Use of Dicranum polysetum extract against Paenibacillus larvae causing American Foulbrood under in vivo and in vitro conditions.

Recent research shows that Dicranum species can be used to ameliorate the negative effects of honeybee bacterial diseases and that novel compounds isolated from these species may have the potential to treat bacterial diseases. This study aimed to investigate the efficacy of Dicranum polysetum Sw. against American Foulbrood using toxicity and larval model. The effectiveness of D. polysetum Sw. ethanol extract in combating AFB was investigated in vitro and in vivo. This study is important in finding an alternative treatment or prophylactic method to prevent American Foulbrood disease in honey bee colonies. Spore and vegetative forms of Paenibacillus larvae PB31B with ethanol extract of D. polysetum were tested on 2040 honey bee larvae under controlled conditions. Total phenolic and flavonoid contents of D. polysetum ethanol extracts were determined as 80.72 mg/GAE(Gallic acid equivalent) and 303.20 µg/mL, respectively. DPPH(2,2-diphenyl-1-picrylhydrazyl) radical scavenging percent inhibition value was calculated as 4.32%. In Spodoptera frugiperda (Sf9) and Lymantria dispar (LD652) cell lines, the cytotoxic activities of D. polysetum extract were below 20% at 50 µg/mL. The extract was shown to considerably decrease infection in the larvae, and the infection was clinically halted when the extract was administered during the first 24 h after spore contamination. The fact that the extract contains potent antimicrobial/antioxidant activity does not reduce larval viability and live weight, and does not interact with royal jelly is a promising development, particularly regarding its use to treat early-stage AFB infection.

Biological Activity and Phytochemical Analysis of Dicranum scoparium against the Bacterial Disease for Honey Bee.

Bacterial diseases, such as American Foulbrood (AFB) and European Foulbrood (EFB), are known to have catastrophic effects on honey bees (if left to spread, can wipe out entire colonies), leading to severe financial losses in the beekeeping industry. The aim of this study was to evaluate the pharmacological properties of methanol extract and its fractions (ethyl acetate, hexane, water) derived from Dicranum scoparium Hedw., which could be utilized as a potential drug to prevent the bacterial diseases (AFB and EFB) affecting the honey bees. For this purpose, crude methanol extract and ethyl acetate/hexane/water fractions were prepared from the aerial part of D. scoparium, collected from Trabzon province. Bio-guided fractionation of the extract and its fractions led to the first-time isolation of five compounds. The structure of all compounds was elucidated by nuclear magnetic resonance (NMR) spectroscopy, ultraviolet (UV) spectral analysis, Fourier-transform infrared spectroscopy (FT-IR), liquid chromatography quadrupole time-of-flight mass spectroscopy (LC-QToF-MS), and by comparison of their NMR data with that of literature. The analysis of these compounds revealed significant antibacterial and sporicidal activities against bacteria causing larval diseases in honey bees. The antibacterial activity of these compounds ranged from 0.6 to 60 µg/mL against AFB and EFB causing bacteria. Therefore, the natural raw extract and fractions of D. scoparium could be used as potential therapeutic agents against bacterial agents affecting honey bees.

Phytochemicals, antimicrobial, and sporicidal activities of moss, Dicranum polysetum Sw., against certain honey bee bacterial pathogens.

Beekeeping is an important agricultural and commercial activity globally practiced. Honey bee is attacked by certain infectious pathogens. Most important brood diseases are bacterial including American Foulbrood (AFB), caused by Paenibacillus larvae (P. larvae), and European Foulbrood (EFB) by Melissococcus plutonius (M. plutonius) in addition of secondary invaders, e.g. Paenibacillus alvei (P. alvei) and Paenibacillus dendritiformis (P. dendritiformis). These bacteria cause the death of larvae in honey bee colonies. In this work, antibacterial activities of extracts, fractions, and isolated certain compounds (nominated 1-3) all originated from moss, Dicranum polysetum Sw. ( D. polysetum), were tested against some honey bee bacterial pathogens. Minimum inhibitory concentration, minimum bactericidal concentration, and sporicidal values ​​of methanol extract, ethyl acetate, and n-hexane fractions ranged between 10.4 and 18.98, 83.4-303.75 & 5.86-18.98 µg/mL against P. larvae, respectively. Antimicrobial activities of the ethyl acetate sub-fractions (fraction) and the isolated compounds (1-3) were tested against AFB- and EFB-causing bacteria. Bio-guided chromatographic separation of ethyl acetate fraction, a crude methanolic extract obtained from aerial parts of D. polysetum resulted in three natural compounds: a novel one, i.e. glycer-2-yl hexadeca-4-yne-7Z,10Z,13Z-trienoate (1, dicrapolysetoate; given as trivial name), in addition to two known triterpenoids poriferasterol (2), and γ-taraxasterol (3). Minimum inhibitory concentration ranges were 1.4-60.75, 8.12-65.0, 2.09-33.44 & 1.8-28.75 µg/mL for sub-fractions, compounds 1, 2, and 3, respectively.

