Enhancement of immune responses by vaccine potential of three antigens, including ROP18, MIC4, and SAG1 against acute toxoplasmosis in mice

Toxoplasma gondii (T. gondii) causes considerable financial losses in the livestock industry and can present serious threats to pregnant women, as well as immunocompromised patients. Therefore, it is required to design and produce an efficient vaccine for controlling toxoplasmosis. The present study aimed to evaluate the protective immunity induced by RMS protein (ROP18, MIC4, and SAG1) with Freund adjuvant, calcium phosphate nanoparticles (CaPNs), and chitosan nanoparticles (CNs) in BALB/c mice. The RMS protein was expressed in Escherichia coli (E. coli) and purified using a HisTrap HP column. Thereafter, cellular and humoral immunity was assessed by injecting RMS protein on days 0, 21, and 35 into four groups [RMS, RMS-chitosan nanoparticles (RMS-CNs), RMS-calcium phosphate nanoparticles (RMS-CaPNs), and RMS-Freund]. Phosphate buffered saline (PBS), CNs, CaPNs, and Freund served as the four control groups. The results displayed that vaccination with RMS protein and adjuvants significantly elicited the levels of specific IgG antibodies and cytokines against toxoplasmosis. There were high levels of total IgG, IgG2a, and IFN-γ in vaccinated mice, compared to those in the control groups, especially in the RMS-Freund, indicating a Th-1 type response. The vaccinated and control mice were challenged intraperitoneally with 1 × 103 tachyzoites of the T. gondii RH strain four weeks after the last injection, and in RMS-Freund and RMS-CaPNs groups, the highest increase in survival time was observed (15 days). The RMS can significantly increase Th1 and Th2 responses; moreover, multi-epitope vaccines with adjuvants can be a promising strategy for the production of a vaccine against toxoplasmosis

Toxoplasma gondii: A possible etiologic agent for Alzheimer's disease

Toxoplasma gondii (T. gondii) is one of the most pervasive neurotropic pathogens causing different lesions in a wide variety of mammals as intermediate hosts, including humans. It is estimated that one-third of the world population is infected with T. gondii; however, for a long time, there has been much interest in the examination of the possible role of this parasite in the development of mental disorders, such as Alzheimer's disease (AD). T. gondii may play a role in the progression of AD using mechanisms, such as the induction of the host's immune responses, inflammation of the central nervous system (CNS), alteration in the levels of neurotransmitters, and activation of indoleamine-2,3-dyoxigenase. This paper presents an appraisal of the literature, reports, and studies that seek to the possible role of T. gondii in the development of AD. For achieving the purpose of the current study, a search of six English databases (PubMed, ScienceDirect, Web of Science, Scopus, ProQuest, and Google Scholar) was performed. The results support the involvement of T. gondii in the induction and development of AD. Indeed, T. gondii can be considered a risk factor for the development of AD and requires the special attention of specialists and patients. Furthermore, the results of this study may contribute to prevent or delay the progress of AD worldwide. Therefore, it is required to carry out further studies in order to better perceive the parasitic mechanisms in the progression of AD

Neospora caninum infection in aborting bovines and lost fetuses: A systematic review and meta-analysis.

BackgroundNeospora caninum (N. caninum) is known to be a major cause of reproductive failure in cattle herds around the world. Therefore, the current comprehensive study was performed to estimate the global prevalence of N. caninum infection in bovines that had an abortion and aborted fetuses.MethodsIn this study, PubMed, ScienceDirect, Web of Science, Scopus, and ProQuest databases were systematically searched for relevant studies up until November 4, 2021. Pooled prevalence and corresponding 95% confidence intervals (CI) were estimated using a random effect model. Other analyzes performed on the data of this study include sensitivity analysis, publication bias test, and quality assessment.ResultsThe final analyses included 71 studies conducted on 2965 abortive cattle and 4805 aborted fetuses. The overall prevalence rates of N. caninum infection in bovines that had an abortion were 47% and 1% using serological and molecular methods. Furthermore, overall prevalence rates of N. caninum infection in bovine aborted fetuses globally were 35% (95% CI: 8%-62%) and 43% (95% CI: 35%-52%) using serological and molecular methods.ConclusionsThe results of this study showed the high prevalence of N. caninum infection in bovines that had an abortion and aborted fetuses. It is hoped that the results of this study will help prevent abortion in bovines around the world and encourage further studies to determine the impact of this parasite on the occurrence of abortion that may help reduce the economic damage caused by abortion worldwide

Evaluation of Anti-amoebic Activity of Peganum harmala Ethanolic Extract on Acanthamoeba In vitro

