scholarly journals Diagnosis of Toxoplasmosis in Ruminants Aborted Fetuses in Northern Iran Using Molecular and Bioassay Techniques

2021 ◽  
Author(s):  
Kaveh Azimi ◽  
Afsaneh Amouei ◽  
Mehdi Sharif ◽  
Shahabeddin Sarvi ◽  
Nemat Shams ◽  
...  
Keyword(s):  
Control Strategies ◽  
Protozoan Parasite ◽  
Economic Losses ◽  
Conventional Pcr ◽  
Pcr Assay ◽  
Mazandaran Province ◽  
Northern Iran ◽  
Wide Range ◽  
Intracellular Protozoan Parasite

Background: Toxoplasma gondii is a zoonotic obligatory intracellular protozoan parasite that infects a wide range of warm-blooded species. This study aimed to obtain further information on the role of T. gondii infection in ruminant abortion (sheep, goats and cattle) using bioassay and PCR methods in Mazandaran province, northern Iran. Methods: Overall, 104 aborted fetuses (52 bovine, 48 ovine, 4 caprine) were collected at different stages of gestation during the lambing seasons in various parts of Mazandaran Province from Mar 2016 to May 2017. Brains of 104 aborted fetuses were bioassayed in female BALB/c mice. DNA was extracted from all brain samples using phenol-chloroformisoamyl Alcohol instructions. RE gene was used for detection all of T. gondii DNA by conventional PCR assay. Results: The results of the bioassayed samples were negative because no tachyzoites or cyst were observed in the peritoneal and brain specimens of the mice. The detection of T. gondii DNA was confirmed by observation of a 529 bp band in 15 out of 104 fetuses (14.4%). The highest prevalence rate of T. gondii detected from sheep (16.6%) followed by cattle (13.4%) and goats (0%). The highest prevalence of the infection was observed in east area, while the lowest prevalence of the infection was observed in west area. Conclusion: T. gondii infection may partly be responsible for abortion and economic losses in livestock husbandry in this region. Therefore, further additional researches such as genotyping T. gondii and designing control strategies for improving management in livestock flocks are necessary.

scholarly journals Species diversity and insecticide resistance within the Anopheles hyrcanus group in Ubon Ratchathani Province, Thailand

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Anchana Sumarnrote ◽  
Hans J. Overgaard ◽  
Vincent Corbel ◽  
Kanutcharee Thanispong ◽  
Theeraphap Chareonviriyaphap ◽  
...  
Keyword(s):  
Insecticide Resistance ◽  
Control Strategies ◽  
Resistance Mechanism ◽  
Abundant Species ◽  
Malaria Vectors ◽  
Pcr Assay ◽  
Insecticide Susceptibility ◽  
Human Landing ◽  
Early Peak

Abstract Background Members of the Anopheles hyrcanus group have been incriminated as important malaria vectors. This study aims to identify the species and explore the insecticide susceptibility profile within the Anopheles hyrcanus group in Ubon Ratchathani Province, northeastern Thailand where increasing numbers of malaria cases were reported in 2014. Methods Between 2013 and 2015, five rounds of mosquito collections were conducted using human landing and cattle bait techniques during both the rainy and dry seasons. Anopheles mosquitoes were morphologically identified and their insecticide susceptibility status was investigated. Synergist bioassays were carried out with An. hyrcanus (s.l.) due to their resistance to all insecticides. An ITS2-PCR assay was conducted to identify to species the Hyrcanus group specimens. Results Out of 10,361 Anopheles females collected, representing 18 taxa in 2 subgenera, 71.8% were morphologically identified as belonging to the Hyrcanus Group (subgenus Anopheles), followed by An. barbirostris group (7.9%), An. nivipes (6.5%), An. philippinensis (5.9%) and the other 14 Anopheles species. Specimens of the Hyrcanus Group were more prevalent during the rainy season and were found to be highly zoophilic. Anopheles hyrcanus (s.l.) was active throughout the night, with an early peak of activity between 18:00 h and 21:00 h. ITS2-PCR assay conducted on 603 DNA samples from specimens within the Hyrcanus Group showed the presence of five sisters species. Anopheles peditaeniatus was the most abundant species (90.5%, n = 546), followed by An. nitidus (4.5%, n = 27), An. nigerrimus (4.3%, n = 26), An. argyropus (0.5%, n = 3), and An. sinensis (0.2%, n = 1). All An. hyrcanus (s.l.) specimens that were found resistant to insecticides (deltamethrin 0.05%, permethrin 0.75% and DDT 4% and synergist tests) belonged to An. peditaeniatus. The degree of resistance in An. peditaeniatus to each of these three insecticides was approximately 50%. Addition of PBO (Piperonyl butoxide), but not DEF (S.S.S-tributyl phosphotritioate), seemed to restore susceptibility, indicating a potential role of oxidases as a detoxifying enzyme resistance mechanism. Conclusions A better understanding of mosquito diversity related to host preference, biting activity and insecticide resistance status will facilitate the implementation of locally adapted vector control strategies.

