasis patients, along with 15/73 samples previously negative by ITS1 qPCR despite strong clinical suspicion for angiostrongyliasis. Intermediate hosts (gastropods) and an accidental host, a symptomatic horse, were also tested with similar improvement in detection observed. AcanR3990 qPCR did not cross-react in 5 CSF from patients with proven neurocysticercosis, toxocariasis, gnathostomiasis, and baylisascariasis. AcanR3990 qPCR failed to amplify genomic DNA from the other related Angiostrongylus species tested except for Angiostrongylus mackerrasae (Am), a neurotropic species limited to Australia that would be expected to present with a clinical syndrome indistinguishable from Ac., Conclusion: These results suggest AcanR3990 qPCR assay is highly sensitive and specific with potential wide applicability as a One Health detection method for Ac and Am., (© The Author(s) 2020. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Angiostrongylus cantonensis constitutes a major etiologic agent of eosinophilic meningoencephalitis. The detection methods for angiostrongyliasis mainly depend on morphology or immunology. A firmer diagnosis could be reached by directly detecting the parasite in the cerebrospinal fluid or through laboratory assays that are specific for Angiostrongylus -induced antibodies or the parasite's DNA. A. cantonensis detection could be carried out by larva release from the tissue upon pepsin digestion. However, the procedure requires live mollusks, which might complicate the analysis of large amounts of samples. Since morphological assays are limited, multiple molecular techniques have been put forward for detecting A. cantonensis , including PCR amplification of targets followed by fragment length or DNA sequence analysis. This allows rapid and accurate identification of A. cantonensis for efficient infection management and epidemiological purposes. In this study, we reviewed the current methods, concepts, and applications of molecular approaches to better understand the genetic characterization, molecular detection methods, and practical application of molecular detection in A. cantonensis .

A 67-year-old man presented with headache, middle back pain that radiated to both legs, and paresthesia in the right leg for 1 day. He had eaten raw shrimp 1 week previously. Over the next week after admission, he developed urinary retention and weakness in both legs. The numbness in his right leg expanded to below the umbilicus. Magnetic resonance imaging of the spinal cord showed myelopathy with minimal cord swelling at T9 to the conus medullaris and a hemorrhagic lesion from T10 to T11. A complete blood count on day 28 after the onset of symptoms revealed leukocytosis without eosinophilia and no white blood cells in his cerebrospinal fluid. Results of an immunochromatographic test kit were positive for Angiostrongylus cantonesis but negative for Gnathostoma spinigerum. After a 4-week course of albendazole combined with a tapering dose of dexamethasone, he achieved nearly complete recovery.

Angiostrongylus costaricensis is a zoonotic parasitic nematode described for the first time in 1971 by Pedro Morera and Rodolfo Céspedes in Costa Rica. This parasite causes an infection known as abdominal angiostrongyliasis, affecting mainly school-aged children and young adults. Infection with A. costaricensis has been associated with a myriad of rodent and mollusk species in the Americas and the Caribbean, as its natural hosts and reservoirs. In this commemorative review, we highlight the extensive research collected through a 50-year journey, which includes ecological, pathological, and molecular studies on A. costaricensis and its implicated disease. We also identify major knowledge gaps in its evolutionary history, the ecological role of imported and invasive mollusk species, and immune response. We propose that the advent of -omics analyses will allow us to gather novel information regarding A. costaricensis biology and infection dynamics, as well as to promote the design of much-needed sensitive and specific diagnostic tools., (© 2021. The Author(s).)

Angiostrongylus vasorum is an emerging parasitic nematode of canids and causes respiratory distress, bleeding, and other signs in dogs. Despite its clinical importance, the molecular toolbox allowing the study of the parasite is incomplete. To address this gap, we have sequenced its nuclear genome using Oxford nanopore sequencing, polished with Illumina reads. The size of the final genome is 280 Mb comprising 468 contigs, with an N50 value of 1.68 Mb and a BUSCO score of 93.5%. Ninety-three percent of 13,766 predicted genes were assigned to putative functions. Three folate carriers were found exclusively in A. vasorum, with potential involvement in host coagulopathy. A screen for previously identified vaccine candidates, the aminopeptidase H11 and the somatic protein rHc23, revealed homologs in A. vasorum. The genome sequence will provide a foundation for the development of new tools against canine angiostrongylosis, supporting the identification of potential drug and vaccine targets., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

This is a short report describing the clinical presentation, investigations, treatment and outcome of a patient living in the Jamaican countryside who was diagnosed with a live, motile subfoveal helminth identified as Angiostrongylus cantonensis on optical coherence tomography at the University Hospital of the West Indies Eye Clinic. This is the second documented case in Jamaica. Early recognition of angiostrongyliasis is important as it may manifest as eosinophilic meningitis which can be fatal without prompt treatment.

