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      Prevalence of Feline Immunodeficiency Virus and Toxoplasma gondii in Feral Cats on St. Kitts, West Indies

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          Abstract

          Toxoplasma gondii ( T. gondii) is a cosmopolitan protozoan parasite that infects all warm-blooded species including humans. The definitive hosts of T. gondii are felid vertebrates including the domestic cat. Domestic cats shed oocysts for approximately two weeks in their feces after the primary infection. It has been shown that feline immunodeficiency virus (FIV) positive cats have a higher prevalence of and a higher titer of antibodies to T. gondii than those of FIV-negative cats. The main purposes of this study were to determine FIV prevalence and to investigate the oocysts shedding in FIV-positive and FIV-negative feral cats on St. Kitts. Fecal samples were collected from feral cats while their FIV statues were determined using a commercial SNAP kit. Total fecal DNA of each cat was tested for the presence of T. gondii DNA using a polymerase chain reaction (PCR) consistently detecting one genome equivalent. A FIV-positive status was detected in 18 of 105 (17.1%, 95% confidence interval (CI): 9.9%−24.3%) feral cats sampled. Furthermore, males were three times more likely to be FIV positive than females ( p = 0.017) with an odds ratio of 3.93 (95% CI: 1.20–12.89). Adults were found to have at least twice the prevalence of FIV compared to cats younger than one year of age ( p = 0.056) with an odds ratio of 3.07 (95% CI: 0.94–10.00). Toxoplasma gondii DNA was not detected in the feces of any of the 18 FIV-positive (95% CI: 0%−0.18%) and 87 FIV-negative cats (95% CI: 0%−0.04%). A follow-up study with a much bigger sample size is needed to prove or disprove the hypothesis that FIV-positive cats have a higher prevalence of shedding T. gondii oocysts than FIV-negative cats.

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          How to Calculate Sample Size for Different Study Designs in Medical Research?

          Calculation of exact sample size is an important part of research design. It is very important to understand that different study design need different method of sample size calculation and one formula cannot be used in all designs. In this short review we tried to educate researcher regarding various method of sample size calculation available for different study designs. In this review sample size calculation for most frequently used study designs are mentioned. For genetic and microbiological studies readers are requested to read other sources.
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            Advances in the life cycle of Toxoplasma gondii.

            J.P. Dubey (1998)
            This paper reviews recent studies on the life cycle of Toxoplasma gondii. Tachyzoites, bradyzoites, and sporozoites are the three infectious stages of T. gondii. Humans and animals become infected mainly by ingesting bradyzoites or oocytes. After ingestion, both bradyzoites and sporozoites convert to tachyzoites inside tissues. The conversion of tachyzoites to bradyzoites and bradyzoites to tachyzoites is of biological and clinical significance because bradyzoites are less susceptible to chemotherapy and reactivation of bradyzoites to tachyzoites is considered the cause of fatal toxoplasmosis in AIDS patients. Of all the methods currently available to assess stage conversion of T. gondii, feeding infective stages to cats is the most reliable method. Felidae, the definitive hosts of T. gondii excrete oocysts 3-10 days after ingesting tissue cysts/bradyzoites, > or = 18 days after ingesting oocysts, and > or = 13 days after ingesting tachyzoites.
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              Comparison of two DNA targets for the diagnosis of Toxoplasmosis by real-time PCR using fluorescence resonance energy transfer hybridization probes

              Background Toxoplasmosis is an infectious disease caused by the parasitic protozoan Toxoplasma gondii. It is endemic worldwide and, depending on the geographic location, 15 to 85% of the human population are asymptomatically infected. Routine diagnosis is based on serology. The parasite has emerged as a major opportunistic pathogen for immunocompromised patients, in whom it can cause life-threatening disease. Moreover, when a pregnant woman develops a primary Toxoplasma gondii infection, the parasite may be transmitted to the fetus and cause serious damnage. For these two subpopulations, a rapid and accurate diagnosis is required to initiate treatment. Serological diagnosis of active infection is unreliable because reactivation is not always accompanied by changes in antibody levels, and the presence of IgM does not necessarily indicate recent infection. Application of quantitative PCR has evolved as a sensitive, specific, and rapid method for the detection of Toxoplasma gondii DNA in amniotic fluid, blood, tissue samples, and cerebrospinal fluid. Methods Two separate, real-time fluorescence PCR assays were designed and evaluated with clinical samples. The first, targeting the 35-fold repeated B1 gene, and a second, targeting a newly described multicopy genomic fragment of Toxoplasma gondii. Amplicons of different intragenic copies were analyzed for sequence heterogeneity. Results Comparative LightCycler experiments were conducted with a dilution series of Toxoplasma gondii genomic DNA, 5 reference strains, and 51 Toxoplasma gondii-positive amniotic fluid samples revealing a 10 to 100-fold higher sensitivity for the PCR assay targeting the newly described 529-bp repeat element of Toxoplasma gondii. Conclusion We have developed a quantitative LightCycler PCR protocol which offer rapid cycling with real-time, sequence-specific detection of amplicons. Results of quantitative PCR demonstrate that the 529-bp repeat element is repeated more than 300-fold in the genome of Toxoplasma gondii. Since individual intragenic copies of the target are conserved on sequence level, the high copy number leads to an ultimate level of analytical sensitivity in routine practice. This newly described 529-bp repeat element should be preferred to less repeated or more divergent target sequences in order to improve the sensitivity of PCR tests for the diagnosis of toxoplasmosis.
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                Author and article information

                Journal
                Vet Sci
                Vet Sci
                vetsci
                Veterinary Sciences
                MDPI
                2306-7381
                21 January 2021
                February 2021
                : 8
                : 2
                : 16
                Affiliations
                [1 ]Departments of Biomedical Sciences, Ross University School of Veterinary Medicine, P.O. Box 334, Basseterre 00265, Saint Kitts and Nevis; xinyuc.20@ 123456intl.zju.edu.cn (X.C.); kxfang@ 123456zju.edu.cn (K.F.)
                [2 ]Departments of One Health Center for Zoonoses and Tropical Veterinary Medicine, Ross University School of Veterinary Medicine, P.O. Box 334, Basseterre 00265, Saint Kitts and Nevis; lkoster@ 123456utk.edu (L.K.); jevanchristie@ 123456gmail.com (J.C.)
                [3 ]Departments of Clinical Sciences, Ross University School of Veterinary Medicine, P.O. Box 334, Basseterre 00265, Saint Kitts and Nevis
                Author notes
                [* ]Correspondence: chyao@ 123456rossvet.edu.kn ; Tel.: +1-732-645-3120; Fax: +1-869-465-1203
                [†]

                X.C. and K.F. were interns of Ross University School of Veterinary Medicine from Zhejiang University, Hangzhou 310058, China.

                [‡]

                Current address: Small Animal Clinical Sciences, C247 Veterinary Medical Center, University of Tennessee, 2407 River Drive, Knoxville, TN 37996-4500, USA.

                [§]

                Current address: The Animal Hospital, Murdoch University, 90 South Street, Murdoch, WA 6150, Australia.

                Author information
                https://orcid.org/0000-0002-8143-7129
                https://orcid.org/0000-0003-2578-6604
                Article
                vetsci-08-00016
                10.3390/vetsci8020016
                7909821
                33494205
                7769fd90-f290-486f-998e-7f3b0ae137d2
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 03 December 2020
                : 16 January 2021
                Categories
                Communication

                toxoplasma gondii,oocyst shedding,domestic cat,feral cat,feline immunodeficiency virus,feline leukemia virus

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