The effects of aging on the central nervous system steroid profiles and myelin basic protein in rats.

The aim of this study was to investigate the effects of aging on the central nervous system steroid and myelin basic protein (MBP) profiles. Forty-seven male Sprague-Dawley rats (newborn, 1, 6, 12 and 24-monthsold) were studied. Tissues were obtained from the cerebellum and parietal, frontal, temporal cortex of the central nervous system of the rats for steroid extraction. The estradiol, progesteron, testosterone and dehydroepiandrosterone (DHEA) levels were measured by Enzyme Linked Immunosorbent Assay (ELISA). The average levels of estradiol (pg/g), progesteron (ng/g), DHEA (ng/g) and testosterone (ng/g) in the brain tissues were respectively 24.29, 4.59, 0.27, 0.92 in the newborn-rats; 4.18 ± 1.10, 1.54 ± 0.30, 0.28 ± 0.01, 0.57 ± 0.10 in the 1 month-old-rats; 11.02 ± 1.10, 2.96 ± 0.30, 0.27 ± 0.01, 0.61 ± 0.10 in the 6 month-old-rats; 15.80 ± 1.10, 4.80 ± 0.30, 0.28 ± 0.10, 0.67 ± 0.10 in the 12 monthold- rats; 20.07 ± 1.10, 4.12 ± 0.30, 0.28 ± 0.01, 0.55±0.10 in the 24 month-old-rats. The myelin basic protein levels were determined by immunohistochemical staining and an elevation was observed in conjunction with the aging process. The results of the study indicate that the alterations in MBP, DHEA, progesterone, testosterone and estrodiol concentrations in the central nervous system of the rats during aging can be considered fundamental for future animal and human studies.

Effects of three types of honey on cutaneous wound healing.

UNLABELLED:  Aim. To evaluate the effects of three types of honey (chestnut, blossom, and rhododendron) on the healing of full-thickness wounds. METHODS: Twenty-four (24) New Zealand White female rabbits were used. Four 1.5 cm x 1.5 cm full-thickness skin wounds were created on the back of each animal and treated with pure honey or sterile saline, respectively. Wounds were assessed by wound measurements and collection of samples at 7, 14, and 21 days post wounding to evaluate the healing process. Variables of interest were hydroxyproline concentration and gross and microscopic morphological characteristics reflective of wound healing. Wounds of the honey-treated groups healed much faster than the control group. RESULTS: On day 7, the formation of granulation tissue, epithelization, angiogenesis, and fibroplasia levels increased in the honey-treated groups (P <0.05). A statistical difference between the honeys was not detected. CONCLUSION: The present results suggest that honey accelerates the inflammatory reaction and initiates healing early on in the treatment process. .

Biochemical evaluation of the therapeutic effectiveness of honey in oral mucosal ulcers.

The aim of this study was to describe and quantify the therapeutic value of honey in oral mucosal ulcers healing in comparison with Glyceroloxytriester (TGO). We also aimed to biochemically evaluate the healing effects of honey which had been collected from the Blacksea region flora on mucosal ulcers resulting in material loss.Thirty wistar rats (240+/-30 g) were enrolled in this study. Excisional wounds were performed in all rats for animal oral mucosal ulcer model. They were randomly allocated to three groups: group 1 was treated with Apitherapeutic agent or honey (0,1 ml, 2x1), group 2 was treated with TGO (0,1ml, 2x1) locally, Group 3 served as the control group.Following the surgical procedure on day 7, biopsy specimens were taken from right buccal mucosa and on day 14 biopsy specimens were taken from left bucal mucosa in all rats. Afterwards, hydroxy pyroline levels were measured. Data were analyzed statistically.There was no statistically significant difference between Group 1 and 2, and also between Group 2 and 3, but there was statistically significant difference between Group 1 and 3 on day 7. There was no statistically significant difference between Group 1, 2 and 3 on day 14.

