First molecular subtyping and zoonotic significance of Blastocystis sp. in Dromedary (C. dromedarius) and Bactrian (C. bactrianus) camels in Iran: A molecular epidemiology and review of available literature.

BACKGROUND: Blastocystis sp. is a zoonotic protozoan parasite, and there is limited information about its molecular prevalence and subtypes (STs) distribution in camels globally, especially in Iran. OBJECTIVES: This study aimed to examine the prevalence, STs distribution, and zoonotic potential of Blastocystis sp. in one-humped and two-humped camels in Ardabil province, northwestern Iran. METHODS: A PCR-sequencing tool using the SSU rRNA gene was employed to examine the occurrence and genetic variation of Blastocystis sp. in 150 faecal samples from Bactrian (Camelus bactrianus, 50 samples) and Dromedary (Camelus dromedarius, 100 samples) camels in Ardabil province. RESULTS: The overall prevalence of Blastocystis sp. in camels was determined to be 12% (18/150) through microscopy and PCR analyses. Phylogenetically, this study identified three distinct zoonotic STs: ST7, ST10, and ST14. ST10 was the most prevalent, comprising 50% (9/18) of the isolated STs from camels. ST14 closely followed with 38.9% (7/18), while ST7 made up 11.1% (2/18) of the total STs. In brief, ST10, ST14, and ST7 represented 50% (7/14), 35.7% (5/14), and 14.3% (2/14) of the Blastocystis-positive cases in one-humped camels, respectively. Further, each of the ST10 and ST14 accounted for 50% (2/4) of the Blastocystis-positive samples in two-humped camels. An analysis of the available data reveals that out of the 37-44 identified Blastocystis STs, 15 (ST1-ST7, ST10, ST14, ST15, ST21, ST24, ST25, ST26, and ST30) have been reported in camels. The predominant STs observed are ST10 and ST14. Furthermore, among the 15 zoonotic STs (ST1-ST10, ST12-ST14, ST16, and ST23) of Blastocystis reported thus far, nine zoonotic STs (ST1-ST7, ST10, and ST14) have been found in camels. CONCLUSIONS: These findings indicate that camels serve as a proper reservoir for a diverse array of Blastocystis STs and thereby can play a significant role in the transmission of this protozoan infection to humans, animals, and water reservoirs.

Review of osmotic dehydration: Promising technologies for enhancing products' attributes, opportunities, and challenges for the food industries.

Osmotic dehydration (OD) is an efficient preservation technology in that water is removed by immersing the food in a solution with a higher concentration of solutes. The application of OD in food processing offers more benefits than conventional drying technologies. Notably, OD can effectively remove a significant amount of water without a phase change, which reduces the energy demand associated with latent heat and high temperatures. A specific feature of OD is its ability to introduce solutes from the hypertonic solution into the food matrix, thereby influencing the attributes of the final product. This review comprehensively discusses the fundamental principles governing OD, emphasizing the role of chemical potential differences as the driving force behind the molecular diffusion occurring between the food and the osmotic solution. The kinetics of OD are described using mathematical models and the Biot number. The critical factors essential for optimizing OD efficiency are discussed, including product characteristics, osmotic solution properties, and process conditions. In addition, several promising technologies are introduced to enhance OD performance, such as coating, skin treatments, freeze-thawing, ultrasound, high hydrostatic pressure, centrifugation, and pulsed electric field. Reusing osmotic solutions to produce innovative products offers an opportunity to reduce food wastes. This review explores the prospects of valorizing food wastes from various food industries when formulating osmotic solutions for enhancing the quality and nutritional value of osmotically dehydrated foods while mitigating environmental impacts.

Development of New PCR Protocols to Detect Genetic Diversity in the Metronidazole Metabolism Genes in Susceptible and Refractory Clinical Samples of Giardia duodenalis.

