Parasites & Vectors最新文献

Ticks are well-known vectors of pathogens, including filarial nematodes, which can affect animal and human health. This review synthesizes current knowledge of filarial nematodes that utilize ticks as vectors, focusing on the Onchocercidae. Five genera of onchocercid filarial nematodes (Cercopithifilaria, Cherylia, Cruorifilaria, Monanema, and Yatesia) have demonstrated vector-parasite relationships with ticks, while Acanthocheilonema has only demonstrated molecular detection without confirmed vector competence. Vector competence has been experimentally proven for several species through documented development to infective stages. However, our understanding of tick-borne filarial nematodes remains limited owing to insufficient detection efforts and methodological challenges. Detection methods include traditional dissection, advanced imaging techniques such as microcomputed tomography (micro-CT), and molecular approaches, each with distinct advantages and limitations. To address knowledge gaps, future research should focus on conducting large-scale surveys of tick populations and wildlife to investigate vector competence, health impacts, and coinfection dynamics. Multi-disciplinary collaboration and community involvement in tick surveillance will advance our understanding of tick-borne filarial nematode ecology and epidemiology. Future research priorities include confirming vector competence where only molecular detection exists, investigating natural transmission patterns, and understanding the mechanisms influencing successful development in tick vectors. By addressing these knowledge gaps through systematic surveillance and experimental studies, we can better understand these complex host-parasite-vector relationships and their implications for animal health.

Background: Neospora caninum is an intracellular parasitic protozoon that can infect pregnant animals and cause symptoms such as miscarriage, stillbirth and mummified fetuses. It is one of the main causes of miscarriage in bovines. Apical membrane antigen (AMA) and dense granule protein (GRA) are two major antigenic proteins of N. caninum.

Methods: In this study, NcGRA9 recombinant subunit vaccine and Ad5-NcAMA1 recombinant adenovirus vaccine were prepared and used to immunize C57BL/6 mice and Yanbian yellow cattle.

Results: IgG, IgG1, IgG2a, IgG2b, IgA and IgE antibodies and interferon (IFN-γ), interleukin (IL-4) and tumor necrosis factor (TNF-α) cytokines were significantly higher in immunized mice than in the control group (P < 0.0001). The biochemical indexes showed that vaccination had no effect on hepatic and renal functions. The survival rate was 70% in mice immunized with NcGRA9 vaccine, 75% in mice immunized with Ad5-NcAMA1 vaccine, 85% in the combined immunization group and 10% in the control group. The parasite load in the brain and liver tissues of the immunized groups was significantly lower than in the control group, as detected by fluorescence quantitative PCR (P < 0.0001). In cattle, IgG, IgG2a, IgG2b, IgA, IgM and IgE antibodies and IFN-γ, IL-4 and TNF-α cytokines were significantly higher in the immunized groups than in the control group (P < 0.0001). Combined immunization with NcGRA9 + Ad5-NcAMA1 was significantly better than immunization with either vaccine alone.

Conclusions: The biochemical indexes showed that the vaccine had no effect on the liver and kidney functions of cattle. Our results indicate that combined immunization with NcGRA9 + Ad5-NcAMA1 may be a candidate for bovine neosporosis vaccination.

Background: Phlebotomine sand flies are the key vectors for phleboviruses (order Hareavirales and family Phenuiviridae), of which some are associated with febrile diseases and nervous system infections. In the Mediterranean Basin, Toscana virus (TOSV; Phlebovirus toscanaense) and sandfly fever Sicilian viruses (SFSV; Phlebovirus siciliaense) are important human pathogens, and their endemicity has been known for decades, particularly in the Balkan countries. While the circulation of both viruses is highly evident among humans and livestock in the Central Balkan country Kosovo, data from companion animals are scarce; however, it might help to further assess the distribution of both viruses in the country.

Methods: Sera of dogs from all seven districts of Kosovo were screened for TOSV and SFSV antibodies by seroneutralization assays.

Results: Altogether, 45 of 288 (15.6%) samples showed anti-Phlebovirus antibodies, of which 36 (12.5%) were against TOSV, 11 (3.8%) were against SFSV, and 2 (0.7%) were positive for antibodies against both viruses.

Conclusions: Phlebovirus seroprevalence was observed in all seven districts of the country, generally being higher for TOSV compared with SFSV. Our study presents the first assessment of neutralization-based seroprevalence of two medically important phleboviruses among dogs in the Republic of Kosovo. Although healthy dogs are unsusceptible to Phlebovirus infection, dogs with leishmaniasis can be potential amplifying hosts. Given the high number of stray dogs, frequent uncontrolled spreading of phleboviruses in dogs, and potential spillover in populated regions of the country, these findings should be taken into consideration.

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 (X² = 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.