Purpose: The literature refers that Angiostrongylus vasorum should always be considered in the differential diagnosis of respiratory diseases in captive red panda (Ailurus fulgens) from endemic areas, and the importance of undertaking a careful diagnostic process and timely medical treatment are crucial when the disease is suspected. The authors think that the description of this clinical case can help other colleagues in the deworming, clinical and anesthesiologic management of infected subjects., Methods: A red panda was referred to the Veterinary Teaching Hospital of the University of Milan in Lodi, due to a diagnosis of A. vasorum formulated in May 2015. The diagnosis was made after the detection of both first-stage larvae by Baermann technique and antigens by serological rapid in-clinic assay. In addition, haemochromocytometric and blood chemistry tests, echocardiography and a CT examination were carried out., Results: The subject was successfully treated by oral administration of milbemycin oxime and praziquantel (Milbemax, Novartis, Italy), respectively, at the weekly dose of 12.5 mg/subject and 125 mg/subject for three consecutive weeks, alternated with 20 days of suspension. Treatment continued with the same scheme until clinical examination carried out in Lodi in December 2018., Conclusion: The follow-up of the described clinical case demonstrates how appropriate management of the infection and the subsequent prophylaxis can correctly eliminate the parasite, thus avoiding the spread of the nematode and the onset of severe and lethal lung forms as described in the literature.

A subcommittee of the Hawaii Governor's Joint Task Force on Rat Lungworm Disease developed preliminary guidelines for the diagnosis and treatment of neuroangiostrongyliasis (NAS) in 2018 (Guidelines, 2018). This paper reviews the main points of those guidelines and provides updates in areas where our understanding of the disease has increased. The diagnosis of NAS is described, including confirmation of infection by real-time polymerase chain reaction (RTi-PCR) to detect parasite DNA in the central nervous system (CNS). The treatment literature is reviewed with recommendations for the use of corticosteroids and the anthelminthic drug albendazole. Long-term sequelae of NAS are discussed and recommendations for future research are proposed.

The principal aim of this study was to optimize the diagnosis of canine neuroangiostrongyliasis (NA). In total, 92 cases were seen between 2010 and 2020. Dogs were aged from 7 weeks to 14 years (median 5 months), with 73/90 (81%) less than 6 months and 1.7 times as many males as females. The disease became more common over the study period. Most cases (86%) were seen between March and July. Cerebrospinal fluid (CSF) was obtained from the cisterna magna in 77 dogs, the lumbar cistern in f5, and both sites in 3. Nucleated cell counts for 84 specimens ranged from 1 to 146 150 cells μL-1 (median 4500). Percentage eosinophils varied from 0 to 98% (median 83%). When both cisternal and lumbar CSF were collected, inflammation was more severe caudally. Seventy-three CSF specimens were subjected to enzyme-linked immunosorbent assay (ELISA) testing for antibodies against A. cantonensis; 61 (84%) tested positive, titres ranging from <100 to ⩾12 800 (median 1600). Sixty-one CSF specimens were subjected to real-time quantitative polymerase chain reaction (qPCR) testing using a new protocol targeting a bioinformatically-informed repetitive genetic target; 53/61 samples (87%) tested positive, CT values ranging from 23.4 to 39.5 (median 30.0). For 57 dogs, it was possible to compare CSF ELISA serology and qPCR. ELISA and qPCR were both positive in 40 dogs, in 5 dogs the ELISA was positive while the qPCR was negative, in 9 dogs the qPCR was positive but the ELISA was negative, while in 3 dogs both the ELISA and qPCR were negative. NA is an emerging infectious disease of dogs in Sydney, Australia.

Serological tests may yield false-negative results for specific antibodies detection before or at the early seroconversion phase. Tests that detect circulating antigens of Angiostrongylus cantonensis would therefore be of value in diagnosis to distinguish current or past infection. Here, a quick, easy to perform, portable and inexpensive diagnostic device for detection of 31-kDa A. cantonensis specific antigens had been developed. This sandwich dot-immunogold filtration assay (AcDIGFAAg), for detecting active angiostrongyliasis was produced using anti-A. cantonensis polyclonal antibody dotted on the nitrocellulose membrane as a capture agent and colloidal gold-labelled anti-31 kDa A. cantonensis antibody as a detection agent. A well-defined pink dot, indicating positivity, was seen readily by naked eye within 10-15 min. The AcDIGFAAg detected A. cantonensis-specific antigens in cerebrospinal fluid samples from 4 out of 10 serologically confirmed angiostrongyliasis cases and 2 out of 5 suspected cases with negative anti-A. cantonensis antibodies. Among the 19 patient sera with A. cantonensis infection, 2 showed positive reaction by AcDIGFAAg. No positive AcDIGFAAg reaction was observed in all the serum samples with other parasitic diseases, and the healthy controls. The present 'AcDIGFAAg' enables rapid qualitative detection of the specific 31-kDa antigens of A. cantonensis in clinical samples with potential for application even under resource-limited settings.

