Mannas, unique products of a dynamic insect-plant interaction: Biodiversity, conservation and ethnopharmacological considerations.

Biodiversity keeping increases the flexibility, interaction and adaptation of different ecosystems with the environment which benefits all organisms, including humans. This diversity can be maintained through different paths including co-evolution between insects and plants. One of these unique interactions leads to the production of "Mannas" in Iran, which have biological and ethno-medicinal importance. Considering the crises of biodiversity and the rapid extinction of species, in this research, we aimed to review the latest scientific findings about mannas and their biological, medicinal and bio perspective considerations. Until August 2023, all English publications in Web of Science, Science Direct, PubMed, Google Scholar and Scopus, as well as Persian databases such as Magiran, Iranmedex, Medlib, SID were surveyed using various search terms such as manna, angabin, sweet secretions and plant-insect interference. Articles that studied manna production from plants or provided a comprehensive description of host plants and manna producing insects were included in the study. In total, out of more than 180 reviewed articles, 113 articles met the inclusion criteria and 7 known mannas (Bidkhesht, Gaz-e-Alafi, Gaz-e-Khansar, Gaz-e-Shahdad, Shirkhesht, Shekartigal, and Taranjebin) have been explained here. This review deals with different aspects of special interactions between insects and plants that lead to the production of manna and presents different uses of manna from medicinal, ethnobotanical, health, conservation and bio perspective points of view. In addition, the changes in the population of manna-breeding insects and host plants are taken into consideration and influencing factors including loss of their growth conditions and climatic conditions, animal breeding in the region, inappropriate exploitation methods and host preference phenomenon which increase the risk of destruction of these natural products have been given. These mannas cannot be produced in any way, and maintaining their production conditions requires environmental care and providing necessary training.

Nature׳s gifts to medicine: The metabolic effects of extracts from cocoons of Larinus hedenborgi (Coleoptera: Curculionidae) and their host plant Echinops cephalotes (Asteraceae) in diabetic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Trehala manna (TM), the edible cocoons of several weevil species, e.g. Larinus hedenborgi Boheman, 1845 (Coleoptera: Curculionidae) and their host plant, i.e. Echinops cephalotes DC. (EC) (Asteraceae), are traditionally used to treat pain, inflammation, infectious diseases, as well as respiratory, renal, reproductive and metabolic disorders. AIM OF THE STUDY: This study investigated the metabolic effects of aqueous extracts from TM and EC on diabetic male Wistar albino rats. MATERIALS AND METHODS: Animals were orally gavaged with the extracts (75, 150, and 300 mg/kg), normal saline, and glibenclamide (Glbn), for 28 days. The serum levels of glucose, insulin, lipid profile, and hepatic enzymes, plus the body weight of rats were measured at the beginning and the end of study. The proximate composition of the extracts was determined, additionally. The antioxidant and cytotoxic potency of the extracts were evaluated by radical scavenging/ferric reducing and viability assays, respectively. RESULTS: Treatment of diabetic rats with the extracts significantly altered metabolic biomarkers compared with diabetic, control and Glbn-treated groups, but not in a dose-dependent manner. However, the antihyperglycemic effects of TM75/EC300, the antiobesity effects of EC150, and the hepatoprotective effects of TM150/EC150 were even stronger than those of Glbn. TM/EC-treated groups represented normal cell architecture in the pancreatic and renal tissues. Nutrient analysis displayed that TM is rich in sugar and magnesium, whereas EC is abundant in protein, sodium, potassium, and calcium. The extracts showed no antioxidant and cytotoxic effects, as compared to the control groups. CONCLUSIONS: The findings suggest that active ingredients in the extracts evaluated are responsible for the metabolic effects by lowering blood sugar and restoring the damaged islets of Langerhans. The close trophic relationship of the TM-producing beetle with the host thistle justifies the overlaps of the bioactivity of the TM and EC extracts.

Anti-Proliferative Activity of A Hydrophilic Extract of Manna from Fraxinus angustifolia Vahl through Mitochondrial Pathway-Mediated Apoptosis and Cell Cycle Arrest in Human Colon Cancer Cells.

