Anti-Inflammatory, Wound Healing, and Anti-Diabetic Effects of Pure Active Compounds Present in the Ryudai Gold Variety of Curcuma longa.

Turmeric (Curcuma longa) contains curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC). Nevertheless, curcumin is the most researched active ingredient for its numerous pharmacological effects. We investigated the impact of these curcuminoids found in Ryudai gold, an approved cultivar of Curcuma longa, on wound healing, inflammation, and diabetes. Sub-planter injections of carrageenan induced acute paw inflammation in rats. The wound-healing ability of 1% curcuminoids was examined by making a 6 mm round wound on the shaved dorsum of the mice with a biopsy punch. A single intraperitoneal injection of streptozotocin (50 mg/kg) was used to induce diabetes in mice. Curcuminoids at a dose rate of 100 mg/kg body weight were used with feed and as a gastric gavage to treat diabetes and inflammation in experimental animals. Paw thickness was measured at 1, 3, and 6 h following carrageenan injection. After three hours, mean paw volume was 58% in carrageenan-injected mice, which was 35%, 37%, and 31% in the curcumin, DMC, and BDMC groups, respectively. Histopathology of the paw tissue demonstrated severe infiltration of inflammatory cells and thickening of the dermis, which were remarkably improved by the curcuminoids. The wound-healing abilities were significantly higher in the curcumin- (95.0%), DMC- (93.17%), and BDMC-treated (89.0%) groups, in comparison to that of the control (65.09%) group at day nine. There were no significant differences in wound-healing activity among the groups treated with 1% curcuminoids throughout the study. Streptozotocin-induced diabetes was characterized by an increased blood glucose (552.2 mg/dL) and decreased body weight (31.2 g), compared to that of the control rats (145.6 mg/dL and 46.8 g blood glucose and body weight, respectively). It also caused an increase in serum alanine aminotransferase (ALT; 44.2 U/L) and aspartate aminotransferase (AST; 55.8 U/L) compared to that of the control group (18.6 U/L and 20.1 U/L, respectively). Histopathological examination of the liver showed that diabetes caused hepatic cellular necrosis, congestion of the central vein, and parenchymatous degeneration. However, all three curcuminoids significantly decreased blood glucose levels, ALT, and AST and improved the histopathological score of the liver. These results evidenced that not only curcumin but also DMC and BDMC have potent anti-inflammatory, wound healing, and anti-diabetic efficacy, and the Ryudai gold variety of turmeric could be used as a functional food supplement.

Effects of Roasting Conditions on Antibacterial Properties of Vietnamese Turmeric (Curcuma longa) Rhizomes.

Processing with heat treatment has been reported to alter several therapeutic effects of turmeric. In Vietnamese traditional medicine, turmeric has been long used for bacterial infections, and roasting techniques are sometimes applied with this material. However, there have been no studies investigating the effects of these thermal processes on the plant's antibacterial properties. Our study was therefore performed to examine the changes that roasting produced on this material. Slices of dried turmeric were further subjected to light-roasting (80 °C in 20 min) or dark-roasting (160 °C in 20 min) processes. Broth dilution and agar-well diffusion methods were applied to examine and compare the effects of ethanol extracts obtained from non-roasted, light-roasted and dark-roasted samples, on a set of 6 gram-positive and gram-negative bacteria. In both investigations, dark-roasted turmeric was significantly less antibacterial than non-roasted and light-roasted materials, as evident by the higher values of minimum inhibitory concentrations and the smaller diameters of induced inhibitory zones. In addition, dark-roasting was also found to clearly reduce curcumin contents, total polyphenol values and antioxidant activities of the extracts. These results suggest that non-roasting or light-roasting might be more suitable for the processing of turmeric materials that are aimed to be applied for bacterial infections.

Evaluation of hematoprotective, hepatoprotective, and anti-inflammatory potentials of chia seed (Salvia hispanica L.) extract in rats.

This study was conducted to evaluate the effects of chia seed extract on CCl4-induced hepatotoxicity, hematological profile, and carrageenan-induced inflammation in rats. Water-ethanol-acetone extract of chia seeds at the doses of 200 and 400 mg/kg body weight/day were applied to evaluate the comparative protective roles. Hematological profile and serum biochemical parameters were measured to evaluate the hematoprotective, and hepatoprotective effects of chia seed extract. Paw thickness and motility level were assessed at 0, 1, 3, 5, and 7 h after sub-planter injection of carrageenan to evaluate the anti-inflammatory potential. Tissue histopathology was performed in both cases. Chia seed extract reduced the elevated level of serum AST and ALT significantly in a dose-dependent manner following intra-peritoneal injection of CCl4. Histopathological study of the liver tissue exhibited acute impairment of the hepatocytes and liver parenchyma following CCl4 exposure, which was markedly regenerated by the chia seed extract treatment. Protective effects of the extracts were also evidenced by the RBC count, Hb (%), PCV (%), ESR, and neutrophil count. Chia seed extract was found to inhibit the carrageenan-induced paw edema and increase motility level in a dose-oriented fashion. Histological examination of the paw tissue revealed severe inflammation characterized by massive infiltration of inflammatory cells in the carrageenan group, which was significantly reduced by chia seed extract treatment. The higher dose of chia seed extract showed significant increases in bodyweight gain and feed efficiency ratio but decrease in visceral fat deposition. These results suggest that chia seeds possess potentials for hematoprotective, hepatoprotective, and anti-inflammatory activities.

