scholarly journals Trichinella spp. infection in European polecats (Mustela putorius Linnaeus, 1758) from Romania

2021 ◽  
Vol 58 (3) ◽  
pp. 323-327
Author(s):  
Z. Boros ◽  
A. M. Ionică ◽  
G. Deak ◽  
A. D. Mihalca ◽  
G. Chișamera ◽  
...  
Keyword(s):  
Molecular Biology ◽  
Multiplex Pcr ◽  
Parasite Species ◽  
Trichinella Spiralis ◽  
Wide Distribution ◽  
Mustela Putorius ◽  
Species Determination ◽  
Human Settlements ◽  
Tissue Samples ◽  
Artificial Digestion

Summary The European polecat (Mustela putorius Linnaeus, 1758) is in decline in Romania, often living near human settlements, from mountains to lowlands. They feed on a wide variety of small animals, including rodents, such as mice or rats. The occurrence of this parasite in polecats from Romania was mentioned only once in 1991, but the parasite species was not confirmed by molecular biology. The study aimed to investigate the occurrence of Trichinella spp. in European polecats from Romania and to identify the parasite species by molecular tools. A total of 75 wild European polecats were examined by trichinoscopy and artificial digestion. A large number of animals were examined because of their wide distribution in Romanian territory and their presence near human settlements. For species determination, the positive muscle samples and the larvae recovered from artificial digestion were collected for DNA isolation and further processed by means of Multiplex PCR. Only two polecats from southern Romania tested positive for Trichinella spp. infection. During trichinoscopy examination, 48 (in a polecat from Giurgiu County) and 78 (in a polecat from Ialomița County) cysts were found in the tested (56 samples/animal) tissue samples. Artificial digestion revealed infection with 2466 larvae/100 g of muscle in the polecat from Ialomița and 254/100 g in the polecat from Giurgiu. The Multiplex PCR indicated the occurrence of Trichinella spiralis in the polecat from Giurgiu and a co-infection with T. spiralis and T. britovi in the polecat from Ialomița. The current study confirms through molecular biology, the occurrence of T. spiralis and T. britovi, as well as the occurrence of co-infection with these two Trichinella species in European polecats from Romania.

Methods for the Detection of Trichinellosis in Horses

1996 ◽  
Vol 59 (4) ◽  
pp. 420-425 ◽  
Author(s):  
H. RAY GAMBLE ◽  
ALVIN A. GAJADHAR ◽  
MORSE B. SOLOMON
Keyword(s):  
Enzyme Immunoassay ◽  
Trichinella Spiralis ◽  
Larval Density ◽  
Human Health Risk ◽  
Dependent Manner ◽  
Sample Sizes ◽  
Tissue Samples ◽  
Artificial Digestion ◽  
Dose Dependent ◽  
Digestion Methods

Twelve horses were infected with various doses of Trichinella spiralis and then tested for infection using direct (artificial digestion) and indirect (enzyme immunoassay) methods. Horses became infected in a dose-dependent manner. Larvae accumulated preferentially in the tongue, followed by the masseter, neck, supraspinatus, trapezius, and diaphragm. At lower infection levels, the tongue harbored several times more parasites than were found in other tissues. The sensitivity of artificial digestion methods for detecting infections was directly related to sample size. One-gram samples were not reliable for detecting infection levels of <3 larvae per g (LPG). In sample sizes of 5 or 10 g the technique allowed infections as low as 1 LPG to be detected. The enzyme immunoassay (EIA) detected all infected horses; the times following infection at which horses became seropositive varied in a dose-dependent manner, but 11 of 12 horses were positive in the EIA by 4 weeks postinoculation. One horse, with a larval density in the tongue of 0.39 LPG, did not become seropositive until 7 weeks postinoculation. The results suggest that artificial digestion of horse carcasses for trichinae should concentrate on tissue samples from the tongue or masseter muscles. Sample sizes should be a minimum of 5 g using pooled-sample digestion methods to assure detection of all infections which might pose a human health risk. The EIA is a potential substitute for artificial digestion methods and could also be useful for antemortem testing and for epidemiological studies.