Background: Malaria is the parasitic disease with the highest morbidity and mortality worldwide. Angola is one of the five sub-Saharan African countries with the highest malaria burden. Real-time PCR diagnosis in endemic areas has not been implemented due to its high cost and the need for adequate infrastructure. Dried blood spots (DBSs) are an alternative for collecting, preserving, and transporting blood samples to reference laboratories. The objective of the study was to assess the efficacy of DBS as a sampling method for malaria research studies employing real-time PCR.

Methods: The study was divided into two phases: (i) prospective study at the Hospital Universitario Vall d'Hebron (HUVH) to compare real-time PCR from whole blood or DBS, including 12 venous blood samples from patients with positive real-time PCR for Plasmodium spp. and 10 quality control samples (nine infected samples and one negative control). Samples were collected as DBSs (10, 20, 50 µl/circle). Samples from both phases of the study were analyzed by generic real-time PCR (Plasmodium spp.) and the subsequent positive samples underwent species-specific real-time PCR (Plasmodium species) and (ii) cross-sectional study conducted at the Hospital Nossa Senhora da Paz, Cubal (Angola), including 200 participants with fever. For each patient, a fresh capillary blood specimen [for thin and thick blood films and rapid diagnostic test (RDT)] and venous blood, collected as DBSs (two 10-µl circles were combined for a total volume of 20 µl of DBS), were obtained. DBSs were sent to HUVH, Barcelona, Spain.

Results: (i) Real-time PCR from whole blood collection was positive for 100% of the 21 Plasmodium spp.-infected samples, whereas real-time PCR from DBSs detected Plasmodium spp. infection at lower proportions: 76.19% (16/21) for 10 µl, 85.71% (18/21) for 20 µl, 88.24% (15/17) for 50 µl and 85.71% (18/21) for 100 µl DBSs. (ii) Field diagnosis (microscopy and/or RDT) showed a 51.5% (103/200) positivity rate, while 50% (100/200) of the DBS samples tested positive by real-time PCR. Using field diagnosis as the reference method, the sensitivity of real-time PCR in DBS samples was 77.67% with a specificity of 79.38%. Plasmodium species were identified in 86 samples by real-time PCR: 81.40% (16/86) were caused by Plasmodium falciparum, 11.63% (10/86) were coinfections of P. falciparum + P. malariae, 4.65% (4/86) were P. falciparum + P. ovale, and 2.33% (2/86) were triple coinfections.

Conclusions: The DBS volume used for DNA extraction is a determining factor in the performance of real-time PCR for Plasmodium DNA detection. A DBS volume of 50-100 µl appears to be optimal for malaria diagnosis and Plasmodium species determination by real-time PCR. DBS is a suitable method for sample collection in Cubal followed by real-time PCR analysis in a reference laboratory.

Background: Anopheles stephensi is a competent malaria vector mainly present in southern Asia and the Arabian Peninsula. Since 2012, it has invaded several countries of eastern Africa, creating an emerging risk of urban transmission. Urgent efforts are required to develop novel and more efficient strategies for targeted vector control. CRISPR/Cas9-based homing gene drives have been proposed as attractive alternative strategies. Gene drives have the potential to spread a desired trait through a population at higher rates than via normal Mendelian inheritance, even in the presence of a fitness cost. Several target genes have been suggested and tested in different mosquito vector species such as Anopheles gambiae and Aedes aegypti. Several promising suppression drives have been developed in An. gambiae that target the sex determination gene doublesex (dsx).

Methods: In this study, a geographically confineable gene drive system targeting dsx was developed (dsx^gRNA). Here, a transgenic line which expresses Cas9 under the control of the endogenous zpg promoter was generated. Separately a transgenic line which expresses a gRNA targeting the female specific exon of dsx was inserted into that same target site. The reproductive fitness of males and females heterozygous and homozygous for this element was determined. A series of experimental crosses was performed to combine the two elements and assess the homing rate of the dsx element in a split drive system.

Results: The drive was able to home in a super-Mendelian rate comparable to those obtained by an autonomous drive in this species. Although inheritance rates as high as 99.8% were observed, potentially providing very potent gene drive, dominant effects on male and female fertility were observed, which would be sufficient to hinder spread of such a drive. Molecular analysis indicated that the gRNA expressing insertion disrupted normal splicing of dsx.

Conclusions: These results should be considered when proposing the viability of dsx as a target gene for a population suppression gene drives in Anopheles stephensi. Although high homing rates were observed, the fitness defects found in both males and females carrying the transgene would likely prohibit this drive from functioning in the field.