The reality of the use of growth hormones in fish (Rui (Labeo rohita), Catla (Catla catla), and Monosex Tilapia (Oreochromis niloticus) production.

The unobservable use of hormones in fish production is becoming an alarming issue worldwide. To reveal the fact in Bangladesh, 144 fish samples (rui (Labeo rohita), catla (Catla catla), and monosex tilapia (Oreochromis niloticus)) were collected from different fish farms and markets of Mymensingh district. The market samples had two sources (Mymensingh and Rajshahi district). The steroid hormonal (testosterone, estrogen, and progesterone) residue was analyzed by HPLC-UV detection. A standard questionnaire survey was conducted where most farmers (80%) denied using the hormone in fish production. Among the analyzed samples of all three fishes, hormonal residues were detected in approximately 98% of samples, and around 92% contained residues above the ADI. Among the contaminated samples, 70% of samples had a single residue and 30% had multiple residues. The testosterone and progesterone hormonal residue was detected in all three fishes in both farm and market samples and ranged (above ADI) from 2.1 to 16.96 µg/kg and 31.47-731.57 µg/kg (p < 0.05) respectively. The estrogen hormone residue was only detected in market samples (Rajshahi district) of rui and catla and no residue was detected in tilapia fish and the hormone level (above ADI) ranged from 8.23 to 40.13 µg/kg. This study revealed that the use of hormones varies on the attitude of farmers based on the local culture pattern as estrogen hormone residue was only detected in market samples. The consumption of contaminated fish at such concentrations may cause many health hazards in humans, especially in children. Thus, this study reveals a new alarming fact to focus on, and an effective monitoring system should be implemented as soon as possible for public health concerns.

Habu snakes (Protobothrops flavoviridis) show variation in thoracic aortic vasoreactivity between adjacent Japanese islands.

Habu snakes (Protobothrops flavoviridis) are pit vipers found in the geographically adjacent but ecologically divergent islands of Tokunoshima and Amami-Oshima in southwestern Japan. Abiotic factors can cause variation in animal populations between the two islands, and Habu snakes may show such intraspecific physiological variation. We therefore evaluated the vasoreactivity in aortas isolated from the Habu of both islands. Tokunoshima Habu showed significantly greater contractile responses to angiotensin (Ang) II, acetylcholine (ACh) and noradrenaline, and significantly higher affinities (pEC50) for Ang II and ACh, than Amami-Oshima Habu. ACh caused contractions in aortas from both populations, a finding previously unreported in snakes. Our findings indicate that vasoreactivity may differ between Tokunoshima and Amami-Oshima Habu.

Pesticides in vegetable production in Bangladesh: A systemic review of contamination levels and associated health risks in the last decade.

This paper reviewed the published data on the levels of different pesticide residues in vegetables (tomato, eggplant, beans, gourds, cauliflower, cabbage, cucumber, potato, carrot, onion, red chilli, red amaranth, lady's finger, spinach, coriander, and lettuce) from Bangladesh in the last decade. Vegetable production in Bangladesh has increased tremendously (37.63%) compared to the last decades, along with its pesticide use. The most observed pesticide groups used in vegetable production were organophosphorus, pyrethroids, carbamate, organochlorine, nereistoxin analogue group, and neonicotinoids. More specifically, chlorpyrifos, dimethoate, diazinon, and malathion were the most used pesticides. More than 29% of the vegetable samples (1577) were contaminated with pesticide residue; among the contaminated samples (458), most cases (73%) exceeded the maximum residue limits (MRLs). The pesticide-contaminated vegetables were cucumber (51%), tomato (41%), cauliflower (31%), miscellaneous vegetables (36%), eggplant (29%), beans (23%), cabbage (18%), and gourds (16%). Among the pesticide-contaminated samples, vegetables with above MRL were gourds (100%), beans (92), tomato (78%), eggplant (73%), miscellaneous vegetables (69%), cucumber (62%), cabbage (50%), cauliflower (50%) (p < 0.05). It was also observed that a single vegetable was often contaminated with multiple pesticides, and farmers did not follow a proper withdrawal period while using pesticides. Hazard quotation (HQ>1) was observed in adolescents and adults in tomato, eggplant, beans, cauliflower, cabbage, cucumber, lady's finger, lettuce, and coriander. There was no health risk observed (HQ<1) in gourds, potato, carrot, onion, red chilli, red amaranth, spinach, and okra. The highest acute and chronic HQ (aHQ, cHQ) was observed for cypermethrin (bean) in adolescents (aHQ=255, cHQ= 510) and adults (aHQ=131, cHQ=263). It was also observed that these pesticides harmed air, soil, water, and non-target organisms. Nevertheless, the review will help the government develop policies that reduce pesticide use and raise people's awareness of its harmful effects.

