scholarly journals Molecular detection of Apicomplexa protozoa in tissues from Alouatta guariba clamitans

2021 ◽  
Vol 41 ◽  
Author(s):  
Aline Ludwig ◽  
Laurete Murer ◽  
Helton F. dos Santos ◽  
Adriana Ludwig ◽  
Luis Antonio Sangioni ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Neospora Caninum ◽  
Positive Sample ◽  
Primate Species ◽  
Howler Monkey ◽  
Rrna Gene ◽  
Tissue Samples ◽  
Genetic Sequencing ◽  
Sarcocystis Spp ◽  
Alouatta Guariba Clamitans

ABSTRACT: The brown howler monkey (Alouatta guariba clamitans) is a primate species widely distributed in South America. Infections by protozoa are common in primates. However, studies on protozoa in primates in Brazil are scarce, so the goal of this study was to investigate DNA from the apicomplexan protozoa Neospora caninum, Sarcocystis spp. and Toxoplasma gondii in tissues of A. guariba clamitans. DNA extraction was performed on tissue samples from the heart, brain, liver, spleen, lung and intestine of six A. guariba clamitans from Santa Maria, Central Region of Rio Grande do Sul, Brazil. Conventional PCR was performed using 18S rRNA gene general primers for Apicomplexa and also specific primers to amplify Neosporaspp. and Toxoplasma gondii DNA. All animals were positive in the 18S PCR and the genetic sequencing confirmed the presence of Sarcocystis spp. DNA in the tissues of four animals belonging to at least two species (S. neurona and S. gigantea) and T. gondii DNA in the other two animals. One positive sample for T. gondii was genotypically characterized as atypical by the restriction fragment length polymorphism technique. N. caninum DNA was not detected in the tested samples. The presence of Apicomplexa protozoan DNA in the tissues of the six animals tested in this study highlights the importance of howler monkeys as maintainers of these pathogens in nature.

scholarly journals Primatology in southern Brazil: a transdisciplinary approach to the conservation of the brown-howler-monkey Alouatta guariba clamitans (Primates, Atelidae)

2010 ◽  
Vol 100 (4) ◽  
pp. 403-412 ◽  
Author(s):  
Leandro Jerusalinsky ◽  
Fernanda Zimmermann Teixeira ◽  
Luisa Xavier Lokschin ◽  
André Alonso ◽  
Márcia Maria de Assis Jardim ◽  
...  
Keyword(s):  
Urban Expansion ◽  
Research Management ◽  
Conservation Education ◽  
Basic Knowledge ◽  
Primate Species ◽  
Howler Monkey ◽  
Conservation Strategies ◽  
Southern Brazil ◽  
Alouatta Guariba Clamitans ◽  
The Impact

Human interventions in natural environments are the main cause of biodiversity loss worldwide. The situation is not different in southern Brazil, home of five primate species. Although some earlier studies exist, studies on the primates of this region began to be consistently carried out in the 1980s and have continued since then. In addition to important initiatives to study and protect the highly endangered Leontopithecus caissara Lorrini & Persson, 1990 and Brachyteles arachnoides E. Geoffroy, 1806, other species, including locally threatened ones, have been the focus of research, management, and protection initiatives. Since 1993, the urban monkeys program (PMU, Programa Macacos Urbanos) has surveyed the distribution and assessed threats to populations of Alouatta guariba clamitans (Cabrera, 1940) in Porto Alegre and vicinity. PMU has developed conservation strategies on four fronts: (1) scientific research on biology and ecology, providing basic knowledge to support all other activities of the group; (2) conservation education, which emphasizes educational presentations and long-term projects in schools near howler populations, based on the flagship species approach; (3) management, analyzing conflicts involving howlers and human communities, focusing on mitigating these problems and on appropriate relocation of injured or at-risk individuals; and finally, (4) Public Policies aimed at reducing and/or preventing the impact of urban expansion, contributing to create protected areas and to strengthen environmental laws. These different approaches have contributed to protect howler monkey populations over the short term, indicating that working collectively and acting on diversified and interrelated fronts are essential to achieve conservation goals. The synergistic results of these approaches and their relationship to the prospects for primatology in southern Brazil are presented in this review.

scholarly journals Serological and molecular detection of Neospora caninum and Toxoplasma gondii in human umbilical cord blood and placental tissue samples