Abstract Background: Acanthamoeba is an opportunistic protozoan pathogen that is known to infect the cornea to produce eye keratitis and the central nervous system to produce lethal granulomatous encephalitis. The overall aim of the present study was to determine the anti-amoebic potential of natural compound Peganum harmala against the trophozoites and cysts of Acanthamoeba in vitro. Materials and Methods: In this experimental study, a clinical isolate of Acanthamoeba was cultured and genotyped. The ethanolic extract of Peganum harmala was prepared. The trophozoites and cysts were collected by washing in page's saline. Various concentrations (1.25, 2.5, 5, and 10 mg/ml) of the ethanolic extract and polyhexanide 0.02% drop as positive control were tested at three different times (24, 48 and 72 h) on trophozoites and cysts of Acanthamoeba in vitro. The viability of trophozoites or cysts was tested by eozin method, MTT, and flowcytometry analysis. Results: The results revealed that alcoholic extract had remarkable inhibitory effect on the proliferation of Acanthamoeba cysts as compared to non-treated control, and the inhibition was time and dose dependent. In the presence of 10 mg/ml ethanolic extract in medium culture after 72 h, no viable trophozoites were determined and 21.10% cysts of Acanthamoeba were viable. Percentage of trophozoites and cysts viability after adding polyhexanide 0.02% drop in medium culture after 72 hours was 0% and 23.71%, respectively. Conclusion: Ethanolic extracts of Peganum harmala could be considered a new natural compound against the Acanthamoeba trophozoites and cysts. Further works are required to evaluate the exact effect of this extract on Acanthamoeba agents in animal models

The global seroprevalence of anti-Toxoplasma gondii antibodies in women who had spontaneous abortion: A systematic review and meta-analysis.

BACKGROUND:Toxoplasma gondii (T. gondii) is an intracellular pathogen that can lead to abortion in pregnant women infected with this parasite. Therefore, the present study aimed to estimate the global seroprevalence of anti-T. gondii antibodies in women who had spontaneous abortion based on the results of published articles and evaluate the relationship between seroprevalence of anti-T. gondii antibodies and abortion via a systematical review and meta-analysis. METHODS:Different databases were searched in order to gain access to all studies on the seroprevalence of anti- T. gondii antibodies in women who had spontaneous abortion and association between seroprevalence of anti-T. gondii antibodies and abortion published up to April 25th, 2019. Odds ratio (OR) and the pooled rate seroprevalence of T. gondii with a 95% confidence interval (CI) were calculated using the random effects model. RESULTS:In total, 8 cross-sectional studies conducted on 1275 women who had abortion in present pregnancy, 40 cross-sectional studies performed on 9122 women who had a history of abortion, and 60 articles (involving 35 cross-sectional studies including 4436 women who had spontaneous abortion as case and 10398 as control and 25 case-control studies entailing 4656 cases and 3178 controls) were included for the final analyses. The random-effects estimates of the prevalence of anti-T. gondii IgG antibody in women who had abortion in present pregnancy and women who had a history of abortion were 33% (95% CI: 17%-49%) and 43% (95% CI: 27%-60%), respectively. In addition, the pooled OR for anti-T. gondii IgG antibody in cross-sectional and case-control studies among women who had spontaneous abortion were 1.65 (95% CI: 1.31-2.09) and 2.26 (95% CI: 1.56-3.28), respectively. Also, statistical analysis showed that the pooled OR of the risk of anti-T. gondii IgM antibody 1.39 (95% CI: 0.61-3.15) in cross-sectional and 4.33 (95% CI: 2.42-7.76) in case-control studies. CONCLUSION:Based on the results of the current study, T. gondii infection could be considered a potential risk factor for abortion. It is recommended to carry out further and more comprehensive investigations to determine the effect of T. gondii infection on abortion to prevent and control toxoplasmosis among pregnant women around the world

Intestinal parasites among migrant barn swallows (Hirundo rustica) in the central region of Mazandaran Province, Northern Iran

Aim: Swallows are a family of migratory birds found worldwide except Antarctica. Annually, a number of species of swallows migrate to Iran. As they make their nests close to human living places, this may be a potential risk for public health. Conversely, no study has been conducted on intestinal parasitic infections of these birds so far. Therefore, the objective of this study was to determine the prevalence of intestinal parasites in migratory swallows (Hirundo rustica) in the central region of Mazandaran Province, Northern Iran. Materials and Methods: In this cross-sectional study, 205 feces samples from two districts (Sari and Qaemshahr) in the central region of Mazandaran were randomly collected during the summer and spring sessions of 2016-2017. The collected samples were examined using the routine direct fecal examination and formalin-ethyl acetate concentration. In addition, the samples were examined by cold acid-fast staining method to detect possible Cryptosporidium oocysts. Results: The results of this study indicated that 38 samples (18.5%) were infected with intestinal parasites. Among the helminthic parasites, eight genera and species were identified as follows: Ascaridia galli, Syngamus trachea, Raillietina, Toxocara spp., Choanotaenia, Taenia spp., Ascaridia spp., and Moniezia spp. In addition, among protozoan parasites, only the Coccidia spp. oocysts were identified. Conclusion: Our findings showed a relatively high prevalence of parasitic infections in migratory barn swallows in Mazandaran Province. Given the presence of zoonotic parasites in the samples, further investigations are needed to identify all parasites fauna, particularly zoonotic species among swallows in the region