Involvement of beta3-adrenergic receptors in relaxation mediated by nitric oxide in chicken basilar artery.

The response of basilar arteries to noradrenaline varies among many animal species, but remains little studied in poultry. Accordingly, we aimed to characterize the adrenergic receptor (AR) subtypes that modulate vascular response in basilar arteries in the chicken, with isometric recording of arterial ring tension using an organ bath. We demonstrated the presence of both alpha and beta (α and ß) receptor subtypes through evaluating the response to noradrenaline, with and without a range of ß-AR and α-AR antagonists. The concentration-dependent relaxations then induced by a range of ß-AR agonists indicated a potency ranking of isoproterenol > noradrenaline > adrenaline > procaterol. We then investigated the effects of ß-AR antagonists that attenuate the effect of isoproterenol (propranolol for ß1,2,3-ARs, atenolol for ß1-ARs, butoxamine for ß2-ARs, and SR 59230A for ß3-ARs), with Schild regression analysis, ascertaining multiple ß-AR subtypes, with neither the ß1-AR nor the ß2-AR as the dominant subtype. SR 59230A was the only antagonist to yield a pA2 value (7.52) close to the reported equivalent for the relevant receptor subtype. Furthermore, treatment with SR 58611 (a ß3-AR agonist) induced relaxation, which was inhibited (P < 0.01) by L-NNA and SR 59230A. Additionally, treating basilar arterial strips (containing endothelium) with SR 58611 induced nitric oxide (NO) production, which was inhibited (P < 0.01) by L-NNA and SR 59230A. Based on this first characterization of AR subtypes in chicken basilar arteries (to our knowledge), we suggest that α- and ß-ARs are involved in contraction and relaxation, and that ß3-ARs, especially those on the endothelium, may play an important role in vasodilation via NO release.

Characterization of Vasoreactivity in a Semi-Arboreal Snake, the Tokara Habu (Protobothrops tokarensis).

Vasoreactivity is relatively well documented in terrestrial snakes but has previously been investigated in only one semi-arboreal snake species. Consequently, the extent to which vasoreactivity is common across snake taxa or varies by habitat is unclear. The Tokara habu (Protobothrops tokarensis) is a semi-arboreal snake endemic to only two small adjacent Japanese islands, and hence a useful species for further investigation of vasoreactivity. We evaluated responses to known vasoactive substances in thoracic aortas isolated from Tokara habu. Under resting tension, noradrenaline and angiotensin II induced concentration-dependent contraction, but acetylcholine, serotonin (5-hydroxytriptamine; 5-HT), and isoproterenol induced relaxation followed by contraction. Histamine and rattlesnake bradykinin had no effect. Experiments with receptor-specific antagonists suggest that M1 and M3 receptors are involved in the acetylcholine-induced response; 5-HT1, 5-HT2, and 5-HT7 receptors in the serotonin-induced response; and ß1 and ß2 adrenoceptors in isoproterenol-induced relaxation. This is the first report on such response patterns in snakes (including serotonin- and isoproterenol-induced relaxation). Nitric oxide may be involved in acetylcholine-induced relaxation but not in the responses to serotonin or isoproterenol. In contrast to the uniform vasoreactivity observed in terrestrial snakes, the vasoreactivity of semi-arboreal snakes may be governed by diverse regulatory mechanisms.

Reduced Nitric Oxide Synthase Involvement in Aigamo Duck Basilar Arterial Relaxation.

The basilar arterial endothelium mediates blood vessel relaxation partly through the release of nitric oxide (NO). Apoptosis of cerebrovascular endothelial cells is linked to a high mortality rate in chickens infected with the highly pathogenic avian influenza virus, but interestingly, ducks exhibit a greater resistance to this virus. In this study, we examined the responsiveness of duck basilar arteries (BAs) to various vasoactive substances, including 5-hydroxytryptamine (5-HT), histamine (His), angiotensin (Ang) II, noradrenaline (NA), acetylcholine (ACh), and avian bradykinin ornithokinin (OK), aiming to characterize the receptor subtypes involved and the role of endothelial NO in vitro. Our findings suggest that arterial contraction is mediated with 5-HT1 and H1 receptors, while relaxation is induced with ß3-adrenergic and M3 receptors. Additionally, OK elicited a biphasic response in duck BAs, and Ang II had no effect. Endothelial NO appears to be crucial in relaxation mediated with M3 and OK receptors but not ß3-adrenergic receptors in the duck BA. The reduced endothelial NO involvement in the receptor-mediated relaxation response in duck BAs represents a clear difference from the corresponding response reported in chicken BAs. This physiological difference may explain the differences in lethality between ducks and chickens when vascular endothelial cells are infected with the virus.