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Pâmella Oliveira Duarte ◽  
Leandra Marla Oshiro ◽  
Namor Pinheiro Zimmermann ◽  
Bárbara Guimarães Csordas ◽  
Doroty Mesquita Dourado ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Molecular Detection ◽  
Neospora Caninum ◽  
Placental Tissue ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Tissue Samples

scholarly journals Antibodies against Neospora caninum, Sarcocystis spp. and Toxoplasma gondii detected in buffaloes from Rio Grande do Sul, Brazil

2016 ◽  
Vol 36 (10) ◽  
pp. 947-950 ◽  
Author(s):  
Luiza P. Portella ◽  
◽  
Gustavo C. Cadore ◽  
Marcelo de Lima ◽  
Luís A. Sangioni ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Neospora Caninum ◽  
Fluorescent Antibody ◽  
Rio Grande ◽  
Antibody Test ◽  
Public Health Issue ◽  
Rio Grande Do Sul ◽  
Serum Samples ◽  
Caninum Infection ◽  
Sarcocystis Spp

ABSTRACT: The presence of antibodies against Neospora caninum, Sarcocystis spp. and Toxoplasma gondii was evaluated in buffaloes (Bubalus bubalis) from Rio Grande do Sul state (RS), southern Brazil. Serum samples (n=220) were analyzed for antibodies by indirect fluorescent antibody test (IFAT). Antibody presence was considered when the titers were equal or higher than 100 for these protozoa. A total of 60.5% (133/220) buffalo serum samples were positive for at least one of the protozoa evaluated in this study. Antibodies for N. caninum, Sarcocystis spp. and T. gondii were found in 36.4% (80/220), 25.5% (56/220) and 16.8% (37/220) of the buffaloes respectively, indicating a higher frequency of N. caninum infection (p=0.0133). The IFAT is a suitable method to diagnose N. caninum, Sarcocystis spp. and T. gondii infection in buffaloes for detecting IgG antibodies. This study demonstrates the presence of these three protozoa in buffalo herds in RS, Brazil, which may be source of infection to other animals. The high frequency of animals positive for N. caninum is important and could be related to reproductive problems. Additionally, the presence of Sarcocystis spp. and T. gondii in buffaloes can be a possible public health issue.

Evaluation of frequency of antibodies against Toxoplasma gondii,Neospora caninum and Sarcocystis spp. and transmission routes in sheep from Humid Pampa, Argentina

2018 ◽  
Vol 63 (2) ◽  
pp. 416-421 ◽  
Author(s):  
Yanina P. Hecker ◽  
Fernando Mogaburu Masson ◽  
Joaquín I. Armendano ◽  
Juan Cora ◽  
Carlos Flores Olivares ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Neospora Caninum ◽  
Transmission Route ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Adult Sheep ◽  
Sarcocystis Spp ◽  
Serological Status ◽  
The Impact

AbstractThe aim of this study was to describe the frequency of ovine specific antibodies toToxoplasma gondii,Neospora caninumandSarcocystisspp. and to estimate different transmission routes of these infections. One hundred and thirty Texel sheep and their 117 Texel lambs were included in the study. Serum samples were tested for antibodies toT.gondii,N.caninumandSarcocystisspp. using IFAT.Toxoplasma gondiiseroprevalence was 10.00% in sheep (IC95%: 4.80–15.20%), being higher in adult sheep (≥12 year) than in younger sheep (OR 1.30; 95% CI, 1.10–1.50).N.caninumandSarcocystisspp. seroprevalences were 1.54% (IC95%: 0.00–5.70) and 72.09% (IC95%: 67.70–82.70), respectively, with no association between age and seropositivity in sheep (P>0.05).T.gondiiseroprevalence in lambs was 4.27% (IC95%: 0.61–7.94). No association betweenT.gondiiserological status in sheep and their lambs was detected (P= 0.07). TwoT.gondiiandSarcocystisspp. seropositive lambs were euthanized andT.gondiiandSarcocystisspp. DNA was detected by PCR in their tissues. In conclusion, the increase ofT.gondiiseropositivity in relationship with sheep age and the lack of association between sheep-lamb serological status, suggest that horizontal infection is the main transmission route in this flock as reported before. Due to the low number ofN.caninum-seropositive ewes no assumptions can be done about the impact of this parasite in this flock. According with previous reports, the main transmission route forSarcocystisspp. in this species in the present study was horizontal.