Prospective enzymes for omega-3 PUFA biosynthesis found in endoparasitic classes within the phylum Platyhelminthes

2020 ◽  
Vol 94 ◽  
Author(s):  
D. Babaran ◽  
M.T. Arts ◽  
R.J. Botelho ◽  
S.A. Locke ◽  
J. Koprivnikar
Keyword(s):  
Parasite Species ◽  
De Novo ◽  
Genomic Data ◽  
Wide Distribution ◽  
Omega 3 ◽  
Free Living ◽  
Chain Pufa ◽  
Pufa Biosynthesis ◽  
Aquatic Food ◽  
Long Chain

Abstract The free-living infectious stages of macroparasites, specifically, the cercariae of trematodes (flatworms), are likely to be significant (albeit underappreciated) vectors of nutritionally important polyunsaturated fatty acids (PUFA) to consumers within aquatic food webs, and other macroparasites could serve similar roles. In the context of de novo omega-3 (n-3) PUFA biosynthesis, it was thought that most animals lack the fatty acid (FA) desaturase enzymes that convert stearic acid (18:0) into ɑ-linolenic acid (ALA; 18:3n-3), the main FA precursor for n-3 long-chain PUFA. Recently, novel sequences of these enzymes were recovered from 80 species from six invertebrate phyla, with experimental confirmation of gene function in five phyla. Given this wide distribution, and the unusual attributes of flatworm genomes, we conducted an additional search for genes for de novo n-3 PUFA in the phylum Platyhelminthes. Searches with experimentally confirmed sequences from Rotifera recovered nine relevant FA desaturase sequences from eight species in four genera in the two exclusively endoparasite classes (Trematoda and Cestoda). These results could indicate adaptations of these particular parasite species, or may reflect the uneven taxonomic coverage of sequence databases. Although additional genomic data and, particularly, experimental study of gene functionality are important future validation steps, our results indicate endoparasitic platyhelminths may have enzymes for de novo n-3 PUFA biosynthesis, thereby contributing to global PUFA production, but also representing a potential target for clinical antihelmintic applications.

scholarly journals Identification of Mycobacterium genavense natural infection in a domestic ferret

2018 ◽  
Vol 31 (1) ◽  
pp. 133-136 ◽  
Author(s):  
Bérengère Dequéant ◽  
Quentin Pascal ◽  
Héloïse Bilbault ◽  
Elie Dagher ◽  
Maria-Laura Boschiroli ◽  
...  
Keyword(s):  
Natural Infection ◽  
Abdominal Ultrasound ◽  
Needle Aspiration ◽  
Mesenteric Lymph Nodes ◽  
Mustela Putorius ◽  
Tissue Samples ◽  
Acid Fast Bacilli ◽  
Mycobacterium Genavense ◽  
History Of ◽  
Progressive Weight Loss

A 6-y-old neutered male ferret ( Mustela putorius furo) was presented because of a 1-mo history of progressive weight loss, chronic cough, and hair loss. On clinical examination, the animal was coughing, slightly depressed, moderately hypothermic, and had bilateral epiphora. Thoracic radiography was suggestive of severe multinodular interstitial pneumonia. Abdominal ultrasound examination revealed hepatosplenomegaly and mesenteric and pancreaticoduodenal lymphadenopathy. Fine-needle aspiration of the pancreaticoduodenal lymph node, followed by routine Romanowsky and Ziehl–Neelsen stains, revealed numerous macrophages containing myriad acid-fast bacilli, leading to identification of mycobacteriosis. Autopsy and histologic examination confirmed the presence of disseminated, poorly defined, acid-fast, bacilli-rich granulomas in the pancreaticoduodenal and mesenteric lymph nodes, intestines, and lungs. Destaining of May-Grünwald/Giemsa–stained slides with alcohol, and then restaining with Ziehl–Neelsen, revealed acid-fast rods and avoided repeat tissue sampling without affecting the Ziehl–Neelsen stain quality and cytologic features. Tissue samples were submitted for a PCR assay targeting the heat shock protein gene ( hsp65) and revealed 100% homology with Mycobacterium genavense. We emphasize the use of special stains and PCR for identification of this potential zoonotic agent.

