scholarly journals The Role of Mustelids in the Transmission of Sarcocystis spp. Using Cattle as Intermediate Hosts

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 822
Author(s):  
Petras Prakas ◽  
Linas Balčiauskas ◽  
Evelina Juozaitytė-Ngugu ◽  
Dalius Butkauskas
Keyword(s):  
American Mink ◽  
Single Species ◽  
Meles Meles ◽  
Cox1 Gene ◽  
Intermediate Hosts ◽  
Specific Pcr ◽  
Sarcocystis Spp ◽  
Species Specific ◽  
Pcr Targeting

There is a lack of research on the role of mustelids in the transmission of various Sarcocystis spp. In the present study we tested the hypothesis that widespread mustelids in Lithuania could be involved in the transmission of Sarcocystis spp. using cattle as intermediate hosts. In 2016–2020, intestinal samples of 84 mustelids were examined. Sarcocystis spp. were identified by species-specific PCR targeting the cox1 gene and subsequent sequencing. Under a light microscope, oocysts/sporocysts of Sarcocystis spp. were observed in 40 samples (47.6%), while using molecular methods, they were detected in 75 animals (89.3%). Four Sarcocystis spp. were identified in the intestinal samples of American mink (Neovisonvison), Beech marten (Martes foina), European pine marten (Martes martes), European badger (Meles meles) and European polecat (Mustela putorius). The prevalence of predominant Sarcocystis spp., S. bovifelis (89.3%) and S. cruzi (73.8%) was significantly higher than that of S. hirsuta (3.6%) and S. hominis (1.2%). In an individual sample, most frequently two Sarcocystis spp. were identified (69.0%), then a single species (15.5%) and three species (4.8%). The present study provides strong evidence that mustelids serve as definitive hosts for Sarcocystis spp. using cattle as intermediate hosts.

scholarly journals The Role of Birds of the Family Corvidae in Transmitting Sarcocystis Protozoan Parasites

Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3258
Author(s):  
Evelina Juozaitytė-Ngugu ◽  
Saulius Švažas ◽  
Donatas Šneideris ◽  
Eglė Rudaitytė-Lukošienė ◽  
Dalius Butkauskas ◽  
...  
Keyword(s):  
Light Microscope ◽  
Sarcocystis Species ◽  
Protozoan Parasites ◽  
Intermediate Hosts ◽  
Specific Pcr ◽  
Sarcocystis Spp ◽  
The Family ◽  
Species Specific ◽  
Pcr Targeting

Members of the family Corvidae are ecologically flexible omnivorous birds, particularly adaptive to urban habitats, and living in proximity to humans; these birds may serve as definitive hosts (DH) for Sarcocystis spp., but research about this is lacking. In the present study, intestinal samples from 91 corvids collected in Lithuania were molecularly tested by species-specific PCR targeting the ITS1 and cox1 genes and subsequently sequenced for the presence of Sarcocystis spp. Under a light microscope, oocysts of Sarcocystis spp. were observed in 43 samples (47.3%), while molecular methods, detected Sarcocystis spp. in 77 birds (84.6%). Eleven Sarcocystis spp. (S. columbae, S. cornixi, potentially pathogenic S. halieti, S. kutkienae, S. lari, S. turdusi, S. wobeseri, S. arctica, S. lutrae, S. ovalis, and S. oviformis) were identified in the intestinal samples from six corvid species from Lithuania. Infections with multiple Sarcocystis spp. were detected in 79.2% of the infected corvid birds. Three of the identified Sarcocystis spp. use corvids as intermediate hosts (IH); therefore, corvids may serve as IH and DH of the same Sarcocystis species. Based on molecular results and on corvid diet, omnivorous corvids may play an important role in transmitting Sarcocystis spp.

scholarly journals Molecular detection of cattle Sarcocystis spp. in North-West Italy highlights their association with bovine eosinophilic myositis

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Selene Rubiola ◽  
Tiziana Civera ◽  
Felice Panebianco ◽  
Davide Vercellino ◽  
Francesco Chiesa
Keyword(s):  
18S Rdna ◽  
Inflammatory Myopathy ◽  
Molecular Techniques ◽  
Diaphragm Muscle ◽  
Economic Losses ◽  
Intermediate Hosts ◽  
Slaughter Cattle ◽  
Significant Difference ◽  
Pcr Targeting