Effect of knowledge of informal poultry drug prescribers on their attitude and practice toward antimicrobial use, residues, and resistance in Bangladesh.

Background and Aim: Informal prescribers (IPs) significantly contribute to the development of antimicrobial resistance and in disseminating pathogens from poultry to humans and other animals through the food chain, posing a serious global health threat. Therefore, this study aimed to assess whether the knowledge of IPs has an impact on their attitude and practice toward antimicrobial use, antibiotic residues, and antimicrobial resistance. Materials and Methods: In this cross-sectional study, we conducted a pre-tested and questionnaire-based survey to investigate the knowledge, attitude, and practice of IPs in selected parts of the Mymensingh division, Bangladesh. Then, we used the linear regression model test with R-squared (R2) to measure the association between the study variables. Results: Our investigation revealed that 70% of the IPs knew about antibiotics and 75% had good knowledge about antibiotic resistance, whereas only 50% were aware of withdrawal periods. Informal prescribers also displayed good attitudes toward the use and sale of antibiotics with withdrawal periods and completion of medication (50%). Analysis of their practice on the sale and prescription of antibiotics showed that 70% and 30% of IPs use antibiotics against bacterial infections and other conditions, respectively. Most of them do not consult a veterinarian before selling or prescribing antibiotics, although 80% claim to do so. This is because 75% of IPs gave other options regarding their consultations. However, 95% of IPs uses antibiotics only for therapeutic purposes. Furthermore, only 10% sell antibiotics based on a veterinarian's recommendation. Approximately 45% of IPs use single antibiotics at a time, while the rest use multiple antibiotics, individually or combined. Approximately 15% use antibiotics monthly, while 85% use them whenever the need arises. The knowledge and attitude of IPs are significantly affected by their age (p ≤ 0.025). The district of domicile also impacted their knowledge. Surprisingly, IPs from Jamalpur had significantly better knowledge compared to those from Mymensingh and Sherpur (p ≤ 0.01). The attitude of IPs from Jamalpur and Netrokona also differed significantly (p ≤ 0.001) from that of Mymensingh and Sherpur. The knowledge of IPs influenced their attitude up to 80.5% (r2 = 0.628) and their practice up to 75.4% (r2 = 0.545). Conclusion: The knowledge of IPs greatly influenced their attitude and practice, while sociodemographics also influenced their knowledge and attitude toward antimicrobial use, antibiotic residues, and antimicrobial resistance.

Screening and quantification of antibiotic residues in poultry products and feed in selected areas of Bangladesh.

Background and Aim: Antibiotic residues in livestock farming have been identified as a potential cause of antimicrobial resistance in humans and animals. This study aimed to determine whether antibiotic residues were present in the chicken meat, eggs, feces, and feed collected from all four districts in the Mymensingh division of Bangladesh. Materials and Methods: To detect antibiotic residues in the collected samples, qualitative thin-layer chromatography (TLC) and quantitative high-performance liquid chromatography (HPLC) were used. A total of 230 samples were analyzed for antibiotic residues of commonly used 11 antibiotics. Out of these, 40 meat and 40 feces samples were collected from broilers and layers, 30 egg samples from ducks and layers, and 120 feed samples from broilers and layers from the study area. Thin-layer chromatography was used to screen the presence of antibiotic residues; TLC-positive samples were then subjected to further HPLC analysis to determine the residue concentrations. Results: Thin-layer chromatography analysis revealed that 23.5% of the tested samples contained residues from six different antibiotic classes (tetracyclines, quinolones, beta-lactams, sulfonamides, aminoglycosides, and macrolides). Thin-layer chromatography analysis showed that 35% and 25% of the meat samples were positive for residues from the broiler and layer, respectively. About 15% and 30% of layer and duck egg samples had positive residues, respectively. Out of 120 feed samples analyzed, about 15.8% had various antibiotic residues. In addition, feces samples from broilers and layers had 50% and 35% antibiotic residues, respectively. A total of 2.5% meat and 3.3% egg samples had antibiotic residues above the maximum residue limit (MRL). Based on the findings of this study, the highest percentage of oxytetracycline, followed by doxycycline and ciprofloxacin, were detected in feed samples, and oxytetracycline was detected in meat and egg samples. Conclusion: This study clearly showed the misuse of antibiotics in the poultry sector in Bangladesh. Although antibiotic residues below the MRL level are suitable for human consumption, they may result in antimicrobial drug resistance to pathogens.