Sensitivity of Artificial Digestion and Enzyme Immunoassay Methods of Inspection for Trichinae in Pigs

1998 ◽  
Vol 61 (3) ◽  
pp. 339-343 ◽  
Author(s):  
H. R. GAMBLE
Keyword(s):  
Sample Size ◽  
Enzyme Immunoassay ◽  
Trichinella Spiralis ◽  
Serological Testing ◽  
Digestion Method ◽  
Artificial Digestion ◽  
Eu Directives ◽  
Digestion Methods ◽  
Pooled Sample ◽  
The Eu

Forty-seven pigs were infected with varying doses of Trichinella spiralis and tested for evidence of infection by serology, using an enzyme immunoassay (EIA), and by artificial digestion methods. Using a 1-g sample, as prescribed in accordance with European Union (EU) directives, the sensitivity of the pooled-sample artificial digestion method was between three and five larvae per gram (LPG) of tissue. Using a 5-g sample size, in accordance with methods described in the U.S. Code of Federal Regulations, and as required for the inspection of horses exported to the EU, the sensitivity of the test was increased to approximately 1 LPG. Serological testing by EIA detected pigs with as few as 0.02 LPG, but detection times varied from 4 to 8 weeks after infection. Mean postinoculation times for detection by serology were 32 to 42 days. On the basis of these results, it is clear that digestion testing using a 5-g sample size is the only method of those tested here that is completely reliable for detection of trichinae infection at a level that will protect public health. Both digestion testing using a 1-g sample and EIA have drawbacks. However, EIA remains a highly effective tool for epidemiological purposes and for monitoring trichinae infection on the farm.

Immunodiffusion Technique for the Identification of Animal Species

1971 ◽  
Vol 54 (5) ◽  
pp. 1152-1156 ◽  
Author(s):  
H Guy Fugate ◽  
Shelton R Penn
Keyword(s):  
Species Identification ◽  
Animal Species ◽  
Ring Test ◽  
Species Determination ◽  
Agar Gel ◽  
Simple Method ◽  
Tissue Samples ◽  
Immunodiffusion Test ◽  
Meat Animal ◽  
Antigen Antibody

Abstract An agar-gel immunodiffusion technique was developed for the identification of meat animal species. A pattern of wells and troughs was cut from agar plates. The wells and troughs contained antigens and antisera, respectively. The diffusion of the antigens and antisera through the agar results in the formation of precipitin lines when optimum antigen-antibody conditions exist. The interpretation of the reactions depends upon the position of the formed precipitin lines in relation to each other. Eleven of 12 mixed tissue samples submitted to the authors’ laboratory for species determination were identified correctly by the agar-gel immunodiffusion test. The test is a relatively rapid and simple method of confirming the results of the tube precipitin ring test for animal species identification.

scholarly journals Genome Characterization, Prevalence, and Transmission Mode of a Novel Picornavirus Associated with the Threespine Stickleback Fish (Gasterosteus aculeatus)

2019 ◽  
Vol 93 (9) ◽  
Author(s):  
Megan A. Hahn ◽  
Nolwenn M. Dheilly
Keyword(s):  
Gasterosteus Aculeatus ◽  
Rna Virus ◽  
Wide Distribution ◽  
Model System ◽  
Threespine Stickleback ◽  
Tissue Samples ◽  
Content Type ◽  
Threespine Sticklebacks ◽  
The Impact