PURPOSE: Investigating the genetic variation in thioredoxin reductase (TrxR) and nitroreductase (NR) genes in both treatment-resistant and -sensitive Giardia duodenalis isolates can provide valuable information in identifying potential markers of resistance to metronidazole. The rapid increase in metronidazole treatment failures suggests the presence of genetic resistance mechanisms. By analyzing these genes, researchers can gain insights into the efficacy of metronidazole against G. duodenalis and potentially develop alternative treatment strategies. In this regard, four G. duodenalis isolates (two clinically sensitive and two clinically resistant to metronidazole) were collected from various hospitals of Shiraz, southwestern Iran. METHODS: Parasitological methods including sucrose flotation and microscopy were employed for the primary confirmation of G. duodenalis cysts in stool samples. Microscopy-positive samples were approved by SSU-PCR amplification of the parasite DNA. All four positive G. duodenalis specimens at SSU-PCR were afterward analyzed utilizing designed primers based on important metronidazole metabolism genes including TrxR, NR1, and NR2. RESULTS: Unlike TrxR gene, the results of NR1 and NR2 genes showed that there are non-synonymous variations between sequences of treatment-sensitive and -resistant samples compared to reference sequences. Furthermore, the outcomes of molecular docking revealed that there is an interaction between the protein sequence and spatial shape of treatment-resistant samples and metronidazole in the position of serine amino acid based on the NR1 gene. CONCLUSION: This issue can be one of the possible factors involved in the resistance of Giardia parasites to metronidazole. To reach more accurate results, a large sample size along with simulation and advanced molecular dynamics investigations are needed.

Identification of Blastocystis spp. in Urban Rodents of Different Districts in Southwestern Iran: Subtype Distribution and Possible Zoonotic Potential.

PURPOSE: Rodents are one of the most abundant and diverse species of mammals and have recently been identified as carriers of numerous human pathogens. The current study was conducted to assess the prevalence, subtype (STs) distribution, and zoonotic potential of Blastocystis spp. in various species of rodents in Shiraz, southwestern Iran. METHODS: For this aim, a total of 120 fresh fecal samples were collected from Mus musculus (n = 40), Rattus norvegicus (n = 40), and Rattus rattus (n = 40) in various municipality districts of Shiraz (6 out of 10 districts) between February and November 2020. Upon detecting parasites using light microscopy, a DNA fragment of the Blastocystis SSU rDNA gene was amplified using conventional PCR. RESULTS: By employing direct wet mount examination, 8 out of 120 fecal samples (6.7%; 2 from house mice, 3 from black rats, and 3 from brown rats) tested positive. Similarly, 5% (2/40) of house mice, 7.5% (3/40) of black rats, and 7.5% (3/40) of brown rats tested positive using the molecular method. Phylogenetic analysis revealed that the Blastocystis infecting different rodent species in Shiraz belonged to two potentially zoonotic STs (ST1 and ST4). Accordingly, rodents should not be overlooked as potential reservoirs of zoonotic Blastocystis infections. Different sampled urban districts and their statistical association with reported prevalence rates were analyzed separately. CONCLUSION:  Overall, the issue of the frequency and ST distribution of Blastocystis in urban rodents of Iran is still open to question and for a proper understanding, wider and more comprehensive studies are needed.

Diabetic cardiomyopathy in rats was attenuated by endurance exercise through the inhibition of inflammation and apoptosis.

Diabetic cardiomyopathy (DCM), as a ventricular dysfunction, is one of the main causes of death in diabetic patients. Former evidence revealed the beneficial effects of exercise on cardiovascular complications of diabetes. We aimed to investigate the effects of high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT) on DCM. Male Wistar rats were divided into control, diabetic, metformin (300 mg/kg), HIIT, MICT, metformin + HIIT, and metformin + MICT diabetic groups. Serum biochemical, inflammatory, and oxidative stress indicators, gene expression of BCL2 and BAX, and histopathologic changes of cardiac tissue were assessed. Our analysis revealed an increase in fasting blood sugar (FBS), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH), and aspartate aminotransferase (AST) in diabetes. Also, the superoxide dismutase (SOD) and catalase (CAT) activity, and the total thiol were decreased, in contrast, malondialdehyde (MDA) levels increased in the cardiac tissue of the diabetic group. All of these changes were significantly ameliorated in diabetic animals treated with exercise and metformin + exercise. The level of tumor necrosis factor-α (TNF-α) and Interleukin-1ß (IL-1ß), as well as the infiltration of inflammatory cells, were decreased in the heart of all exercise training groups. Up-regulation of BCL2 and down-regulation of BAX gene expressions were observed in the cardiac tissue of all exercise-treated groups. In conclusion, HIIT and MICT exercises are effective in preventing DCM development. Exercise training, besides improving oxidative stress and inflammation in cardiac tissue, alleviates cardiac damage by modulating the apoptotic gene expression in diabetic rats.