Abstract Background Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa because of the evidence supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based on either morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis. Methods To reach our aim, individual striated muscle samples from BEM condemned carcasses (N = 54) and diaphragm muscle samples from randomly sampled carcasses (N = 59) were obtained from Northwest Italy slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S rDNA and cox1 genes. PCR products amplified using the genus-specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis were sequenced to achieve species identification. Results Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species in slaughter cattle (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses (42.6% and 1.8%, respectively) and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in two carcasses. Conclusions Our study contributes to updating the data on the prevalence of the different Sarcocystis spp. in cattle in Italy, highlighting the presence of three Sarcocystis spp., S. cruzi, S. hominis and S. bovifelis, in BEM lesions and allowing us to speculate on the possible role of S. hominis and S. bovifelis as the major sarcosporidian species involved in bovine eosinophilic myositis. Graphic Abstract

Investigating the role of wild carnivores in the epidemiology of bovine neosporosis

Parasitology ◽  
2012 ◽  
Vol 140 (3) ◽  
pp. 296-302 ◽  
Author(s):  
PETER STUART ◽  
ANNETTA ZINTL ◽  
THEO DE WAAL ◽  
GRACE MULCAHY ◽  
CONALL HAWKINS ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
American Mink ◽  
Protozoan Parasite ◽  
Meles Meles ◽  
Pcr Analysis ◽  
Red Foxes ◽  
Bovine Neosporosis ◽  
Low Prevalence ◽  
Local Ecology

SUMMARYNeospora caninumis a protozoan parasite, primarily associated with bovine abortion. The only definitive hosts discovered to date are carnivores. This study aimed to identify the role of mammalian carnivores in the epidemiology of bovine neosporosis. A sample bank of serum, fecal and brain samples was established: American mink (Mustela vison), red foxes (Vulpes vulpes), pine martens (Martes martes), badgers (Meles meles), stoats (Mustela erminea), otters (Lutra lutra) and feral ferrets (Mustela putorius). Approximately 1% of mink and 1% of fox samples were positive by IFAT. According to PCR analysis of DNA extracted from brain tissue, 3% of the mink, 4% of the otters and 6% of the foxes examined were infected withN. caninum.All fecal samples tested negative forN. caninumDNA (n = 311), suggesting that the species that tested positive were intermediate not definitive hosts. This is the first time that tissues from mustelids have tested positive forN. caninum. The need to test 2 relatively large (∼200 mg) targeted parts of the brain to avoid false negatives was also identified. The relatively low prevalence ofN. caninumin Irish carnivores suggests that the local ecology of a species has an important influence on its epidemiological role.

scholarly journals Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China

2020 ◽  
Author(s):  
Jing Pan ◽  
Chunli Ma ◽  
Zhumei Huang ◽  
Yulong Ye ◽  
Hongxia Zeng ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
18S Rdna ◽  
Sequence Similarity ◽  
Rpob Gene ◽  
28S Rdna ◽  
Ground Substance ◽  
Molecular Characteristics ◽  
Cox1 Gene ◽  
Intermediate Hosts ◽  
Sarcocystis Spp

Abstract Background: There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China.Methods: Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, ITS1 region, the mitochondrial cox1 gene and the apicoplastic rpoB gene, were amplified from each sarcocyst, sequenced and analyzed.Results: Only S. wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and the sequences deposited in GenBank. At 18S rDNA, ITS1 and cox1, the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e. 99.9–100%, 98.1–98.5% and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS1, cox1 and rpoB) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g. 99.8%, 99.0–99.2%, 89.3-89.7%, 98.5%, and 97.5%, respectively, with S. anasi). Phylogenetic analysis based on four of the loci (18S rDNA, cox1, rpoB and ITS1) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that utilize geese or ducks as intermediate hosts and canines as the known or presumed definitive host.Conclusions: To our knowledge, the sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S. wenzeli might be responsible for the neurological disease in chickens, and ITS1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks.

Phenotypic characteristics and population genetics ofEnterococcus faecaliscultured from patients in Tehran during 2000–2001

2003 ◽  
Vol 49 (10) ◽  
pp. 645-649 ◽  
Author(s):  
Mohammad Mehdi Feizabadi ◽  
Atusa Aliahmadi ◽  
Fatemeh Mobasheri ◽  
Ahmad Asgharzadeh ◽  
Soroor Asadi ◽  
...  
Keyword(s):  
Population Genetics ◽  
Enterococcus Faecalis ◽  
Enzyme Electrophoresis ◽  
Bacterial Isolates ◽  
Phenotypic Characteristics ◽  
Considerable Heterogeneity ◽  
Specific Pcr ◽  
Species Specific ◽  
High Level ◽  
Pcr Targeting