scholarly journals New insights into the prevalence and phylogenetic diversity of Cysticercus ovis isolates in sheep from Sulaymaniyah, Iraq

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Aram Ahmad Mohammed
Keyword(s):  
Sequence Analysis ◽  
Phylogenetic Diversity ◽  
Nucleotide Diversity ◽  
Control Strategies ◽  
Economic Losses ◽  
Cox1 Gene ◽  
Conventional Pcr ◽  
Single Clade ◽  
First Time ◽  
Cysticercus Ovis

Abstract Introduction Although ovine cysticercosis is not a zoonotic problem, it results in substantial economic losses due to the condemnation of infected tissues or entire carcasses. This study aimed to record preliminary data on the prevalence, and phylogenetic diversity of Cysticercus ovis isolates from slaughtered sheep in the province of Sulaymaniyah, Iraq. Material and Methods From January to September 2020, 6, 411 slaughtered sheep were examined for C. ovis by routine meat inspection. The amplification and sequence analysis of the COX1 gene for up to 35 specimens of C. ovis was performed using conventional PCR. Results The overall prevalence rate was 1.3%, and the prevalence was significantly higher in older sheep (>1 year) than younger ones (<1 year) (P< 0.05). The cardiac muscle showed a higher tendency to carry C. ovis infection compared to other examined muscles. Sequence analysis of the COX1 gene revealed six haplotypes, and the level of pairwise nucleotide diversity between individual haplotypes was 1–2%. Five out of six of the Taenia ovis haplotypes recovered could have been recorded for the first time globally. Phylogenetic interpretation indicated that all the T. ovis haplotypes clustered in a single clade, and it also indicated an extremely close similarity to Iranian and New Zealand isolates. Conclusions Globally, this report adds new data on C. ovis genetic diversity, which provide an extremely useful molecular background with regard to future preventive as well as control strategies.

scholarly journals Evaluation of Helminth Infections Identified in Slaughtered Livestock in Iran, 2015-2019

2020 ◽  
Author(s):  
Behzad Kiani ◽  
Christine M. Budke ◽  
Ebrahim Shams Abadi ◽  
Soheil Hashtarkhani ◽  
Amene Raouf Rahmati ◽  
...  
Keyword(s):  
Control Strategies ◽  
Health Impact ◽  
Direct Costs ◽  
Economic Losses ◽  
Infection Prevalence ◽  
Northern Iran ◽  
Helminth Infections ◽  
High Infection ◽  
Northwestern Iran ◽  
Direct Economic Losses

Abstract Introduction: Helminth infections of livestock can result in considerable economic losses. This study aims to evaluate the spatial frequency of cystic echinococcosis (CE), dicrocoeliasis, and fascioliasis in livestock slaughtered in Iran during the years 2015-2019 and estimate direct costs associated with organ condemnation due to these parasites.Methods: Abattoir data from all 31 Iranian provinces were collected from the Iran Veterinary Organization. Infection prevalence was calculated per year at the province level. The Local Moran's I statistic was performed to evaluate spatial autocorrelation of animals positive at slaughter for the years 2015-2019. Direct costs associated with condemned livers were calculated for each parasitic condition. Results: Overall prevalence values for the study timeframe were as follows: sheep and goat fascioliasis (1.5%, 910,282/58,393,349), cattle fascioliasis (3.8%, 23,3175/6,038,419), sheep and goat dicrocoeliasis (4.6%; 270,1274/58,393,349), cattle dicrocoeliasis (3.1%; 186,009/6,038,419), sheep and goat CE (5.3%; 3,108,767/58,393,349), and cattle CE (7.2%; 438,534/6,038,419). Northwestern Iran had the highest prevalence of CE and fascioliasis. High infection areas for Dicrocoelium spp. included the provinces of Zanjan, Gilan, Qazvin, and Tehran, which are located in northern Iran. Direct economic losses for sheep and goat fascioliasis, dicrocoeliasis, and CE for the study period were US$13,841,826, US$41,768,472, and US$22,801,296, respectively. Direct economic losses for cattle fascioliasis, dicrocoeliasis, and CE for the study period were US$1,989,582, US$1,669,289, and US$2,656,535, respectively.Conclusion: Our findings provide valuable data for future monitoring of these important parasitic diseases in Iranian livestock. Disease control strategies are required to reduce the economic and public health impact of these helminths.

