Dataset Information

Molecular Detection of Gurltia paralysans by Semi-Nested PCR in Cerebrospinal Fluid and Serum Samples from Domestic Cats (Felis catus).

ABSTRACT: Gurltia paralysans is an angio-neurotropic metastrongyloid nematode that infects domestic and wild cats, invading the veins of the subarachnoid space of the spinal cord and mainly causing progressive paralysis of the pelvic limbs. The definitive diagnosis of feline gurltiosis can only be achieved by post-mortem examination that reveals the presence of the nematode in the spinal cord vein vasculature. An early diagnosis with conclusive results is required since laboratory and imaging findings are not sufficient. Therefore, the purpose of this study was to detect the presence of G. paralysans, via semi-nested PCR, in samples of cerebrospinal fluid (CSF) and the sera of domestic cats naturally infected with the parasite. A total of 12 cats with a diagnosis suggestive of feline gurltiosis were selected, and they underwent a complete neurological and imaging examination. DNA samples were analysed by semi-nested PCR, with universal (AaGp28Sa1/AaGp28Ss1) and specific (Gp28Sa3/Aa28Ss2) primers, for G. paralysans (G. paralysans 18S rRNA gene, partial sequence; ITS 1, 5.8S rRNA gene, and ITS 2, complete sequence; and 28S rRNA gene, partial sequence) and Aelurostrongylus abstrusus, obtaining amplifications of 356 and 300 bp, which indicated the presence or absence of nematode DNA, respectively. The presence of G. paralysans was detected in the CSF of four out of nine cats, and the sera of seven out of seven cats. In the sera analysis of five out of seven cats, a mixed infection with A. abstrusus was found, despite no alterations of the respiratory tract being observed during the necropsies. It is proposed that serum samples could be more effective than CSF in detecting the parasite by PCR analysis. Sequencing analysis showed high percentages of identity with G. paralysans, which indicated the feasibility of detection and the sensitivity/specificity of the method used, suggesting the implementation of semi-nested PCR as a routine diagnostic test for early and timely detection of feline gurltiosis.

SUBMITTER: Lopez-Contreras F

PROVIDER: S-EPMC7401636 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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Molecular Detection of Gurltia paralysans by Semi-Nested PCR in Cerebrospinal Fluid and Serum Samples from Domestic Cats (Felis catus).

López-Contreras Freddy F Rojas-Barón Lisbeth L Gómez Marcelo M Morera Francisco F Sepúlveda Paulina P Moroni Manuel M Muñoz Pamela P Acosta-Jammett Gerardo G Mieres Marcelo M Hirzmann Jörg J Hermosilla Carlos C Taubert Anja A

Animals : an open access journal from MDPI 20200709 7

Gurltia paralysans is an angio-neurotropic metastrongyloid nematode that infects domestic and wild cats, invading the veins of the subarachnoid space of the spinal cord and mainly causing progressive paralysis of the pelvic limbs. The definitive diagnosis of feline gurltiosis can only be achieved by post-mortem examination that reveals the presence of the nematode in the spinal cord vein vasculature. An early diagnosis with conclusive results is required since laboratory and imaging findi ...[more]