White-tailed deer (Odocoileus virginianus) are a ubiquitous species in North America. Their high reproductive potential leads to rapid population growth, and they exhibit a wide range of biological adaptations that influence their interactions with vectors and pathogens. This review aims to characterize the intricate interplay between white-tailed deer and the transmission cycles of various tick- and mosquito-borne pathogens across their range in the eastern United States and southeastern Canada. The first part offers insights into the biological characteristics of white-tailed deer, their population dynamics, and the consequential impacts on both the environment and public health. This contextual backdrop sets the stage for the two subsequent sections, which delve into specific examples of pathogen transmission involving white-tailed deer categorized by tick and mosquito vectors into tick-borne and mosquito-borne diseases. This classification is essential, as ticks and mosquitoes serve as pivotal elements in the eco-epidemiology of vector-borne diseases, intricately linking hosts, the environment, and pathogens. Through elucidating these associations, this paper highlights the crucial role of white-tailed deer in the transmission dynamics of tick- and mosquito-borne diseases. Understanding the interactions between white-tailed deer, vectors, and pathogens is essential for effective disease management and public health interventions.

Background: The intestinal protozoa Entamoeba spp. can infect humans and various animals, including donkeys, causing diarrhea and malabsorption and presenting significant risks to animal husbandry and public health. Most Entamoeba species are not pathogenic except for Entamoeba histolytica. China has among the highest rates of donkey farming worldwide. Donkey (Equus asinus) farming is increasingly important in China because of their draft and medicinal value; however, epidemiological data on Entamoeba spp. in donkeys remains limited globally. This study aimed to investigate the prevalence of Entamoeba in donkeys in Shanxi Province, North China, and assess associated risk factors using a molecular approach.

Methods: Fecal samples of 815 donkeys from three representative geographical locations in Shanxi Province were collected to investigate the presence of Entamoeba spp. A portion of the small-subunit rRNA gene (SSU rRNA) was amplified and sequenced to determine the prevalence and species/genotypes of Entamoeba spp. Statistical analysis of possible risk factors was performed using Statistical Product and Service Solutions (SPSS) 26.0 software. The phylogenetic relationship of Entamoeba spp. was reconstructed using the neighbor-joining (NJ) method in Molecular Evolutionary Genetics Analysis (Mega) 7.0 software.

Results: The overall prevalence of Entamoeba spp. in donkeys in Shanxi Province was 7.12% (58/815). Two species (Entamoeba sp. RL9 and Entamoeba equi) were identified by sequence analysis; of these, Entamoeba sp. RL9 was the most prevalent species in donkeys in this study. Statistical analysis revealed that the donkeys' sex, region, age, and altitude are the risk factors associated with Entamoeba spp. prevalence (P < 0.05). Phylogenetic analysis indicated that the sequences of Entamoeba sp. RL9 and E. equi isolated from donkeys in this study were clustered with previously reported animal-derived Entamoeba sp. RL9 and E. equi sequences, respectively.

Conclusions: This study reports the occurrence and prevalence of Entamoeba spp. in donkeys worldwide for the first time to our knowledge. This not only expands the geographical distribution but also broadens the host range of Entamoeba spp., addressing the knowledge gap regarding the prevalence of Entamoeba spp. in donkeys, providing baseline data for carrying out prevention and control of Entamoeba spp. in donkeys in China.

Background: Jingmen tick virus (JMTV) is a novel tick-borne virus detected for the first time in Rhipicephalus (Boophilus) microplus in China. To date, there is no information regarding the circulation of JMTV in ticks collected from livestock in Cameroon. As part of the surveillance for arboviral circulation, this study aimed to assess the presence of JMTV in ticks collected from livestock (cattle and sheep) in an area of the Akonolinga health district, Center Region, Cameroon.

Methods: A cross sectional study was carried out during the dry season between 5 and 14 March 2024. Ticks were collected from cattle and sheep in six sampling sites in an area approximately 30 km long and 18 km wide along the Nyong River, in central Cameroon. Ticks were identified morphologically and molecularly. Total RNA/DNA was extracted from tick pools and screened for JMTV RNA using a segment 2 RT-qPCR system. Positive JMTV pools were sequenced for partial JMTV-Segment 1 and full genome analyses.

Results: A total of 622 ticks, organized into 251 pools were collected from 155 cattle and nine sheep. They consisted of five species covering three genera: R. (B.) microplus (472; 75.9%), Amblyomma variegatum (118; 19.0%), Hyalomma truncatum (13; 2.1%), Hyalomma rufipes (2; 0.3%), and other Rhipicephalus spp. (17; 2.7%). The quantitative reverse transcription polymerase chain reaction (qRT-PCR) screening of 251 tick pools yielded 61 JMTV-positive pools, of which 58 corresponded to R. (B.) microplus. Multiple sequence analysis revealed that JMTV from the Akonolinga area shared > 95% identity with strains from Guinea, and that these strains clustered phylogenetically together.

Conclusions: We provide molecular evidence of the presence of JMTV in R. (B.) microplus and A. variegatum collected from cattle and sheep from an area not yet recognized as endemic for this virus, confirming its wide geographical distribution.