scholarly journals Epichromatin is conserved in Toxoplasma gondii and labels the exterior parasite chromatin throughout the cell cycle

Parasitology ◽  
2013 ◽  
Vol 140 (9) ◽  
pp. 1104-1110 ◽  
Author(s):  
LAURA VANAGAS ◽  
MARIA C. DALMASSO ◽  
JEAN F. DUBREMETZ ◽  
ENRIQUE L. PORTIANSKY ◽  
DONALD E. OLINS ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Human Fibroblasts ◽  
Protozoan Parasite ◽  
Nuclear Fraction ◽  
Mitotic Chromosomes ◽  
Interphase Nuclei ◽  
The Face ◽  
Surface Covering ◽  
Intracellular Protozoan Parasite

SUMMARYToxoplasma gondii is an apicomplexan intracellular protozoan parasite responsible for toxoplasmosis, a disease with considerable medical and economic impact worldwide. Toxoplasma gondii cells never lose the nuclear envelope and their chromosomes do not condense. Here, we tested the murine monoclonal antibody PL2-6, which labels epichromatin (a conformational chromatin epitope based on histones H2A and H2B complexed with DNA), in T. gondii cultured in human fibroblasts. This epitope is present at the exterior chromatin surface of interphase nuclei and on the periphery of mitotic chromosomes in higher eukaryotes. PL2-6 reacted with T. gondii H2A and H2B histones in Western blot (WB) assays. In addition, the antibody reacted with the nuclear fraction of tachyzoites, as a single band coincident with H2B histone. In the T. gondii tachyzoite stage, PL2-6 also had peripheral nuclear localization, as observed by epifluorescence/confocal microscopy and immunoelectron microscopy. Confocal analysis showed that epichromatin is slightly polarized to one face of the parasite exterior chromatin surface. In replicating tachyzoites, PL2-6 also labels the exterior chromatin surface, covering the face of both segregating nuclei, facing the plasma membrane of the mother cell. The possible role of epichromatin in T. gondii is discussed.

scholarly journals Toll-Like Receptor 3–TRIF Pathway Activation by Neospora caninum RNA Enhances Infection Control in Mice

2019 ◽  
Vol 87 (4) ◽  
Author(s):  
Vanessa dos Santos Miranda ◽  
Flávia Batista Ferreira França ◽  
Mylla Spirandelli da Costa ◽  
Vanessa Resende Souza Silva ◽  
Caroline Martins Mota ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Parasitophorous Vacuole ◽  
Protozoan Parasite ◽  
Interferon Response ◽  
Economic Losses ◽  
Toll Like Receptor ◽  
Content Type ◽  
Pathway Activation ◽  
Th1 Immune Responses

ABSTRACT Neospora caninum is a protozoan parasite closely related to Toxoplasma gondii and has been studied for causing neuromuscular disease in dogs and abortions in cattle. It is recognized as one of the main transmissible causes of reproductive failure in cattle and consequent economic losses to the sector. In that sense, this study aimed to evaluate the role of Toll-like receptor 3 (TLR3)–TRIF-dependent resistance against N. caninum infection in mice. We observed that TLR3−/− and TRIF−/− mice presented higher parasite burdens, increased inflammatory lesions, and reduced production of interleukin 12p40 (IL-12p40), tumor necrosis factor (TNF), gamma interferon (IFN-γ), and nitric oxide (NO). Unlike those of T. gondii, N. caninum tachyzoites and RNA recruited TLR3 to the parasitophorous vacuole (PV) and translocated interferon response factor 3 (IRF3) to the nucleus. We also observed that N. caninum upregulated the expression of TRIF in murine macrophages, which in turn upregulated IFN-α and IFN-β in the presence of the parasite. Furthermore, TRIF−/− infected macrophages produced lower levels of IL-12p40, while exogenous IFN-α replacement was able to completely restore the production of this key cytokine. Our results show that the TLR3-TRIF signaling pathway enhances resistance against N. caninum infection in mice, since it improves Th1 immune responses that result in controlled parasitism and reduced tissue inflammation, which are hallmarks of the disease.