Conventional bacteriology techniques were used to identify enterococci isolates cultured from patients at different hospitals in Tehran during 2000–2001. The identification was confirmed using species-specific PCR targeting the D-alanyl-D-alanine ligase gene. A total of 59 isolates of Enterococcus faecalis were identified. The rates of resistance to different antibiotics were in the following order: penicillin 84%, ciprofloxacin 42%, high-level gentamicin 30%, nitrofurantoin 14%, imipenem 4%, and chloramphenicol 2%. Resistance to ampicillin was found to be rare among the Iranian isolates of E. faecalis. Multi-locus enzyme electrophoresis was then used to analyze the strains. Forty-five electrophoretic types were obtained when 10 enzyme loci were screened. Although the collection of bacterial isolates was limited in time and location, considerable heterogeneity was found. Analysis of strains for linkage disequilibrium demonstrated that the studied population is not clonal, since the index of association was not significantly different from zero (Ia= 0.0296). Enterococcus faecalis isolates recovered from patients in Tehran were genetically diverse and seemed to possess a high potential for genetic recombinations, though none were resistant to vancomycin.Key words: Enterococcus faecalis, population genetics, MEE analysis, nosocomial infections.

scholarly journals Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China

2020 ◽  
Author(s):  
Jing Pan ◽  
Chunli Ma ◽  
Zhumei Huang ◽  
Yulong Ye ◽  
Hongxia Zeng ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
18S Rdna ◽  
Sequence Similarity ◽  
28S Rdna ◽  
Ground Substance ◽  
Molecular Characteristics ◽  
Cox1 Gene ◽  
Intermediate Hosts ◽  
Rdna Its ◽  
Sarcocystis Spp

Abstract Background: There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China. Methods: Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, the ITS-1 region ( ITS-1 ), the mitochondrial cox1 gene ( cox1 ), and the apicoplastic rpoB gene ( rpoB ), were amplified from each sarcocyst, sequenced and analyzed. Results: Only S . wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and deposited in GenBank. At 18S rDNA, ITS-1 and cox1 , the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e., 99.9–100%, 98.1–98.5%, and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS-1 , cox1 and rpoB ) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g., 99.8%, 99.0–99.2%, 89.3-89.7%, 98.5%, and 97.5%, respectively, with S . anasi ). Phylogenetic analysis based on three of the loci (18S rDNA, cox1 , and rpoB ) revealed that S . wenzeli formed an independent clade with Sarcocystis spp. that employ geese or ducks as intermediate hosts and canines as the known or presumed definitive host. Conclusions: The sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S . wenzeli might be responsible for the neurological disease in chickens, and ITS-1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S . wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks.

Orphan genes are involved in drought adaptations and ecoclimatic-oriented selections in domesticated cowpea

2019 ◽  
Vol 70 (12) ◽  
pp. 3101-3110 ◽  
Author(s):  
Guojing Li ◽  
Xinyi Wu ◽  
Yaowen Hu ◽  
Maria Muñoz-Amatriaín ◽  
Jie Luo ◽  
...  
Keyword(s):  
Agronomic Traits ◽  
Germplasm Collection ◽  
Single Species ◽  
High Rate ◽  
Soil Drought ◽  
Nucleotide Polymorphisms ◽  
Orphan Genes ◽  
Specific Pcr ◽  
New Genes ◽  
Species Specific

Abstract Orphan genes (OGs) are genes that are restricted to a single species or a particular taxonomic group. To date, little is known about the functions of OGs in domesticated crops. Here, we report our findings on the relationships between OGs and environmental adaptation in cowpea (Vigna unguiculata). We identified 578 expressed OGs, of which 73.2% were predicted to be non-coding. Transcriptomic analyses revealed a high rate of OGs that were drought inducible in roots when compared with conserved genes. Co-expression analysis further revealed the possible involvement of OGs in stress response pathways. Overexpression of UP12_8740, a drought-inducible OG, conferred enhanced tolerance to osmotic stresses and soil drought. By combining Capture-Seq and fluorescence-based Kompetitive allele-specific PCR (KASP), we efficiently genotyped single nucleotide polymorphisms (SNPs) on OGs across a 223 accession cowpea germplasm collection. Population genomic parameters, including polymorphism information content (PIC), expected heterozygosity (He), nucleotide diversity (π), and Tajima’s D statistics, that were calculated based on these SNPs, showed distinct signatures between the grain- and vegetable-type subpopulations of cowpea. This study reinforces the idea that OGs are a valuable resource for identifying new genes related to species-specific environmental adaptations and fosters new insights that artificial selection on OGs might have contributed to balancing the adaptive and agronomic traits in domesticated crops in various ecoclimatic conditions.

scholarly journals Prevalence of mixed Trypanosoma congolense infections in livestock and tsetse in KwaZulu-Natal, South Africa