Molecular survey for cyst-forming coccidia (Toxoplasma gondii, Neospora caninum, Sarcocystis spp.) in Mediterranean periurban micromammals

2020 ◽  
Vol 119 (8) ◽  
pp. 2679-2686
Author(s):  
Mercedes Fernández-Escobar ◽  
Javier Millán ◽  
Andrea D. Chirife ◽  
Luis Miguel Ortega-Mora ◽  
Rafael Calero-Bernal
Keyword(s):  
Toxoplasma Gondii ◽  
Neospora Caninum ◽  
Molecular Survey ◽  
Sarcocystis Spp

scholarly journals Molecular characterization of Toxoplasma gondii and Sarcocystis spp. in raw kibbeh and other meat samples commercialized in Botucatu, Southeastern Brazil

2021 ◽  
Vol 30 (2) ◽  
Author(s):  
Helio Langoni ◽  
Diego Generoso ◽  
Ênio Yoshinori Hayasaka ◽  
Karine Bott Mantovan ◽  
Benedito Donizete Menozzi ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Meat Products ◽  
Positive Sample ◽  
Southeastern Brazil ◽  
Economic Losses ◽  
Base Pairs ◽  
Sarcocystis Spp ◽  
Commercial Markets ◽  
Meat Samples ◽  
Pcr Targeting

Abstract Toxoplasmosis occurs worldwide causing economic losses to the animal production and problems to the public health. The study aimed to detect Toxoplasma gondii and Sarcocystis spp.in 141 meat products from commercial meat cuts of pork, beef, and kibbeh sold in commercial markets from Botucatu, SP, Brazil. Samples were bioassayed in mice to isolate the parasite, and the parasite DNA detected by PCR targeting the 529 base pairs repeat element region (PCR-529-bp). All samples resulted negative on bioassay, whereas PCR positive for 9 (6,38%), distributed as 5/48 beef, 3/49 pork, and 1/44 kibbeh. PCR-positive were investigated for the the parasite genotype using multiplex-, nested-, and RFLP-PCR for 11 markers (SAG1, 5’-3’SAG2, alt.SAG2, SAG3, B-TUB, GRA6, L358, c22-8, c29-6, PK1, Apico). Complete genotype was determined on just one PCR-positive sample that matched MAS, TgCkBr89 and TgCkBr147 isolates already identified. In addition, nested- and RFLP-PCR targeting 18S rRNA was run for all PCR-positive samples and, the products, sequenced and aligned to the GenBank at NCBI website. Four samples showed 100% homology with T. gondii (GenBank #L37415.1), three with Sarcocystis hominis (GenBank #AF006471.1), two Sarcocystis cruzi (GenBank #AF176934.1), and one Sarcocystis hirsuta (GenBank #AF006469.1), indicating the circulation of T. gondii and Sarcocystis spp.

Rapid and sensitive identification ofNeospora caninumbyin vitroamplification of the internal transcribed spacer 1

Parasitology ◽  
1996 ◽  
Vol 112 (2) ◽  
pp. 177-182 ◽  
Author(s):  
O. J. M. Holmdahl ◽  
J. G. Mattsson
Keyword(s):  
Internal Transcribed Spacer ◽  
Neospora Caninum ◽  
Neuromuscular Disorders ◽  
Rrna Gene ◽  
Tissue Samples ◽  
Rdna Unit ◽  
Coccidian Parasites ◽  
Base Pair Fragment ◽  
Polymerase Chain ◽  
Pair Fragment

SummaryNeospora caninumandN. caninum-like organisms are cyst-forming coccidian parasites known to cause neuromuscular disorders in dogs and abortion in cattle. In this article we report on the use of the polymerase chain reaction (PCR) for the detection of DNA fromN. caninum. After determining the sequence of the internal transcribed spacer 1 (ITSl) ofN. caninumandToxoplasma gondii, and part of the sequences for & species ofSarcocystis, we designed a primer set for the amplification of a 279-base-pair fragment of ITSl fromN. caninum. The PCR system made possible the specific detection of 5N. caninumorganisms and no amplification was observed from any of the other cyst-forming coccidia tested, including the closely relatedT. gondii. Furthermore, we were also able to demonstrate the presence ofN. caninumin brain and lung tissue samples from experimentally infected mice. Our data also link the 5-8S rRNA gene forT. gondiiandN. caninumto the 16S-like rRNA gene, within the rDNA unit.