First Molecular Identification and Subtyping of Blastocystis sp. in the Most Consumed Edible Marine Fish of Iran: A Foodborne Concern.

PURPOSE: The presence of Blastocystis sp. is commonly observed in humans and different animals, displaying a wide range of genetic variations with the discovery of multiple subtypes (STs). However, the prevalence and distribution of these STs in edible marine fish and marine mammals remain uncertain. This study marks the first survey conducted in Iran and the second global molecular investigation to examine the occurrence and STs distribution of Blastocystis in various species of edible marine fish. METHODS: This study screened 200 fresh intestinal contents from 10 well-known fish species (Narrow-barred mackerel, Indo-pacific king mackerel, Tigertooth croaker, Silver pomfret, Black pomfret, Longtail tuna, John's snapper, Blackspotted croaker, Four-finger threadfin, and Javelin grunter) in southern Iran, caught in the Persian Gulf. All collected samples were evaluated by microscopy and SSU-PCR methods. RESULTS: Based on both microscopy and PCR, the overall prevalence of Blastocystis sp. in evaluated fish species was 2% (4/200). In brief, Blastocystis sp. was reported from Narrow-barred mackerel [10% (2/20)], Silver pomfret [5% (1/20)], and Tigertooth croaker [5% (1/20)]. Interestingly, among infected fish species three zoonotic STs (ST1, ST2, and ST7) were identified. ST2 was the most predominant ST [50% (2/4)], followed by ST1 and ST7, one sample each [5% (1/20)]. CONCLUSION: Overall, the prevalence and STs distribution of Blastocystis in edible marine fish along with the possibility of its zoonotic transmission are still open to question and require extensive and more detailed studies.

Comparative molecular epidemiology, subtype distribution, and zoonotic potential of Blastocystis sp. in Equus animals (horses, donkeys, and mules) in northwestern Iran.

A total of 500 fecal samples were collected from Equus animals in six different cities (Ardabil, Namin, Nir, Meshginshahr, Germi, and Khalkhal) of Ardabil Province, northwestern Iran, with 200 samples from horses, 200 from donkeys, and 100 from mules. Of the horse samples, 100 were from racing horses under special monitoring and care, while the remaining 100 were from non-racing horses, including those used for herding or in rural areas. All fecal samples were examined for the presence of Blastocystis sp. using PCR amplification of the SSU rRNA gene's barcode region after DNA extraction. The molecular prevalence of Blastocystis infection in Equus animals was 7.6% (38/500). Blastocystis was more common in horses [11.5% (23/200)] than in donkeys [5.5% (11/200)] and mules [4% (4/100)] (P > 0.05). Compared to racing horses [3% (3/100)], non-racing/rural horses [20% (20/100)] exhibited a substantially higher prevalence of Blastocystis (P < 0.05). The prevalence of Blastocystis in diarrheal samples and younger animals was remarkably higher than in formed samples and older animals, respectively (P < 0.05). No significant difference in Blastocystis infection prevalence was found between the genders of examined animals (P > 0.05). In Equus animals, 38 Blastocystis isolates included eight STs: ST10 [31.6% (12/38)], ST1 [21.1% (8/38)], ST2 [15.8% (6/38)], ST3 [10.5% (4/38)], ST4 [7.9% (3/38)], ST7 [5.2% (2/38)], ST14 [5.2% (2/38)], and ST6 [2.6% (1/38)]. These results suggest that Equus animals act as a proper reservoir for numerous Blastocystis STs, consequently playing a crucial part in the spread of this protozoan infection to humans, animals, and water reservoirs.

First molecular characterization of Blastocystis subtypes from domestic animals (sheep and cattle) and their animal-keepers in Ilam, western Iran: A zoonotic concern.