Evaluation of a SYBR Green Real-Time PCR Assay for Specific Detection of Pasteurella multocida in Culture and Tissue Samples from Sheep

2020 ◽  
Author(s):  
Jyoti Kumar ◽  
G. G. Sonawane ◽  
Fateh Singh ◽  
S. Jegaveera Pandian ◽  
Rajiv Kumar
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pasteurella Multocida ◽  
Bacterial Species ◽  
Sybr Green ◽  
Economic Losses ◽  
Conventional Pcr ◽  
Specific Detection ◽  
Pcr Assay ◽  
Tissue Samples

Pasteurella multocida is one of the bacterial species involved in cases of ovine respiratory complex that has been implicated to cause significant economic losses in sheep production system worldwide. The present study was undertaken with the aim of evaluating a SYBR Green dye based real time PCR assay targeting KMT1 gene for the detection of P. multocida. The analytical specificity and sensitivity of the PCR primers were evaluated. The test showed ten-fold more sensitivity than conventional PCR and detected down to 275.5 fg/ µl of genomic DNA concentration, equivalent to 100 copies of KMT1 gene of P. multocida. The real-time PCR was found to be specific for KMT1 gene of P. multocida, as no cross reactivity was detected with a variety of known bacterial isolates. A total of 52 ovine lung tissue samples were screened for P. multocida, which showed improved level of detection as compared to conventional PCR. It is concluded that, this assay may be used as a valuable diagnostic tool for the rapid and specific detection of P. multocida. By virtue of its high throughput format and its ability to accurately identify as well as quantify the bacterial DNA, the method may be useful in large scale epidemiological studies and clarification of pathogenesis.

scholarly journals Duplex PCR Methods for the Molecular Detection ofEscherichia fergusoniiIsolates from Broiler Chickens

2014 ◽  
Vol 80 (6) ◽  
pp. 1941-1948 ◽  
Author(s):  
Karen Simmons ◽  
Heidi Rempel ◽  
Glenn Block ◽  
Vincenzo Forgetta ◽  
Rolland Vaillancourt ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Broiler Chickens ◽  
Hypothetical Protein ◽  
Cellulose Synthase ◽  
Conventional Pcr ◽  
Bacterial Concentration ◽  
Pcr Assay ◽  
Wide Range ◽  
Fraser Valley

ABSTRACTEscherichia fergusoniiis an emerging pathogen that has been isolated from a wide range of infections in animals and humans. Primers targeting specific genes, includingyliE(encoding a conserved hypothetical protein of the cellulose synthase and regulator of cellulose synthase island),EFER_1569(encoding a hypothetical protein, putative transcriptional activator for multiple antibiotic resistance), andEFER_3126(encoding a putative triphosphoribosyl-dephospho-coenzyme A [CoA]), were designed for the detection ofE. fergusoniiby conventional and real-time PCR methods. Primers were screened byin silicoPCR against 489 bacterial genomic sequences and by both PCR methods on 55 reference and field strains. Both methods were specific and sensitive forE. fergusonii, showing amplification only for this bacterium. Conventional PCR required a minimum bacterial concentration of approximately 102CFU/ml, while real-time PCR required a minimum of 0.3 pg of DNA for consistent detection. Standard curves showed an efficiency of 98.5%, with anR2value of 0.99 for the real-time PCR assay. Cecal and cloacal contents from 580 chickens were sampled from broiler farms located in the Fraser Valley (British Columbia, Canada). PresumptiveE. fergusoniiisolates were recovered by enrichment and plating on differential and selective media. Of 301 total presumptive isolates, 140 (46.5%) were identified asE. fergusoniiby biochemical profiling with the API 20E system and 268 (89.0%) using PCR methods.E. fergusoniidetection directly from cecal and cloacal samples without preenrichment was achieved with both PCR methods. Hence, the PCR methods developed in this work significantly improve the detection ofE. fergusonii.

scholarly journals Comparative Evaluation of Four Potent Neospora caninum Diagnostic Antigens Using Immunochromatographic Assay for Detection of Specific Antibody in Cattle

2021 ◽  
Vol 9 (10) ◽  
pp. 2133
Author(s):  
Ragab Fereig ◽  
Hanan Abdelbaky ◽  
Yoshifumi Nishikawa
Keyword(s):  
Current Knowledge ◽  
Neospora Caninum ◽  
Control Strategies ◽  
Acute Infection ◽  
High Sensitivity ◽  
Protozoan Parasite ◽  
Dense Granule ◽  
Caninum Infection ◽  
Kappa Value ◽  
Intracellular Protozoan Parasite