Canine angiostrongylosis is an emergent cardio-pulmonary gastropod-borne helminthic infection caused by the metastrongyloid nematode Angiostrongylus vasorum. Clinically, it is characterized by a wide spectrum of non-specific signs and the red fox serves as the most important reservoir for dog infections. In Italy, this disease has been well documented both in northwestern and central-southern regions, whereas it is apparently poorly recognized in the northeastern area of the country. This report describes the diagnostic findings of two autochthonous cases of fatal canine angiostrongylosis and of one case in a wild red fox detected in northeastern Italy. Reporting cases is relevant to clinicians in order to increase their awareness for the prompt diagnosis of a potentially life-threatening disease that may go unnoticed or misdiagnosed., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Canine angiostrongylosis caused by Angiostrongylus vasorum is a life-threatening disease which is emerging in regions of Europe. Thus, there is the merit for a continuous epidemiological surveillance in dog populations. This is the first description of a clinical autochthonous case of canine angiostrongylosis in Greece. A 7-month-old, male, mixed-breed dog was presented with progressively worsening anorexia, respiratory distress, coughing, bleeding diathesis and succumbed four days post admission. Gross post mortem examination revealed numerous nodular fistulated lesions in the lungs, and pulmonary cytology and histopathology showed a verminous pyogranulomatous pneumonia. The definitive diagnosis was based on the morphological identification of first stage larvae (L1) retrieved in the feces by the Baermann method, the detection of the circulating antigen by an in-clinic test and the molecular identification of L1. This report underlines the epidemiological and clinical implications, as well as the infection risks when the index of clinical suspicion is low and the disease is unexpected in a country., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Eosinophilic meningitis (EM) is a rare clinical syndrome caused by both infectious and noninfectious diseases. In tropical pacific countries, Angiostrongylus cantonensis is the most common cause. However, the EM definition varies in the literature, and its relation to parasitic meningitis (PM) remains unclear., Methodology/principal Findings: Adult and adolescent patients of 13 years old or above with suspected central nervous system (CNS) infections with abnormal CSF findings were prospectively enrolled at a tertiary referral hospital in Hanoi, Vietnam from June 2012 to May 2014. Patients with EM or suspected PM (EM/PM) were defined by the presence of either ≥10% eosinophils or an absolute eosinophil cell counts of ≥10/mm3 in the CSF or blood eosinophilia (>16% of WBCs) without CSF eosinophils. In total 679 patients were enrolled: 7 (1.03%) had ≥10% CSF eosinophilia, 20 (2.95%) had ≥10/mm3 CSF eosinophilia, and 7 (1.03%) had >16% blood eosinophilia. The patients with ≥10% CSF eosinophilia were significantly younger (p = 0.017), had a lower body temperature (p = 0.036) than patients with ≥10/mm3 CSF eosinophilia among whom bacterial pathogens were detected in 72.2% (13/18) of those who were tested by culture and/or PCR. In contrast, the characteristics of the patients with >16% blood eosinophilia resembled those of patients with ≥10% CSF eosinophilia. We further conducted serological tests and real-time PCR to identify A. cantonensis. Serology or real-time PCR was positive in 3 (42.8%) patients with ≥10% CSF eosinophilia and 6 (85.7%) patients with >16% blood eosinophilia without CSF eosinophils but none of patients with ≥10/mm3 CSF eosinophilia., Conclusions: The etiology of PM in northern Vietnam is A. cantonensis. The eosinophil percentage is a more reliable predictor of parasitic EM than absolute eosinophil count in the CSF. Patients with PM may present with a high percentage of eosinophils in the peripheral blood but not in the CSF., Competing Interests: The authors have declared that no competing interests exist.