A total of 360 fecal samples were randomly collected from 150 cattle, 150 sheep, and 60 humans (30 people with close animal contact and 30 individuals without close animal contact) at 10 farms in Ilam, western Iran from June 2022 to August 2023. All samples were directly examined for Blastocystis by zinc sulfate flotation, followed by microscopic observation. Positive samples were further subtyped using conventional PCR and sequencing methods. A mean prevalence of 5.3% (16/300) was estimated for Blastocystis infection among examined animals, with 6% and 4.7% for cattle and sheep, respectively. Among the people who had close and non-close animal contact, 16.7% (5/30) and 3.3% (1/30) were infected with Blastocystis, respectively (p < 0.05). All 22 positive samples were successfully sequenced at the SSU rRNA locus. Accordingly, Blastocystis isolates infecting domestic animals in Ilam belonged to the four STs (ST1-ST3, and ST10). Of the 16 animal isolates, nine sequences (four ST10, three ST3, and two ST1) were related to cattle, and seven sequences (three ST10, two ST3, and two ST2) were isolated from sheep. Among the six human isolates, ST3 was the most predominant ST, followed by STs 1, 2, 6, and 7 (one case each). Of note, ST1-ST3 were isolated in various farms both from animals and their breeders, which indicates the possible circulation of these STs between animal and human populations.

Facile preparation, characterization, and investigation of mechanical strength of Starchy NaCl-binder as a lightweight construction material.

Sodium chloride (NaCl), commonly known as salt, is a substance that is utilized in a variety of businesses, including the tourism and construction industries. Therefore, the main purpose of this article is to accommodate a salt-based building material called NaCl-binder for tourist and industrial applications. By utilizing salt mortar with varying grain sizes, food-grade corn starch as an exclusive binder agent (without using any non-starch binder), and water under microwave-cured conditions, environmentally friendly hydrophobic hybrid NaCl-binder samples with low bulk density were successfully produced. The fabrication of these samples involved an inventive utilization of small quantities of starch. This study evaluated the impact of microwave exposure time on the strength of salt samples, particle interconnectivity and chemical composition using SEM, XRD, and XRF analyses. The compressive strength of the samples showed a remarkable increase, with a 600% improvement when using 0 to 1% corn starch, and a 137% increment when using 1 to 10% corn starch, indicating a lower rate of increment with higher starch consumption. A key aspect of this research is the significant reduction in starch consumption compared to other corn starch-based materials during the manufacturing process of the incorporated materials, highlighting its novelty and importance.

Physical and physiological characteristics, as well as miRNA concentrations, are affected by the storage time of tomatoes.

This study aims to investigate the potential of miRNA measurements in indicating tomato quality during transportation and storage. The impact of storage temperature, duration, and mechanical loading on tomato senescence, carotenoid content, total soluble solids, fruit firmness, and relevant miRNA concentrations were examined. Significant two-way or three-way interactions were observed between storage conditions and physical/physiological characteristics (excluding carotenoids). Remarkably, significant three-way interactions were found between storage conditions and miRNA concentrations. Strong correlations were observed between the physiological characteristics of the tomatoes and their miRNA concentrations. These findings suggest that measuring miRNAs could serve as a convenient and portable method for evaluating postharvest fruit quality, reducing reliance on labor-intensive laboratory techniques.

Deciphering the level of genetic diversity in some aegilops species using CAAT box-derived polymorphism (CBDP) and start codon target polymorphism (SCoT) markers.

BACKGROUND: Maintaining genetic diversity is of the most essential principle for a long-term conservation of plant genetic resources and could play a crucial role in their management. The genus Aegilops is one important member of wheat germplasm, and there are evidences that novel genes of this genus' species can be studied/utilized as ideal sources for the wheat cultivar improvement. The objective of this study was to dissect the genetic diversity and population structure among a set of Iranian Aegilops using two gene-based molecular markers. METHODS AND RESULTS: This study investigated the level of genetic diversity among 157 Aegilops accessions consisting of Ae. tauschii Coss. (DD genome), Ae. crassa Boiss. (DDMM genome), and Ae. cylindrica Host. (CCDD genome) belonging to NPGBI using two sets of CBDP and SCoT markers. The SCoT and CBDP primers yielded 171 and 174 fragments, out of which 145 (90.23%) and 167 (97.66%) fragments were polymorphic, respectively. The average of polymorphism information content (PIC)/ marker index (MI)/resolving power (Rp) for SCoT and CBDP markers were 0.32/3.59/16.03 and 0.29/3.01/16.26, respectively. Results of AMOVA revealed the genetic variability within species was greater than the variation observed among them (SCoT: 88% vs. 12%; CBDP: 72% vs. 28%; SCoT + CBDP: 80% vs. 20%). Based on the information obtained from both markers, the higher level of genetic diversity was found in Ae. tauschii as compared to other species. The grouping patterns obtained by Neighbor-joining algorithms, principal coordinate analysis (PCoA), and Bayesian-model-based structure were consistent with each other and resulted in grouping all studied accessions according to their genomic constitutions. CONCLUSION: The results of this study revealed a high level of genetic diversity among Iranian Aegilops germplasm. Moreover, SCoT and CBDP marker systems were efficient in deciphering DNA polymorphism and classification of Aegilops germplasm.