Neospora caninum is an intracellular protozoan parasite responsible for numerous abortion outbreaks and neonatal abnormalities in cattle. Rapid and accurate diagnosis is critical for N. caninum control owing to the lack of vaccine or drug-based control strategies. Herein, we evaluated the performance of four frequently used antigens in the diagnosis of N. caninum infection using immunochromatographic tests (ICTs) as a rapid, affordable, and field applicable tool. These antigens included recombinant proteins of N. caninum surface antigen 1 (NcSAG1), dense granule proteins 7 (NcGRA7) and 6 (NcGRA6), in addition to native Neospora lysate antigen (NLA). Our study revealed the utility of all antigen-based ICTs for detection of specific antibodies to N. caninum. However, the NcSAG1-based ICT was the best for detection of all control N. caninum-infected mouse or cattle sera, while NcGRA7 and NcGRA6-based ICTs exhibited specific ability to detect samples from acute and sub-acute infection in mice and cattle, respectively. Analyses of the NcSAG1-based ICT against enzyme-linked immunosorbent assays (ELISAs) of the same antigen revealed its efficiency in detection of field cattle samples as observed in high sensitivity (84.2%), specificity (93.5%), agreement (90%), and kappa value (0.78). The current knowledge provides an efficient platform for N. caninum control through on-site diagnosis of infected cattle.

scholarly journals TLR3-TRIF pathway activation by Neospora caninum RNA enhances infection control

2018 ◽  
Author(s):  
Vanessa dos Santos Miranda ◽  
Flávia Batista Ferreira França ◽  
Mylla Spirandelli da Costa ◽  
Vanessa Resende Souza Silva ◽  
Caroline Martins Mota ◽  
...  
Keyword(s):  
Immune Responses ◽  
Neospora Caninum ◽  
Parasitophorous Vacuole ◽  
Parasite Burden ◽  
Protozoan Parasite ◽  
Economic Losses ◽  
Pathway Activation ◽  
Ifn Γ ◽  
Th1 Immune Responses

AbstractNeospora caninum is a protozoan parasite closely related to Toxoplasma gondii and has been studied for causing neuromuscular disease in dogs and abortions in cattle. It is recognized as the major cause of economic losses in bovine products. In that sense, this study aimed to evaluate the role of TLR3-TRIF dependent resistance against N. caninum infection. We observed that TLR3−/− and TRIF−/− mice presented higher parasite burden, increased inflammatory lesions and reduced production of IL-12p40, TNF, IFN-γ, and NO. Differently from T. gondii, N. caninum tachyzoites and its RNA recruited TLR3 and IRF3 to the parasitophorous vacuole (PV). We observed that N. caninum upregulated the expression of TRIF in macrophages, which by its turn upregulated IFN-α and IFN-β in the presence of the parasite. Furthermore, TRIF−/− infected macrophages produced lower levels of IL-12p40 and IFN-α replacement was able to completely restore the production of this key cytokine. Our results have shown that TLR3-TRIF signaling pathway enhances resistance against N. caninum infection, since it improves Th1 immune responses that control parasitism and tissue inflammation, which are hallmarks of the disease.

scholarly journals From Signaling Pathways to Distinct Immune Responses: Key Factors for Establishing or Combating Neospora caninum Infection in Different Susceptible Hosts

Pathogens ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 384 ◽  
Author(s):  
Ragab M. Fereig ◽  
Yoshifumi Nishikawa
Keyword(s):  
Host Cell ◽  
Neospora Caninum ◽  
Protozoan Parasite ◽  
Survey Method ◽  
Economic Losses ◽  
Key Factors ◽  
Caninum Infection ◽  
Successful Infection ◽  
Comprehensive Survey

Neospora caninum is an intracellular protozoan parasite affecting numerous animal species. It induces significant economic losses because of abortion and neonatal abnormalities in cattle. In case of infection, the parasite secretes numerous arsenals to establish a successful infection in the host cell. In the same context but for a different purpose, the host resorts to different strategies to eliminate the invading parasite. During this battle, numerous key factors from both parasite and host sides are produced and interact for the maintaining and vanishing of the infection, respectively. Although several reviews have highlighted the role of different compartments of the immune system against N. caninum infection, each one of them has mostly targeted specific points related to the immune component and animal host. Thus, in the current review, we will focus on effector molecules derived from the host cell or the parasite using a comprehensive survey method from previous reports. According to our knowledge, this is the first review that highlights and discusses immune response at the host cell–parasite molecular interface against N. caninum infection in different susceptible hosts.

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