ABSTRACT The complete genome sequence of an RNA virus was assembled from RNA sequencing of virus particles purified from threespine stickleback intestine tissue samples. This new virus is most closely related to the Eel picornavirus and can be assigned to the genus Potamipivirus in the family Picornaviridae. Its unique genetic properties are enough to establish a new species, dubbed the Threespine Stickleback picornavirus (TSPV). Due to their broad geographic distribution throughout the Northern Hemisphere and parallel adaptation to freshwater, threespine sticklebacks have become a model in evolutionary ecology. Further analysis using diagnostic PCRs revealed that TSPV is highly prevalent in both anadromous and freshwater populations of threespine sticklebacks, infects almost all fish tissues, and is transmitted vertically to offspring obtained from in vitro fertilization in laboratory settings. Finally, TSPV was found in Sequence Reads Archives of transcriptome of Gasterosteus aculeatus, further demonstrating its wide distribution and unsought prevalence in samples. It is thus necessary to test the impact of TSPV on the biology of threespine sticklebacks, as this widespread virus could interfere with the behavioral, physiological, or immunological studies that employ this fish as a model system. IMPORTANCE The threespine stickleback species complex is an important model system in ecological and evolutionary studies because of the large number of isolated divergent populations that are experimentally tractable. For similar reasons, its coevolution with the cestode parasite Schistocephalus solidus, its interaction with gut microbes, and the evolution of its immune system are of growing interest. Herein we describe the discovery of an RNA virus that infects both freshwater and anadromous populations of sticklebacks. We show that the virus is transmitted vertically in laboratory settings and found it in Sequence Reads Archives, suggesting that experiments using sticklebacks were conducted in the presence of the virus. This discovery can serve as a reminder that the presence of viruses in wild-caught animals is possible, even when animals appear healthy. Regarding threespine sticklebacks, the impact of Threespine Stickleback picornavirus (TSPV) on the fish biology should be investigated further to ensure that it does not interfere with experimental results.

The use of chicory for parasite control in organic ewes and their lambs

Parasitology ◽  
2006 ◽  
Vol 134 (2) ◽  
pp. 299-307 ◽  
Author(s):  
S. ATHANASIADOU ◽  
D. GRAY ◽  
D. YOUNIE ◽  
O. TZAMALOUKAS ◽  
F. JACKSON ◽  
...  
Keyword(s):  
Parasite Species ◽  
Experimental Period ◽  
Cichorium Intybus ◽  
Parasite Control ◽  
Species Determination ◽  
Predominant Species ◽  
Teladorsagia Circumcincta ◽  
Potential Benefits ◽  
Intestinal Worm ◽  
Certified Organic

The aim of the present study was to investigate the potential benefits of grazing lactating ewes and their lambs on chicory (Cichorium intybus). Fifty-six certified organic twin-rearing ewes were either drenched with an anthelmintic or not, within 2 days after parturition and were grazed upon either grass/clover or chicory pastures. Around 12 weeks after parturition a subset of 12 lambs per treatment was slaughtered for worm number and parasite species determination. The faecal egg counts of lambs from undrenched ewes grazing on chicory were significantly lower than those of lambs from undrenched ewes grazing on grass. Lambs grazing on chicory had similar abomasal worm counts as those grazing on grass at 12 weeks of age; the predominant species wasTeladorsagia circumcincta. There was no difference between the intestinal worm counts in lambs grazing on grass or chicory, withTrichostrongylus vitrinusbeing the predominant species. Liveweight gains over the 126-day experimental period were significantly higher in lambs from drenched than those from undrenched ewes. Lambs from undrenched ewes grazing on chicory had higher liveweight gains compared to those from undrenched ewes grazing on grass. Although chicory grazing did not affect ewe nematode egg excretion, it resulted in lower egg counts in lambs and improved their liveweight gains to the same level as those deriving from drenched ewes.