The effects of endurance exercise and metformin on memory impairment caused by diabetes.

OBJECTIVES: Diabetes has a negative effect on learning and memory performance, and it is a risk factor for Alzheimer's disease and dementia development. The present study aims to investigate the effects of two kinds of endurance exercise including high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT) as well as metformin on impaired memory and learning related to streptozotocin (STZ) induced diabetes in rats. METHODS: Forty adult male rats (250 ± 20 g weight) were divided into five groups (n=8), including control, diabetic, as well as diabetic rats treated with metformin (300 mg/kg), and HIIT (20 m/min), and MICT (15 m/min) exercises. Diabetes was induced by STZ (60 mg/kg, i.p.). Serum glucose concentration and oxidative stress markers (SOD, CAT, thiol, and MDA) in the cortex and hippocampus were determined by colorimetric assay. Behavioral tests were performed with a passive avoidance test. RESULTS: The diabetic groups treated with metformin and both HIIT, and MICT exercises improved the latency and the staying time in the darkroom and lightroom. The entrance frequency into the darkroom also was restored (p<0.01-p<0.001). In both HIIT and MICT exercises as well as metformin groups the oxidative stress induced by diabetes has been reversed and attenuation of the serum glucose level has been observed compared to non-treated diabetic ones (p<0.05-p<0.001). CONCLUSIONS: The results of the present study revealed both HIIT and MICT exercises had protective effects against oxidative stress and behavioral impairments induced by diabetes and these effects were comparable to the effects of metformin.

Global molecular prevalence of Giardia duodenalis in pigs (Sus domesticus): A systematic review and meta-analysis.

Giardia duodenalis is one of the common intestinal parasites causing diarrhea in humans and livestock, including pigs. Thus, a healthy livestock would result in a clean environment, which benefits humans. In the present study, the global molecular prevalence of G. duodenalis infection was determined in pig populations, through systematic exploration of 4 international databases (MEDLINE/PubMed, Scopus, Web of Science, and Google Scholar) until March 4th, 2022. A random-effects meta-analysis model was used to estimate the overall and subgroup-based pooled prevalence of G. duodenalis, and I 2 index was used for the evaluation of the heterogeneity. Altogether, 42 datasets from 18 papers examined 7272 pigs across 12 nations, showing a 9.1% (95% CI: 5.6-14.3%) pooled molecular prevalence. Sensitivity analysis demonstrated no remarkable variation in the reported total prevalence upon removing individual studies. It was found that 6 Giardia assemblages (A-F) are capable to infect pigs around the world, including assemblage E [16 datasets, 41.1% (95% CI: 24.8-59.6%)], B [8 datasets, 28.2% (95% CI: 12.2-52.6%)], D [3 datasets, 16.2% (95% CI: 10.6-24.1%)], C [3 datasets, 11.6% (95% CI: 7.3-17.9%)], and A [11 datasets, 9.9% (95% CI: 5.6-16.9%)]. Of note, assemblage F was only reported in one study. Meta-regression analysis showed that publication year was not significantly associated with the Giardia prevalence in swine population, in contrast to the sample size. Substantially, animals in weaner and fattener stages were more prone to giardiasis. Assemblages A and B are of utmost zoonotic significance for humans, while assemblages C, D and F have, also, been found in dogs and cats. Still, little is known on the prevalence and distribution of Giardia assemblages in pigs and requires more extensive and detailed studies.

Protective effects of cinnamon on acetylcholinesterase activity and memory dysfunction in diabetic rats.