Determination of Quality Criteria that Allow Differentiation Between Honey Adulterated with Sugar and Pure Honey.

This study used various parameters of honey to develop a potentially more robust approach to the detection of adulterated honey. For this purpose, 25 multifloral, natural honey samples and 20 samples of adulterated honey produced by bees that had been fed supplementary sucrose syrup were analysed. The mean total phenolic content of the natural honeys was considerably higher than in the adulterated honeys at 157 ± 13 and 35.2 ± 7.3 mg GAE/100 g, respectively. Similarly, considerable variation was determined between natural and adulterated honeys in terms of total flavonoids (3.3 ± 0.3 and 2.1 ± 0.4 mg QE/100 g, respectively), antiradical activity (87.9 ± 12 and 163 ± 11 mg/mL, respectively) and proline content (202 ± 26 and 71.1 ± 21.6 mg/kg, respectively.) The potassium, phosphorus, calcium and magnesium contents of natural honeys were also higher than in adulterated honeys (P < 0.01). In conclusion, the determination of the proline level, phenolic content, antioxidant activity and mineral profile may collectively provide a more holistic method approach to the differentiation of natural and adulterated honey, and also for comparing their food values.

Effects of trichophytosis on serum zinc levels in calves.

The purpose of this study was to determine the zinc levels in calves with trichophytosis and to research the importance of zinc for fungi. The sera of 20 calves with trichophytosis and 10 healthy calves were used in this study. Zinc levels of the sera were measured by the atomic absorption spectrophotometer method. Serum zinc levels of diseased and healthy animals were found to be 42.0+/-16.6 microg/dL and 75.8+/-5.9 microg/dL, respectively. Serum zinc levels of diseased calves were lower than healthy ones and this difference were found to be important statistically (p<0.001), whereas there is no statistical difference on the levels of lymphocyte, monocyte, granulocyte, erythrocyte, hemoglobin, hematocrit, and mean corpuscular volume between groups. These parameters were not influenced by low zinc levels.

Effects of electromagnetic field (1.8/0.9 GHz) exposure on growth plate in growing rats.

The purpose of this study was to determine the effects of whole-body electromagnetic field (EMF) exposure on growth plates in growing male rats. Two groups of rats were exposed to either 900 MHz EMF or 1800 MHz EMF 2 h/day for 90 days. Sham control rats were kept under similar conditions without exposure to the EMF. The rats in the EMF group experienced a more rapid weight gain and increase in length (p < 0.05). Calcium, growth hormone, estradiol and testosterone levels in the EMF groups were higher (p < 0.05). The Safranin O staining density of femoral growth plate was lowest in the reserve zone of rats exposed to 1800 MHz and was increased in the proliferative zone of the control group (p < 0.05). The trabecular zone was thinnest among all zones and the reserve and proliferative zones were thicker (p < 0.05) than other zones in 1800 MHz group.In conclusion, 1800 MHz and 900 MHz EMF may cause prolong the growth phase in growing rats.

Therapeutic role of curcumin in oxidative DNA damage caused by formaldehyde.