2010 ◽  
Vol 81 (4) ◽  
Author(s):  
K. Gillingwater ◽  
M.V. Mamabolo ◽  
P.O.A. Majiwa
Keyword(s):  
South Africa ◽  
Trypanosoma Congolense ◽  
Tsetse Flies ◽  
Mixed Infections ◽  
Disease Manifestation ◽  
Blood Samples ◽  
Specific Pcr ◽  
Commercial Farm ◽  
Species Specific ◽  
Pcr Targeting

Trypanosoma congolense causes the most economically important animal trypanosomosis in Africa. In South Africa, a rinderpest pandemic of the 1890s removed many host animals, resulting in the near-eradication of most tsetse species. Further suppression was achieved through spraying with dichlorodiphenyltrichloroethane (DDT); however, residual populations of Glossina austeni and G. brevipalpis remained in isolated pockets. A total of 506 of these tsetse flies were captured in the Hluhluwe-iMfolozi Park, the St Lucia Wetland Park and Boomerang commercial farm. The polymerase chain reaction (PCR) was used to determine the infection rate and frequency of mixed infections of these flies. Additionally, 473 blood samples were collected from cattle at communal diptanks and a commercial farm in the area and each one examined by the haematocrit centrifugation technique (HCT). Furthermore, buffy coats from these blood samples were spotted onto FTA Elute cards and the DNA extracted from each one tested using 3 separate PCRs. The HCT revealed the presence of trypanosomes in only 6.6 % of the blood samples; by contrast, species-specific PCR detected trypanosome DNA in 50 % of the samples. The species-specific PCR detected trypanosome DNA in 17 % of the tsetse flies, compared with the nested PCR targeting rDNA which detected trypanosome DNA in only 14 % of the samples. Over time, the transmission of Savannah-type T. congolense and Kilifi-type T. congolense as mixed infections could have an impact on disease manifestation in different hosts in the area.

scholarly journals Molecular Detection of Cattle Sarcocystis Spp. In North-West Italy Targeting Cox1 And 18S Genes Highlights Its Association With Bovine Eosinophilic Myositis.

2021 ◽  
Author(s):  
Selene Rubiola ◽  
Tiziana Civera ◽  
Felice Panebianco ◽  
Davide Vercellino ◽  
Francesco Chiesa
Keyword(s):  
Inflammatory Myopathy ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Intermediate Hosts ◽  
Prevalence Studies ◽  
North West ◽  
Slaughter Cattle ◽  
Significant Difference ◽  
Pcr Targeting

Abstract Background: Cattle are intermediate hosts of six Sarcocystis species, among which Sarcocystis hominis and Sarcocystis heydorni can infect humans through the consumption of raw or undercooked meat. In addition to the zoonotic potential, there is increasing interest in these protozoa due to the evidences supporting the role of Sarcocystis spp. in the occurrence of bovine eosinophilic myositis (BEM), a specific inflammatory myopathy which leads to carcass condemnation and considerable economic losses. Actually, all the prevalence studies carried out on cattle in Italy have been based either on morphological or 18S rDNA-based molecular techniques, most likely leading to misidentification of closely related species. Therefore, there is a strong need for new data on the prevalence of the different Sarcocystis spp. in cattle in Italy and their association with bovine eosinophilic myositis.Methods: To reach our aim, individual cattle samples from BEM condemned carcasses (N=54) and randomly sampled carcasses (N=59) were obtained from Piedmont slaughterhouses. Genomic DNA was extracted and analyzed by multiplex-PCR targeting 18S and cox1 genes. PCR products amplified using the genus specific primer set in absence of the specific fragment for S. hirsuta, S. cruzi, S. hominis or S. bovifelis, were sequenced to achieve species identification.Results: Sarcocystis DNA was detected in 67.8% of the samples from slaughter cattle and in 90.7% of the samples from BEM condemned carcasses. S. cruzi was identified as the most prevalent species (61%), followed by S. bovifelis (10.2%), S. hominis (8.5%) and S. hirsuta (1.7%). Notably, among the different Sarcocystis spp. detected, the presence of S. bovifelis and S. hominis was significantly higher in samples isolated from BEM condemned carcasses (46.3% and 40.7% respectively), while there was no statistically significant difference between the presence of S. cruzi or S. hirsuta in BEM condemned carcasses and randomly sampled carcasses. Furthermore, DNA sequence analysis revealed the presence of a putative new species in 2 carcasses.Conclusions: Our study contributes to update the data on the prevalence of the different Sarcocystis spp. in cattle in Italy and emphasize the role of S. hominis and S. bovifelis as the major sarcosporidian species involved.

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