OBJECTIVES: Regarding neurocognitive and immunomodulatory properties of cinnamon (Cinn) we aimed to investigate whether cinnamon regulates acetylcholinesterase (AChE) activity, and oxidative abnormalities with concomitant memory dysfunction in streptozotocin (STZ)-induced diabetes. METHODS: Forty-seven male adult rats were divided into seven groups (n=8 animals): Control group: in these non-diabetic rats only saline 0.9% NaCl was gavaged, Diabetic (Dia) group: diabetic rats in them saline 0.9% NaCl was gavaged for six weeks. Dia-Cinn 100, Dia-Cinn 200, and Dia-Cinn 400, Dia-Met groups: in these diabetic rats the extract (100, 200, 400 mg/kg respectively) or metformin (300 mg/kg) was gavaged for six weeks. Passive avoidance performance, AChE enzyme activity, and oxidative indicators were examined among the groups. RESULTS: Vs. the control group, blood glucose level and stay time in the dark were remarkably increased in Dia group whereas the latency time was decreased. Meanwhile, antioxidant levels (superoxide dismutase, catalase, and thiols) noticeably decreased in the Dia group compared to the Control group. On the other hand, Cinn extract espicailly at the highest dose recovered the changes similar to those found in the metformin-treated group. CONCLUSIONS: These findings proposed that the cinnamon hydro-ethanolic extract promotes memory recovery in diabetic conditions through the atteuation of the AChE activity and oxidative injury.

Cardiac hypertrophy and fibrosis were attenuated by olive leaf extract treatment in a rat model of diabetes.

The key role of fibrosis and hypertrophy processes in developing diabetes-induced heart injury has been demonstrated. Considering the known hypoglycemic effects of olive leaf extract (OLE), we decided to investigate its potential effect and associated mechanisms on cardiac fibrosis and myocardial hypertrophy in streptozotocin (STZ)-induced diabetic rats. Eight groups were included in this study: control, diabetic, diabetic-OLEs (100, 200 and 400 mg/kg), diabetic-metformin (300 mg/kg), diabetic-valsartan (30 mg/kg), and diabetic-metformin/valsartan (300/30 mg/kg). After a treatment period of 6 weeks, echocardiography was used to assess cardiac function. Heart-to-body weight ratio (HW/BW) and fasting blood sugar (FBS) were measured. Myocardial histology was examined by Masson's trichrome staining. Gene expressions of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), ß-myosin heavy chain (ß-MHC), TGF-ß1, TGF-ß3, angiotensin II type 1 receptor (AT1), alpha-smooth muscle actin (α-SMA), and collagen were evaluated by the quantitative real-time PCR in heart tissue. A reduction in the FBS level and HW/BW ratio in the extract groups was obvious. The improvement of left ventricular dysfunction, cardiac myocytes hypertrophy, and myocardial interstitial fibrosis was also observed in treated groups. A lowering trend in the expression of all hypertrophic and fibrotic indicator genes was evident in the myocardium of OLE treated rats. Our data indicated that OLE could attenuate fibrosis and reduce myocardial hypertrophy markers, thus improving the cardiac function and structure in the STZ-induced diabetic rats. PRACTICAL APPLICATIONS: This study demonstrates that olive leaf extract in addition to lowering blood glucose levels and the heart-to-body weight ratio (HW/BW) may also improve cardiac function and reduce cardiac hypertrophy and fibrosis in cardiac tissue, which leads to inhibition of diabetic heart damage. Thus it is possible that including olive leaf extracts in the diets of individuals with diabetes may assist in lowering cardiovascular disease risk factors.

Blastocystis sp. in Small Ruminants: A Universal Systematic Review and Meta-analysis.