Direct evidence of nitric oxide production induced by lactoferrin and its enhancement by magnesium ions in cultured endothelial cells.

Bovine lactoferrin (BLF) reportedly lowers blood pressure and induces vasorelaxation, but its effect on nitric oxide (NO) production has not been established. Accordingly, we aimed to determine whether BLF induces NO production in bovine aortic endothelial cells, and the effects of extracellular free magnesium (Mg) ion concentrations on this NO production. BLF induced NO production time-dependently. NO production was markedly inhibited by the NO synthase inhibitor, NG-nitro-L-arginine methyl ester, in an effect abolished by L-arginine, but not D-arginine. NO production was suppressed at low concentrations, and enhanced at high concentrations, of Mg ions in culture medium. These results suggest that BLF has an important role in hypotensive effects. Mg ions may affect BLF-induced NO production.

Isolation of multidrug-resistant Escherichia coli and Salmonella spp. from sulfonamide-treated diarrheic calves.

Background and Aim: The bovine industry is threatened by one of the most serious and deadly enteric diseases, calf diarrhea, particularly in developing nations like Bangladesh. In this context, bacterial resistance to antimicrobial drugs and its detrimental consequences have become a critical public health issue that is difficult to address globally. This study aimed to isolate and identify Escherichia coli and Salmonella spp. with their antibiogram and antibiotic resistance gene detection from sulfonamide-treated diarrheic calves. Materials and Methods: Twelve diarrheic calves suffering from calf diarrhea in a dairy farm were selected and a total of 36 fecal samples were aseptically collected directly from rectum before, during, and at the end of treatment for each calf to determine the total viable count, total E. coli count and total Salmonella count. A polymerase chain reaction was used for the specific detection of E. coli and Salmonella genus targeting fliC and invA genes, respectively. Antibiotic sensitivity test of the isolated E. coli and Salmonella spp. were performed by the disk diffusion method for eight commonly used antibiotics. Results: A total of 36 E. coli (100%) and 12 Salmonella spp. (33%) were isolated from the samples and were confirmed by polymerase chain reaction. Total viable count was found to be ranged from 35 × 107 to 99 × 1010 colony-forming unit (CFU)/g fecal sample before starting sulfonamide treatment, 34 × 105 to 25 × 1010 CFU/g during treatment with sulfonamide, and 48 × 103 to 69 × 1010 CFU/g immediately after completion of sulfonamide treatment. Total E. coli count was found to be ranged from 4 × 104 to 36 × 1010 CFU/g, 24 × 104 to 23 × 108 CFU/g, and 13 × 104 to 85 × 1010 CFU/g, whereas total Salmonella count was found to be ranged from 16 × 106 to 18.5 × 1011 CFU/g, 15 × 104 to 44 × 107 CFU/g, and 13.2 × 105 to 21 × 1010 CFU/g fecal sample before starting sulfonamide treatment, during treatment with sulfonamide immediately after completion of sulfonamide treatment, respectively. The in vitro antibiotic sensitivity test showed that all the E. coli and Salmonella spp. isolated from diarrheic calves (100%) contained multidrug-resistant (MDR) phenotypes. Escherichia coli isolates were found 100% resistant to amoxicillin (AMX), cefuroxime, cephalexin (CN), erythromycin (ERY), and tetracycline (TET); whereas 94.4%, 86.1%, and 77.8% isolates were resistant to doxycycline (DOX), moxifloxacin (MOF), and gentamycin (GEN), respectively. In case of Salmonella isolates, all were found 100% resistant to AMX, CN, and ERY; whereas 91.7% of resistance was observed for DOX, MOF, cefuroxime, GEN, and TET. Based on the molecular screening of the antibiotic resistance genes, tetA gene was present in 83.3% of the isolated E. coli and 75% of the isolated Salmonella strains, whereas 83.3% E. coli and 79.2% Salmonella isolates contained blaTEM gene. Conclusion: These findings suggest that MDR E. coli and Salmonella spp. might be responsible for calf scouring, which is challenging to treat with antibiotics or sulfonamide drugs alone. Therefore, it is important to check the antibiotic sensitivity pattern to select a suitable antibiotic for the treatment of calf scoring. A suitable antibiotic or combination of an antibiotic and sulfonamide could be effective against E. coli and Salmonella spp. responsible for calf scouring.

A Survey on Knowledge, Attitude, and Practices of Large-Animal Farmers towards Antimicrobial Use, Resistance, and Residues in Mymensingh Division of Bangladesh.