PURPOSE: Formaldehyde is a common environmental contaminant that causes oxidative DNA damage in cells by increasing the production of reactive oxygen species. The aim of this study was to investigate the amount of 8-hydroxy-deoxyguanosine (8-OhdG), tumor protein 53(TP53), beta-amyloid[Aß(1-42), Aß (1-40)], total antioxidant capacity (TAC) and malondialdehyde (MDA) and the therapeutic role of curcumin in rat cells with oxidative DNA damage caused by formaldehyde. METHOD: The control group was given physiological saline for 15 days (i.p.) and the second group was given 37% formaldehyde (i.p.) at a dose of 9 mg/kg group every other day. The third group was given 9 mg/kg formaldehyde (i.p.) every other day and treated therapeutically with 100 mg/kg curcumin every day by gavage. At the end of the trial period, urine, blood, and brain tissue was collected from the rats. RESULTS: The levels of MDA in sera were increased and the TAC, TP53, and Aß (1-40) levels were reduced in the formaldehyde-treated group with respect to the control group (p<0.005). After treatment with curcumin, the levels of sera MDA were significantly reduced, the TAC, TP53, and Aß (1-40) levels were significantly increased (P < 0.05). The levels of whole brain Aß (1-42) and 8-OhdG were increased in the formaldehyde-treated group and reduced after treatment with curcumin (P < 0.05). Urinary 8-OhdG excretion increased in the formaldehyde-treated group (P < 0.05) and decreased after treatment with curcumin (P > 0.05). CONCLUSIONS: In conclusion, the oxidative stress caused by formaldehyde exposure was reduced with the application of curcumin.

Effects of exposure to electromagnetic field (1.8/0.9 GHz) on testicular function and structure in growing rats.

The aim of our study was to evaluate the possible effects of whole-body electromagnetic field (EMF) exposure on reproduction in growing male rats. Male albino Wistar rats (2 days old) were exposed to EMF 1800 and 900 MHz for 2 h continuously per day for 90 days. Sham control was kept under similar conditions except that the field was not applied for the same period. After blood samples were collected, the animals were sacrificed 24 h after the last exposure and the tissues of interest were harvested. The mean plasma total testosterone showed similarity among the two study groups and was significantly higher than the sham control rats. The percentage of epididymal sperm motility was significantly higher in the 1800 MHz group (P<0.05). The morphologically normal spermatozoa rates were higher and the tail abnormality and total percentage abnormalities were lower in the 900 MHz group (P<0.05). Histopathologic parameters in the 1800 MHz group were significantly higher (P<0.05). In conclusion, the present study indicated that exposure to electromagnetic wave caused an increase in testosterone level, epididymal sperm motility (forward), and normal sperm morphology of rats. As a consequences, 1800 and 900 MHz EMF could be considered to be a cause of precocious puberty in growing rats.

Evaluation of bee venom and hyaluronic acid in the intra-articular treatment of osteoarthritis in an experimental rabbit model.

The aim of this study was to investigate bee venom and hyaluronic acid in the intra-articular treatment of osteoarthritis in an experimental rabbit model. Forty-five rabbits were used and they were randomly divided into three groups (BVI, BVII, and HA) and each group was divided to two subgroups to evaluate the radiologic, magnetic resonance imaging, histopathologic, and biochemical evaluation in post treatment second week (a) and twelfth week (b). Radiologically, a significant difference was observed in the HA group (P<0.05). The MRI evaluation of at any time in group BVI(b) was found to be different. No significant differences were seen between the groups, biochemically. Histopathologically, cellularity, and orthochromasia was evident with Safranin-O in the BVI(b) and BVII(a); adhesions were seen in the BVII(a) group and clustering of chondrocyte in the HA(b) group were found to be different. Consequently, intra-articular application of HA and BV for experimental model of osteoarthritis has no significant influence upon recovery after therapy.

Effect of fluoride exposure on serum glycoprotein pattern and sialic acid level in rabbits.

This study describes the effects of fluoride exposure on the protein profile, glycoprotein pattern, and total sialic acid concentration of serum in rabbits. For this aim; 20 healthy New Zealand rabbits were used. The rabbits were divided into two equal groups each with ten animals according to their weighing: control group and experimental group. The rabbits in control group were given drinking tap water containing 0.29 mg/l sodium fluoride and experimental group received the same tap water to which was added 40 mg/l sodium fluoride for 70 days. Blood samples were taken from each rabbit on day 70. Serum fluoride concentrations were measured by a fluoride-specific ion electrode in serum. The fluoride levels in the serum were found as 18.4 (+/-1.58) microg/L in control and 301.3 (+/-52.18) microg/L in fluoride exposed rabbits. The sialic acid levels were found as 69.2 (+/-0.32) mg/dL in control and 43.4 (+/-0.13) mg/dL in fluoride exposed group. The electrophoretic patterns of serum proteins, glycoproteins, and total sialic acid concentration were determined. Fifteen different protein fractions with molecular weights ranging from 22 to 249 kDa were displayed in the serum protein electrophoretic gel of both groups. The raw concentrations of the protein fractions decreased in fluoride exposed rabbits as compared with the control rabbits. The serum glycoprotein pattern revealed seven major protein bands from 47 to 167 kDa in experimental and control groups. The slight decrease of raw concentration of the protein bands in glycoprotein pattern of serum was observed in fluoride toxication comparing to control. The results suggest that serum TSA determination and serum protein electrophoresis can be used to evaluate prognosis of fluoride exposure as a supplementary laboratory test in combination with clinical and other laboratory findings of fluorosis.