Background: Intermediate hosts are key organisms in maintaining parasite life cycles, because they can act as amplifiers in the transmission from natural reservoirs to humans. One of the most important groups of intermediate hosts for zoonotic nematode infections are gastropods,slugs and snails. These are essential organisms in the larval development of Angiostrongylus species., Objectives: The objective of this paper is to review reports of Angiostrongylus spp. in naturally infected gastropods from the Americas, taking into account the diagnostic methods used in their identification, to be able to provide more accurate list of their intermediate hosts. We also discuss the factors that aid the dispersion of Angiostrongylus spp. in the Americas., Methods: This study reviews scientific publications and book sections on Angiostrongylus spp. in the Americas, including original works assessing larvae of Angiostrongylus in intermediate hosts. The eligible reports were classified accordingly to their geographical location, year of first record, and the larvae identification methodologies used. Digital repositories were used for the search. The bioecological characteristics of the main intermediate hosts are summarised., Findings: A total of 29 gastropod species that are naturally infected with Angiostrongylus spp. have been reported as intermediate hosts, 16 of which are land snails, two are freshwater snails, and 11 land slugs., Main Conclusions: This study highlights the importance of integrative studies, analysing both the etiological agent and its transmission dynamic in the environment, the biological and ecological characteristics of the hosts, and the impact on host populations. It is necessary to increase interdisciplinary studies to determine the potential epidemiological health risk of angiostrongyliasis in the Americas, and thus be able to establish prevention, monitoring and contingency strategies in the region.

Angiostrongylus cantonensis is the main causative agent of eosinophilic meningoencephalitis (EoM) in humans. Molecular diagnostic methods are essential since the identification of larvae in cerebrospinal fluid (CSF) is extremely rare. To date, the detection of a 31 kDa antigen by Western blotting has been the primary immunodiagnostic method for EoM caused by A. cantonensis. However, cross-reactivity with other parasites has been observed. Therefore, we conducted a comparative analysis using sera from individuals with angiostrongyliasis. We also characterized proteins isolated from different cellular sources of A. cantonensis, Toxocara canis, Schistosoma mansoni, and Strongyloides stercoralis with mass spectrometry. A total of 115 cross-reactive proteins were identified. Three of these proteins, heat shock protein, an intermediate filament protein, and galectin 1, represent potential markers for cross-reactivity. In addition, synthetic peptides were generated from previously identified diagnostic targets and tested against sera from individuals infected with several other parasites. As a result, two other markers of cross-reactivity were identified: peptide #4 derived from the 14-3-3 protein and peptide #12 derived from the Lec-5 protein. In contrast, 34 proteins were exclusively present in the Angiostrongylus extracts and represent promising diagnostic molecules for specific identification of A. cantonensis infection. In particular, cytochrome oxidase subunit I is of great interest as a possible immunodiagnostic target for angiostrongyliasis., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Cyathostomins infect virtually all horses, and concomitant infections with 10 or more species per horse is standard. Species-specific knowledge is limited, despite potential species bias in development of disease and anthelmintic resistance. This is the first meta-analysis to examine effects of geographical region and cyathostomin collection method on reported composition of cyathostomin communities., Methods: Thirty-seven articles published in English in 1975 or later, in which adults of individual species were systematically enumerated, were included. Seven regions; North America, South America, eastern Europe, western Europe, northern Europe, southern Africa, and Oceania, and three cyathostomin collection methods; (i) standard necropsy recovery from the large intestine, (ii) critical test collection from post-treatment feces and necropsy, and (iii) diagnostic deworming recovery solely from post-treatment feces, were considered. Generalized mixed linear models analyzed the effects of region and collection method on species-specific prevalence and relative abundance. Species richness was analyzed by mixed linear models., Results: Definitively, the most prevalent and relatively abundant species were Cylicocyclus nassatus (prevalence = 93%, relative abundance = 20%), Cylicostephanus (Cys.) longibursatus (93%, 20%), and Cyathostomum catinatum (90%, 16%). A bias toward horses with high infection intensities and cyathostomin collection from feces resulted in North American critical tests and eastern European diagnostic deworming overestimating the species-specific prevalence and underestimating the relative abundance of rare/uncommon species compared to respective intra-regional standard necropsies. North American critical tests underestimated species richness due partially to identification key errors. Inter-regional standard necropsy comparisons yielded some species-specific regional differences, including a significantly higher Cys. longibursatus prevalence and relative abundance in North America (92%, 33%) than in eastern Europe (51%, 7%) (P > 0.0001). Localization of critical tests to North America and diagnostic deworming to Eastern Europe precluded expansive 'region by collection method' interaction analyses., Conclusion: We provide substantial data to inform study design, e.g. effect and study size, for cyathostomin research and highlight necessity for method standardization and raw data accessibility for optimal post-factum comparisons.