PURPOSE: The present review was done to evaluate the prevalence and subtype distribution of Blastocystis infection among small ruminants, at a global perspective. METHODS: Systematic search was performed in PubMed, Scopus, Google Scholar, and Web of Science until 30th January 2022 and total estimates along with 95% confidence intervals (CIs) were computed using a random-effects model. RESULTS: Ultimately, the required data were extracted from 25 papers including 19 datasets for each animal. Among 3125 sheep, the Blastocystis prevalence was 25.3% (95% CI 16.1-37.4%) (10 countries), being lower in comparison to that in 2869 examined goats [20.5% (95% CI 11-35.1%)] (12 countries). Regarding STs distribution, fourteen genetically diverse STs of Blastocystis (ST1-ST5, ST7, ST10, ST12, ST14, ST15, ST21, ST23, ST24, ST26) have been reported in sheep, and the highest pooled prevalence was related to ST10 [11 datasets, 57.8% (95% CI 43.7-70.8%)], followed by ST14 [8 datasets, 28.4% (95% CI 20.2-38.4%)], and ST7 [2 datasets, 21.1% (95% CI 4.5-60.3%)]. Compared to sheep, more STs (ST1, ST3-ST7, ST10, ST12, ST14, ST21, ST23-ST26, and ST32) were reported from goats, and the highest weighted frequency was related to ST10 [6 datasets, 45.1% (95% CI: 25.6-66.2%)], followed by ST7 [2 datasets, 40.4% (95% CI 30-51.7%)], and ST14 [4 datasets, 29% (95% CI 15.5-47.7%)]. Out of ten known zoonotic STs reported for Blastocystis (ST1-ST9, and ST12), 7 were isolated from sheep (ST1-ST5, ST7, and ST12) and 7 were reported from goats (ST1, ST3-ST7, ST12). CONCLUSIONS: Overall, Blastocystis epidemiology in sheep and goats is yet to be elucidated and demands more comprehensive studies.

Distribution, genetic diversity, and zoonotic significance of Blastocystis subtypes in pet dogs.

The common intestinal protist, Blastocystis sp., is a highly frequent parasitic agent in human and animal fecal samples, globally. The current study evaluated the prevalence of this parasite and involved subtypes (STs) in examined pet dogs in Shiraz, southwestern Iran. A number of 100 fresh fecal specimens were randomly collected from pet dogs referred to the veterinary clinics and pet shops in Shiraz during July 2021-January 2022. The genomic DNA was extracted from fecal samples and a conventional PCR was performed. Next, sequencing of the positive samples was done based on the small subunit ribosomal rRNA gene. The results showed that 5 % (5 out of 100) of the examined samples were positive for Blastocystis. As anticipated, three known Blastocystis subtypes with zoonotic concern, ST1 (60 %, n = 3), ST2 (20 %, n = 1) and ST3 (20 %, n = 1), were characterized. Based on phylogenetic analysis, these subtypes showed 100% identity with their reference counterparts. In conclusion, the zoonotic subtypes found in pet dogs in Shiraz can pose a drastic challenge to their pet owners and other domestic animals. Albeit the role of dogs as a potential natural reservoir of Blastocystis sp. stays unknown, it appears that these animals could represent possible reservoirs of zoonotic transmission of Blastocystis sp. Still, there is paucity of data on the epidemiology of Blastocystis and its subtypes in dogs in Iran.

Towards the First Multiepitope Vaccine Candidate against Neospora caninum in Mouse Model: Immunoinformatic Standpoint.

Neospora caninum is an economically significant parasite among livestock, particularly in dairy cattle herds, causing storm abortions. Vaccination seems necessary to limit the infection and its harsh consequences. This is the first steps towards developing a multiepitope vaccine candidate against N. caninum using in silico approaches. High-ranked mouse MHC-binding and shared linear B-cell epitopes from six proteins (SRS2, MIC3, MIC6, GRA1, IMP-1, and profilin) as well as IFN-γ-inducing epitopes (from SAG1) were predicted, screened, and connected together through appropriate linkers. Finally, RS-09 protein (TLR4 agonist) and histidine tag were added to N- and C-terminal of the vaccine sequence, yielding 486 residues in length. Physicochemical properties showed a stable (instability index: 27.23), highly soluble, antigenic (VaxiJen score: 0.9554), and nonallergenic candidate. Secondary structure of the multiepitope protein included 58.85% random coil, 20.99% extended strand, and 20.16% alpha helix. Also, the tertiary structure was predicted, and further analyses validated a stable interaction between the vaccine model and mouse TLR4 (binding score: -1261.6). Virtual simulation of immune profile demonstrated potently stimulated humoral (IgG+IgM) and cell-mediated (IFN-γ) responses upon multiepitope vaccine injection. Altogether, a potentially immunogenic vaccine candidate was developed using several N. caninum proteins, with the capability to elicit IFN-γ upsurge and other components of cellular immunity, and can be used in prophylactic purposes against neosporosis.

Current global status, subtype distribution and zoonotic significance of Blastocystis in dogs and cats: a systematic review and meta-analysis.