PMID: 32660139

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Project description:Gurltia paralysans is a metastrongyloid nematode which belongs to the Angiostrongylidae family and presents tropism for veins of the subarachnoid space in vivo of domestic and wild felids causing a progressive and chronic clinical manifestation of paraparesis/paraplegia. The geographic distribution of G. paralysans includes rural and periurban areas of South America and was recently reported in Europe. To date, a definitive diagnosis of feline gurltiosis is still conducted by post-mortem examination of the spinal cord in affected animals. A presumptive diagnosis of feline gurltiosis can also be achieved based on data of compatible clinical signs and associated epidemiological risk factors. The aim of this preliminary study was to evaluate the commercial serological test Angio Detect TM® (IDEXX Laboratories) as a possible diagnostic method of feline gurltiosis in vivo. For the study, 10 domestic felines (Felis catus) which originated from a high endemic area of Southern Chile, were analyzed. All felines presented chronic paraparesis or severe paraplegia. Subsequently, commercial Angio Detect TM® serological tests for the detection of closely related Angiostrongylus vasorum in canids were performed according to manufacturer's instructions. Conducted serological tests were positive in seven of ten felines showing paraplegia/paraparesis and presumably infected with G. paralysans, and four of them were additionally necropsied, and presented macroscopic findings compatible with feline gurltiosis. Furthermore, the presence of adult female and male G. paralysans specimens at the level of the subarachnoid vasculature in affected spinal cord segments were observed during necropsy. Histopathology demonstrated severe eosinophilic meningomyelitis, coagulopathies with thrombosis in G. paralysans-parasitized leptomeningeal veins. Results of this preliminary study suggest a cross-reaction between A. vasorum-specific antigens, which also parasitize blood vessels in vivo, and G. paralysans when using an Angio Detect TM® test, which suggests its helpful use as a new diagnostic method for feline gurltiosis in live domestic felines. Additional specific antigen research will be required in order to better understand the sensitivity and specificity of A. vasorum antigens used in this test and for existing cross-reactivity with G. paralysans-derived antigens for future a suitable intra vitam immunodiagnosis of neglected feline gurltiosis.

Project description:Cats are obligate carnivores and under most circumstances eat only animal products. Owing to the pseudogenization of one of two subunits of the sweet receptor gene, they are indifferent to sweeteners, presumably having no need to detect plant-based sugars in their diet. Following this reasoning and a recent report of a positive correlation between the proportion of dietary plants and the number of Tas2r (bitter receptor) genes in vertebrate species, we tested the hypothesis that if bitter perception exists primarily to protect animals from poisonous plant compounds, the genome of the domestic cat (Felis catus) should have lost functional bitter receptors and they should also have reduced bitter receptor function. To test functionality of cat bitter receptors, we expressed cat Tas2R receptors in cell-based assays. We found that they have at least 7 functional receptors with distinct receptive ranges, showing many similarities, along with some differences, with human bitter receptors. To provide a comparative perspective, we compared the cat repertoire of intact receptors with those of a restricted number of members of the order Carnivora, with a range of dietary habits as reported in the literature. The numbers of functional bitter receptors in the terrestrial Carnivora we examined, including omnivorous and herbivorous species, were roughly comparable to that of cats thereby providing no strong support for the hypothesis that a strict meat diet influences bitter receptor number or function. Maintenance of bitter receptor function in terrestrial obligate carnivores may be due to the presence of bitter compounds in vertebrate and invertebrate prey, to the necessary role these receptors play in non-oral perception, or to other unknown factors. We also found that the two aquatic Carnivora species examined had fewer intact bitter receptors. Further comparative studies of factors driving numbers and functions of bitter taste receptors will aid in understanding the forces shaping their repertoire.

Project description:BACKGROUND:The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemotropic mycoplasmas in client-owned cats in Romania. METHODS:Blood samples from 51 unhealthy cats, originating from Timi?oara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. PCR-positive samples were subsequently analyzed by phylogenetic and population genetic analysis. RESULTS:Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations (p?>?0.05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, feline leukemia virus/feline immunodeficiency virus positivity of cats, or the sampling season. However, outdoor access was positively associated (p?=?0.049) with infection and could be considered a risk factor (OR?=?4.1) in acquiring feline hemotropic mycoplasmas. Phylogenetic analysis revealed that our sequences clustered with those selected from the GenBank database in two distinct clades. The registered population genetic indices were strongly supportive of the great variance in sequences between the recorded Mycoplasma species. CONCLUSIONS:The findings support the occurrence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

Dataset Information

Molecular Detection of Gurltia paralysans by Semi-Nested PCR in Cerebrospinal Fluid and Serum Samples from Domestic Cats (Felis catus).

Publications

Molecular Detection of <i>Gurltia paralysans</i> by Semi-Nested PCR in Cerebrospinal Fluid and Serum Samples from Domestic Cats (<i>Felis catus</i>).

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