The widespread and indiscriminate use of antimicrobials in food animals is a key contributor to antimicrobial resistance and antimicrobial residue, which have become a growing public and animal health concern in developing countries such as Bangladesh. This study was aimed to assess the knowledge, attitude, and practices (KAP) of large-animal farmers towards antimicrobial use (AMU), antimicrobial resistance (AMR), and antimicrobial residue (AR) with their correlation. A cross-sectional survey was conducted with a structured and pretested questionnaire in the Mymensingh division of Bangladesh. A total of 212 large-animal farmers (dairy, beef fattening, buffalo, sheep, and goat farmers) were surveyed. Results showed that most of the farmers are male (85.8%) and belong to the 18-30 age group (37.3%). About 20.3% had no formal education, and nearly half of the participants (48.1%) received training regarding antibiotic use and resistance. Penicillin is the most common class of antibiotic used (61.8%) in the study area, followed by other antimicrobials. Only 37.7% of the farmers used antimicrobials on the recommendation of their veterinarian. Overall, 41.5%, 42.5%, and 21.7% of farmers possess adequate knowledge and a satisfactory attitude and perform desirable practices, respectively. Farmers in the 31-40 age group have adequate knowledge, attitude, and ability to implement desired practices compared to farmers in the 18-30 age group. Farmers having a graduate or post-graduate degree scored better in relation to knowledge, attitude, and practice than other farmers. Analysis revealed that farmers who received training on AMU and AMR had 10.014 times (OR = 10.014, 95% CIs: 5.252-19.094), 9.409 times (OR = 9.409, 95% CIs: 4.972-17.806), and 25.994 times (OR = 25.994, 95% CIs: 7.73-87.414) better knowledge, attitude, and performance, respectively, compared to their counterparts. A significant proportion of farmers (97.2%) dispose of leftover antibiotics inappropriately. The findings of the present study will be used to intervene in the education and training of the farmers, which will help to limit the indiscriminate and irrational use of antimicrobials, leading to reducing the chances of developing AMR.

Potency of spirulina (Spirulina platensis) on arsenic-induced lipid peroxidation in rat.

OBJECTIVE: Natural substances found in dietary sources and medicinal plants have attracted considerable attention in recent years as chemopreventive agents. Spirulina is a blue-green alga that possesses chemopreventive properties. The purpose of this study was to determine the effect of spirulina on rats with inorganic arsenic (As) [sodium arsenite (NaAsO2)]-induced lipid peroxidation. MATERIALS AND METHODS: 120 rats were randomly assigned to 10 groups and designated T0, T1, T2, T3, T4, T5, T6, T7, T8, and T9. One group was kept as a control (T0) that received no treatment. The seven groups received 3.0 mg of NaAsO2/kg body weight in drinking water and were given spirulina ad libitum. T1 was treated with NaAsO2 but not with spirulina. Two groups of rats (T2 and T3), on the other hand, were treated with spirulina without receiving any As (NaAsO2). T2 received agro-based spirulina (Ab-Sp; grown in 1.5% soybean meal media and harvested on day 12 of seed inoculation) at 2.0 gm/kg feed, whereas T3 received commercially available spirulina (Com-Sp) at 2.0 gm/kg feed. T4, T5, and T6 were concurrently treated with Ab-Sp at 1.0, 1.5, and 2.0 gm/kg of feed. On the other hand, T7, T8, and T9 induced by NaAsO2 were concurrently treated with Com-Sp at 1.0, 1.5, and 2.0 gm/kg feed. All groups received treatment for 90 days. RESULTS: The efficacy of both spirulina in preventing lipid peroxidation caused by As was determined quantitatively by measuring the rats' serum malondialdehyde (MDA). The results indicated that As supplementation increased serum MDA levels, whereas both types of spirulina significantly decreased them. The highest dose of Ab-Sp (2.0 gm/kg feed) was found to be the most effective in preventing lipid peroxidation in rats treated with inorganic As. CONCLUSION: Ab-Sp could be a natural, cost-effective, and safe measure to mitigate As toxicity.

Vasomotor effects of noradrenaline, 5-hydroxytryptamine, angiotensin II, bradykinin, histamine, and acetylcholine on the bat (Rhinolophus ferrumequinum) basilar artery.