Morphological and molecular analyses of larval taeniid species in small mammals from contrasting habitats in Denmark

2013 ◽  
Vol 89 (1) ◽  
pp. 112-117 ◽  
Author(s):  
M.N.S. Al-Sabi ◽  
P.M. Jensen ◽  
M.U. Christensen ◽  
C.M.O. Kapel
Keyword(s):  
Multiplex Pcr ◽  
Small Mammals ◽  
Habitat Type ◽  
Prevalence Rates ◽  
Intermediate Hosts ◽  
Morphological Examination ◽  
Species Determination ◽  
North East ◽  
Significant Difference ◽  
Species Specific

AbstractTaeniid infections in intermediate hosts manifest themselves as extraintestinal larval stages which, in early development, lack species-specific characteristics. The inability to distinguish infections of zoonotic importance such as Echinococcus multilocularis from other taeniid infections that have mainly veterinary significance stimulated the development of species-specific molecular diagnostics. In this study, the prevalence of taeniid infections in potential intermediate hosts was evaluated using both morphological diagnosis and a newly described multiplex polymerase chain reaction (PCR) for species determination. Small mammals (N= 719) were trapped in three different types of habitats in north-east Zealand, Denmark. The sensitivity of the multiplex PCR (90.5%) exceeded that of morphological examination (57.9%) for identifying 95 taeniid infections. The use of the multiplex PCR resulted in higher prevalence rates due to improved detection of immature liver infections with Hydatigera taeniaeformis and Versteria mustelae, but did not affect the observed prevalence rates of peritoneal metacestodes of Taenia polyacantha. The prevalence of taeniid infections showed a significant difference according to habitat type, potentially identifying a ‘sylvatic’ transmission and an ‘urban’ transmission, with marked variation among different taeniid species. Versteria mustelae and T. polyacantha were more prevalent in rural forests, while infections with H. taeniaeformis were dominant in urban parks/forests and in residential and farm gardens. The multiplex PCR facilitated a better utilization of wildlife samples by yielding a higher number of definitive diagnoses of ambiguous taeniid infections in liver lesions, allowing for more accurate epidemiological data and, hence, a more accurate risk assessment.

scholarly journals Detection and quantification of house mouse Eimeria at the species level – challenges and solutions for the assessment of Coccidia in wildlife

2019 ◽  
Author(s):  
Víctor Hugo Jarquín-Díaz ◽  
Alice Balard ◽  
Jenny Jost ◽  
Julia Kraft ◽  
Mert Naci Dikmen ◽  
...  
Keyword(s):  
House Mouse ◽  
Parasite Species ◽  
Host Population ◽  
Species Level ◽  
Free Living ◽  
Additional Indication ◽  
Tissue Samples ◽  
Species Specific ◽  
Apicoplast Genome ◽  
Detection And Quantification

AbstractDetection and quantification of coccidia in studies of wildlife can be challenging. Therefore, the prevalence of coccidia is often not assessed at the parasite species level in non-livestock animals. Parasite species-specific prevalences are especially important when studying evolutionary questions in wild populations. We tested whether increased host population density increases the prevalence of individual Eimeria species at the farm level, as predicted by epidemiological theory. We studied free-living commensal populations of the house mouse (Mus musculus) in Germany and established a strategy to detect and quantify Eimeria infections. We show that a novel diagnostic primer targeting the apicoplast genome (Ap5) and coprological assessment after flotation provide complementary detection results increasing sensitivity. Genotyping PCRs confirm detection in a subset of samples and cross-validation of different PCR markers does not indicate a bias towards a particular parasite species in genotyping. We were able to detect double infections and to determine the preferred niche of each parasite species along the distal-proximal axis of the intestine. Parasite genotyping from tissue samples provides an additional indication for the absence of species bias in genotyping amplifications. Three Eimeria species were found infecting house mice at different prevalences: Eimeria ferrisi (16.7%; 95% CI 13.2 – 20.7), E. falciformis (4.2%; 95% CI 2.6 – 6.8) and E. vermiformis (1.9%; 95% CI 0.9 – 3.8). We also find that mice in dense populations are more likely to be infected with E. falciformis and E. ferrisi.We provide methods for the assessment of prevalences of coccidia at the species level in rodent systems. We show and discuss how such data can help to test hypotheses in ecology, evolution and epidemiology on a species level.

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