BACKGROUND: Blastocystis is a common intestinal protozoa found in animal and human fecal samples, with over 1 billion individuals infected worldwide. Since domestication, dogs and cats have had a close bond with humans. However, their close proximity poses a potential health risk since they may harbor several zoonotic agents. A global estimate of Blastocystis infection and subtype (ST) distribution in dogs and cats would therefore be of great health importance to humans. METHODS: We performed a comprehensive systematic search of four English-language databases (PubMed, Scopus, Google Scholar, Web of Science) for relevant articles up to 8 November 2021. The random-effects model was used to make pooled estimates with confidence intervals (CIs). RESULTS: In total, we identified 49 publications that met our inclusion criteria and subsequently analyzed the 65 datasets in these articles, of which 23 and 42 datasets were on cats and dogs, respectively. Among the 2934 cats included in the 23 datasets, which involved 16 countries, the prevalence rate of Blastocystis infection was 9.3% (95% CI 5.3-15.9%). The prevalence of Blastocystis infection was slightly lower [7%, 95% CI 4.7-10.4%) among the 7946 dogs included in the 42 datasets, involving 23 countries. The sensitivity analysis showed that no remarkable variation in the estimates upon the stepwise removal of each dataset. Higher ST diversity was found among the examined dogs (ST1-8, ST10, ST23, ST24) than among cats (ST1-4, ST10, ST14). Among dogs, ST3 was the most frequent ST (41.3%), followed by ST2 (39.3%), ST1 (30.9%), ST4 (13.4%), ST8 (12.7%), ST10 (11%) and ST5 (8.1%). Also among dogs, each of ST6, ST7, ST23 and ST24 was observed in only one study. Of the ST found in the cats examined, ST4 (29.5%), followed by ST10 (22.5%), ST1 (19.8%) and ST3 (17.6%) were the most common. A single study also reported the presence of both ST2 and ST14 in cats. With respect to zoonotic Blastocystis STs (ST1-ST9 and ST12), eight were reported from dogs (ST1-ST8) and four were isolated from cats (ST1-ST4), showing the implication of dog and cats in zoonotic transmission. CONCLUSIONS: Taken together, our results show that elucidation of the true epidemiology and ST distribution of Blastocystis in dogs and cats demands more comprehensive studies, particularly in the negelected regions of the world.

Anti-inflammatory, anti-oxidant and anti-apoptotic effects of olive leaf extract in cardiac tissue of diabetic rats.

OBJECTIVES: Inflammatory process and apoptosis are involved in the pathogenesis of cardiac injury and oxidative damage caused by diabetes mellitus. The cardioprotective effects of standardized aqueous ethanolic olive leaf extract (OLE), metformin (as a cardiovascular protective agent) and valsartan (as an angiotensin receptor blocker) in the streptozotocin-induced diabetic rats were evaluated. METHODS: Wistar rats divided into control, diabetic, OLE-treated (100, 200 and 400 mg/kg), metformin (300 mg/kg)-treated, valsartan (30 mg/kg)-treated and metformin/valsartan-treated diabetic groups. Biochemical parameters, including malondialdehyde (MDA) levels, superoxide dismutase (SOD) and catalase (CAT) activates, and the total contents of thiol were measured, and histopathological and gene expression studies were done on cardiac tissues. Fasting blood sugar (FBS) and cardiac injury markers were examined in serum. KEY FINDINGS: FBS; the serum levels of lactate dehydrogenase (LDH), creatine kinase-muscle/brain (CK-MB), aspartate aminotransferase (AST); and heart tissue MDA levels due to diabetes were significantly alleviated by OLE treatment (effect size; ηp2 = 0.934, 0.888, 0.848, 0.888 and 0.879, respectively), and SOD and CAT activity and the thiol content in heart tissue were significantly increased (effect size; ηp2 = 0.770, 0.749 and 0.753, respectively). Interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α) and the number of infiltrating inflammatory cells were reduced in cardiac tissues of OLE-treated groups compared with the diabetic rats (effect size; ηp2 = 0.969 and 0.949, respectively). OLE up-regulated BCL2 gene expression and down-regulated BAX gene expression in cardiac tissue (effect size; ηp2= 0.490 and 0.522, respectively). CONCLUSION: OLE in a dose-dependent manner ameliorates cardiac damage in diabetic cardiomyopathy, perhaps through attenuating inflammation, oxidative stress and apoptosis.