The responsiveness of the basilar artery to intrinsic vasoactive substances is species-specific and can be a unique characteristic. We investigated the responsiveness of the bat (Rhinolophus ferrumequinum) basilar artery to noradrenaline (NA), 5-hydroxytryptamine (5-HT), angiotensin (Ang) II, bradykinin (BK), histamine (His), and acetylcholine (ACh). NA, 5-HT, Ang II, and BK induced contraction, whereas His and ACh induced relaxation, in a concentration-dependent manner. The NA cumulative concentration-response curve was shifted to the right in parallel with phentolamine (an α-antagonist). However, propranolol, a ß-antagonist, had no significant effect. The 5-HT curve was shifted to the right in parallel by ketanserin (a 5-HT2 antagonist) and methiothepin (a 5-HT1 and 5-HT2 antagonist). Losartan (an AT1 antagonist) shifted the Ang II curve to the right, whereas PD123319 (an AT2 antagonist) had no significant effect. L-NA, indomethacin, and des-Arg9-[Leu8]-BK (a B1 antagonist) did not significantly affect BK-induced contractions. HOE140 (a B2 antagonist) shifted the BK concentration-response curve to the right. The His curve was shifted to the right weakly by diphenhydramine (an H1 antagonist) and strongly by cimetidine (a H2 antagonist). ACh-induced relaxation was significantly inhibited by L-NA, atropine, and pFHHSiD (a muscarinic M3 antagonist), whereas pirenzepine and methoctramine (muscarinic M1 and M2 antagonists, respectively) showed no significant effects. At a resting vascular tone, L-NA-induced contraction and indomethacin induced relaxation. These results suggest that α-adrenergic, 5-HT1, 5-HT2, AT1, and B2 receptors might be important in arterial contraction, whereas M3 and H2 (>H1) receptors might modify these contractions, inducing relaxation.

Epidemiology and antimicrobial resistance profiles of Salmonella in chickens, sewage, and workers of broiler farms in selected areas of Bangladesh.

INTRODUCTION: A cross-sectional study was conducted to determine the prevalence and AMR pattern of Salmonella isolated from broiler chickens, farm sewage, and farm workers. This study also aimed at identifying the risk factors for Salmonella infection in chickens. METHODOLOGY: Cloacal swabs (n = 50) from broiler chickens, farm sewage (n = 50), and hand washed water of farm workers (n = 50) were collected along with data on farm management and antimicrobial usage from 50 broiler farms of Mymensingh and Gazipur districts. All samples were analysed for the presence of Salmonella using selective media and PCR assay. Antimicrobial susceptibility test was done for ten antimicrobials by disk diffusion test. Risk factor analyses were carried out by multivariable logistic regression using SPSS. RESULTS: The overall Salmonella prevalence was 66% (99/150). Salmonella prevalence were 82% and 72% in cloacal swabs and farm sewage samples, respectively. From hand washed water, 44% of the samples were positive for Salmonella. Salmonella infection in broiler chickens was significantly associated with farming experience (≤ 5 years) and age of birds (≥ 11 days). Similar pattern of antimicrobial resistance was observed in Salmonella isolated from three types of samples, and high resistances were observed to colistin, doxycycline, ciprofloxacin and ceftazidime. Moreover, isolates from all the three sources showed high percentage of multidrug-resistance (80.6% to 97.6%). CONCLUSIONS: The findings of this study reveal that antimicrobial-resistant Salmonella are prevailing at animal-human-environment interface, which needs an integrated approach to limit further spread of AMR.

Vasomotor effects of 5-hydroxytryptamine, histamine, angiotensin II, acetylcholine, noradrenaline, and bradykinin on the cerebral artery of bottlenose dolphin (Tursiops truncatus).

From an evolutionary aspect, dolphins share a very close phylogenetic relationship with pigs. Previously, we characterized porcine cerebral artery responsiveness to intrinsic vasoactive substances. Therefore, here, we investigated dolphin (Tursiops truncatus) cerebral artery responsiveness to 5-hydroxytryptamine (5-HT), histamine (His), angiotensin (Ang) II, acetylcholine (ACh), noradrenaline (NA), and bradykinin (BK) to characterize their related receptor subtypes. We also compared dolphin cerebral artery responsiveness with porcine cerebral artery responsiveness. We found that 5-HT and His induced concentration-dependent contraction of the dolphin cerebral artery. Ketanserin (a 5-HT2 antagonist) and methiothepin (a 5-HT1 and 5-HT2 antagonist) shifted the concentration-response curve for 5-HT to the right. Although diphenhydramine (an H1 antagonist) shifted the concentration-response curve for His to the right, cimetidine (an H2 antagonist) had no such effect. Ang II and ACh did not produce any vasomotor actions. NA induced concentration-dependent relaxation. Propranolol (a ß antagonist) shifted the concentration-response curve for NA to the right, whereas phentolamine (an α antagonist) had no significant effect. BK induced relaxation followed by contraction in pre-contracted arteries with intact endothelium. HOE140 (a B2 antagonist) shifted the concentration-response curve for BK to the right, whereas des-Arg9-[Leu8]-BK (a B1 antagonist) had no significant effect. These results suggest that 5-HT1, 5-HT2, and H1 receptor subtypes are important in arterial contraction and that ß and B2 receptor subtypes modify these contractions to relaxations. The responsiveness of the dolphin cerebral artery is very similar to that of porcine cerebral artery, supporting their evolutionary linkage.