Manna is produced from the spontaneous solidification of the sap of some Fraxinus species, and, owing its content in mannitol, is used in medicine as a mild laxative. Manna is also a rich source of characteristic bio-phenols with reducing, antioxidant and anti-inflammatory properties. This study assesses the activity of a hydrophilic extract of manna (HME) on cellular and molecular events in human colon-rectal cancer cells. HME showed a time- and concentration-dependent anti-proliferative activity, measured by MTT assay, in all the cell lines examined, namely Caco-2, HCT-116 and HT-29. The amounts of HME that caused 50% of cell death after a 24 h treatment were 8.51 ± 0.77, 10.73 ± 1.22 and 28.92 ± 1.99 mg manna equivalents/mL, respectively; no toxicity was observed in normally differentiated Caco-2 intestinal cells. Hydroxytyrosol, a component of HME known for its cytotoxic effects on colon cancer cells, was ineffective, at least at the concentration occurring in the extract. Through flow-cytometric techniques and Western blot analysis, we show that HME treatment causes apoptosis, assessed by phosphatidylserine exposure, as well as a loss of mitochondrial membrane potential, an intracellular formation of reactive oxygen species (ROS), increases in the levels of cleaved PARP-1, caspase 3 and Bax, and a decrease in Bcl-2 expression. Moreover, HME interferes with cell cycle progression, with a block at the G1/S transition. In conclusion, the phytocomplex extracted from manna exerts an anti-proliferative activity on human colon cancer cells through the activation of mitochondrial pathway-mediated apoptosis and cell cycle arrest. Our data may suggest that manna could have the potential to exert chemo-preventive effects for the intestine.

The efficacy of camel milk and Tarangabin (manna of Alhagi maurorum( combination therapy on glomerular filtration rate in patients with chronic kidney disease: A randomized controlled trial.

OBJECTIVE: This study was designed to investigate the effect of camel milk and Tarangabin (manna of Alhagi maurorum) combination therapy in addition to conventional treatments in patients with chronic kidney disease (CKD). MATERIALS AND METHODS: Forty-four patients of 15 to 70 years old, with CKD due to hypertension or diabetes, and estimated glomerular filtration rate (eGFR) of 15-60 ml/min per 1.73 m2, were enrolled in this trial. The patients were randomized to receive either 400 cc of camel milk with 10 cc of Tarangabin syrup orally in two divided daily doses for 3 months plus conventional therapy or conventional therapy alone. The conventional treatment included diabetes medications and angiotensin converting enzyme inhibitors or angiotensin receptor blockers. RESULTS: The baseline characteristics of patients were similar in the two groups. Serum levels of creatinine (p=0.01), blood levels of urea nitrogen (p=0.0001), triglyceride (p=0.02), and potassium (p=0.05), and diastolic blood pressure (p=0.0001) decreased, while eGFR (p=0.001) improved in intervention group significantly. CONCLUSION: It seems that the therapeutic protocol used in this study can improve renal function in patients with CKD through regulating glucose and anti-inflammatory, laxative, and immunostimulatory properties.

Astragalus fascicolifolius manna abortifacient risk and effects on sex hormones in BALB/c mice.

BACKGROUND: Astragalus fascicolifolius manna is used to treat different diseases. Because pregnant women tend to use Astragalus. fascicolifolius and Iranian traditional medicine emphasizes the abortifacient potential of this plant, this study aimed to investigate Astragalus fascicolifolius manna abortifacient property and effects on estrogen, progesterone, LH and FSH levels in BALB/c mice. METHOD: This experimental study was conducted with 70 female BALB/c mice assigned to seven groups: Nonpregnant, untreated; nonpregnant, Astragalus. fascicolifolius extract (400 mg/kg)-treated; pregnant, Astragalus. fascicolifolius extract (400, 800 and 1200 mg/kg)-treated; and two pregnant control groups. On 18 and 19 days of pregnancy, cesarean section performed on mice, resorbed embryos counted; then Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), estrogen and progesterone levels were measured by the ELISA. RESULTS: Astragalus. fascicolifolius extract caused a significant increase abortion in mice. The levels of progesterone, FSH and LH were significantly different among the groups such that mean progesterone level was lower and mean LH and FSH levels were higher in the Astragalus. fascicolifolius extract-treated groups than the pregnant, untreated group. CONCLUSION: This extract has abortifacient properties and this plant can be used cautiously in pregnancy. Decreasing progesterone, increasing FSH and LH feedback in response to decreased progesterone by this extract is one of the potential mechanisms involved in abortion.

Comparison of Purgative Manna Drop and Phototherapy with Phototherapy Treatment of Neonatal Jaundice: A Randomized Double-Blind Clinical Trial.

OBJECTIVES: Herbal remedies such as purgative manna are used to treat neonatal jaundice. In this study Bilineaster drop (purgative manna) and phototherapy, and phototherapy treatment alone were compared by assessing phototherapy duration and number of days in hospital. METHODS: There were 150 consecutive term neonates with jaundice, weighting from 2,500 g to 4,000 g enrolled in this randomized double blind clinical trial. The neonates were randomly assigned to intervention and control groups. The control patients received only phototherapy and the intervention group underwent phototherapy treatment and purgative manna drop (5 drops per kg of body weight, 3 times a day). Direct and total measurements of bilirubin concentration in the serum were measured and the reduction in concentration of bilirubin was calculated. RESULTS: There were 28% of patients whose hospital duration following phototherapy was 2 days, for Bilineaster and phototherapy treatment this was 49.3% of patients. At 48 hours and 72 hours the reduction in the concentration of total bilirubin in the serum was statistically significantly different across groups (p < 0.05) but at 24 hours and 96 hours there were no significant differences between groups (p > 0.05). The reduction in direct bilirubin concentration in the serum was significantly different between groups at 72 hours and 96 hours (p > 0.001). CONCLUSION: Purgative manna and phototherapy, can statistically significantly reduce total bilirubin concentration at 48 hours and 72 hours compared with phototherapy alone, and reduce the length of hospital stay for jaundiced neonates at 2 days compared with phototherapy treatment.