scholarly journals Antibodies against Apicomplexa protozoa and absence sarcocysts in heart tissues from horses in southern Brazil

2017 ◽  
Vol 26 (1) ◽  
pp. 100-103 ◽  
Author(s):  
Luiza Pires Portella ◽  
Gustavo Cauduro Cadore ◽  
Luis Antonio Sangioni ◽  
Luiz Fernando Vilani Pellegrini ◽  
Rafael Fighera ◽  
...  
Keyword(s):  
Life Cycle ◽  
Toxoplasma Gondii ◽  
Myocardial Tissue ◽  
Southern Brazil ◽  
Serum Samples ◽  
Positive Serology ◽  
Tissue Samples ◽  
Sarcocystis Spp ◽  
Direct Microscopic Examination ◽  
Paired Serum

Abstract Sarcocystis spp., Neospora spp., and Toxoplasma gondii are Apicomplexa protozoa that can infect horses. This study aimed to investigate the occurrence of antibodies against Sarcocystis spp., Neospora spp., and T. gondii in horses slaughtered in southern Brazil. The presence of histological lesions, tissue cysts, and Sarcocystis spp. DNA in the hearts of these horses was also investigated. A total of 197 paired serum and heart samples were evaluated by serology and direct microscopic examination; 50 of these samples were subjected to histopathological and PCR analyses. Antibodies against at least one of the protozoa were detected in 146 (74.1%) of the serum samples. The frequencies of positive serology were: 36% (71/197) against Sarcocystis spp., 39.1% (77/197) against Neospora spp., and 47.2% (93/197) against T. gondii. No cysts, Sarcocystis spp. DNA, or histopathological lesions were observed in myocardial tissue samples. The frequencies of antibody seropositivity against Sarcocystis spp., Neospora spp., and T. gondii showed that horses are frequently infected by these parasites in southern Brazil. The absence of sarcocysts in horse tissues is compatible with their role as aberrant/accidental hosts in the life cycle of Sarcocystis spp..

scholarly journals Comparative aspects of laboratory testing for the detection of Toxoplasma gondii and its differentiation from Neospora caninum as the etiologic agent of ovine abortion

2020 ◽  
Vol 32 (6) ◽  
pp. 898-907
Author(s):  
Nicola Meixner ◽  
Marie F. Sommer ◽  
Nelly Scuda ◽  
Kaspar Matiasek ◽  
Matthias Müller
Keyword(s):  
Toxoplasma Gondii ◽  
Real Time ◽  
Real Time Pcr ◽  
Neospora Caninum ◽  
Placental Tissue ◽  
Etiologic Agent ◽  
Detection Methods ◽  
Tissue Samples ◽  
Specificity And Sensitivity ◽  
Histologic Evaluation

Histologic examination of aborted material is an essential component in the diagnosis of ovine toxoplasmosis. However, the detection of Toxoplasma gondii in histologic sections, and its differentiation from the closely related protozoan Neospora caninum, is challenging. We developed a chromogenic in situ hybridization (ISH) assay for the identification of T. gondii in paraffin-embedded tissue samples. We examined retrospectively the archived placental tissue of 200 sheep abortion submissions for the presence of T. gondii by immunohistochemistry (IHC), ISH, and real-time PCR (rtPCR). All placental samples that tested positive for T. gondii by rtPCR (9 of 200) were also positive by IHC, with inconclusive IHC staining in an additional 7 rtPCR-negative cases. Further testing for N. caninum of all 200 placentas by rtPCR revealed 7 Neospora-positive cases. T. gondii ISH was positive in 4 of 9 IHC-positive samples and 1 of the 7 N. caninum rtPCR-positive samples. Real-time PCR was used as the reference standard for specificity and sensitivity calculations regarding placenta samples. Specificity of ISH and IHC was 99% and 96–100%, respectively. The sensitivity of ISH (44%) was quite low compared to IHC (100%). The exclusive use of ISH for the detection of T. gondii, and thus for the diagnosis of ovine toxoplasmosis, was not acceptable. However, combined with rtPCR, both ISH and IHC can be useful detection methods to improve histologic evaluation by visualizing the parasite within tissue sections.

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