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. Graphical

Leishmaniasis is a neglected tropical disease that seriously influences global public health. Among all the parasitic diseases, leishmaniasis is the third most common cause of morbidity after malaria and schistosomiasis. Circular RNAs (circRNAs) are a new type of noncoding RNAs that are involved in the regulation of biological and developmental processes. However, there is no published research on the function of circRNAs in leishmaniasis. This is the first study to explore the expression profiles of circRNAs in leishmaniasis. GO and KEGG analyses were performed to determine the potential function of the host genes of differentially expressed circRNAs. CircRNA–miRNA–mRNA (ceRNA) regulatory network analysis and protein–protein interaction (PPI) networks were analyzed by R software and the STRING database, respectively. A total of 4664 significant differentially expressed circRNAs were identified and compared to those in control groups; a total of 1931 were up-regulated and 2733 were down-regulated. The host genes of differentially expressed circRNAs were enriched in ubiquitin-mediated proteolysis, endocytosis, the MAPK signaling pathway, renal cell carcinoma, autophagy and the ErbB signaling pathway. Then, five hub genes (BRCA1, CREBBP, EP300, PIK3R1, and CRK) were identified. This study provides new evidence of the change of differentially expressed circRNAs and its potential function in leishmaniasis. These results may provide novel insights and evidence for the diagnosis and treatment of leishmaniasis.

(1) Background: As a prominent zoonotic parasitic disease, fascioliasis threatens the sustainable development of animal husbandry and public health. Current research focuses mainly on individual species (parasite or intermediate host), neglecting systematic evaluation of the transmission chain and exposure risks to animal husbandry. Thus, comprehensive studies are urgently needed, especially in the ecologically fragile alpine region of the Qinghai-Tibet Plateau; (2) Methods: Distribution data of Radix spp. and Fasciola hepatica in the Qinghai-Tibet Plateau and adjacent areas were gathered to establish a potential distribution model, which was overlaid on a map of livestock farming in the region; (3) Results: The key environmental factors influencing Radix spp. distribution were temperature seasonality (21.4%), elevation (16.4%), and mean temperature of the driest quarter (14.7%). For F. hepatica, the main factors were elevation (41.3%), human footprint index (30.5%), and Precipitation of the driest month (12.1%), with all AUC values exceeding 0.9. Both species exhibited extensive suitable habitats in Qinghai and Tibet, with higher F. hepatica transmission risk in Qinghai than Tibet; (4) Conclusions: The significant transmission risk and its impacts on the livestock industry in the Qinghai-Tibet Plateau highlight the need for proactive prevention and control measures. This study provides a scientific foundation for targeted alpine diseases control, establishes an interdisciplinary risk assessment framework, fills gaps in high-altitude eco-epidemiology, and offers insights for ecological conservation of the plateau.

Background: The detection of Angiostrongylus spp. larvae in intermediate host snails is a critical epidemiological investigation, essential for the effective control of disease outbreaks. Compared to molecular biological detection methods, lung microscopy, a traditional pathogen morphological detection approach, is susceptible to oversights and exhibits relatively lower sensitivity. However, we posit that lung microscopy offers irreplaceable advantages in the context of large-scale field surveys and can serve as a vital foundation for use in conjunction with other diagnostic technologies. Methods: In this study, 348 Achatina fulica samples were examined using lung microscopy, PCR, and AcanITS1 qPCR. Statistical analysis was conducted to compare detection rates and sensitivities among these methods. DNA from a snail confirmed positive by lung microscopy was diluted and tested using PCR and AcanITS1 qPCR to assess the diagnostic efficacy of the molecular assays. Finally, we combined the highly sensitive AcanITS1 qPCR with lung microscopy for identifying Angiostrongylus spp. larvae in Achatina fulica for the first time to our knowledge and compared its diagnostic efficacy with that of individual testing methods. Results: The lung microscopy, PCR, AcanITS1 qPCR, and combined test yielded detection rates of 29.31%, 32.18%, 38.22%, and 38.51%, respectively. These differences were statistically significant (X ² = 9.565, p < 0.05). Notably, AcanITS1 qPCR demonstrated superior sensitivity with a detection threshold of 10 pg/μl, outperforming the PCR with a threshold of 10 ng/μl. When PCR was utilized as the gold standard, the sensitivities for lung microscopy, AcanITS1 qPCR, and the combined test were determined to be 88.39%, 97.32%, and 98.21%, respectively. Correspondingly, the specificities were 98.73%, 89.83%, and 89.83%, respectively. Conclusions: This novel straregy, the combined test for the detection of Angiostrongylus spp. larvae in Achatina fulica exhibited superior positive detection rates and sensitivity compared to each of the three individual methods. We believe that this novel strategy is not only applicable to large-scale field investigations of Achatina fulica and Pomacea canaliculata but also has potential application value for monitoring the infection of these snails sold at the local farmers’ markets with Angiostrongylus spp. larvae. Of course, while qPCR is exceptionally sensitive, the potential for false negatives remains a consideration. Repeated experimentation is also essential to maximize the reliability and accuracy of the outcomes. Graphical