Preventive effect of purgative manna on neonatal jaundice: A double blind randomized controlled clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Cotoneaster nummularioides Pojark manna (Shir-e-Khesht) is popular in Persian medicine. Different effects of some Cotoneaster species manna include antibacterial, antioxidant, anticancer, and hepatoprotective effects, as well as bilirubin serum levels reduction. Cotoneaster species manna is used in many parts of Iran as a laxative and accelerates the passage of meconium. Neonatal jaundice has relatively costly and sometimes invasive therapeutic interventions, which its prevention from becoming severe cases can be a priority in neonatal medicine. AIM OF THE STUDY: The aim of this study was to evaluate the effectiveness of an herbal product (Purgative Manna, native to Iran and Asian countries) in preventing severe cases of jaundice and reducing total bilirubin levels in neonates. MATERIALS AND METHODS: This randomized double-blind controlled clinical trial included full-term babies. Four hundred and forty-five (445) eligible neonates were assigned to two groups using the block balanced randomization method; 222 neonates received the Purgative Manna product as drops, and 223 neonates received placebo drops. The neonates received a dose of 5 drops per kilogram of neonatal weight (divided into three doses per day) for three days. The treatment period was three days, and a 24-h (three times) examination was performed to measure the initial outcome of the trial (i.e., the total serum bilirubin level). The secondary outcomes of this trial were the need for hospitalization due to jaundice and/or phototherapy from 4 to 14 days after birth, the frequency of defecation within 24 h, and the triple complications of diarrhoea, dehydration symptoms, and abdominal colic. RESULTS: In this study, 220 neonates in the Purgative Manna product group and 222 neonates in the placebo group completed their interventions within the predicted period of the study. At the end of study, the total bilirubin level in the Purgative Manna treated group was significantly lower than that of the placebo group. The difference between the mean total bilirubin levels of the two groups was approximately 2.1 mg/dl on the third day after treatment, with an effect size of 0.79 (95% CI: 0.06-0.98). The relative risk for reducing the need for hospitalization or phototherapy in the group treated with Purgative Manna drops was 0.26, compared with the placebo group. The risk of occurrence of severe jaundice or phototherapy in the Purgative Manna group was 75% lower than that of the placebo group. The median frequency of defecation in the intervention group at three time intervals in the first, second and third days after treatment was 1-2 times more than that of the comparison group (p < 0.001). CONCLUSIONS: Meanwhile, considering the fact that one in every eight neonates who used the product avoided having a severe and high-risk case of jaundice or the need for phototherapy intervention (even through an exchange transfusion), the use of Purgative Manna drops can be recommended; however, further study is necessary.

Comparison of Purgative Manna Drop and Phototherapy with Phototherapy Treatment of Neonatal Jaundice: A Randomized Double-Blind Clinical Trial

OBJECTIVES: Herbal remedies such as purgative manna are used to treat neonatal jaundice. In this study Bilineaster drop (purgative manna) and phototherapy, and phototherapy treatment alone were compared by assessing phototherapy duration and number of days in hospital. METHODS: There were 150 consecutive term neonates with jaundice, weighting from 2,500 g to 4,000 g enrolled in this randomized double blind clinical trial. The neonates were randomly assigned to intervention and control groups. The control patients received only phototherapy and the intervention group underwent phototherapy treatment and purgative manna drop (5 drops per kg of body weight, 3 times a day). Direct and total measurements of bilirubin concentration in the serum were measured and the reduction in concentration of bilirubin was calculated. RESULTS: There were 28% of patients whose hospital duration following phototherapy was 2 days, for Bilineaster and phototherapy treatment this was 49.3% of patients. At 48 hours and 72 hours the reduction in the concentration of total bilirubin in the serum was statistically significantly different across groups (p 0.05). The reduction in direct bilirubin concentration in the serum was significantly different between groups at 72 hours and 96 hours (p > 0.001). CONCLUSION: Purgative manna and phototherapy, can statistically significantly reduce total bilirubin concentration at 48 hours and 72 hours compared with phototherapy alone, and reduce the length of hospital stay for jaundiced neonates at 2 days compared with phototherapy treatment.

Immunomodulatory Properties of Taranjebin (Camel's Thorn) Manna and Its Isolated Carbohydrate Macromolecules.