Detection and Molecular Characterization of Potentially Pathogenic Free-Living Amoebae from Recreational and Public Soils in Mazandaran, Northern Iran

Background: Free-living amoeba (FLA) belonging to Acanthamoeba spp., Naegleria, and Balamuthia mandrillaris are the soil-born protozoa. This study aimed to survey the occurrence of FLA, including Acanthamoeba spp., B. mandrillaris, Vermamoeba spp., and Naegleria spp., in soil samples collected from various districts of Mazandaran Province (Northern Iran) from July to December 2018. Methods: Overall, 118 soil samples from the recreational and public places were surveyed for the existence of Acanthamoeba spp., Vermamoeba, Naegleria, and B. mandrillaris using both morphological key and molecular tools with genus-specific primers of JDP1, NA, ITS1, and Bal, respectively. To verify the taxonomic status of isolated amoeba, the phylogenetic tree was made based on sequences of 18S rRNA by MEGA (5.05) software with the maximum likelihood model. Results: Overall, 61/118 samples (51.6%) were contaminated with FLA, and based on the sequencing data, 29 isolates were successfully sequenced. Among the samples, all isolated Acanthamoeba (52.4%) belonged to the T4 genotype with amplification of the DF3 region (18S rRNA gene). Internal transcribed spacer (ITS) sequencing revealed the presence of one strain of Naegleria americana. Twenty-eight V. vermiformis were also confirmed based on Nuclear SSU rDNA. Morphological survey and PCR assay did not show any positive samples for B. mandrillaris. Conclusion: The present study indicates the occurrence of FLA in soil sources of the recreational and public places in Mazandaran province that it can be a severe risk to human health. Thus, more studies are expected to survey the infection source in patients with FLA-related diseases

Protective efficacy by a novel multi-epitope vaccine, including MIC3, ROP8, and SAG1, against acute Toxoplasma gondii infection in BALB/c mice

Toxoplasma gondii is an intracellular apicomplexan parasite, which can cause a serious infectious disease in pregnant women and immunocompromised individuals. Therefore, the development of a polyvalent vaccine consisting of all stages of the parasite life cycle using the epitopes from tachyzoites, bradyzoites, and sporozoites is likely to be required for complete protective immunity. In this study, we designed protein vaccine candidate based on the prediction of specific epitopes (i.e., B cell and T cell) from three Toxoplasma gondii antigens. The MRS protein (MIC3: 30–180, ROP8: 85–185, and SAG1: 85–235) was expressed in Escherichia coli, and purification was performed using a HisTrap HP column and then we evaluated immunogenicity and protective property in BALB/c mice. Seventy-two mice were randomly divided into six groups, including three vaccinations (i.e., MRS, MRS-Freund, and MRS-Calcium Phosphate Nanoparticles (MRS-CaPNs)) and three control (i.e., Phosphate-buffered saline, Freund, and CaPNs) groups. All groups were immunized three times via subcutaneous injection within three-week intervals. In the vaccination groups, the BALB/c mice were injected with 20 μg of MRS protein for the first time and 10 μg of MRS for the next two times. Antibodies, cytokines, and splenocytes proliferation in the immunized mice were assayed using the enzyme-linked immunosorbent assay. Protective efficacy was analyzed by challenging the immunized mice with T. gondii of RH strain. Antibody, cytokine, and lymphocyte proliferation assays showed that the mice immunized with MRS induced stronger humoral and T helper type 1 cell-mediated immune responses, compared to the control mice. However, co-immunization with adjuvants (i.e., Freund and CaNPs) resulted in impaired immune responses. Effective protection against the parasite achieved an increase in survival time in the immunized mice, especially in the MRS-CaNPs group. The obtained results of the present study demonstrated that multi-epitope protein vaccination, MRS, is a potential strategy against toxoplasmosis infection. In addition, the vaccine co-delivered with CaPNs could provide an important key for vaccine candidate to control T. gondii infection. © 2021 Elsevier Ltd Author keywords Calcium phosphate; Freund; Multi-epitope; Protein vaccine; Toxoplasma gondi

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

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-chloroform-isoamyl 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