Effect of discriminate and indiscriminate use of oxytetracycline on residual status in broiler soft tissues.

AIM: This study aimed to evaluate the effects of discriminate and indiscriminate use of oxytetracycline on hematological parameters, residual status in soft tissue of broiler and of thermal effect on oxytetracycline residual status. MATERIALS AND METHODS: Eighteen, day-old male broiler chickens were purchased and were divided into three different groups (control group, discriminate group, and indiscriminate group). The control group received no antibiotics. The discriminate group received oxytetracycline 1 g/L drinking water for 5 consecutive days, and 10 days' withdrawal period was maintained before sacrifice. The indiscriminate group received oxytetracycline 1 g/L drinking water till the sacrificed day. Blood samples were collected before sacrificing for hematological analysis. After sacrificing liver, kidney, spleen, and muscle samples were collected for analysis of oxytetracycline residues in raw soft tissues. Since meat is used to cook by traditional method in Bangladesh before consumption that is why positive meat samples were cooked by traditional cooking method to evaluate the thermal effect on oxytetracycline residual status as well. Thin-layer chromatography (TLC) was done for screening of oxytetracycline residues in soft tissues. RESULTS: Mean differences of total erythrocyte count (million/mm3), hemoglobin estimation (gm%), and packed cell volume (%) estimation were not statistically significant among the groups. TLC analysis of raw samples showed 100% positive results of all samples collected from the indiscriminate group. In contrast, samples collected from the discriminate group were negative for oxytetracycline residues. In the control group, all samples were negative for oxytetracycline residue. There was a significant (p<0.05) relationship of oxytetracycline residues among three different groups for liver, kidney, spleen, and muscle samples. Positive liver and muscle samples from the indiscriminate group were subjected to thermal treatment by traditional cooking method of Bangladesh. Oxytetracycline residues had found in cooked meat, liver, and juice part, suggesting that antibiotic residues disseminated to juice part from flesh part after cooking. CONCLUSION: Evidence suggests that proper maintenance of withdrawal period would minimize oxytetracycline residues in broiler soft tissues, whereas antibiotics retained in soft tissues of broiler in case of indiscriminate use. Traditional cooking does not change oxytetracycline residual status in edible tissues. Therefore, awareness regarding the proper maintenance of withdrawal period after antibiotic treatment of broiler is one of the best strategies which may positively reduce the risk of antimicrobial drugs residue in meat.

Antioxidant activity of different species and varieties of turmeric (Curcuma spp): Isolation of active compounds.

There are >80 species of turmeric (Curcuma spp.) and some species have multiple varieties, for example, Curcuma longa (C. longa) has 70 varieties. They could be different in their chemical properties and biological activities. Therefore, we compared antioxidant activity, total phenolic and flavonoid content of different species and varieties of turmeric namely C. longa [variety: Ryudai gold (RD) and Okinawa ukon], C. xanthorrhiza, C. aromatica, C. amada, and C. zedoaria. The antioxidant activity was determined using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, oxygen radical absorbance capacity (ORAC), reducing power and 2-deoxyribose (2-DR) oxidation assay. Our results suggested that RD contained significantly higher concentrations of total phenolic (157.4 mg gallic acid equivalent/g extract) and flavonoids (1089.5 mg rutin equivalent/g extract). RD also showed significantly higher DPPH radical-scavenging activity (IC50: 26.4 µg/mL), ORAC (14,090 µmol Trolox equivalent/g extract), reducing power absorbance (0.33) and hydroxyl radical scavenging activity (IC50: 7.4 µg/mL). Therefore, RD was chosen for the isolation of antioxidant compounds using silica gel column, Toyopearl HW-40F column, and high-performance liquid chromatography. Structural identification of the compounds was conducted using 1H NMR, 13C NMR, and liquid chromatography-tandem mass spectrometry. The purified antioxidant compounds were bisabolone-9-one (1), 4-methyllene-5-hydroxybisabola-2,10-diene-9-one (2), turmeronol B (3), 5-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)-1-hepten-3-one (4), 3-hydroxy-1,7-bis(4-hydroxyphenyl)-6-hepten-1,5-dione (5), cyclobisdemethoxycurcumin (6), bisdemethoxycurcumin (7), demethoxycurcumin (8) and curcumin (9). The IC50 for DPPH radical-scavenging activity were 474, 621, 234, 29, 39, 257, 198, 47 and 18 µM and hydroxyl radical-scavenging activity were 25.1, 24.4, 20.2, 2.1, 5.1, 17.2, 7.2, 3.3 and 1.5 µM for compound 1, 2, 3, 4, 5, 6, 7, 8 and 9, respectively. Our findings suggested that the RD variety of C. longa, developed by the University of the Ryukyus, Okinawa, Japan, is a promising source of natural antioxidants.