Biomphalaria pfeifferi is a key intermediate host for Schistosoma mansoni transmission in Sudan. In total, 27 complete mitochondrial genomes from seven B. pfeifferi populations in Gezira State, Sudan, were sequenced for the first time to investigate their population structure and phylogenetic relationships. This involved comparing the nucleotide composition, codon usage, rRNAs, and tRNAs of the East Gezira (EG), South Gezira (SG), Hasahisa (HA), Greater Wad Medani (GW), Managil (MA), and North Umelgura (NU1, NU3) populations. All 27 mitogenomes (13,688–13,696 bp) contained 37 genes with conserved AT/GC content (76.7/23.4%). Phylogenetic analysis revealed that although samples clustered within the same clade, B. pfeifferi from EG, SG, NU1, and NU3 grouped closely with B. pfeifferi from Kenya, whereas HA and GW samples formed distinct ancestral lineages. The MA population exhibited unique genetic characteristics, supported by phylogenetic trees and nucleotide/amino acid identity, suggesting the potential presence of a distinct B. pfeifferi subspecies that warrants further investigation. All protein-coding genes evolved under negative selection, with the amino acids of nad1 and nad6 being highly conserved, while nad3 exhibited some variation. Further research on the mitogenomic diversity of B. pfeifferi and other Biomphalaria species in Sudan and across Africa is needed in order to better understand the population structure and evolutionary history of Biomphalaria.

Angiostrongylus cantonensis (A. cantonensis) is the primary causative agent of human angiostrongyliasis and is widely distributed in Southeast Asia and China, with increasing reports from the Americas. Achatina fulica (A. fulica), Pomacea canaliculata (P. canaliculata), and slugs constitute established intermediate hosts of A. cantonensis, whereas Camaena hainanensis (C. hainanensis) has been newly reported as a host species in Hainan. A TaqMan quantitative PCR (qPCR) method assay targeting a novel genomic region of A. cantonensis was developed to detect infection in 150 snails collected from Hainan Province, China. The assay was employed to detect the parasite larvae across various snail tissues (lung sac, mucus, and foot), and its performance was compared with conventional lung sac microscopy. Out of the 120 A. fulica examined, 75 tested positive using the qPCR assay, yielding a significantly higher detection rate than lung-sac examination (p < 0.05). Significant differences were also observed in the positivity rates across the three snail tissues (lung sac, mucus, and foot) (p < 0.05), with the lung sac showing the highest rate of infection. Importantly, the detection of A. cantonensis DNA in snail mucus highlights its potential for development as a non-invasive diagnostic sample. Additionally, C. hainanensis was identified as a new host of A. cantonensis in Hainan, suggesting its possible contribution to parasite transmission. The newly developed qPCR assay demonstrated superior sensitivity (reflected by lower Ct values) compared with previously published TaqMan qPCR methods. The established qPCR method provides a sensitive and non-invasive tool for detecting A. cantonensis in snails, and can be applied for monitoring and early warning of parasite prevalence and transmission.