Serum protein alterations in dogs naturally infected with Toxoplasma gondii.

We conducted this study to describe the serum electrophoretic pattern in dogs associated with the infection of Toxoplasma gondii (T. gondii). The serum protein pattern of 25 dogs with confirmed T. gondii infection and 15 clinically healthy dogs were evaluated using native polyacrylamide gel electrophoresis. Albumin, alpha-1 globulin, alpha-2 globulin, beta globulin, and gamma globulin bands were seen from the serum electrophoresis of infected and healthy dogs. Compared to the control group, significant decreases in the mean percentages of albumin (from 46.1+/-7.2 to 40.8+/-4.5%, P<0.05), alpha-1 globulin (from 3.9+/-0.4 to 0.8+/-0.2%, P<0.001), alpha-2 globulin (from 9.0+/-0.4 to 8.3+/-0.8%, P<0.01), and beta globulin (from 18.4+/-1.2 to 12.1+/-0.6%, P<0.001) in the infected group were determined. In contrast, gamma globulin fraction was significantly higher in infected dogs (38.1+/-4.6%) than in control dogs (22.7+/-7.2%; P<0.001). Moreover, significant correlations were determined between the percentages of the albumin and gamma globulin fractions and liver enzyme tests including aspartate aminotransferase and alanine aminotransferase in infected dogs; however, no correlation was observed for the other protein fractions. In conclusion, marked alterations in serum protein pattern associated with strong modifications of serum protein concentrations are in accordance with the hepatic injury as affirmed by liver enzyme tests that were demonstrated in the canine toxoplasmosis. These findings showed that serum protein electrophoresis can be used in the diagnosis and prognosis of canine toxoplasmosis as a supplementary analysis in combination with serological, clinical, and laboratory findings of this disease.

Biochemical and histopathological findings of N-butyl-2-cyanoacrylate in oral surgery: an experimental study.

OBJECTIVES: The increasing use of cyanoacrylates in dentistry, particularly as an adhesive and sealing glue, has raised concerns regarding its potential toxicity in humans. Several different forms of these compounds including methyl- (MCA), ethyl- (ECA), isobutyl-, isohexyl-, and octyl CA have been developed to eliminate tissue toxicity. N-butyl-2-cyanoacrylate is becoming an increasingly popular method for wound closure under low tension. Despite their increasing use, pharmacologic effects of these substances on liver and kidney functions are not widely known. The objective of the present study was to investigate possible immediate and long-term systemic effects of N-butyl-2-cyanoacrylate in oral surgery. STUDY DESIGN: Ten male Wistar rats weighing 220 to 270 g were used in the study. Straight incisions were made to the buccal mucosa of the animals. N-butyl-2-cyanoacrylate adhesive (Indermil) was applied and wounds were closed primarily. Blood specimens were taken periodically from the vena cava of the animals before the surgical procedure and 2, 14, 21, and 65 days after the surgical procedure. The blood specimens of those taken before the application of the adhesive were defined as the control group; blood specimens that were taken 2, 14, 21, and 65 days from the application were defined as study group. The stored plasma samples were analyzed for blood urea nitrogen (BUN), creatinine (CRE), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBI), total protein (TP), albumin (ALB), and amylase (AML). In addition to biochemical parameters, histopathological examination was performed. Blood parameter values of the control and study groups were statistically compared with the Duncan test (P < .05). RESULTS: There were no significant differences in the values of BUN, CRE, ALT, AST, TBI, TP, ALB, and AML between the control and at 2, 14, 21, and 65 days. CONCLUSION: The present study shows that N-butyl-2-cyanoacrylate is a suitable adhesive applicable in oral surgery.