Taranjebin manna is a substance produced by Poophilus nebulosus Leth. (Aphrophoridae) larva that feed from host plant Alhagi maurorum (Leguminosae). In Persian ethnomedicine, it is used as an antipyretic, antiviral, antimicrobial, demulcent, and adaptogen. But it is contraindicated in acute fever and some infections. This controversy might be due to its immunomodulatory properties. This study evaluated immunomodulatory properties of Taranjebin and its macromolecules. Taranjebin solution was prepared as described in traditional literature. After dialysis and precipitation, the macromolecules were isolated on DEAE Sephadex A-25. The cytotoxic/proliferative properties of Taranjebin and its isolated macromolecules on human Jurkat E6.1 cells were investigated (15.62-1000 µg/mL) using WST-1 reagent. Three of 4 isolated acidic polysaccharides inhibited the proliferation of Jurkat cells in a dose-dependent manner at concentrations higher than 31.25 µg/mL (IC50 range of 44.81-147.97 µg/mL). The crude aqueous Taranjebin solution had proliferative effects. These results indicate the immunomodulatory properties of Taranjebin.

Immunomodulatory properties of Trehala manna decoction and its isolated carbohydrate macromolecules.

ETHNOPHARMACOLOGICAL RELEVANCE: Trehala manna (Shekar tighal) is a cocoon-shaped manna produced by larval activity of Curculionidae family on some Echinops species. In Persian traditional medicine, it is used not only for the treatment of fever and constipation but also for sore throat, influenza and some other viral and bacterial infections which seems to be related to its boosting effects on immune system. The aim of this study was to evaluate immunomodulatory properties of Trehala manna and its isolated polysaccharides. MATERIALS AND METHODS: Several carbohydrate macromolecules were isolated from Trehala manna decoction using solvent-solvent extraction, ion exchange and size-exclusion chromatography. The macromolecules were characterized by gas chromatography-mass spectrometry and spectrophotometric methods. The immunomodulatory effects of Trehala manna decoction and its isolated polysaccharides were evaluated on Jurkat cells at different concentrations using WST-1 reagent. RESULTS: Seven water-soluble polysaccharides were isolated with the structures of rhamnogalacturonan, glucogalacturonan, manno-arabinogalacturonan or homogalacturonan and the molecular weights (MW) of ≤1.67-667 ≤ kDa. Large polysaccharides with MW ≥667 kDa and pectin inhibited the proliferation of Jurkat cells in a dose-dependent manner with the IC50 range of 98.06-429.53 µg/mL. The IC50 for low MW polysaccharides (around 1 kDa) did not show considerable cytotoxicity (IC50>1000 µg/mL), in contrast, they exhibited a dose-dependent enhancement of cell proliferation at concentration range of 15.62-62.50 µg/mL. CONCLUSION: Trehala aqueous decoction and its isolated polysaccharides showed some immunomodulatory effects on Jurkat cell line in a dose-dependent manner and the molecular weight of these polysaccharides also revealed a great impact on their cytotoxic/proliferative properties.

Looking beyond sugars: phytochemical profiling and standardization of manna exudates from Sicilian Fraxinus excelsior L.

Different grades of genuine and counterfeit Fraxinus excelsior exudates, marketed as natural sweeteners or mild laxatives, were evaluated for their proximate composition and for saccharidic, organic acids, lipidic and phenolic profile by means of GC-MS and (1)H NMR. Genuine samples contained mannitol (39-48 g/100 g, according to the grade), fructose (9-16 g/100 g), glucose (2-3.7 g/100 g), sorbitol (0,5-0,6 g/100 g), galactose (0.02-0.74 g/100 g), oligosaccharides as mannotriose (13-22 g/100 g) and stachyose (1-11 g/100 g), and traces of myo-inositol, mannose, sucrose. On the contrary, counterfeit samples contained mostly mannitol and sorbitol, with traces of fructose, glucose and mannose. Differences in ash, total polyphenolic content and fatty acid composition allowed a quick identification of counterfeit products, confirmed by a distinct mono-, oligosaccharidic and phenolic pattern. Elenolic acid (63-1628 mg/kg), tyrosol (15-774 mg/kg), homovanillic acid (2,39-52.8 mg/Kg), dopaol (0.8-63 mg/kg), pinoresinol (4.2-18.5 mg/kg) and fraxetin (0.25-11.64 mg/kg), albeit showing a wide concentration range, were the most abundant substances detected in the phenolic fraction of Fraxinus manna, while esculetin, p-hydroxybenzoic acid, 4-hydroxyphenacetic acid, 3,4 hydroxybenzoic acid, hydroxy-pinoresinol, medioresinol and siringaresinol were present in low amounts. The polyphenolic profile may be used as a marker for authentication and should be considered in the evaluation of nutritional and health properties ascribed to Fraxinus manna.